Abstracts Submitted to the 8th International Congress on the Biology of Fish

Portland, Oregon, USA July 28- August 1, 2008

Compiled by Don MacKinlay

Fish Biology Congress Abstracts 1 HISTOLOGICAL AND HISTOCHEMICAL STUDY OF LIVER AND PANCREAS IN ADULT OTOLITHES RUBER IN PERSIAN GULF Abdi , R. , Sheibani , M and Adibmoradi , M. Symposium: Morphometrics Presentation: Oral Contact: Rahim Abdi, Khoramshahr University of Marine science and Technology Khoramshahr Khozestan 64199-43175 E-Mail: [email protected] Abstract: In this study, the digestive system of 10 adult Otolithes ruber, were removed and the livers and pancreases were put in the formalin 10 % to be fixed. The routine procedures of preparation of tissues were followed and the paraffin blocks were cut at 6 microns, stained with H&E, PAS and Gomori studied under light microscope. The results of microscopic studies showed that liver as the greatest accessory organ surrounds the pancreatic tissue. Liver is a lobulated organ which surrounds the pancreas as an accessory gland among its lobules. Hepatic tissue of this fish is similar to many other osteichthyes. Hepatocytes include glycogen stores and fat vacuoles located around the hepatic sinusoids. Pancreas as a mixed gland microscopically was composed of lobules consisting of serous acini(exocrine portion) and langerhans islets (endocrine portion). However, pancreatic lobules are usually seen as two rows of acini among which there is a large blood vessel.

ECOLOGICAL CONSEQUENCES OF A TRANSPANTING EXOTIC FISH TO FRESHWATER ECOSYSTEMS OF IRAN: A CASE STUDY OF RAINBOW TROUT ONCORHYNCHUS MYKISS (WALBAUM, 1792) Abdoli, A., Patimar, R., Mirdar, J., Rahmani, H., and Rasooli, P. Symposium: Fish Growth Presentation: Poster Contact: Asghar Abdoli, Department of Biodiversity and Ecosystem Management, Environmental Sciences Research Institute (ESRI), Shehid Beheshti University Tehran ------Iran E-Mail: [email protected] Abstract: Abundance and diet of Oncorhynchus mykiss were examined from November 1996 to June 1998 in Madarsoo River in Golestan National Park. A total of 144 specimens investigated (average of Total length= 190mm and Total weight= 80g). Ephemeroptera was the most abundance benthic invertebrates of collected organisms. The Ivelev index showed that Ephemeroptera, Gammaridae and Tricoptera were the most important food items for O.mykiss our result has indicated this species has potentially food competition with fish species, Paracobitis malapterura , Neogobius melanostomus, Neogobius fluviatilis, Albunoides bipunctatus in Golestan National Park stream.

A COMPARISON STUDY ON LENGTH-WEIGHT RELATIONSHIPS OF MUDSKIPPERS PRIPHTHALMUS WALTONI(GOBIIDEA) IN COASTAL AREAS OF HORMOZGAN AND BOSHEHR PROVINCES, IRAN Abdoli,L, Abdoli,A, Kamrani,E, nad ,kiabi,B. Symposium: Fish Growth Presentation: Poster Contact: Leila abdoli, Department of Marine Biology – Faculty of Science – University of Hormozgan –P.O.Box 3995 Bandar Abbas - Iran Bandarabbs hormozgan Iran E-Mail: [email protected] Abstract: This study carried out from January 2007to July 2008 in costal of hormozgan and boshehr , and during this time 320 specimens were catch. Sampling carried out by hand. Specimens preserved in 10% formalin and measured to the nearest mm and weight determined with a digital balance. The relation of weight to length was calculated applying the exponential regression equationw=al b, where W is total weight, L is total length ,a & b the parameters to be estimated(Ricker,1975).Length-weight relationship was calculated for specimens hormozgan province by using Ln weight = - 11.87+3.07ln length for males and Ln weight = -11.51+ 2.99 Ln length for females, and for specimens boshehr , Ln weight = - 10.41+2.76 Ln length for males and Ln weight = - 11.62+3.03 Ln length for females . the slopes (b values) of the length weight regression was not significantly different between sexes.(ANCOVA, P< 0.00)

THE SPAWNING MIGRATION OF KISSING LOACH (LEPTOBOTIA CURTA) BETWEEN RIVERS AND FLOOD AREAS: POSSIBLE GENERAL TRIGGER FOR FISH MIGRATIONS IN RIVERS Abe, T., Soma, Y., Kobayashi, I., Kon, M., and Sakamoto, T. Symposium: Stress Effects and Response Presentation: Oral Contact: Tsukasa Abe, Ushimado Marine Laboratory, Graduate School of Natural Science and Technology, Okayama University, 130-17, Kashino, Ushimado, Setouchi, 701-4303, Japan Setouchi Okayama 701-4303 Japan

Fish Biology Congress Abstracts 2 E-Mail: [email protected] Abstract: We have observed that kissing loach and some other freshwater fishes migrate from the main stream of rivers to spawn in flood areas. These flood areas are unstable and ephemeral, however, they contain few aquatic predators and a large amount of food suitable for the development of embryos/juveniles. The present study provides the first evidence that the spawning migration of kissing loach is stimulated by both rainfall and the subsequent rise in water level, as well as the new moon. Since similar observations have been reported for downstream migrations of salmonids, these stimuli may serve as general cues for migrations of fishes in rivers. Furthermore, the spawning of kissing loach is rigidly limited spacio-temporally after the formation of flood areas. Recently we have successfully demonstrated that changes in water quality directly induce migration and spawning behavior, suggesting some particular odoriferous substances may be the environmental proximate factors.

EFFECTS OF SUBLETHAL CONCENTRATION OF DIAZINON ON PLASMA BIOCHEMICAL PROFILE OF THE AFRICAN CLARIAS GARIEPINUS Adedeji, O.B., Symposium: Environmental Pollution Presentation: Oral Contact: B. Adedeji Olufemi, Faculty of Veterinary Medicine University of Ibadan, Nigeria Ibadan Oyo Nigeria E-Mail: [email protected] , [email protected] Abstract: Pesticides and drugs used in agriculture and veterinary medicine may end up in aquatic environments and bioaccumulate in the food chain, thus posing serious problems for fauna and human health. This study determined the acute effects of organophosphate pesticides on biochemical changes in the African catfish Clarias. gariepinus as a result of exposure to sublethal concentrations of diazinon. The values obtained from the experimental fish were comp aired with normal values and subjected to statically analysis. The results showed that acute effect of diazinon on plasma biochemistry varies. It causes significant increase in some enzymes, (p<0.05) CHE, LDH, blood glucose, plasma protein, K+ concentration and significant decrease, (p<0.05) in Ca2+ and P. This shows that plasma biochemical profile can be used to monitor pesticides toxicity in fish. Proper surveillance and environmental monitoring of pesticides is therefore recommended in aquaculture for fish health and food safety.

EFFECTS OF DIAZINON ON BLOOD PARAMTERS IN THE AFRICAN CATFISH (CLARIAS GARIEPINUS) Adedeji, O.B., Adedeji, O.A., Adeyemo O.K., and Agbede S.A., Symposium: Environmental Pollution Presentation: Poster Contact: Olufemi Adedeji, Faculty of Veterinary Medicine University of Ibadan, Nigeria Ibadan Oyo Nigeria E-Mail: [email protected] , [email protected] Abstract: Effects of diazinon (aquatic pesticides pollutant) on cultured and wild African catfish (Clarias gariepinus) was assessed, based on results of acute toxicity tests and on a comparison of results of haematological examination of a control and an experimental group exposed to DiazintolR pesticide preparation (active substance 162mg.l-1of diazinon). The 96hr acute toxicity test was performed semistatically on the catfish . Examination of erythrocyte and leucocyte profile was performed on 16 control and 20 experimental specimens. The experimental group showed significantly lower values (p < 0.05) of erythrocyte count , haemoglobin content and haematocrit . Values of MCV, MCH and MCHC were comparable in both groups. There was a significant decrease in leucocyte count (p < 0.05). Changes in values of both the erythrocyte and leucocyte profile after exposure to diazinon-based preparation may be referred to disruption of haematopoiesis as well as to a decrease on non-specific immunity of the fish.

BIODIVERSITY OF FRESHWATER FISHES OF MARATHWADA REGION FROM INDIA Ahirrao, S.D. Symposium: Contributed Papers Presentation: Poster Contact: Anand Manwar, dept of fisheries, shri shivaji college parbhani maharashtra 430401 india E-Mail: [email protected] Abstract: Marathwada is one of the important region of india with high fish production and large numbers of rivers,reservoirs,lakes and ponds. it plays imp role in indian economy as will as the economy of the present area by contrubuting share in the total fish production similarly it has a great fish diversity. in the present paper the ichthyofauna has observed for last five years and taxonomically identified and verious types of fishes such as food fishes,culurable fishes,migretory fishes,predatory fishes,weed fishes,thretened fishes larvaevorous fishes medicinal value fishes has been described in the present paper.

Fish Biology Congress Abstracts 3 SOME OF REPRODUCTIVE PROPERTIES OF NEOGOBIUS CASPIOUS FROM THE SOUTH EAST COASTS OF THE CASPIAN SEA(GUILAN OFF SHORE-IRAN) Alinaghi,S.S.,Ali,S.,Shabanali,N.,Anni,Ch.,Bin Saad,Ch.,and Gholamreza.Gh. Symposium: Contributed Papers Presentation: Oral Contact: Gholamreza Ghasemzadeh, Ayatollah Taleghani Ave.,Ghaziyan,Anzali Port,Fisheries Departement of Guilan province, Anzali Guilan 166 Iran E-Mail: [email protected] Abstract: N.caspious is an endemic species of the Caspian sea Gobiidae that ecologically play an important role in predator fish food chain .The chief objective of this research is focused on some of the reproductive properties of N.caspious in south east coasts of the Caspian sea.Sampling were carried out with a bottom trawler in three stations (Astara,Anzali and Chabuksar)and three depths in each station(0-5,5-10 and 10-15 meter)from 2004 October until to the 2005 September. N.caspious species were identified based on morphometric and meristic count parameters. according to the high valid references.mmaximum Gonadosomatic index were recorded for male(7.52%) and female(100.12%) in June.A sharp decreased in GSI value from June to the end of September in both sex.and spowning takes place in several times ,hence this species considered as a later spowner.Sex ratio was 1.47:1 female to male.Absolute fecundity ranged between 212 to 1234 eggs and relative fecundity fluctuated between 18.73 and 29.89 egg per each gram of the body weight.Correlation between absolute and relative fecundity with age, total length and full weight was positive and negative respectively.Egg diameter ranged between 0.06 to 0.23 mm. Correlation between egg diameter, age and total length was positive.

ION AND OSMOREGULATION IN GREEN STURGEON; SCIENCE AND FUN WITH JOE Allen, P.J., Cole, S., Nicholl, M.M., Hobbs, J.A., Kültz, D., Doroshov, S.I., and Cech, J.J., Jr. Symposium: Tribute to Joe Cech, Jr. Presentation: Oral Contact: Peter Allen, Dept of Biological Sciences, University of Manitoba, Duff Roblin Building Canada E-Mail: [email protected] Abstract: The green sturgeon, Acipenser medirostris, migrates from freshwater to seawater relatively early in its life history, however little is known about its age, size or the mechanisms that facilitate this transition. Initially, the timing of this transition was investigated through examination of its salinity tolerance, swimming performance, and hormonal (T3, T4, cortisol) and enzymatic changes (gill Na+, K+, ATPase) with increasing age and size. Later, we acclimated 3 sizes/ages of juvenile sturgeon to 0, 10, and 33‰ and measured MO2 to determine if their was an energetic advantage indicating the likely timing of seawater entry. Furthermore, we examined ion and osmoregulation, chloride cell abundance and distribution, and gill, kidney, and gut Na+, K+, ATPase. Finally, laser ablation-inductively coupled plasma-mass spectrometry was used in pectoral fin rays to determine the timing of initial seawater entry from comparisons between fish held in freshwater and seawater and those collected from the wild.

REPRODUCTIVE-BASED CHANGES IN CALCIUM REGULATION IN A FRESHWATER, CARTILAGINOUS FISH Allen, P.J., Guerreiro, P.M., Fuentes, J., Canario, A.V.M., Peake, S.J., Anderson, W.G. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Peter Allen, Dept. of Biological Sciences, University of Manitoba Winnipeg MB R3T 2N2 Canada E-Mail: [email protected] Abstract: During the female reproductive cycle, increasing demands for calcium are placed on the body. Fishes are able to compensate through external uptake from the environment as well as internal mobilization from calcified tissues. However, if environmental calcium concentrations are low, and internal stores are limited, the reproductive cycle may strain a fish’s ability to regulate calcium. To examine this is a natural context, we investigated calcium regulation in a wild population of lake sturgeon, Acipenser fulvescens. Blood plasma, urine and gonadal samples were collected to analyze calcium and associated hormone concentrations. In vitellogenic females, plasma total calcium peaked at 8.7 ± 0.6 mM, before returning to pre-vitellogenic concentrations near 4 mM. Plasma parathyroid hormone-related protein (PTHrP) concentrations in vitellogenic fish (0.10 ± 0.02 ng/ml, n=13) decreased as compared to non-vitellogenic fish (0.36 ± 0.10 ng/ml, n=18). Research supported by Manitoba Hydro, NSERC (CRDPJ 321520-05) and the Canadian Foundation for Innovation.

USING TRACE ELEMENT ANALYSES TO ASSESS MIGRATIONAL MOVEMENTS IN ANADROMOUS STURGEON Allen, P.J., Hobbs, J.A., Cech, J.J., Jr., Van Eenennaam, J.P., and Doroshov, S.I. Symposium: Contributed Papers Presentation: Poster

Fish Biology Congress Abstracts 4 Contact: Peter Allen, University of Manitoba, Z320 Duff Roblin Building Winnipeg MB R3T 2N2 Canada E-Mail: [email protected] Abstract: Many sturgeon species move between waters of different salinities at some point in their life history. Our goal was to assess the ability to detect this transition in a calcified structure that would not require terminal sampling, due to the rarity of this family of fishes. We used laser ablation-inductively coupled plasma-mass spectrometry on green sturgeon (Acipenser medirostris) pectoral fin rays, for the examination of initial seawater entry. In control, hatchery fish (n=6), reared 1-2 years in fresh water and 1-3 years in seawater, we found a significant increase in strontium (Sr)/calcium (Ca), decrease in barium (Ba)/Ca, and increase in Sr/Ba (16.5-fold) in fresh water versus seawater calcified growth zones. Sr/Ca, Ba/Ca and Sr/Ba changes were also assessed in wild adults from the Klamath River, CA. The results demonstrate the usefulness of trace element analyses of sturgeon pectoral fin rays, and an early age of green sturgeon entry into seawater.

ASSESSING THE SWIMMING BEHAVIOUR OF UPRIVER MIGRATING SEA LAMPREY WITH ELECTROMYOGRAM TELEMETRY Almeida, P.R.; Mateus, C.S. and Quintella, B.R. Symposium: Biology of Lampreys Presentation: Oral Contact: Pedro Raposo de Almeida, Department of Biology, University of Évora, Apartado 94 Évora Évora 7002-554 Portugal E-Mail: [email protected] Abstract: A primary goal in fish behaviour studies, particularly those with respect to migrants, is to obtain direct measures of the nature and locomotor activity of their journey in the wild. Conventional telemetry has been used to study the spawning migration of the sea lamprey (Petromyzon marinus L.) but these studies provided limited information on the swimming behaviour of this species particularly when passing weirs or riffles and other complex rapid flow reaches. With the advent of EMG radio telemetry, it has become possible to better interpret the activity and behaviour of migrating fish, by permitting to monitor remotely individual fish in their natural environment. This presentation resumes a 6 year experience whereby EMG telemetry was used to monitor sea lampreys’ spawning migration. This technique provided important information of relevance to this species conservation and habitat rehabilitation. Particular reference is made to sea lampreys’ behaviour when moving on river stretches with different hydraulic characteristics.

DIFFERENTIAL GENE EXPRESSION IN COLOSSOMA MACROPOMUM EXPOSED TO CRUDE OIL AND HYPOXIA. Almeida-Val, V.M.F., Braga, L.R., Val, A.L. and Nozawa, S.R. Symposium: Tropical Fish Adaptations Presentation: Oral Contact: Vera Val, INPA - LEEM. Av. André Araujo, 2936. Manaus AM 69.083-000 E-Mail: [email protected] Abstract: Differential Display RT-PCR was used to identify differentially expressed genes in juveniles of the Amazonian fish Colossoma macropomum, locally known as tambaqui, exposed to petroleum, anoxia and hypoxia. Four genes were tracked: (i) RAG2 (recombination-activating genes); (ii) ERK (mitogen-activated protein kinase); (iii) GR (glutathione reductase); e (iv) CFTR (Cystis fibrosis transmembrane conductance regulator). These later three genes are up- or down-regulated when detoxification is needed, while the first regulates cell synthesis of the immune system. The gene RAG-2 was sequenced and the stress-regulated differential expression was quantified by Real Time qRT-PCR. Using the constitutive 18S gene as reference, high levels of expression of RAG-2 were detected in control (5.4-fold), while animals exposed to hypoxia (2.7-fold), anoxia (2.5-fold) and petroleum (2.9-fold) presented lower expression. We propose that exposure to environmental stressors causes an immunodeficiency in tambaqui that may be related to the decrease in RAG-2 gene expression (CNPq, FAPEAM, PRONEX).

BISPHENOL A IMPACTS EMBRYOGENESIS AND GROWTH BY DISRUPTING THE SOMATOTROPIC AXIS IN RAINBOW TROUT. Aluru, N., Leatherland, J.F., Vijayan, M.M. Symposium: Environmental Pollution Presentation: Oral Contact: Neel Aluru, Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, Guelph, Ontario. Guelph Ontario N1G 2W1 Canada E-Mail: [email protected] Abstract: We investigated the impact of maternal exposure to Bisphenol A (BPA), a xenoestrogen, on development and growth in rainbow trout (Oncorhynchus mykiss). Oocytes were exposed to 0, 30 and 100 micro grams/ml of BPA for 3 h in the

Fish Biology Congress Abstracts 5 ovarian fluid to mimic in utero exposure, and this was followed by fertilization. The higher vitellogenin gene expression in the BPA groups confirmed xenoestrogen exposure. High BPA treatment delayed hatching, yolk absorption, time to first feed and growth rate compared to the other two groups. BPA-mediated developmental abnormality correlated with lower IGF-2 but not IGF-1 mRNA levels. The lower growth rate post-hatching in the high BPA group corresponded with suppressed IGF-1 and GH-receptor mRNA abundance. Together, these results for the first time suggest that maternal exposure to BPA leads to developmental and longer-term growth effects in . The mode of action of BPA involves disruption of the IGF/GH axis functioning in trout.

APPLYING METHODS AND CONCEPTS OF PSYCHOLOGICAL RESEARCH TO FISH PASSAGE Anderson, J. J. Symposium: Swimming Performance Presentation: Oral Contact: James Anderson, 1325 4th ave, suite 1820 seattle wa 98101 USA E-Mail: [email protected] Abstract: From the perspective of psychology, the trajectories of fish through a passage system involved a series distinct information processing steps: stimulus detection, stimulus processing and finally decision. I will briefly review research relevant to each step and discuss how the concepts might be applied to passage studies. Concepts include the Webber-Fechner law of signal detection, the leaky-integrator model of stimuli habituation, the linear-nonlinear-Poisson and Take the Best choice models, and the Strain-Velocity-Pressure hypothesis.

TOXICITY ANALYSIS OF MILKFISH EXPOSURE TO THREE HEAVYMETAL POLLUTANTS: PB, HG, CD. Archana Devi, C, M. Pradhap and A. K.Kumaraguru Symposium: Environmental Pollution Presentation: Oral Contact: C. Archana Devi, Field lab - Centre for Marine and Coastal studies, Madurai Kamaraj University, Pudhumadam, Ramanathapuram district,Tamilnadu, India.pin: 623 524, 2.Dept. of Zoology, Annamalai university, chidambaram, chennai Tamilnadu 623524 India E-Mail: [email protected], [email protected] Abstract: The marine ecosystem is a highly complex environment and natural fluctuations in species abundance and distribution are a feature of the normal way it functions. Toxicity of Heavy metals in marine organisms can be determined by acute toxicity tests. Acute toxicity tests or bioassay have historically played an important role in assaying the effects of human activities; on animals and such tests have wide applicability when evaluating the toxicities of various types and mixtures of pollutants to fish and other aquatic species. The parameters of short-term mortality have been the most common measure of toxicity. A variety of marine invertebrates have been examined to test the lethal effects of several toxicants on aquatic biota. The present study is aimed to investigate the acute toxicological effects of the three heavy metals – Pb, Hg and Cd in the marine fish. Chanos chanos (Milkfish) is the only species in the Family Chanidae. Its distribution is restricted to either low latitude tropics or the subtropical northern hemisphere along continental shelves and around islands, where temperatures are greater than 20 C. The median lethal concentration (LC50) of Fish to heavy metals was also investigated in 24, 48, 72 and 96 hrs static bioassays. The test result revealed that mercury was more sensitively toxic, when compared to cadmium and lead.

DISEASE SUSCEPTIBILITY OF JUVENILE RAINBOW TROUT ONCORHYNCHUS MYKISS EXPOSED TO A DIETARY MIXTURE OF HIGH MOLECULAR WEIGHT POLYCYCLIC AROMATIC HYDROCARBONS Arkoosh, M.R., Bravo, C.F., Curtis, L.R., Myers, M.S., Meador, J., Johnson, L., Buzitis, J., Morrow, J.D., Laetz, C., Loge, F.J., and Collier, T.K. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Mary Arkoosh, Environmental Conservation Division, Northwest Fisheries Science Center, 2030 South Marine Science Drive Newport Or 97365 USA E-Mail: [email protected] Abstract: High molecular weight polycyclic aromatic hydrocarbons (HMW PAHs) contaminate marine and fresh water environments, but few studies have examined the biological effects of environmentally relevant PAH concentrations on fish. The main source of HMW PAHs is pyrogenic or derived from incomplete combustion of fossil fuels. In this study, juvenile rainbow trout were fed a diet containing an environmentally relevant mixture of ten HMW PAHs at a dose of 0.66 or 7.82 µg PAH g fish -1 day-1 or a control diet with no PAHs for 50 days. Disease susceptibility to the pathogen Aeromonas salmonicida was examined after dietary exposure to PAHs. An increase in mortality was observed due to the

Fish Biology Congress Abstracts 6 pathogen in fish fed either 0.66 or 7.82 µg PAH g fish -1 day-1 relative to control. Additionally, biomarkers of exposure and oxidative stress were examined.

ACUTE BIOCHEMICAL EFFECTS OF ROUNDUP TO PROCHILODUS LINEATUS Assmann-Modesto, K. and Martinez, C.B.R. Symposium: Tropical Fish Adaptations Presentation: Poster Contact: Kathya Assmann Modesto, Carlos Gomes street, number 244, ap. 02 Cornélio Procópio PR 86300-000 Brazil E-Mail: [email protected] Abstract: Prochilodus lineatus represents a native fish of economic importance in Southern Brazil, where the herbicide Roundup is widely used in agriculture. To evaluate the effects of this herbicide to Neotropical fish, juveniles of P. lineatus were exposed to 10mg.L-1 of Roundup (RDP) or only to water (control). After 96h exposure fish were sampled. Liver, brain and white muscle were removed for biochemical analyses: liver activity of glutathione-S-transferase (GST), superoxide dismutase (SOD), glutathione peroxidase (GPX) and catalase; liver content of reduced glutathione (GSH) and lipid peroxidation (FOX); brain and muscle acetylcholinesterase (AChE) activity. RDP fish showed increased GST and GSH and reduced GPx in relation to control. Liver SOD, catalase and FOX did not vary significantly between RDP and control groups. Brain and muscle AChE were significantly reduced in RDP fish. These results demonstrated that Roundup interferes in the antioxidant defenses of P. lineatus and also acts as an AChE inhibitor.

THE THERMAL TOLERANCE OF ATLANTIC COD (GADUS MORHUA L.) JUVENILES HELD UNDER NORMOXIC AND MODERATELY HYPOXIC CONDITIONS Bailey, J.A., Busby, C.D., Tosh, J., Murphy, L., and Gamperl, A.K. Symposium: Fish Living on the Edge Presentation: Poster Contact: Jason Bailey, Ocean Sciences Centre, Memorial University of Newfoundland St. John's NL A1C5S7 Canada E-Mail: [email protected] Abstract: We examined the tolerance of 14 cod families to acute (~ 2oC • h-1) and chronic (1oC every 4 days) temperature increases under both normoxia and moderate hypoxia (75% O2 saturation). Cod challenged with an acute temperature increase had a CTM (Critical Thermal Maximum) of 22.5 ± 0.7 oC (mean ± s.d.), and exposure to hypoxia decreased their CTM only slightly (to 21.8 ± 0.7 oC). In the chronic challenge, the normoxic CTM was 21.9 ± 1.0 oC, a value 0.6 oC lower than that measured acutely. However, hypoxia had no effect on the cod’s chronic CTM or its temperature-food consumption (FC) relationship; FC decreasing by ~ 50% from 17-18oC, and ceasing altogether by 20oC. Large differences in family means for acute CTM (up to 1.8 oC) were observed, but reduced fish numbers precluded family analysis in the chronic temperature challenge. This data suggests that thermal tolerance may be heritable, and thus could be selected for in a breeding program.

PHYSIOLOGICAL MECHANISMS OF CO2 TOLERANCE IN THE PRIMITIVE FISH, THE WHITE STURGEON (ACIPENSER TRANSMONTANUS) Baker, D.W., Rummer, J., Hueng, K.T., Wilson, J.M., Morgan, J.D., Matey, V., and Brauner, C.J. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Daniel Baker, Department of Zoology, University of British Columbia Vancouver BC V6T 1Z4 Canada E-Mail: [email protected] Abstract: White sturgeon are among the most tolerant fish species to hypercarbia (elevated environmental CO2). Physiological responses typically observed in teleosts (i.e., net plasma Cl-/HCO3- exchange) can only compensate for the acidosis at a PCO2 below 15 mm Hg. White sturgeon exposed to a PCO2 of 15 mmHg, exhibited blood pH (pHe) compensation associated with net plasma Cl-/HCO3- exchange, and a decrease in gill mitochondrial rich cell (chloride cell) apical surface area, consistent with that observed in fishes. During exposure to more severe hypercarbia (20-45 mm Hg), pHe was not compensated (pH decreased 0.3-0.7), but intracellular pH (pHi) was protected in heart, liver, brain and white muscle. Only at extremely high CO2 tensions (60+ mm Hg) was pHi significantly reduced. The exceptional CO2 tolerance of sturgeon is likely associated with pHi protection due to active cellular pH regulation. (This research was supported by NSERC Discovery to CJB)

EVALUATING THE IMPACTS OF CURRENT USE PESTICIDES ON THE PHYSIOLOGY, ECOLOGY, AND PRODUCTIVITY OF POPULATIONS Baldwin, D.H., Spromberg, J.A., Macneale, K.H., Laetz, C.A., and Scholz, N.L. Symposium: Environmental Pollution Presentation: Oral

Fish Biology Congress Abstracts 7 Contact: David Baldwin, NOAA Fisheries, Northwest Fisheries Science Center, 2725 Montlake Blvd E Seattle WA 98112 USA E-Mail: [email protected] Abstract: While acknowledged as a potential factor in the decline of salmon populations, the impacts of pesticide exposures on the health of individual salmon and the viability of salmon populations are poorly understood. Our studies focus on insecticides that target acetylcholinesterase, an enzyme critical for brain and muscle function in salmon. We are examining the impacts at several biological scales to link suborganismal effects (e.g. acetylcholinesterase inhibition), organismal effects (e.g. reduced feeding and growth) and population effects (e.g. reduced productivity). We measured reductions in acetylcholinesterase activity and correlated reductions in swimming and feeding behaviors of juvenile coho salmon and modeled reductions in insect abundance to build a model linking pesticide exposures to reductions in juvenile growth. Size- dependent survival rates were incorporated into a population model estimating long-term changes in salmon population productivity. Since pesticides frequently occur together, we assessed the cumulative impact of mixtures on the acetylcholinesterase activity of juvenile salmon.

RISK OF HIGHER TEMPERATURE ON REPRODUCTIVE PERFORMANCE OF A TROPICAL DAMSELFISH Bapary, M.A.J., Fainuulelei, P., and Takemura, A. Symposium: Tropical Fish Adaptations Presentation: Poster Contact: Takemura, A., Tropical Biosphere Research Center, University of the Ryukyus, 3422 Sesoko Motobu Okinawa 905-0227 Japan E-Mail: [email protected] Abstract: The aim of the present study was to examine the involvement of photoperiod and temperature in ovarian development in the sapphire devil Chrysiptera cyanea, which is a reef-associated damselfish of tropical origin. Rearing the fish during the resting phase under a long photoperiod (LD14:10) at 25 or 31oC resulted in an increase in gonadosomatic index (GSI) and induction of vitellogenic oocytes. Higher temperature initially accelerated but finally suppressed the ovarian development. On the other hand, a short photoperiod (LD10:14) at the same temperatures failed to increase GSI and induce oocytes at the yolk stage. These results suggest that photoperiod and temperature are essential environmental factors for the reproductive activity of the sapphire devil. Since tropical fishes are likely to possess a suitable range of temperature in habitats, the increase in temperature by global warming may influence their reproductive success in tropical regions.

LITTLE BUGS FROM FISH WITH SMALLER, NASTIER BUGS: PRELIMINARY STUDIES ON THE ROLE OF SEA LICE AS A VECTOR OF BACTERIAL PATHOGENS TO SALMON IN BRITISH COLUMBIA, CANADA. Barker, D.E., Braden, L.M., Coombs, M.P. and Boyce, B. Symposium: Contributed Papers Presentation: Oral Contact: Duane Barker, Fisheries and Aquaculture Department, Vancouver Island University, 900 Fifth St., Nanaimo, British Columbia, Canada, V9R 5S5 Nanaimo British Columbia V9R 5S5 Canada E-Mail: [email protected] Abstract: Parasite vector-pathogen associations (e.g., malaria from mosquitoes), have been well described from humans and domestic animals but have rarely been examined within a marine ecosystem (e.g., ectoparasites of fishes). Farmed salmon in B.C. provide an almost ideal system to examine because of the local abundance of wild and farmed salmon and the potential for reciprocal exchange of ectoparasites such as sea lice (Lepeophtheirus salmonis). Using standard bacteriological screening protocols, we sampled the carapace and internal viscera of motile sea lice collected from farmed Atlantic salmon from May 2007 to April 2008. Our preliminary results include the first isolation of three pathogenic bacteria (Tenacibaculum maritimum, Pseudomonas fluorescens and Vibrio sp.) from sea lice and their salmon hosts. Spatiotemporal variation among bacteria prevalence was evident from external (58-100%) and internal (12.5-100%) samples. From such intriguing preliminary results, we propose to examine and describe this association within an ecological context to identify the significant geographic, seasonal and biological influences.

THE EFFECT OF DIETARY RATION AND FASTING ON GLOBAL PROTEOMIC EXPRESSION IN RAINBOW TROUT (ONCORHYNCHUS MYKISS) LIVER, SERUM, AND MUSCLE. Baumgarner, B.L., Bharadwaj, A. S., Pei, Z., and Brown, P.B. Symposium: Fish Growth Presentation: Oral Contact: Bradley Baumgarner, 715 West State Street West Lafayette IN 47907 USA E-Mail: [email protected]

Fish Biology Congress Abstracts 8 Abstract: The goal of this study was to examine the effect dietary ration has on the proteome of important metabolic tissues that control and respond to growth stimuli. Eighteen rainbow trout were stocked into eighteen individual 30L tanks provided with flow thru well water. Each fish was randomly assigned to one of three treatments. Six fish were fed to satiation twice daily, another six were pair fed to half-satiation, and the remaining six were fasted during the trial. Each fish was sacrificed and sampled to remove liver, serum, and muscle. Tissues from all fish were identically processed and assayed to determine total protein in each 1 ml sample volume. Equal amounts of protein were loaded on to 1D gel strips and separated based on isoelectric point. Each gel strip was then equilibrated and proteins were run onto a 2D gel and separated based on size. Proteomic expression and tissue profile data will be presented in full detail.

TEMPORAL SEQUENCE OF GROWTH-RELATED ENDOCRINE CHANGE FOLLOWING AN ANABOLIC STIMULUS IN COHO SALMON Beckman, B.R., and Shimizu, M. Symposium: Fish Growth Presentation: Oral Contact: Brian Beckman, NWFSC, NOAA, 2725 Montlake Blvd E Seattle WA 98112 USA E-Mail: [email protected] Abstract: We conducted an experiment to assess the effects of a meal, following a period of fasting, on the endocrine growth axis. Juvenile coho salmon were fasted for one day, one week or three weeks and then fed a meal. Blood was sampled at four hour intervals, starting at six hours before the meal, and continuing for another 26 hours. Plasma levels of IGF-I, insulin, growth hormone, IGFBP1 and 41 kDa IGFBP were measured from each time point. Insulin levels rapidly increased following the meal and plasma levels were significantly higher than controls throughout the time course. Changes in plasma IGF-I were minor in fed fish and non-significant in fasted fish while a meal resulted in significantly decreased plasma GH levels in fasted fish. Plasma 41 kDa IGFBP levels increased following the meal while IGFBP1 levels decreased. These results present insights into the endocrine regulation of metabolism and growth in fishes.

GROWTH FACTORS OF TELEOST MACROPHAGES Belosevic, M. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Mike Belosevic, CW-405 Biological Sciences Building, University of Alberta Edmonton AB T6G2E9 Canada E-Mail: [email protected] Abstract: Macrophage growth factors (MGFs) regulate mononuclear cell proliferation, differentiation and survival. Their functions are well documented in mammals, however, little is known about MGFs of lower vertebrates. I report on the molecular and functional analysis of three macrophage growth factors and their receptors that we recently identified in the goldfish: macrophage colony stimulating factor-1 (CSF-1) and its receptor CSF-1R, granulin and leukemia inhibitory factor (LIF) and its receptor LIFR. Collectively, the teleost growth factors and their receptors were found to be highly expressed in the hematopoietic tissues of the goldfish and developing embryos and adult zebrafish. Recombinant MGFs produced in a variety of expression systems, induced proliferation, differentiation and pro-inflammatory responses in different subpopulations of developing teleost macrophages in a dose-dependent manner. Although the baseline MGF-controlled functions appear to be highly conserved there are differences in the regulation of MGF-induced responses between mammals and teleosts. [Supported by NSERC Canada]

IDENTIFICATION AND CHARACTERIZATION OF A FC RECEPTOR HOMOLOG IN THE CHANNEL CATFISH (ICTALURUS PUNCTATUS) Bengten, E., Nayak, N., Stafford, J.L., Quiniou, S., Miller, N.W., and Wilson, M. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Eva Bengten, Department of Microbiology, University of Mississippi Medical Center, 2500 North State Street Jackson MS 39216 USA E-Mail: [email protected] Abstract: A catfish Fc receptor homolog (IpFcRI) was identified by using mammalian FcR sequences to mine catfish expressed sequence tag databases. The single copy IpFcRI gene encodes for a protein of three immunoglobulin (Ig) domains, lacking transmembrane and cytoplasmic regions. IpFcRI-related genes are also present in pufferfish, Fugu rubripes, and rainbow trout, Onchorynchus mykiss. The encoded Ig domains are phylogenetically and structurally related to mammalian FcRs and the putative Fc-binding region appears conserved. IpFcRI is highly expressed in lymphoid tissues and peripheral blood leukocytes. In contrast, catfish clonal leukocyte cell lines express lower levels of message. Importantly, IpFcRI binds catfish IgM as assessed by co-immunoprecipitation and cell transfection studies and the native receptor was detected in

Fish Biology Congress Abstracts 9 catfish plasma using a polyclonal antibody. Mapping studies demonstrate that IpFcRI binds IgM within Cmu3 and Cmu4 domains. Thus, IpFcRI appears to function as a secreted FcR akin to the soluble FcRs identified in mammals.

SELENIUM SUPPLEMENTATION INFLUENCES BEHAVIOR AND GENE EXPRESSION IN ZEBRAFISH (DANIO RERIO) Benner, M.J., Drew, R.E., Hardy, R.W., Robison, B.D. Symposium: Social Behavior Models Presentation: Poster Contact: Maia Benner, University of Idaho, Department of Biological Sciences Moscow Idaho 83844-3051 USA E-Mail: [email protected] Abstract: Selenium is critical to normal brain function, and can influence patterns of behavior. We tested the effect of two forms of dietary selenium supplementation on place preference behaviors and expression of key selenoprotein genes in the brains of adult zebrafish. We used four distinct strains in the study and tested whether behavioral and transcriptional responses varied among populations. When fed a normal diet, the strains differed significantly in their behavior and in expression levels of glutathione peroxidase 3, thioredoxin reductase 1, and selenoprotein P 1a. Expression of deiodinase 2 was highly elevated in male brains relative to female brains. Dietary supplementation with selenium significantly increased observer preference behavior and expression of tRNA selenocysteine associated protein 1. Supplementation also affected depth preference behavior and expression of selenoprotein P 1a, but these effects varied by strain. These data are the first evidence that behavioral effects of selenium supplementation may differ among populations.

THE ROLE OF MYOSTATIN IN THE GROWTH AXIS OF INDETERMINATE AND DETERMINATE GROWING FISH. Biga, P.R. and Meyer, J. Symposium: Fish Growth Presentation: Oral Contact: Peggy Biga, North Dakota State University, 1340 Bolley Drive, Stevens Hall 218 Fargo ND 58105 USA E-Mail: [email protected] Abstract: Myostatin (MSTN) is known to function as a negative regulator of muscle growth in mammals. Removing functional MSTN results in increased muscle mass and altered energy partitioning in rodents. It is still unclear if MSTN exhibits similar functions in fish. However, evidence suggesting a similar role in muscle growth regulation in fish is appearing. It has been demonstrated that growth hormone treatment in rainbow trout decreases muscle MSTN-2 expression (74%), suggesting a negative regulatory role. Also, in trout and giant danio primary myoblast cells, MSTN gene expression is lowest during rapid cellular proliferation and increases as cell differentiate into myofibers. In determinate and indeterminate growing species that are closely related, zebrafish and giant danio respectively, MSTN is differentially expressed in response to GH. These results suggest that MSTN plays a role in muscle growth regulation and might be important in satellite cell fate in indeterminate growing fish.

MORE THAN MEETS THE EYE: THE SENSORY BASIS OF LIGHT AVOIDANCE IN UPSTREAM MIGRATING SEA LAMPREYS (PETROMYZON MARINUS) Binder, T.R., and McDonald, D.G. Symposium: Biology of Lampreys Presentation: Oral Contact: Tom Binder, Department of Integrative Biology, University of Guelph Guelph Ontario N1G 2W1 Canada E-Mail: [email protected] Abstract: Lampreys possess dermal photoreceptors that are critical to light avoidance in blind ammocoetes, but which seem redundant in adults because adult lampreys also have well-developed, fully functional eyes. We conducted a series of field and laboratory experiments to examine the relative contributions of the eyes and dermal photoreceptors to light avoidance in upstream migrating sea lampreys. Blinded lampreys showed no reduction in their willingness or ability to avoid light, in either the laboratory or field. Furthermore, their response to sudden tail illumination matched the response to whole-body illumination, suggesting that the tail photoreceptors continue to mediate light avoidance in adults. This was confirmed in subsequent experiments, which found that temperature-induced changes in light avoidance corresponded with a reduction in dermal photosensitivity. We conclude that light avoidance in upstream migrants continues to be mediated by dermal photoreceptors, rather than the eyes. This indicates that there is functional specialization between the two light detecting systems in lampreys.

MINIMAL EFFECTS OF THE LAMPRICIDE 3-TRIFLUOROMETHYL-4-NITROPHENOL (TFM) ON ION BALANCE IN THE RAINBOW TROUT (ONCORHYNCHUS MYKISS) AND LARVAL SEA LAMPREYS (PETROMYZON MARINUS)

Fish Biology Congress Abstracts 10 Birceanu, O., McClelland, G. B. and Wilkie, M. P. Symposium: Biology of Lampreys Presentation: Poster Contact: Oana Birceanu, Wilfrid Laurier University, 75 University Ave E Waterloo ON N2L 3C5 Canada E-Mail: [email protected] Abstract: Despite its effectiveness for sea lamprey population control in the Great Lakes, the mode(s) of toxicity for 3- trifluoromethyl-4-nitrophenol (TFM) remain unclear. One hypothesis states that by disrupting gill-mediated ion uptake, TFM exposure may deplete internal ions (Na+,Cl-) and lower plasma osmolarity, leading to circulatory collapse and death. A second hypothesis is that TFM inhibits mitochondrial ATP production via oxidative phosphorylation, causing fuel depletion and death. We tested each hypothesis in larval sea lampreys, and non-target rainbow trout. Exposure to their respective 12h LC50s for TFM resulted in a 60% decrease in Na+-uptake by trout, but not lamprey. However, whole body and plasma ion concentrations, haemoglobin and haematocrit, and mean cell haemoglobin concentration were unaffected. Greater blood and tissue lactate concentrations in both species suggested that TFM interfered with fuel production. We conclude that disturbances to ion balance do not explain TFM toxicity in either trout or lamprey.

DIGESTIVE ENZYME ACTIVITIES IN THE WOOD-EATING CATFISH, NOCTURNUS Bittong, R.A., German, D.P., Evans, D.H., and Miles, R.D. Symposium: Tropical Fish Adaptations Presentation: Oral Contact: Donovan German, Department of Zoology, University of Florida, 321 Bartram Hall Gainesville FL 32611-8525 USA E-Mail: [email protected] Abstract: We measured the activities of nine digestive enzymes in the guts of wild-caught Panaque nocturnus, a wood-eating catfish from Peru. Enzyme activities were measured from the hepatopancreas (representing fish enzymes), pelleted gut contents (representing microbial enzymes), gut fluid (representing fish and microbial enzymes), and rinsed gut wall (representing fish enzymes). Cellulolytic and xylanolytic activities were low and largely restricted to the pelleted gut contents of the proximal intestine. Beta-glucosidase activity was found in the pelleted gut contents and in the gut wall of the fish. However, the Michaelis-Menten constant (Km) of beta-glucosidase from the gut wall was an of magnitude lower than that of the gut contents, indicating that the fish more efficiently digest some disaccharides than microbes in the gut contents. The distally decreasing cellulase and xylanase activities in the intestine suggest that these enzymes arise from microbes ingested with wood and detritus rather than an endosymbiotic community.

ONTOGENY AND EXPRESSION OF THE CFTR CHLORIDE CHANNEL IN THE IONOCYTES OF THE EUROPEAN SEA-BASS DICENTRARCHUS LABRAX ACCORDING TO SALINITY. Bodinier, C., Boulo, V., Lorin-Nebel, C., Charmantier, G. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Charlotte Bodinier, UMR 5119, eq. AEO, cc092, UM2, place E. bataillon, Université Montpellier 2 Montpellier 34095 France E-Mail: [email protected] Abstract: The function of CFTR was studied using cellular and molecular approaches in different organs during the ontogeny of the European sea-bass, a euryhaline teleost. At hatch, CFTR is in the digestive tract and tegument, suggesting ion excretion. During ontogeny, the secretion site shifts from tegument to gills. In the kidney, the CFTR expression is transitory before metamorphosis when juvenile-like physiological abilities are acquired. In sea water (SW), in the posterior intestine, the CFTR may facilitate ionic absorption, perhaps in relation to water uptake through aquaporins. Excess ions are excreted through the gills. In fresh water (FW), CFTR is apical. The expression of protein and transcript of CFTR were followed after a transfer from SW to FW. In gills and intestine, both expressions decrease, in relation to a lower chloride efflux. These changes in CFTR expression according to salinity constitute one of the adaptive mechanisms for sea-lagoon migrations.

GENETIC DIVERSITY, ENDEMISM, AND BIOGEOGRAPHY OF THE WESTERN BROOK LAMPREY (LAMPETRA RICHARDSONI) Boguski, D.A., Reid, S.B., Goodman, D.H., and Docker, M.F. Symposium: Biology of Lampreys Presentation: Oral Contact: Dave Boguski, Biological Sciences, University of Manitoba, Duff Roblin Bldg, 190 Dysart Rd. Winnipeg Manitoba R3T 2N2 Canada E-Mail: [email protected] Abstract:

Fish Biology Congress Abstracts 11 Lampreys are among the most ancient vertebrates and the least understood. There are 38 described species of lampreys worldwide and approximately half are non-parasitic, non-migratory brook lampreys. However, the true number of brook lamprey species may be underestimated since isolated brook lamprey populations that occur over broad geographic ranges are often considered the same species. Classification based on morphology alone may be misleading. We therefore compared mitochondrial and nuclear ribosomal DNA sequence among 42 brook lamprey populations distributed along the Pacific coast from Alaska to California (including Western Brook Lamprey, L. richardsoni; Pacific Brook Lamprey, L. pacifica; and Kern Brook Lamprey, L. hubbsi). Pacific and Kern brook lampreys were both genetically distinct, but the greatest sequence divergence (more than 6%) was observed among putative Western Brook Lamprey populations. We will discuss morphological convergence and speciation in brook lampreys and discuss whether such genetic differences warrant species designation.

PROGLYCOGEN AND MACROGLYCOGEN IN CARDIAC, HEPATIC, AND WHITE MUSCLE TISSUE IN MALE AND FEMALE RAINBOW TROUT (ONCORHYNCHUS MYKISS) Bolinger, M.T., and Rodnick, K.J. Symposium: Contributed Papers Presentation: Poster Contact: Ken Rodnick, Dept. of Biological Sciences, 650 Memorial Drive, Idaho State University Pocatello Idaho 83209- 8007 USA E-Mail: [email protected] Abstract: Glycogen appears to be an important source of energy in teleost tissues, and mammalian studies suggest that glycogen exists in two pools - proglycogen (PG) and macroglycogen (MG) - that can be distinguished by acid solubility. PG is smaller, protein-associated, and acid-precipitable whereas MG is larger and acid-soluble. We measured glycogen pools using acid and enzymatic hydrolysis in various tissues from immature rainbow trout of both sexes. Ventricles, livers, and white muscle were sampled from chemically-anesthetized fish due to rapid activation of glycogenolysis in white muscle by contraction. Total glycogen (TG) was much higher in liver than ventricle and white muscle. MG was more abundant than PG in liver, but was similar in ventricles and white muscle. Female ventricles did store proportionately more MG than males. Tissue and sex-specific differences in MG vs. PG pools may reflect differences in rates of anaerobic vs. aerobic metabolism, and alter cardiac contractile activity. Supported by NSF grant IOB-517669.

NRAMP, TNF, TLR5, AND HEPCIDIN EXPRESSION IN RESISTANT AND SUSCEPTIBLE FAMILIES OF CHANNEL CATFISH FOLLOWING CHALLENGE WITH E. ICTLAURI Booth, N. J. and Peterson, B. C. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Natha Booth, 141 Experiment Station Rd., P.O. Box 38 Stoneville MS 38776 USA E-Mail: [email protected] Abstract: Real-time PCR was used to measure gene expression of Nramp, TNF, TLR5, and Hepcidin, in spleen and liver tissue from two families of channel catfish, one resistant and one susceptible to ESC, following challenge with Edwardsiella ictaluri. There were no significant differences in relative copy numbers of the four genes in either of the families at any time point. There were no significant differences in copy number of TNF or Hepcidin in uninfected control fish. TLR5 (P<0.01) and Nramp (P<0.0001) expression was significantly different in uninfected fish only at 6 hours post exposure (PE). Fold induction was similar in both families for Nramp and TLR5; however, Hepcidin demonstrated strong induction in the susceptible family at 72 hours PE, while TNF was strongly induced at 48 hours PE in the resistant fish. These results give insight into innate immune responses and suggest potential candidates for genetic markers of disease resistance.

ENDOCRINOLOGY OF CATCH-UP GROWTH FOLLOWING SEASONAL COLD-BANKING IN HYBRID STRIPED BASS (MORONE CHRYSOPS X M. SAXATILIS) Borski, R.J., Picha, M.E., Strom, C.N., Won, E.T., Baltzegar, D.A., Johnstone, W.M. Symposium: Fish Growth Presentation: Oral Contact: Russell Borski, Department of Zoology, Box 7617 Raleigh NC 27695-7617 USA E-Mail: [email protected] Abstract: Many fish species exhibit compensatory growth (CG), but few studies demonstrate complete catch-up growth. We investigated CG in hybrid striped bass (HSB) following prolonged cold-banking and complete feed restriction. HSB achieved complete catch-up growth upon return to optimal water temperatures and satiation feeding, with the response characterized by hyperphagia, improved feed conversion and growth rates 3.4X higher than controls. Skeletal length in treatment fish was similar to continuously fed controls following cold-banking and feed restriction, suggesting a possible criterion of complete CG. Plasma insulin-like growth factor-I (IGF-I) paralleled changes in growth with both feed

Fish Biology Congress Abstracts 12 restriction and temperature, while circulating GH showed an opposite response, indicating a possible GH resistance or downregulation of GH receptor during catabolism and upregulation during CG. IGF-II may also play a functional role in regulating accelerated growth characteristic of CG. Potential endocrine regulation of hyperphagia and the CG response by leptin will be discussed.

VARIATION IN GENE-EXPRESSION IN TEMPERATURE STRESSED SEXUALLY-SELECTED VERSUS HATCHERY-BRED CHINOOK SALMON Bortoluzzi, R.B., Aykanat, T., Heath, J.A., and Heath, D.D. Symposium: Stress Effects and Response Presentation: Oral Contact: Romina Bortoluzzi, Great Lakes Institute for Environmental Research, University of Windsor, 401 Sunset Avenue, Windsor, Ontario N9B 3P4 Windsor ON N9B 3P4 Canada E-Mail: [email protected] Abstract: Under sexual selection theory, female choice and male-male competition for mates results in non-random mating. Previous studies indicate that sexual selection can increase genetic variance in offspring, possibly through inbreeding avoidance. Our experiment compares gene transcription in unstressed and temperature-stressed fish from sexually selected and randomly hatchery-mated Chinook salmon. The stressed fish were subjected to 12 hours at ~ 6 degrees above ambient temperature, and gill tissue was collected for subsequent RNA extraction. Specific genes were identified using a custom Chinook salmon microarray and quantitative real-time PCR was used for differential expression analyses. Differences in the mean or variance in gene transcription may be attributable to the differential effects of sexual selection versus hatchery breeding, although environmental effects of rearing may also contribute. This information will be useful in Pacific salmon breeding programs, and contribute to our understanding of the evolution of sexual selection in Pacific salmon.

EXPRESSION OF IGF-I E PEPTIDES IN RESPONSE TO FEEDING IN ATLANTIC SALMON (SALMO SALAR) Bower, N.I., Taylor, R., and Johnston, I.A. Symposium: Fish Growth Presentation: Poster Contact: Neil Bower, Gatty Marine Laboratory, School of Biology, University of St Andrews St Andrews Fife KY168LB UK E-Mail: [email protected] Abstract: IGF-I plays an important role in growth, differentiation and metabolism in vertebrates. In salmonids, IGF-I mRNA has been shown to be alternatively spliced, producing four different IGF-I prohormones which differ in their E peptide region. The E peptides of rainbow trout have mitogenic activity in mammalian cell lines, and so their expression patterns in fish muscle warrants attention. We have cloned, sequenced and designed primers that specifically amplify the Ea1, Ea2, Ea3 and Ea4 mRNA sequences from Atlantic salmon (Salmo salar). Using quantitative PCR, we have determined that Ea4 and Ea3 are the most abundant transcripts in salmon muscle and liver. In fasted fish, expression of all muscle transcripts increased in response to feeding with significant differences after 3d. Expression of all transcripts initially declined in liver in response to feeding. The changes in expression suggest that IGF-I signalling in muscle following refeeding acts through autocrine rather than endocrine pathways.

EFFECTS OF POLLUTION ON IMMUNE FUNCTION AND DISEASE SUSCEPTIBILITY IN SALMONID POPULATIONS: A REVIEW OF FIELD AND LABORATORY STUDIES AND FUTURE DIRECTIONS Boylen, D.A., Collier, T.K. and Arkoosh, M.R. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Deborah Boylen, DOC NOAA Fisheries, NW Fisheries Science Center, Environmental Conservation Division, 2032 SE OSU Drive Newport OR 97365 USA E-Mail: [email protected] Abstract: The decline of wild Pacific salmon has been linked to numerous anthropogenic factors including pollution. Exposure to toxic xenobiotics can lead to immunosuppression and increased disease susceptibility in juvenile salmon. Studies of juvenile salmon collected from a contaminated urban estuary, demonstrated that exposure to contaminants suppresses immunological memory. Laboratory studies with intraperitoneal injection of contaminated sediment extracts and model contaminant mixtures indicate that non-lethal xenobiotic exposure increases disease susceptibility in juvenile salmon. Field surveys conducted in numerous estuaries of the Pacific Northwest have shown that both contaminants and salmonid pathogens co-occur in these habitats. These studies show that pollutant-associated immunodysfunction in juvenile salmon

Fish Biology Congress Abstracts 13 may lead to increased susceptibility to infection and disease-induced mortality. Ongoing research focuses on the immunological impacts of environmentally relevant contaminant exposures and the potential synergy of chemical mixtures.

THE ROLE OF MYOSTATIN AND ACTRIIB IN MUSCLE DEVELOPMENT OF FISH Bradley, T., Phelps, M. and Mobley, E. Symposium: Fish Growth Presentation: Oral Contact: Terence Bradley, Dept. of Fisheries, and Veterinary Science, University of Rhode Island, Bldg #14, East Farm, Route 108 Kingston RI 2881 USA E-Mail: [email protected] Abstract: Myostatin (GDF-8) has been demonstrated to be an important regulator of muscle development in mammals but its role in fish remains unclear. To investigate the function of myostatin in fish, transgenic rainbow trout (Oncorhnycus mykiss) expressing a mutated form of the myostatin protein lacking a cysteine at residue 311 (C311Y) or a truncated activin RIIb receptor sans the kinase domain (ActRIIb) were produced. Both constructs were under the control of a muscle specific promoter and expression in muscle tissue was confirmed by QRT-PCR. Individuals expressing high levels of ActRIIB exhibited a unique phenotype, dubbed “six-pack”, in which the abdominal wall and the dorsal region were excessively muscled. Although growth rates of transgenic individuals varied, several transgenic fish were significantly larger than controls raised in the same tank. Comparison of the two transgenic strains suggests that myostatin may not play as significant a role in muscle development and growth in fish as it does in mammals and that other regulatory factors may be of greater importance. This project was supported by National Research Initiative Competitive Grant no. 2003-35206- 13506 from the USDA Cooperative State Research, Education, and Extension Service.

MIGRATION BEHAVIOR AND SURVIVAL OF KELT STEELHEAD (ONCORHYNCHUS MYKISS) Branstetter, R., D.R. Hatch, J. Whiteaker, B. Bosch, D. Fast, and J. Blodgett Symposium: Physiological BioTelemetry Presentation: Oral Contact: Ryan Branstetter, 729 NE Oregon St. Suite 200 Portland OR 97232 USA E-Mail: [email protected] Abstract: All wild steelhead populations originating above Bonneville Dam on the Columbia River are listed under the U.S. Endangered Species Act. A promising approach to effectively increase natural production of wild steelhead is to capitalize on their iteroparous life history strategy by capturing fish during the early stages of their outmigration and transporting them around the migratory barriers. Prior to a full scale implementation of this type of program it is important to assess the migration behavior and estuary residence time to evaluate any potential impacts of releasing adult steelhead into this environment. We evaluated two steelhead management strategies by using hydroacoustic telemetry to track individuals and evaluate survival, travel time, and behavior in the lower 233km of the Columbia River. The experiment consisted of two treatment groups; one group that was collected, and immediately transported and released at rkm 233, and the second group that was collected, placed in a reconditioning facility and fed for 4-6 weeks and then transported and released at rkm 233.

THE DEVELOPMENT OF NITROGEN EXCRETION IN ZEBRAFISH Braun, M.H., Steele, S.L. and Perry, S.F. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Marvin Braun, Department of Biology, University of Ottawa, 30 Marie Curie Ottawa Ontario K1N 6N5 Canada E-Mail: [email protected] Abstract: While most adult teleosts rid themselves of nitrogenous waste by excreting ammonia directly to the environment, a different strategy could be required during embryogenesis, when the chorion may interfere with ammonia diffusion. The zebrafish embryo utilizes urea as the primary waste product early in life, with ammonia becoming more important as development continues. Microscopy and real time PCR reveal that while both ammonia (Rh proteins) and urea (UT) transporters are highly expressed within the embryonic gills, the onset of ammonotely correlates with an increased expression of Rh proteins, specifically Rhcg. The expression of these proteins is plastic and can be modified in response to phloretin or high external ammonia. The functional importance of these proteins can be observed in embryos where expression of the ammonia (Rhag, Rhbg, Rhcg) and urea (UT) transporter proteins have been inhibited; as they exhibit a reduction in the excretion of ammonia and urea, respectively.

CONSERVATION PHYSIOLOGY: SALINITY TOLERANCE OF THE ENDANGERED LAKE QINGHAI SCALELESS CARP, PRZEWALSKII. Brauner, CJ, Richards, JG, Matey, V, Wood, CM, Rogers, J, Murray, B , Chen, X-Q, Du, J-Z , and Wang, Y. Symposium: Tribute to Joe Cech, Jr. Presentation: Oral

Fish Biology Congress Abstracts 14 Contact: Colin Brauner, Department of Zoology, University of British Columbia, 6270 University Blvd. Vancouver BC V6T 1Z4 Canada E-Mail: [email protected] Abstract: Lake Qinghai, the largest lake in China, has historically supported a fishery for scale-less carp. Due to lack of outflow and evaporative water loss, the lake is saline (11 ppt) but due to water diversion for agricultural use, water level is progressively decreasing and salinity is increasing. At present, the fish are isosmotic with lake water, which is metabolically favorable relative to freshwater, but small increases in salinity (2 ppt) resulted in an osmoregulatory disturbance within 24h that was exacerbated over the following 48-72h. This was associated with 10% mortality and an additional 30% loss of equilibrium by 3 days. Minor changes in gill morphology and gill Na+,K+-ATPase were observed at higher salinities indicating that this species has a very limited ability to hypoosmoregulate. These data indicate that small changes in salinity of Lake Qinghai may have large consequences for the scale-less carp population (Funded by NSERC Canada and NSF China).

COMPETITION FOR ANTERIOR KIDNEY PLASMA CELL NICHES DETERMINES THE SUCCESS OF VACCINES Bromage, E.S. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Erin Bromage, University of Massachusetts Dartmouth, Biology Department, 285 Old Westport Road Dartmouth MA 2747 USA E-Mail: [email protected] Abstract: Vaccine trials are often conducted in controlled conditions, using a single immunogen, and the results from these studies can be promising. However, when the same vaccine is used in the field, the results are not as clear. What processes lead to the differences between these two trials? It is believed that competition within the anterior kidney (AK) for plasma cell (PC) survival niches is pivotal to the success or failure of the vaccine. For example, a single high dose vaccination generates many PC and in ‘clean’ laboratory conditions there is little pressure on the AK to harbor other PC. This results in a long-term humoral response due to the persistence of PC within the AK niches. However, in field trials ubiquitous environmental and pathogenic microbes may also induce PC generation which competes for these AK survival niches. This results in reduced magnitude and duration of the vaccine-induced humoral responses.

TRANSCRIPTOMIC PROFILING REVEALS A SURPRISING CAPACITY TO RESPOND TO HEAT IN COLD-ADAPTED ANTARCTIC FISH AND A POTENTIAL NEW BIOMARKER OF THERMAL STRESS. Buckley, B.A. and Somero, G.N. Symposium: Genomics of Adaptation Presentation: Oral Contact: Brad Buckley, Portland State University, Department of Biology, 1719 SW 10th Ave Portland OR 97201 USA E-Mail: [email protected] Abstract: During their evolutionary history spent in the near-freezing waters of the Southern Ocean, some Antarctic fishes have lost the well-conserved cellular response to thermal stress termed the “heat shock response”. Using cDNA microarrays designed from expressed sequence tags from a goby species, we generated transcriptomic profiles for a common Antarctic species, Trematomus bernachii, immediately following and during recovery from heat-shock, to determine what other broadly conserved patterns of heat stress may have been lost or retained in Antarctic fishes. Suprisingly, dozens of genes were found to be responsive to temperature, including genes involved in numerous cellular processes. One induced gene encodes an agent of cell cycle arrest, the transcription factor C/EBP-δ. The role of this gene as a heat-inducible mediator of cell cycle arrest is being explored, both as an potential indicator of thermal stress in the field but also as a possible link between hyperthermia and human cancer therapy.

GENETIC DIVERSITY AND HOST-SPECIFICITY OF KUDOID PARASITES IN FISHES OF THE GREAT BARRIER REEF Burger, M.A.A., and Adlard, R.D. Symposium: Genomics of Adaptation Presentation: Oral Contact: Mieke Burger, University of Queensland, Building 76 Saint Lucia Qld 4072 Australia E-Mail: [email protected] Abstract: The Kudoidae is a major but understudied family of myxosporean parasites that infect a wide range of fishes. They infect many different tissues and may be benign or highly pathogenic. This study surveyed fishes of the Great Barrier Reef to explore the richness of Kudoidae and their host-specificity in that system. The survey concentrated on pomacentrid fishes (185 individuals of 27 species) but incorporated examination of representatives of another five families. Kudoids were

Fish Biology Congress Abstracts 15 found in high prevalence in pomacentrids (up to 100%) and often there was more than one species infecting a single individual. Combined morphological and genetic analysis identified five species of Kudoa, three new. Of the five, two were restricted to pomacentrids; the other three were found in combinations of carangids, chaetodontids, labrids, scarids and serranids. This pattern of varying and unpredictable host-specificity means that sampling within fish populations must be broad if a true sense of host range and overall parasite richness is to be developed.

GILL NA+K+-ATPASE ISOFORM SWITCHING IN SALMONID FISHES DURING SALINITY ACCLIMATION Bystriansky, J.S., Schulte, P.M. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Jason Bystriansky, Department of Zoology, University of British Columbia, #2370-6270 University Blvd. Vancouver British Columbia V6T 1Z4 Canada E-Mail: [email protected] Abstract: The up-regulation of gill Na+K+-ATPase (the sodium pump) activity is pivotal for the successful acclimation of many euryhaline teleost fishes to increased salinity. We have shown in several studies that the observed increase in gill sodium pump activity and protein number during seawater acclimation of salmonid fishes is preceded by an increase in the mRNA expression of a specific isoform of Na+K+-ATPase, a1b, and coincides with the significant decrease in the message for the a1a isoform. We will discuss the importance of this 'isoform switching' in gills of several Salmonid species during acclimation to both seawater and freshwater and comment on how the independent regulation of these two Na+K+-ATPase isoforms may limit or enhance the seawater tolerance of certain species. The evolution of anadromy in the Salmonid family may be related to the expression and independent regulation of these gill Na+K+-ATPase isoforms, which appear to be specialized to work in different environments.

DEFINING ACQUIRED IMMUNITY TO FLAVOBACTERIUM PSYCHROPHILUM WITH IMPLICATIONS FOR DEVELOPMENT OF A COLDWATER DISEASE (CWD) VACCINE Cain, K.D., and LaFrentz, B.R. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Ken Cain, University of Idaho, Department of Fish and Wildlife and the Aquaculture Research Institute, CNR rm 105D Moscow Idaho 83844-1136 USA E-Mail: [email protected] Abstract: Fish immunized with various preparations of Flavobacterium psychrophilum have been shown to mount significant antibody responses. Furthermore, challenge experiments have confirmed that antibody-mediated protection is important and reduces mortality due to coldwater disease (CWD). Based on this, we have utilized anti-F. psychrophilum serum antibodies from trout to identify potentially important antigens for vaccine development. Following isolation of immunogenic bacterial antigens between approximately 70-100 kDa and immunization of trout, it was found that these antigens provided near complete protection with a relative percent survival (RPS) of 94%. Antigens within this region have been further characterized using an immunoproteomic approach, and genes encoding proteins of interest were identified and are being targeted for vaccine development. The above experiments highlight the importance of acquired immunity, but recent vaccination experiments utilizing a live attenuated F. psychrophilum strain suggests that cell-mediated or other immune factors may also contribute to disease protection.

TESTING THE "ADAPTIVE MODULATION HYPOTHESIS" IN A DETRITIVOROUS FISH Callahan, M.N., German, D.P., and Evans, D.H. Symposium: Tropical Fish Adaptations Presentation: Oral Contact: Donovan German, Department of Zoology, University of Florida, 321 Bartram Hall Gainesville FL 32611-8525 USA E-Mail: [email protected] Abstract: We measured the activities of nine digestive enzymes in the digestive tract of Pterygoplichthys disjunctivus, a detritivorous loricariid catfish, consuming three different food types: wild-caught fish on their natural diet, fish fed a sterile-wood diet in the laboratory, and fish fed a commercial algal diet in the laboratory. Thus, we were able to test the “adaptive modulation hypothesis” relating to dietary quality and fiber content. Amylase activities were the highest among all measured enzymes and did not change on the different diets. Xylanase activity disappeared on the wood and algal diets. Laminarinase activity increased on the wood diet. Maltase activities were the highest of all the measured brushborder enzymes, and trypsin activities were relatively high for a non-carnivorous fish. Overall, our results do not support the adaptive modulation

Fish Biology Congress Abstracts 16 hypothesis in P. disjunctivus, and indicate that soluble polysaccharides and proteins are the most important nutrient sources to these fish.

OSMO-IONIC ALTERATIONS IN PROCHILODUS LINEATUS AFTER ACUTE ALUMINUM EXPOSURE IN LOW PH Camargo, M.M.P; Fernandes, M.N. and Martinez, C.B.R. Symposium: Tropical Fish Adaptations Presentation: Oral Contact: Marina Mori Pires de Camargo, Federal University of São Carlos, Dept. of Physiological Sciences, São Carlos, SP, Brazil São Carlos São Paulo 13565-905 Brazil E-Mail: [email protected] Abstract: Recent studies determined values of dissolved Al in the Brazilian rivers higher than 0.1 ppm, determined as the limit in Brazilian guidelines. This metal is very toxic, especially in low pH. Therefore, the effects of Al were evaluated on a neotropical fish. Juveniles of P. lineatus were exposed for 24h to 0.1 ppm of Al in pH 5, to water with pH 5 or only to clean water (control). Plasma was used to measure sodium, chloride and osmolality. Gills Na+-K+/ATPase activity was determined and the imunohistochemistry was carried out to identify chloride cells. Animals exposed to Al showed decreased osmo-ionic parameters, Na+-K+/ATPase activity and chloride cells when compared to control. Results indicate that an acute exposure to Al alters the hyperegulation, probably due to damages in the chloride cells and also by decreasing the ionic uptake in the gills, jeopardizing the survival of fish.

IDENTIFICATION OF NOVEL SNPS IN CHINOOK SALMON AND VARIATION AMONG LIFE HISTORY TYPES Campbell, N.R., and Narum, S.R. Symposium: Genomics of Adaptation Presentation: Poster Contact: Shawn Narum, CRITFC, Hagerman Fish Culture Experiment Station, 3059-F National Fish Hatchery Rd. Hagerman ID 83332 USA E-Mail: [email protected] Abstract: In this study, we used EST sequences from rainbow trout (Onchorhynchus mykiss) to design primers for amplification of Chinook salmon (O. tshawytscha) genomic DNA fragments. Regions flanking the repeat sequence of published microsatellite loci in Chinook salmon were also chosen to examine nucleotide variation. Thirty-two individuals representing the majority of the species range from Russia to California were sequenced at 19 loci to identify SNPs. Fifty- eight polymorphisms were discovered during the screen and 13 were chosen for development into 5’ exonuclease assays (TaqMan assays). Genotypes from TaqMan assays were compared to sequence data from individuals from the ascertainment panel to confirm proper allele designations. Significant allele frequency differences were observed among life history types of Chinook salmon at seven SNP loci, demonstrating the power of these markers to differentiate lineages.

MYOGENIC REGULATORY FACTORS AND MYOSTATIN EXPRESSION IN SKELETAL MUSCLE OF PIRARUCU (ARAPAIMA GIGAS CUVIER 1817: OSTEOGLOSSIDAE, OSTEOGLOSSOIDEI, TELEOSTEI), DURING GROWTH. Carani, F.R., Carvalho, R.F., Almeida, F.L.A., Castan, E.P., Padovani, C.R, Martins, C. Dal-Pai Silva, M Symposium: Tropical Fish Adaptations Presentation: Poster Contact: M. Dal Pai-Silva, Departamneto de Morfologia, Instituto de Biociências de Botucatu, UNESP, Rubião Júnior, S/N Botucatu São Paulo 18618000 Brazil E-Mail: [email protected] Abstract: In most fish skeletal muscle growth occurs by hypertrophy and/or hyperplasia of muscle fibers, processes controlled by the Myogenic Regulatory Factors (MRFs) and the growth factor Myostatin. During giant fish growth stages, MRFs and Myostatin expression may control muscle celullarity and growth mechanisms which directly influence fillet quality. In this work we evaluated MyoD and Myostatin genes expression of skeletal muscle in pirarucu (Arapaima gigas), during alevin and juvenile growth phases. Fish were weighed and then sacrificed. White muscle samples were collected and frozen in liquid nitrogen. Muscle samples were used to determine MyoD and Myostatin genes expression by Polymerase Chain Reaction after Reverse Transcription (RT-PCR) and values normalized by constitutive gene 18S rRNA. MyoD expression in alevin was significantly higher than in juvenile whereas Myostatin expression was similar in both stages. These results reflect the muscle growth characteristics in this specie, where hyperplasia predominates in alevin and hypertrophy in juvenile.

Fish Biology Congress Abstracts 17 CRF1 RECEPTORS INFLUENCE STRESS COPING CHOICE IN A DICHOTOMOUS MODEL OF FEAR CONDITIONING IN ONCORHYNCHUS MYKISS Carpenter, Russ E., and Summers, Cliff H. Symposium: Social Behavior Models Presentation: Oral Contact: Cliff Summers, University of South Dakota, Dept. of Biology, 414 E. Clark St. Vermillion SD 57069 USA E-Mail: [email protected], [email protected] Abstract: Our newly developed model of fear conditioning offers a dichotomous choice for test fish exposed to the unconditioned stimulus (US) of aggression from a large fish. Reliably, half of the trout presented with this US escape through a hole to a neutral chamber, while half remain and become submissive. Presentation of the predictive conditioned stimulus (CS; water off) alone to submissive fish, following 7 days of paired CS-US training, elicits a significant increase in both plasma cortisol and limbic monoamine activity. We also predict that spatial and conditioned learning stimulate gene expression associated with neural plasticity in the dorsolateral and dorsomedial pallia respectively. Furthermore, after four days of training, treatment with the CRF1 receptor antagonist antalarmin reverses the stress coping strategy of submissive fish. That is, fish that remain to be submissive on days 1-3, when treated with antalarmin on day 4, choose to escape from aggression on days 5-7.

A NOVEL SODIUM UPTAKE MECHANISM IN THE BLACKSKIRT TETRA (GYMNOCORYMBUS TERNETZI): INSIGHT INTO ITS PH SENSITIVITY Casciato, A.N., Patrick, M.L., and Gonzalez, R.J. Symposium: Tropical Fish Adaptations Presentation: Poster Contact: Allison Casciato, Department of Biology, University of San Diego, 5998 Alcalá Park San Diego CA 92110 USA E-Mail: [email protected] Abstract: The blackskirt tetra (Gymnocorymbus ternetzi) is native to the ion-poor, acidic Rio Negro. When exposed to pH 4.5, its sodium balance is not disrupted, despite a stimulation of Na+ loss, due to a rapid stimulation of Na+ uptake. Protein expression (immunolocalization, Western blotting) and functional assays (in vivo Na+ flux measurements, ATPase activity assays) indicate that V-type H+ ATPase is neither present in the gill nor contributing to Na+ uptake. Instead, we observe high expression of Na+/K+ ATPase, and shortly after fish are transferred to pH 4.5 the location of this transporter shifts from lamellae to the more protected inter-lamellar spaces on the filaments. Protein expression assays confirm the presence of an epithelial Na+ channel (eNaC), and in vivo Na+ fluxes are quite sensitive to phenamil, an eNaC blocker. We propose a eNaC/-Na+/K+ ATPase coupled mechanism for Na+ uptake, that would explain how Na+ transport is relatively pH- insensitive.

JUVENILE SALMON ENERGETIC ESTIMATES IN A VEGETATED, SIMULATED FLOODPLAIN, VIA TBF – MO2 RELATIONSHIPS Cech, J. J. Jr., Cocherell, D.E., O’Rear, T.A., Soyster, G., Bandeh, H., Chen, Z.Q., Lorenzato, S., Carlon, J., and Kavvas, M.L. Symposium: Tribute to Joe Cech, Jr. Presentation: Oral Contact: Joe Cech, Jr., Department of Wildlife, Fish, and Conservation Biology, University of California, Davis, One Shields Avenue Davis California 95616 USA E-Mail: [email protected] Abstract: To address hypotheses regarding how juvenile Chinook salmon Oncorhynchus tshawytscha use floodplain vegetation at water velocities <1m/s, we conducted (day-time) experiments in a large (23-m) flume, containing a 10-m-long by 1.2-m- wide planted area of sandbar willows Salix interior, with 1.5-m water depth. Fish in the flume were videotaped to determine behavior and their tail-beat frequencies (TBF). Also, in a calibrated, Brett-type recirculating-flow respirometer, we measured the fish’s TBF and oxygen consumption rates (MO2) at relevant water velocities. From these relationships, we estimated the fish swimming velocities (checked with flow meter) and energetics of the videotaped salmon in the flume planted with sandbar willow. Salmon occupied vegetated microhabitats, which minimized swimming velocity and energy demands of holding station in the simulated floodplain at nominal velocities >45 cm/s, arguing for using vegetated floodplains, as potentially energy-saving habitat along their migratory paths. Research supported by CDWR, USEPA, & River Partners.

FISH CONSERVATION IN CALIFORNIA: PHYSIOLOGIAL/BEHAVIORAL APPROACHES Cech, J.J., Jr. Symposium: Plenary Presentation: Oral

Fish Biology Congress Abstracts 18 Contact: Joe Cech, Dept. Wildlife, Fish, and Conservation Biology, University of California Davis, 1 Shields Ave. Davis California 95616 USA E-Mail: [email protected] Abstract: Several stocks of native, California fishes, including salmon and sturgeon, are declining, and many are government-listed as “threatened” or “endangered.” Impounding rivers and diverting water for anthropogenic uses have altered natural flows and temperatures in these environments and have erected barriers to migratory fishes. By investigating effects of flow and temperature on physiological (e.g., swimming performance, muscle contraction/relaxation rates, stress responses) and physiological/behavioral (e.g., fish screen passage, fish ladder passage) variables, my students, colleagues, and I elucidated some of the mechanisms involved in these native, anadromous fishes’ abilities to persist in, or successfully migrate through, altered California rivers. Continued research, along with science-based alterations of water-management practices, will be needed for saving California’s native, anadromous fishes, as we face global-climate-change-related and other environmental challenges. Research funded by the California Bay-Delta Authority (CALFED), the Anadromous Fish Restoration Program (USFWS, USBR), and the California Resources Agency.

GONADOTROPIN-RELEASING HORMONE (GNRH) STIMULATION OF HORMONE SECRETION AND SYNTHESIS INVOLVES COMMON, AS WELL AS DISTINCT, KINASE SIGNALING SYSTEMS IN GOLDFISH PITUITARY CELLS. Chang, J.P., Klausen, C., Booth, M., and Habibi, H.R. Symposium: Fish Growth Presentation: Oral Contact: John P Chang, CW405, Biol. Sci. Bldg., University of Alberta Edmonton Alberta T6G 2E9 Canada E-Mail: [email protected] Abstract: The two endogenous goldfish GnRHs, salmon GnRH and chicken GnRH-II, increase maturational gonadotropin (GTH-II) and growth hormone (GH) release via protein kinase C (PKC)-dependent mechanisms. However, recent results indicate that PKC does not mediate GnRH-induced elevations in GTH subunits and GH mRNA levels. On the other hand, GnRH and PKC activators increased the ratio of phosphorylated to total extracellular signal-regulated kinase (ERK). Application of an ERK kinase (MEK) inhibitor, PD 98059, reduced basal and GnRH-stimulated increase in ERK phosphorylation, as well as GnRH-induced elevation in GTH subunit and GH mRNA level. In addition, PD reduced GnRH-elicited GTH-II, but not GH, secretion. These results indicate that MEK-ERK signaling mediates GnRH effects on GTH and GH gene expression, and GTH-II, but not GH, secretion. Interestingly, while PKC can also activate the MEK-ERK pathway, PKC only mediates GnRH-elicited GTH and GH release. (Supported by NSERC Canada Discovery Grants to JPC and HRH.)

ENVIRONMENTAL FACTORS WHICH MODULATE THE TIMING OF THE SPAWNING PERIOD IN TROPICAL FRESHWATER FISH Chellappa, S., Bueno, R.M.X., Chellappa, T., Val, V.M.F.A. and Chellappa, N.T. Symposium: Tropical Fish Adaptations Presentation: Poster Contact: Sathyabama Chellappa, Post-Graduate Programme in Aquatic Bioecology, Department of Oceanography and Limnology, Centre of Bioscience, Universidade Federal do Rio Grande do Norte (UFRN), Praia de Mãe Luiza, s/n, Natal, RN. Brazil. CEP: 59014-100. Natal Rio Grande do Norte 59078-970 Brazil E-Mail: [email protected] Abstract: The native fish fauna commonly found in the drainage basins of rivers and reservoirs of Latin America are representatives of the Neotropical region, and the ichthyofauna of the semi-arid Northeastern Brazil has not been fully investigated. This work reports on the reproductive ecology of native fish species in relation to rainfall and hydrological variables of the semi- arid reservoirs in Brazil. Pluviometric precipitation, temperature, pH, dissolved oxygen and electrical conductivity of the water were correlated to stages of maturation of fish gonads on a monthly basis over a period of one year. Rainfall seems to be the main environmental factor which modulates the timing of the spawning period of fish in semi-arid region. Construction of reservoirs without adequate facilities for fish migration has an adverse impact on the migratory fish species which are of commercial and ecological importance to semi-arid Northeastern Brazil.

COMPARISONS OF GENE EXPRESSIONS IN ZEBRAFISH (DANIO RERIO) BRAIN AND GILL AFTER COLD ACCLIMATION Chou, MY and Hwang, PP Symposium: Stress Effects and Response Presentation: Poster Contact: Ming-Yi Chou, ICOB, Academia Sinica, Taiwan Taipei 115 Taiwan E-Mail: [email protected] Abstract:

Fish Biology Congress Abstracts 19 Ambient temperature affects organisms comprehensively, however cold responses are different among tissues. Here, we adopt a transcript screening approach to explore and compare the cold responses in zebrafish gills and brain. Zebrafish were exposed to cold and the microarray was used to identify cold-induced genes. Principle component analysis of the gene expression profiles indicated that gills developed different strategies depending on the durations of cold exposure while brain remained more stable. Based on statistic and clustering analysis, gills were higher in protein metabolism and cell activity, while brain were more active in stress responses and detoxification during cold acclimation. We also extended the study on ionocyte- and isotocin neuron-related genes in gills and brain, respectively, and found these genes were broadly stimulated by cold, suggesting that cold activates specific physiological functions in different tissues. Taken together, our results provide molecular evidences to elucidate the differential acclimations to cold in zebrafish.

BRANCHIAL COEXPRESSION OF GILL NHE3 AND RHG IN THE MARINE LONG HORN SCULPIN (MYOXOCEPHALUS OCTODECEMSPINOSUS): DOES NA+/H+ EXCHANGE PLAY A ROLE IN AMMONIA EXCRETION ACROSS THE GILLS? Claiborne, J.B., Kratochvilova, H., Diamanduros, A.W., Phillips, J., Hyndman, K.A., Phillips, M., Hirose, S., Evans, D.H., and Edwards, S.L. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: James Claiborne, Department of Biology, 202 Georgia Avenue, Georgia Southern University Statesboro GA 30459 USA E-Mail: [email protected] Abstract: Na+/H+ exchangers are thought to be involved in acid excretion and Na+ uptake across the gills of fishes. High ambient [Na+] is the likely driving force for transfer of H+ to the water in marine species. Rh glycoproteins are postulated to facilitate the diffusive efflux of ammonia across the gills. We have studied the expression of NHE3 and Rhg’s in the gill of the marine sculpin. Rhag, Rhcg1, and NHE3 are strongly expressed on the apical membrane of gill chloride cells (also high in Na+/K+-ATPase). Rhbg, and Rhcg2 are also apparent in the gill epithelium. Exposure to higher external ammonia or intraperitoneal NH4Cl infusion caused an increase in gill NHE3 protein expression. We hypothesize that apical H+ efflux by NHE3 assists net ammonia excretion by acidifying the water near the cell membrane, thus enhancing concentration gradient driving ammonia efflux via Rhgs. Supported by NSF IOB-0616187 to JBC.

RELATING MIGRATION CHARACTERISTICS OF ADULT PACIFIC LAMPREY, ENTOSPHENUS TRIDENTATUS, TO HIGH SUMMERTIME TEMPERATURES Clemens, B.J., van de Wetering, S., and Schreck, C.B. Symposium: Biology of Lampreys Presentation: Oral Contact: Ben Clemens, Oregon Cooperative Fish and Wildlife Research Unit, Department of Fisheries and Wildlife, U. S. Geological Survey, 104 Nash Hall Oregon State University Corvallis OR 97331-3803 USA E-Mail: [email protected] Abstract: It is likely that the prolonged freshwater residency (~1 yr) of adult Pacific lamprey may make them reliant upon river temperature for cueing migration and maturation. We conducted three collaborative studies on the migration characteristics of adult Pacific lamprey, including 1) an aerial survey to track radiotagged lamprey, 2) a controlled temperature experiment in the laboratory, and 3) monitoring of maturation timing in the wild. Summertime river temperatures > 20 ºC were correlated with 1) cessation of upstream migration and associations with river mouths, 2) expedited maturation vis-à-vis significant decreases in body size in the laboratory, and 3) apparent synchronization of maturation timing to avoid > 20 ºC. We hypothesize that if spatial and temporal aspects of thermal regimes > 20 ºC expand, then the combination of decreased migration capacity and expedited maturation of Pacific lamprey may occur, which could disconnect spawn timing with optimal locations in the upper watershed.

DEVELOPING A MECHANISTIC UNDERSTANDING OF FISH MIGRATIONS BY LINKING TELEMETRY WITH PHYSIOLOGY, BEHAVIOUR, GENOMICS AND EXPERIMENTAL BIOLOGY Cooke, S.J., S.G. Hinch, A.P. Farrell, K. Miller-Saunders, D. Patterson, M. Donaldson, G. Crossin, K. Hanson, K.K. English, D. Welch, M. Shrimpton, G. Van Der Kraak, and R. Thomson Symposium: Physiological BioTelemetry Presentation: Oral Contact: Steven Cooke, Biology Dept., Carleton University, 1125 Colonel By Drive Ottawa ON K1Y 3X4 Canada E-Mail: [email protected] Abstract: Fundamental questions in migration biology are beginning to be addressed through the use of telemetry. However, telemetry alone will not yield a complete understanding of the migration biology of fish or provide solutions to problems such as identifying physical barriers to migration or understanding potential impacts of climate change. Telemetry can be

Fish Biology Congress Abstracts 20 coupled with other tools and techniques to yield new insights into animal biology. Using Fraser River sockeye salmon (Oncorhynchus nerka) as a model, we summarize the advances that we have made in understanding salmonid migration biology through the integration of disciplines including physiology, behaviour, functional genomics, and experimental biology. Only through integrative and interdisciplinary research will it be possible to understand the mechanistic basis of fish migrations and to predict and possibly mitigate the consequences of anthropogenic impacts.

BREATHING IN THROUGH THE GILL AND OUT THROUGH THE GUT - UNUSUAL GAS EXCHANGE AND ACID-BASE BALANCE IN FISH AT HYPERSALINITIES Cooper, C. A. Wilson, J. M. and Wilson, R. W. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Chris Cooper, University of Exeter Exeter Devon EX4 1PS UK E-Mail: [email protected] Abstract: A previously unrecognised mechanism that aids water transport in marine teleost fish results from bicarbonate secretion into the intestine and resultant precipitation of calcium and magnesium carbonates. Intestinal bicarbonate secretion requires matching acid excretion by the gills to maintain acid-base balance, which increases in proportion to external salinity. Furthermore, as bicarbonate is primarily derived from endogenous CO2 we predict that a proportion of the net CO2 excretion will occur via the gut, which challenges the dogma of CO2 excretion being performed almost exclusively by the gill. To understand how fish cope at hypersalinities, European flounder had their gut perfused in vivo with increasing [Ca2+] (10-90mM) or were acclimated to range of salinities (0-60 ppt). Osmoregulatory and acid-base balance related proteins, gas exchange and various blood parameters were measured. From the protein to the whole animal, we show how fish can regulate ionic, respiratory and acid-base disturbances caused by varying salinities.

STREAM QUALITY ASSESSMENT USING REAL-TIME BIOASSAYS, CHEMSITRY, AND MOLECULAR MARKERSOVER A RURAL-URBAN LAND-USE GRADIENT Corsi, S.R., Klaper, R, Weber, D.N., and Bannerman, R Symposium: Environmental Pollution Presentation: Oral Contact: Daniel Weber, Marine & Freshwater Biomedical Sciences Center, University of Wisconsin-Milwaukee, 600 E. Greenfield Ave. Milwaukee WI 53204 USA E-Mail: [email protected] Abstract: To understand causes of stream impairments due to composite anthropogenic influences two in-stream bioassays were used over a range of urban-rural influences. 1) Stream water was pumped to a mobile field station with separate aquaria for minnow male-female breeding pairs; 2) 6 individual minnow breeding pairs were contained in separate flow-through cartridges in a larger flow-through chamber. Sediment and water chemistry, hydrology, and habitat were tested and correlated to reproductive success. Liver (vitellogenin mRNA) and blood (estrogen, testosterone) were collected. Significant differences occurred across streams in levels of female vitellogenin and male reproductive behavior/success. Egg production assays and reproductive biomarkers were compared with water and sediment chemistry as well as biological and land use indices for these watersheds. Low DO was a controlling factor for egg production in many streams with levels falling below 5 mg O2/L in >1/3 of sites. Correlations were evident with some biotic and chemical indices.

PRELIMINARY DATA ON THE USE OF A TWO-CHANNEL IMPLANTABLE DOPPLER FLOW METER FOR RECORDING FLATFISH CARDIAC OUTPUT Costa, I.A.S.F. and Gamperl, A.K. Symposium: Physiological BioTelemetry Presentation: Poster Contact: Isabel Costa, Ocean Sciences Centre, Memorial University of Newfoundland St. John's NL A1C5S7 Canada E-Mail: [email protected] Abstract: Physiological biotelemetry holds great promise as a tool for monitoring fish under aquaculture conditions, and for assessing the impact of human activities on the marine environment given that some species (e.g. flounder) live in close association with the substrate and have very limited home ranges. Thus, we attached a dual-channel Doppler blood flow biotelemetry unit (Endosomatic Systems, Inc.) to the dorsal surface of Atlantic halibut and winter flounder, and measured cardiac output (Q) for up to 8 days post-surgery. With appropriate filtering, clear signals could be obtained in 5 C halibut fitted with 2.5 mm probes, and significant increases in Q and heart rate were apparent following a 15 sec. handling stress (50 and 32%, respectively), and after feeding. In contrast, suitable recordings could only be obtained from flounder fitted with 1.5 mm probes (at 3 C) while anaesthetized on the surgical table. While very promising, these results suggest that the present system has some limitations.

Fish Biology Congress Abstracts 21 ANOXIA TOLERANCE AND OXYGEN DEBT IN THE PACIFIC HAGFISH EPTATRETUS STOUTII Cox, G. and Farrell, T. Symposium: Stress Effects and Response Presentation: Poster Contact: Georgina Cox, H.R. MacMillan 2357 Main Mall UBC Vancouver BC V6T 1Z4 Canada E-Mail: [email protected] Abstract: Hagfish are known to tolerate severe hypoxia and even short term anoxia (Hansen and Sidell, 1983). This is a vital adaptation given that they are routinely exposed to such conditions as a result of their habitat and feeding strategies. However, anoxia tolerance in this species has not been systematically examined previously. Consequently, the purpose of this study was to investigate how long the Pacific hagfish, Eptatretus stoutii can tolerate prolonged anoxia. In addition, critical oxygen tensions during progressive hypoxia were established using respirometry. Lastly, oxygen consumption rate following varying periods of anoxic exposure was examined to assess the levels of oxygen debt.

PREVALENCE OF UROLITHIASIS IN FREE-RANGING DESERT TORTOISES, GOPHERUS AGASSIZII, IN JOSHUA TREE NATIONAL PARK. Crocker, C.E. Symposium: Tribute to Joe Cech, Jr. Presentation: Oral Contact: Carlos Crocker, Western University Of Health Sciences, College of Veterinary Medicine, 309 Second Street Pomona Ca 91766 USA E-Mail: [email protected] Abstract: The main objective of this project is to study the prevalence of urolithiasis (uroliths/cystic calculi; bladder stones) in free- ranging desert tortoises, Gopherus agassizii, in Joshua Tree National Park. The cause of urolith formation in tortoises is not known, but is believed to be intimately tied to hydration status and diet, supplies of both are labile in the desert. The shell of a tortoise is a potential space and the growth of uroliths may continue throughout the course of the animal’s life. Large uroliths (size of a softball) can interfere with normal physiological functioning in tortoises and pose a direct threat to survival. This proposed research will also quantify the spatial and temporal distribution of desert tortoises in Joshua Tree National Park, as well as document the time course for growth of uroliths (when presence is verified), via telemetry and serial radiography of radiotagged tortoises. It is hypothesized that geographical barriers, ambulation problems related to large cystic calculi, and food/water limitations in the park may lead to long-term stress associated with foraging, finding mates and shelter, and predator avoidance. Inability to forage efficiently and/or find suitable mates may negatively affect recruitment of juvenile tortoises into the population and increase mortality of adults.

EST-DERIVED SNPS REVEAL DIVERSIFYING SELECTION IN CANADIAN ATLANTIC SALMON (SALMO SALAR) POPULATIONS. Culling, M.A., Freamo, H., Patterson, K., Berg, P.R., Glebe, B., Lien, S., Moen, T., O’Reilly, P., and Boulding, E.G. Symposium: Genomics of Adaptation Presentation: Oral Contact: Mark Culling, Department of Integrative Biology, University of Guelph, 50 Stone Road E. Guelph Ontario N1G 2W1 Canada E-Mail: [email protected] Abstract: We investigated genetic evidence for local adaptation among Atlantic salmon parr (Salmo salar) populations. Seven wild populations were genotyped for 320 SNP markers, located in transcribed regions, to identify genomic regions potentially under different selection pressures in different rivers. Neutrality tests identified 19 SNP loci as outliers, suggesting that they are located in regions of the genome under divergent selection. Chamcook Lake, and Hammond and Serpentine river populations, had highly significantly different parr mark (camouflage) patterns when compared to populations from the other rivers. Highly significant differences were found in body shape among all populations, reared in a common environment. We found QTLs for early and late juvenile growth rate, condition factor, parr mark number and contrast, and 19/30 morphological traits that quantified body shape and fin position in transatlantic crosses using the same SNP markers. We are now investigating whether QTLs associated with smoltification and size are the same.

ECOPHYSIOLOGY OF THE CENTRAL MUDMINNOW, UMBRA LIMI, AN AIR-BREATHING FISH LIVING IN A VARIABLE ENVIRONMENT Currie, S., DeMille, M., Learner, A., Leblanc, D., Marks, C., Ong, K., Parker, J., Templeman, N., Tufts, B., Bagatto, B. and Wright, P. Symposium: Fish Living on the Edge Presentation: Oral Contact: Suzie Currie, Department of Biology, Mount Allison University Sackville NB E4L 1G7 Canada E-Mail: [email protected]

Fish Biology Congress Abstracts 22 Abstract: The central mudminnow, Umbra limi, inhabits ponds and ditches, submerging in the mud and transitions to air breathing when water and oxygen levels fall. Field measurements revealed diurnal variations in water oxygen (2-140% sat) and temperature (15-35ºC). Aerobic metabolic rate increased steadily and significantly with increasing temperature but decreased transiently in short-term hypoxia. In contrast, metabolism was maintained in air-exposed fish as indicated by constant CO2 excretion over time. Low (15ºC) or high (31ºC) water temperatures significantly increased urea excretion over and above ammonia excretion rates whereas short-term hypoxia resulted in reduced ammonia excretion rates, but not urea excretion. Exposure to high temperatures, aquatic hypoxia and air resulted in a significant induction of heat shock proteins signifying a robust cellular stress response. We conclude that U. limi generally maintain metabolic rate during low oxygen availability; they are ammoniotelic in O2 saturated water of moderate temperatures, but urea excretion dominates in hypoxia and temperature extremes.

FUNCTIONAL EXPRESSION OF AQUAPORIN WATER AND SMALL SOLUTE CHANNELS IN THE EEL (ANGUILLA ANGUILLA) Cutler, C.P., MacIver, B., Yin, J., Hill M.G., Zeidel, M.L. and Hill W.G. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Christopher Cutler, Georgia Southern University, Department of Biology, 202 Georgia Avenue Statesboro GA 30458-8042 USA E-Mail: [email protected] Abstract: Recently data concerning the physiology and immunohistochemical localization of aquaporins in several species of teleost fish has been published. However only a very limited amount of data is currently available concerning aquaporin function, and this is essential to a proper understanding of the role of these proteins. The present study aims to begin to rectify this situation. Four aquaporins (AQP1, AQP1dup, AQP3 and AQPe), previously characterized in the eel, have been cloned into a Xenopus expression vector. Complementary RNA (cRNA) generated from these plasmids was then microinjected in Xenopus oocytes. The function of the expressed eel aquaporins was measured using a swelling assay (H2O) or using radioactive solutes and compared to non-expressing control oocytes. Functions measured included water, glycerol and urea permeability, as well as inhibition of permeability due to incubation of oocytes in various metal ions or by changes in extracellular pH.

RAPID FIELD ASSESSMENT OF FISH MORBIDITY AND DELAYED MORTALITY USING EXTRINSIC REFLEX IMPAIRMENT Davis, M.W. Symposium: Stress Effects and Response Presentation: Oral Contact: Michael W. Davis, Fisheries Behavioral Ecology Program, Alaska Fisheries Science Center, NOAA Fisheries, Hatfield Marine Science Center, 2030 SE Marine Science Dr. Newport Oregon 97365 USA E-Mail: [email protected] Abstract: Fish capture and culture systems can include: commercial and sport capture; aquaculture; aquarium specimen collection, transport, and holding; stock enhancement; and laboratory and field research. Stress, morbidity, and mortality fitness outcomes in fish systems are associated with significant ethical, economic, and ecological consequences. Mitigation of system consequences requires identification of fish stressors and tracking of relationships between stressors and fitness outcomes. Measuring extrinsic reflex impairment in free swimming or restrained fish is a powerful method for predicting fish condition and can be correlated with fitness. Reflex impairment measurements are inexpensive, real time, ecologically relevant predictors of fitness that can be validated in controlled fishery experiments. Reflex impairment is an overlooked experimental system for analysis of the physiological and behavioral effects of stressors. Use of reflex impairment to predict morbidity and mortality could significantly improve monitoring and maintenance of health and production in all types of fish capture and culture systems.

FOOD AVAILABILITY ALTERS HYPOXIA RESISTANCE OF THE AMAZONIAN OSCAR (ASTRONOTUS OCELLATUS). De Boeck, G. Wood, C.M., Iftikar, F., Scott, G., Sloman, K.A., Almeida-Val, V.M.F., Val, A.L. Symposium: Tropical Fish Adaptations Presentation: Oral Contact: Gudrun De Boeck, Ecophysiology, Biochemistry and Toxicology Group, University of Antwerp - Department of Biology Groenenborgerlaan 171 Antwerp B-2020 Belgium E-Mail: [email protected] Abstract:

Fish Biology Congress Abstracts 23 The goal of our study was to unravel the mechanisms allowing a hypoxia tolerant Amazonian fish, the Oscar, to deal with the ion poor soft water under hypoxic conditions, both with or without access to food. Starvation protected against hypoxia, as seen by the lower Pcrit. As expected, Na+ uptake rates are lower in fed fish compared to starved fish, and are rapidly reduced under hypoxia. Contrary to expectations, ammonia excretion was lower in fed fish, as was K+ efflux. Hypoxia reduces ammonia and K+ efflux, indicating that transcellular permeability decreases under hypoxic conditions. Urea efflux was larger in fed fish, but turned down during hypoxia. Also urine flow was reduced under hypoxia, leading to a reduction in ion loss. In conclusion, ion uptake is compromised by hypoxia in Oscars, but successfully counteracted by decreased permeability and reduced ion loss. Starvation leads to increased ion losses, but improves hypoxia resistance.

THE AMAZONIAN CHALLENGE: COPING WITH LOW ION, OXYGEN AND FOOD LEVELS. De Boeck, G., Wood, C.M., Kajimura M., Scott G.R., Sloman K.A., Walsh P.J., Almeida-Val V.M.F., Val A.L. Symposium: Fish Living on the Edge Presentation: Oral Contact: Gudrun De Boeck, Ecophysiology, Biochemistry and Toxicology Group, Department of Biology, Groenenborgerlaan 171 Antwerp B-2020 Belgium E-Mail: [email protected] Abstract: Amazonian fishes face multiple environmental challenges. First, they live in very dilute, ion poor water. Ionoregulation is costly process in all freshwater fishes, and the dilute nature of the Amazonian waters may exacerbate these costs. But these fish commonly encounter hypoxia as well, when they enter the seasonally flooded jungle to feed and reproduce. Gills play an important role in ionoregulation as well as in oxygen uptake, and it can be expected that oxygen deprivation affects the capacity of the mitochondria rich gill cells to take up the necessary ions. These fish exhibit a high degree of parental care as well, protecting their nest vigorously, and it has been suggested that they use their mouth as shelter for their young when danger occurs. Under such conditions, it is not unlikely that food deprivation occurs, at least for limited periods of time. This would further challenge ion homeostasis since food is an important source of ions as well. The goal of our study was to unravel the mechanisms that allow a hypoxia tolerant Amazonian fish, the Oscar (acará-açu, Astronotus ocellatus), to deal with the ion poor soft water under hypoxic conditions, both with or without access to food. It was clear that ion uptake is compromised by hypoxia in Oscars, but successfully counteracted by decreased permeability and reduced ion loss. Starvation leads to increased ion losses, but improves hypoxia resistance (lower Pcrit).

THE PHYSIOLOGICAL IMPACTS OF DECOMPRESSION AND FIZZING ON SMALLMOUTH BASS DURING LIVE-RELEASE TOURNAMENTS DeMille, M.J. and B.L. Tufts Symposium: Stress Effects and Response Presentation: Poster Contact: Matthew DeMille, Queen's University, Department of Biology Kingston ON K0K1V0 Canada E-Mail: [email protected] Abstract: During an angling event, decompression occurs when a fish is rapidly brought to the surface from depth. The rapid change in pressure can cause significant alterations to a fish’s internal physiology, which includes tissue damage, anaerobic disturbance, and swim bladder expansion. Positive buoyancy and loss of equilibrium caused by swim bladder overexpansion can expose fish to a suite of biotic and abiotic stressors. Artificial swim bladder deflation, or fizzing, has been proposed as a technique to release built-up gas pressure within the swim bladder and allow decompressed fish to return to depth. Current experiments are underway to determine the factors that influence decompression during live- release tournaments. This research will also examine the role of fizzing as a possible technique to reduce the physiological disturbance in decompressed fish. Answering these questions will assist fisheries agencies in making sound policy decisions on the merit of fizzing as a management tool.

PROBING CONTAMINANT EFFECTS ON THE HPG AXIS OF LARGEMOUTH BASS BY EXPRESSION PROFILING Denslow, N.D. Symposium: Plenary Presentation: Oral Contact: Nancy Denslow, University of Florida, Department of Physiological Sciences and Center for Environmental and Human Toxicology Gainesville FL 32611 USA E-Mail: [email protected] Abstract: Largemouth bass reproduce seasonally in a semi-synchronous manner, making it possible to isolate each part of their cycle to determine normal ordered gene expression changes of estrogen receptors, LH, FSH, steroidogenic acute regulator protein among other genes. Environmental contaminants that behave as endocrine active compounds can alter expression of

Fish Biology Congress Abstracts 24 critical genes resulting in abnormal levels of endogenous plasma steroid hormones and reproductive dysfunction. Largemouth bass were exposed either subchronically through their diet to p,p’DDE or to dieldrin or to organochlorine pesticides in mesocosms built directly on contaminated muck farms. Alterations in gene expression in the liver and gonad were measured by custom microarrays, using a 15,000 gene array. Pathway analysis indicated that critical pathways involved in steroidogenesis, metabolism, gonad development, sex differentiation, cell growth and innate immune response, among others were altered by exposure to the contaminants. This research suggests exposure to organochlorine pesticides results in severe consequences for reproduction.

MICROARRAY AND PHENOTYPIC ASSESSMENTS OF TRANSGENIC AND DOEMSTICATED SALMONIDS. Devlin, R.H., Sakhrani, D., Tymchuk, W., Raven, P., Sundstrom, F., and Lohmus, M. Symposium: Genomics of Adaptation Presentation: Poster Contact: Robert Devlin, Fisheries and Oceans Canada West Vancouver BC V7V 1N6 Canada E-Mail: [email protected] Abstract: Domestication has been extensively utilized to develop growth-enhanced salmonids for aquaculture purposes. More recently, growth hormone (GH) gene transgenesis has also been explored as an approach to enhance growth rate and feed conversion efficiency. Domestication and transgenesis can affect many physiological and behavioural phenotypes in fish, including enhanced feeding motivation, altered nutrient metabolism, reduced predator awareness, altered swimming performance, and in some cases reduced disease resistance. Whereas we know the genetic basis for these changes in transgenic strains, we have little understanding of the genetic changes that have occurred during domestication, and in both cases it is not well known how physiological pathways have been modified to influence these alterations. We have examined hormone levels and gene expression patterns among wild type, domesticated, and GH transgenic strains using a cDNA microarray (GRASP 16K chip) Q-PCR analysis, and have found that many genes show analogous effects in domesticated and transgenic strains.

LONG-TERM CUTANEOUS IMMUNITY IN CHANNEL CATFISH AGAINST THE PARASITIC CILIATE ICHTHYOPHTHIRIUS Dickerson, H.W., Zhao, Z., and Findly, R.C. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Harry Dickerson, Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, D. W. Brooks Drive Athens GA 30605 USA E-Mail: [email protected] Abstract: Immunity against the parasitic ciliate Ichthyophthirius multifiliis is elicited following either exposure to live organisms or intracoelomic injection of major surface proteins, referred to as immobilization antigens (I-antigens). I-antigen-specific antibodies in the skin and mucus are involved in the adaptive immune response against the parasite. We have shown that antibody-secreting cells (ASC) and LPS-inducible B cells are present in skin of channel catfish. ELISPOT analysis demonstrates that fish immunized over 2 years previously respond to infection by increasing total ASC and I-antigen- specific ASC in skin at Days 7, 14 and 21 following infection. The number of I-antigen-specific ASC also increases in head kidney at Days 14 and 21. This shows that I-antigen-specific ASC are found in skin following infection at least 2 years after immunization. These fish remain protected against parasite challenge and we hypothesize that cutaneous antibodies produced by ASC in the skin are responsible for protection.

STRESS BIOMARKER DISCOVERY IN STURGEON USING PROTEOMICS Dietmar Kueltz Symposium: Tribute to Joe Cech, Jr. Presentation: Oral Contact: Dietmar Kültz, Physiological Genomics Group, Department of Animal Science, Meyer Hall, One Shields Avenue, University of California-Davis Davis CA 95616 USA E-Mail: [email protected] Abstract: California green sturgeon populations are threatened and white sturgeon are of commercial value making these fish important species for scientific study. In particular effects of environmental stress such as salinity, temperature, selenium, and methylmercury are of interest as they determine important life history traits of sturgeon, including longevity and fecundity. We have developed two proteomics approaches for stress biomarker discovery in sturgeon. These techniques are based on two-dimensional fractionation of sturgeon gill proteins, either by 2D gel electrophoresis or by 2D nano- ultraperformance liquid chromatography. Fractionated proteins are then identified and compared between stressed and unstressed samples by offline mass spectrometry. The basis and potential of these approaches is discussed.

Fish Biology Congress Abstracts 25

INCREASED DISEASE SUSCEPTIBILITY OF SUBYEARLING CHINOOK SALMON (ONCORHYNCHUS TSHAWYTSCHA) AFTER EXPOSURE TO A DIET CONTAINING AN ENVIRONMENTALLY RELEVANT CONCENTRATIONS AND CONGENER RATIOS OF THE FLAME RETARDANT POLYBROMINATED DIPHENYL ETHER (PBDES) Dietrich, J.P. Symposium: Immunity and Immunity and Disease Resistance Presentation: Oral Contact: Joseph Dietrich, NOAA Northwest Fisheries Science Center, Newport Research Station, 2032 SE OSU Drive Newport OR 97365 USA E-Mail: [email protected] Abstract: To determine the effects of polybrominated diphenyl ethers (PBDEs) on the immune system, subyearling Chinook salmon (Oncorhynchus tshawytscha) were fed two diets of PBDEs: (1) 1X-PBDE that reflected the concentration of PBDEs (0.1861 μgPBDEs/g food) found in the stomach contents of juvenile subyearling Chinook salmon from Lower Willamette River; and (2) 10X-PBDE that reflected a concentration ten times that found in these fish. The two concentrations of the diet were fed to the salmon for 40 days. Juvenile Chinook salmon fed the lX-PBDE diet were found to be more susceptible to the marine pathogen Listonella anguillarum than fish fed a control diet. The increase in disease susceptibility in the 1X-PBDE diet is hypothesized to be due to the effects of PBDEs on thyroid hormones that regulate immune system. However, fish fed the 10X-PBDE diet were not more susceptible to disease, potential hypotheses for this result will be presented.

FAST-START PERFORMANCE AND MUSCLE GROWTH AMONG GROWTH HORMONE (GH) TRANSGENIC AND WILD COHO SALMON (ONCORHYNCHUS KISUTCH). Dimoulas, P.M., Devlin, R.H., and Shadwick, R.E. Symposium: Fish Growth Presentation: Poster Contact: Peter Dimoulas, University of British Columbia, 3475-6270 University Blvd. Vancouver BC V6T1Z4 Canada E-Mail: [email protected] Abstract: Coho salmon that are transgenic for increased GH production grow significantly faster than wilds, dependent on a supply of sufficient rations. Furthermore, GH coho achieve lower sustained swimming speeds, throughout their life history, which raises the possibility of differences in muscle physiology. We investigated fast-start kinematics to address the maximum mechanical and behavioral performance among GH and wild coho at 15cm and 40cm fork lengths (FL). Since no mechanical differences have been found among muscles of other GH upregulated animals we did not expect mechanical differences among GH and wild coho axial muscle. Indeed, we found no mechanical differences among GH and wild coho at 15cm FL. But are they behaviorally distinct? Do they have they similar quantities or proportions of glycolytic (or even oxidative) muscle? We will report additional (including behavioral) components of fast-starts at 15cm and 40cm FL, as well as the growth patterns of their axial muscle.

PARTITIONING THE EFFECTS OF GROWTH HORMONE (GH) ON MUSCLE PHYSIOLOGY: IN VIVO FINDINGS AMONG GH TRANSGENIC AND WILD COHO SALMON (ONCORHYNCHUS KISUTCH). Dimoulas, P.M., Richards, J.G., Devlin, R.H., and Shadwick, R.E. Symposium: Fish Growth Presentation: Oral Contact: Peter M. Dimoulas, University of British Columbia, 3475-6270 University Blvd. Vancouver BC V6T1Z4 Canada E-Mail: [email protected] Abstract: Coho salmon transgenic for increased GH production grow significantly faster, than wilds, dependent on a supply of sufficient rations. While there are no mechanical differences during fast-starts, indicating similar burst potential among glycolytic axial muscle, they achieved lower maximum sustained swimming speeds than length-matched wilds. Do they differ in their sustained glycolytic or sprinting capacity? We measured kinematics and substrate usage among rested, sprinted (20s), and exhausted GH and wild coho at 20cm and 40cm fork-lengths (FL). We report that GH coho took significantly longer to exhaust at 20cm FL but not at 40cm FL. Moreover, at 20cm, GH coho sprinted and used substrates more slowly, but shared similar substrate levels at rest and exhaustion. We will report additional kinematics (at 40cm FL) and further analysis of their tail-beat. We suggest that glycolytic capacity may not be maintained as a result of the behavioral and physiological consequences of GH upregulation.

ODORANT-INDUCED CHANGES IN OLFACTORY RECEPTOR MRNA EXPRESSION IN SOCKEYE SALMON (ONCORHYNCHUS NERKA) AFTER IMPRINTING. Dittman, A.H., May, D, and Havey, M.A.

Fish Biology Congress Abstracts 26 Symposium: Contributed Papers Presentation: Poster Contact: Andrew Dittman, Northwest Fisheries Science Center, NOAA-Fisheries, 2725 Montlake Boulevard East Seattle WA 98112 USA E-Mail: [email protected] Abstract: Salmon are well known for their extraordinary homing migrations from oceanic feeding grounds back to their river of origin to spawn. These migrations are governed by olfactory discrimination of homestream odors that juvenile salmon learn (imprint to) prior to their seaward migrations. Our previous studies have suggested that one component of imprinting involves long-term sensitization of the peripheral olfactory system to specific odorants. In this current study, we examined changes in odorant receptor expression in the olfactory epithelium of sockeye salmon during imprinting to the odorant L- arginine. Fish exposed to arginine as juveniles demonstrated long-term memory formation for this imprinting odorant as evidenced by behavioral attraction as adults (P≤ 0.05; two-sample t-tests). Treatment groups that successfully imprinted also demonstrated increased expression in the olfactory epithelium (relative to arginine-naïve fish) of a putative arginine receptor mRNA.

INTEGRATING PHYSIOLOGICAL, BEHAVIOURAL, AND ENVIRONMENTAL INDICES TO UNDERSTAND MIGRATION FALL BACK AND MORTALITY DURING THE FRESHWATER PHASE OF THE SOCKEYE SALMON (ONCORHYNCHUS NERKA) SPAWNING MIGRATION IN THE FRASER RIVER, BRITISH COLUMBIA Donaldson, M.R., Hruska, K.A., Hinch, S.G., Patterson, D.A., Farrell, A.P., Shrimpton, J.M., Miller-Saunders, K.M., Robichaud, D., Hanson, K.C., English, K.K., and Cooke, S.J. Symposium: Stress Effects and Response Presentation: Oral Contact: Michael Donaldson, Fish Ecology and Conservation Physiology Laboratory, Ottawa-Carleton Institute of Biology, Carleton University,1125 Colonel By Drive Ottawa ON K1S5B6 Canada E-Mail: [email protected] Abstract: To gain greater insight into the interplay of physiology and behaviour in salmonid spawning migrations, we intercepted sockeye salmon (Onchorhynchus nerka) at the onset of the freshwater phase of their migration in the Fraser River, British Columbia. Individuals were biosampled for physiological indices of stress, osmoregulatory status, and energy content then gastrically implanted with radio telemetry transmitters to assess migration behaviour and fate (en-route mortality versus survival to spawning grounds). We found ~24 % of individuals fell back down-river following capture. Individuals that fell back were characterized by elevated hematocrit and plasma sodium concentrations and had slower migration rates. Individuals that died en-route to spawning grounds had slower migration rates, but the physiological or environmental variables measured here did not affect migration fate. We did not find a relationship between fall back and fate. We conclude that the factors that contribute to migration fate are multifactorial and future targeted research is required to continue to unravel the complex relationship between the physiology and behaviour of Pacific salmonid migration biology.

UNDERSTANDING THE MOLECULAR MECHANISMS BEHIND DIETARY IMMUNOSTIMULATION IN THE RAINBOW TROUT, O. MYKISS, USING PARALLEL TARGETED CDNA MICROARRAY AND ISH ANALYSIS Donate, C., Balasch, J.C., Callol, A., Travesset, R., Bobe, J., MacKenzie, S. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Carmen Donate, Departament Biol. Cel., Fisiol., i Immunologia, Unitat de Fisiologia Animal, Facultat de Ciencias Universitat Autonoma de Barcelona Bellaterra Barcelona 8193 Spain E-Mail: [email protected] Abstract: Immunostimulant diets (IS) are utilized in aquaculture in order to enhance the resistance of fish to disease and stress, however conflicting results have been published about the efficacy of IS. Using a salmonid-specific microarray platform enriched with immune-related genes and in situ hybridization (ISH), we investigated dietary acclimation in 4 immune- relevant organs in the rainbow trout (Oncorhynchus mykiss). IS-diet feeding significantly changed gene expression profiles and gene distribution in a tissue-specific manner: genes and functional GO categories involved in immunity were differently expressed in both head kidney and spleen, whereas at portals of entry, intestine and gills, significant changes in genes and functional groups related to remodeling processes and antigen presentation were observed. Furthermore, localization of hematopoietic, apoptotic, antimicrobial and antigen presenting genes revealed that one of the main effects of IS-diets in trout is the increase of genes involved in antigen recognition in epithelial cells of gills.

MOLECULAR TO ORGANISMAL RESPONSES TO SALINITY CHANGE IN THE LEOPARD SHARK Dowd, W.W., Kültz, D., and Cech, J.J. Jr.

Fish Biology Congress Abstracts 27 Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Wes Dowd, University of California, Davis, Meyer Hall, 1 Shields Ave Davis CA 95616 USA E-Mail: [email protected] Abstract: Juvenile leopard sharks (Triakis semifasciata) inhabit California estuaries throughout much of the year, where they may experience stressful changes in salinity. We exposed leopard sharks to gradual reductions in salinity (to 50-60%, 75-80% and 100% SW) in short-term (48 hours) and long-term (3 weeks) experiments, mimicking conditions likely to be experienced in the wild. We demonstrated a time lag in reduction of ion and urea concentrations, with a peak osmotic gradient between shark and water at 24 hours. Urea, but not ionic, concentrations decreased further by 3 weeks. Behaviorally, sharks increased their activity in the short-term, while in the long-term activity decreased by ~50%. We also examined molecular changes in the composition of the proteome of osmoregulatory tissues that may contribute to these observed patterns. Using proteomics and mass spectrometry, we identified proteins and pathways that are influenced by exposure to decreased salinity.

PROTEOME RESPONSES TO ANOXIA AND HYPOXIC PRECONDITIONING IN THE EPAULETTE SHARK Dowd, W.W., Renshaw, G.M.C., and Kültz, D. Symposium: Fish Living on the Edge Presentation: Oral Contact: Wes Dowd, University of California, Davis, Meyer Hall, 1 Shields Avenue Davis CA 95616 USA E-Mail: [email protected] Abstract: Epaulette sharks (Hemiscyllium ocellatum) withstand severe, episodic hypoxia and even anoxia at tropical temperatures. The reef platform around Heron Island, Australia, serves as a natural hypoxic preconditioning environment. We adopted a discovery-based approach to identify proteins/mechanisms involved in low oxygen tolerance. Using two-dimensional gel electrophoresis, we examined proteome changes in response to hypoxic preconditioning (1 or 2 sessions of ~5% oxygen saturation, 24 hours apart) and anoxia (1 or 2 sessions of <0.3% oxygen saturation, 24 hours apart) in epaulette shark tissues. Proteins that were regulated by anoxia exposure and/or hypoxic preconditioning were identified by MALDI- TOF/TOF mass spectrometry and mapped to molecular pathways using bioinformatics tools. Using these data, we tested the hypotheses that 1. the severity of low oxygen exposure influences patterns of protein abundance in relevant cellular pathways (e.g., cellular stress response, redox balance, metabolism), and 2. preconditioning leads to upregulation of compensatory mechanisms of low oxygen tolerance.

IGF SIGNALING IN A SMALL FISH Duan, C. Symposium: Fish Growth Presentation: Oral Contact: Cunming Duan, Department of Molecular, Cellular and Developmental Biology, University of Michigan, Nat Sci Bldg. Room 3065B, 830 N. Univ. St. Ann Arbor MI 48109 USA E-Mail: [email protected] Abstract: Insulin-like growth factors (IGFs) are conserved peptides that play key roles in regulating growth, development, reproduction, and aging. The biological actions of IGFs are mediated through the IGF-I receptor. In extracellular environment, IGFs are present in complexes with high-affinity binding proteins (IGFBPs). The in vivo physiological functions of these mammalian IGFBPs, however, are poorly understood. In recent years, there has been a remarkable acceleration in our understanding of the IGF signaling system in teleost fishes, most notably in zebrafish. To date, genes encoding for zebrafish IGF ligands, receptors, and several IGFBPs have been characterized, and their spatial and temporal expression patterns mapped. Many of these genes are duplicates in zebrafish and possibly other teleosts. Using genetic and biochemical approaches, the in vivo actions of zebrafish IGF ligands, receptors, and several IGFBPs in regulating embryonic growth and development have been elucidated. These results will be reported and discussed.

LOCALIZATION OF IONOCYTES IN CRUCIAN CARP (CARASSIUS CARASSIUS) GILLS DURING HYPOXIC REMODELLING Dymowska, A.K., Stecyk, J.A.W, Nilsson, G.E. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Agnieszka Dymowska, University of Oslo, Physiology Programme, Dept. of Molecular Biosciences, Blinderveien 31 Oslo N-0316 Norway E-Mail: [email protected] Abstract:

Fish Biology Congress Abstracts 28 In normoxic crucian carp (Carassius carassius), an intralamellar cell mass (ILCM) completely covers the gill lamellae. In hypoxia, the ILCM recedes to increase respiratory surface area. This morphological change could lead to increased fluxes of water and ions across the gills. We investigated how the presence or absence of the ILCM affected the location of gill ionocytes (i.e., mitochondria-rich cells; MRCs), plasma ion balance and blood pH. 8 C-acclimated carp was exposed to hypoxia for 7d (6-8% O2). MRCs were localized with immunolabeling and TEM. We found that MRCs were located on top of the ILCM during normoxia and hypoxia and that the abundance of MRCs increased progressively with hypoxia. Plasma concentrations of Na+, Cl-, K+ and pH were identical in normoxic and 7d hypoxic fish. Experiments are currently being conducted to determine if ATPase activities of MRCs also increase and to quantify the effect of ILCM loss on water influx.

IDENTIFICATION OF FRESH WATER FISHES IN PERMANENT RIVERS BASIN OF LOOT & SIRJAN DESERTS OF PROVINCE IN IRAN Ebrahiumi, M., and Ramin , M., and Afzali, H., and Azadshahraki, F., and Eslami, F. Symposium: Contributed Papers Presentation: Poster Contact: Mahtab Ebrahimi, Research Center of Agriculture and Natural Resource of Kerman Kerman Kerman 7617913739 Iran E-Mail: [email protected] Abstract: This research has been performed in the province of Kerman,sout east of Iran (55 ,25´to33˚of the northern latitiude and 26˚,53´ to 29˚,59´ of the eastern longitud) with five basins , sixteen permanent rivers and hundreds season rivers , hundreds of ranges of quanats and a lot of springs. since 2001-2002 during one years identification fishes of permanent rivers basin of Loot desert ,namely: nesa ,gahoueih , Abbaric, Gishigan , Tahroud , Gorooh , Khamrootek , Adouri , Kouhpaieh , Sirch , Andohjerd , , Dehbacri and Sirjan desert,namely: Tangouieh , ostoor Hossein abad , Sooch. We sampling from above mentioned rivers which were in 1 order 2 family series and 3 species . The identification fishes were :C apoeta damascina and Ciprinion watsoni from family , Nemachilus sargadensis from BALITORIDAE family.

WHAT IS THE ACTIVE CORTICOSTEROID IN AGNATHANS? THE EFFECTS OF CORTICOSTEROIDS ON THE ABILITY OF THE PACIFIC HAGFISH TO OSMOCONFORM FOLLOWING A MGCL2 CHALLENGE. Ede, J.D., Bury, N., Goss, G.G. Symposium: Ion and Acid-base Regulation Presentation: Poster Contact: Nic Bury, Kings College London, Nutritional Sciences Research Division, Department of Biochemistry Franklin-Wilkins Building, 150 Stamford Street London SE1 9NH UK E-Mail: [email protected] Abstract: To help elucidate the evolutionary history of corticosteroid receptors (CRs), the physiological effects of corticosteroids in the primitive agnathan group of fish, the myxini, was investigated. Pacific hagfish (Eptatretus stouti) were injected with a bolus of coconut oil containing either cortisol, corticosterone or 11-deoxycorticosterone (DOC). Plasma cortisol and glucose measurements were made, the latter being indicative of glucocorticoid action in the gnathostomata. Gill tissue ATPase activity was also measured. In an additional experiment, hagfish were injected with 100mg/Kg of either cortisol or 11-deoxycorticosterone and on the fifth day transferred to seawater containing 50mM MgCl2 to assess their ability to osmoconform and regulate Mg - potential evidence of mineralocorticoid action. The fish treated with DOC showed a greater ability to deal with the MgCl2 challenge and also showed evidence of increase plasma glucose concentrations on day seven. These results suggest that DOC, and not cortisol, is the active corticosteroid in hagfish.

WASTE EXCRETION IN AGNATHAN FISHES Edwards, S.L. Fortier, J, Diamanduros, A and Claiborne J.B. Symposium: Fish Living on the Edge Presentation: Oral Contact: Susan Edwards, Department of Biology, Appalachian State University, 527 Rivers St Boone NC 28606 USA E-Mail: [email protected] Abstract: It is widely accepted that in fishes, the excretion of nitrogenous wastes and excess acid and base are accomplished through transepithelial excretion ions across the branchial epithelium. The hagfish and lamprey are the sole survivors of the jawless stage in vertebrate evolution. Our research focuses on the identification of epithelial transport mechanisms involved in branchial waste excretion in agnathan fishes .We have identified 3 Na+/H+ exchanger isoforms (NHE1,3 &8) and a H+ATPase from branchial tissue of two agnathan species Myxine glutinosa and Petromyzon marinus. These transporters have been shown to play a role in maintaining acid/base homeostasis in fishes and higher

Fish Biology Congress Abstracts 29 vertebrates. In addition, we have recently identified several members of the Rh glycoprotein family in the gills of M. glutinosa and postulate that they may play a role in the transepithelial excretion of ammonia.

DIFFERENCES IN THE BODY MORPHOMETRICS AND ASSOCIATED SWIMMING COSTS BETWEEN WILD, HATCHERY, AND FARMED ATLANTIC SALMON Enders, E.C., Boisclair, D., and Roy, A.G. Symposium: Morphometrics Presentation: Oral Contact: Evan Enders, Northwest Fisheries Science Center, 2725 Montlake Blvd. E. Seattle WA 98112-6-2097 USA E-Mail: [email protected] Abstract: We compared morphometry and swimming costs of wild, hatchery (first-generation hatchery progeny of wild progenitors), and farmed (seventh-generation progeny of the Norwegian aquaculture strain) Atlantic salmon (Salmo salar). Spatial coordinates of 18 morphometric landmarks were obtained from individual fish ranging from 4 to 16 g wet mass. Morphology of farmed Atlantic salmon differed significantly from both wild and hatchery fish, with the head being smaller, the body more robust, and rayed fins smaller. In contrast, wild fish had slender body shapes and larger fins than farmed Atlantic salmon. These observed morphological differences may affect the swimming costs. Respirometry experiments were performed to assess swimming costs at a water temperature of 15 C. Swimming costs of wild and hatchery fish were not statistically different. However, farmed fish had swimming costs up to 29.2% higher than wild or hatchery fish. This may be related to farmed fish having deeper bodies and smaller fins.

CHANGING VELOCITY CONDITIONS ENCOUNTERED BY SMOLTS: HOW DO THEY INFLUENCE CHINOOK SALMON BEHAVIOR Enders, E.C., Gessel, M.H. and Williams, J.G. Symposium: Swimming Performance Presentation: Oral Contact: Evan Enders, Northwest Fisheries Science Center, 2725 Montlake Blvd. E. Seattle WA 98112-6-2097 USA E-Mail: [email protected] Abstract: The success of bypass structures on hydropower dams relies strongly on the capacity to guide fish away from turbines towards the bypass. A better understanding of fish behavior to flow environments may help to improve the design of efficient guidance devices to bypass structures. The objectives of this project were to identify reactions and thresholds of Chinook salmon (Oncorhynchus tshawytscha) smolts over a range of flow velocity gradients. In the experiments, we observed a clear avoidance reaction of smolts approaching the steep velocity gradient. Furthermore, the motivation of smolts to pass the velocity gradient changed over the smolt run. Smolts took longer before attempting to pass the velocity gradient at the beginning of the smolt run, and the attempt rate increased as time progresses. Results from this study may provide recommendations to engineers and biologists for designing novel fish guidance structures and modifications to existing structures to provide effective guidance for downstream migrating smolts at hydropower developments.

TELEMETRY, DIDSON CAMERA, AND GIS USED WITH TRADITIONAL MARK-RECAPTURE METHODS TO ESTIMATE THE NUMBER OF GREEN STURGEON THAT SPAWN IN THE ROGUE RIVER, OREGON Erickson, D.L., Lindley, S., Mora, E., Weber, J, Confer, T., Krohn, B., and and Doukakis, P. Symposium: Physiological BioTelemetry Presentation: Oral Contact: Dan Erickson, University of Miami, Pew Intitute for Ocean Science, 126 East 56th St, Level M New York NY 10022 USA E-Mail: [email protected] Abstract: We describe steps taken to design and conduct a mark-recapture study to estimate the abundance of adult green sturgeon that spawn in the Rogue River, Oregon. Estimates were improved by studies to (a) understand the behavior and habits of green sturgeon using sonic and radio telemetry, (b) estimate the total possible number of summer/fall holding sites throughout the river based upon known-holding sites (via telemetry) and using a GIS framework, and (c) verify the presence of sturgeon at potential holding sites using a DIDSON camera before setting gillnets in order to maximize capture rates. Because we now understand the behavior of green sturgeon, and because more than 400 adults are tagged, little effort (< 3 weeks per year) is needed for acquiring annual abundance estimates for Rogue River green sturgeon. Our methods could be used as a template to develop sturgeon-monitoring programs for important spawning rivers

CORTISOL-MEDIATED EXPRESSION OF GLUTAMINE SYNTHETASE IN THE GULF TOADFISH (OPSANUS BETA): A GENOMICS PERSPECTIVE. Esbaugh, A.J. and Walsh, P.J.

Fish Biology Congress Abstracts 30 Symposium: Stress Effects and Response Presentation: Poster Contact: Andrew Esbaugh, Department of Biology, University of Ottawa Ottawa ON K1N 6N5 Canada E-Mail: [email protected] Abstract: Unlike most teleost fish, the gulf toadfish can shift from ammonotely to ureotely as the primary strategy for nitrogen excretion. The regulatory control of this strategic switch is the enzyme glutamine synthetase (GS). Toadfish contain two distinct GS enzymes, one specific to the gill and another general form that is abundant in the liver. Previous experiments have suggested that a cortisol-mediated increase in liver GS activity initiates the switch to ureotely. As such, we undertook a gene promoter analysis of the gill and liver GSases in an effort to ascertain the mechanism of cortisol-mediated expression. We cloned more than 1000 bp of promoter sequence from both genes. The liver GS promoter region contained two imperfect glucocorticoid receptor elements consisting of 15 bp palindromic AGAACA motifs. The gill form contained no such motifs. Studies regarding the effectiveness of these motifs in controlling cortisol-mediated gene expression are currently underway.

MEMBRANE-ASSOCIATED CARBONIC ANHYDRASE IN THE RESPIRATORY SYSTEM OF THE PACIFIC HAGFISH (EPTATRETUS STOUTI). Esbaugh, A.J., Gilmour, K.M. and Perry, S.F. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Andrew Esbaugh, Department of Biology, University of Ottawa Ottawa ON K1N 6N5 Canada E-Mail: [email protected] Abstract: In most fish, red blood cell (RBC) carbonic anhydrase (CA) is responsible for dehydrating plasma bicarbonate for gas exchange and acid-base balance. Hagfish RBCs do not have anion exchange capabilities, meaning the substantial amount of plasma bicarbonate (87.6 ± 1.2 % of whole blood bicarbonate) is not available to RBC CA. Additionally, hagfish haemoglobin does not contribute to plasma buffering (true plasma βnb = -4.83 ± 0.52 mM pH unit-1; separated plasma βnb = -4.92 ± 0.43 mM pH unit-1). We used molecular and biochemical methods to verify the presence of plasma accessible CA in the Pacific hagfish, which could help mobilize plasma bicarbonate for both gas exchange and acid- base balance. Two distinct CA IV-like isozymes were identified using homology cloning techniques, both of which were expressed in the gills. One was also expressed in the RBCs. Biochemical analysis of the membrane fractions from both tissues confirmed the presence of CA IV-like activity.

SPATIAL DISTRIBUCIÓN OF REPRODUCTIVE ACTIVITY OF PERUVIAN HAKE (MERLUCCIUS GAYI PERUANUS) Espinoza, C.; Perea, A.; Buitrón, B.; Cisneros, P.; Vera, V.; Catcoparco, Ch.; Romero, L.E.; Alberro, A.; Rojas, P. and Vizziano, D. Symposium: Contributed Papers Presentation: Oral Contact: Betsy Buitrón, Laboratorio de Biología Reproductiva - Instituto del Mar del Perú, Esquina Gamarra y Gral. Valle s/n, Apartado 22, Callao, Lima-Perú [tel: +51 1 4297630 anex 250. Callao Lima 1 Peru E-Mail: [email protected] Abstract: The present work evaluated the effect of busereline acetate injection on female spawning and male spermiation of Peruvian anchovy. Ripe fishes were injected intraperitoneally with 0,005 μg GnRHa g-1 body weight (b.w.) (H); 0,005 μg GnRHa g-1 b.w. + 0,01 mg DOM g-1 b.w. (HD) or saline solution to 0,7 % (S). The treated fish were evaluated at 0, 12, 24 and 48 hours post-injection (p.i.). There was an inductor effect of the treatments with GnRH and GnRH + DOM on the spawning and spermiation. Maximum percentages of evacuation were at 12 hours post-injection, 85,7 % with H and 75,0 % with HD. The effect of the hormone on the evacuation of the males of the H treatment was immediate. It was observed a considerable reduction of the percentages of spawning and spermiation after injected GnRHa + DOM with regard of the treatment only with GnRHa.

THE ZEBRAFISH COELOMIC CAVITY: A NOVEL MODEL READ-OUT TISSUE FOR STUDIES OF IMMUNE RESPONSES TO BACTERIAL INFECTIONS Evans, D. L., Moss, L. D., Leary, J. H. III, Dougan, S., and Jaso-Friedmann, L. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: D. L. Evans, Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia Athens Ga. 30602 USA E-Mail: [email protected] Abstract:

Fish Biology Congress Abstracts 31 The zebrafish (zf) is an emerging animal model for studies of developmental biology and basic genetics. The usefulness of this species for host resistance and basic immune function studies has been poorly developed. The inability to harvest leukocytes (lymphocytes), antibody or lymphoid organs in sufficient quantity to be utilized in ex vivo or in vitro studies is currently a limiting factor for use of the zf by immunologists. In the current study, the coelomic cavity was developed as a tissue capable of providing an immune read-out for development of an infectious disease model. The coelomic cavity of adult Danio rerio constitutively contains approximately 10^5 cells. The approximate distribution of these cells is granulocytes (60-80%), lymphocytes (10-20%) and nonspecific cytotoxic cells (NCC) (4-10%). Granulocytes were identified by phagocytosis of E. coli and S. aureus; the lymphocyte fraction expressed B- and T-cell mRNA; and NCC were identified based on expression of NCCRP-1 and NCAMP-1. A cellular exudate was produced by intracoelomic (IC) injections of LPS and killed Edwardsiella ictaluri at 2 and 6 hours post-injection respectively. Infection with E. ictaluri likewise produced a coelomic exudate at 24h post-immersion. Studies of the functions of coelomic exudate cells (CEC) determined the presence of cytotoxic cells that killed conventional mouse and human NK cell targets (YAC-1, K562, HL- 60 target cells) at 1:1, 2:1 and 4:1 effector:target cell ratios. Redirected lysis by CEC of hybridoma cells expressing membrane mab 9C9 demonstrated that the protein identified by this mab (i.e. NCAMP-1) is associated with activation of the lytic cycle. These data demonstrated that the zf coelomic cavity is composed of cells that can be upregulated/activated and as such are useful for studies of both innate and adaptive immune responses. Supported by USDA and Veterinary Medical Experiment Station (UGA).

ANNUAL AND SEASONAL VARIATIONS IN GONADAL DEVELOPMENT AND STEROIDS PROFILES IN CATLA CATLA (TELEOSTEI, CYPRINIDAE) Fatima, S., Seher, S. and Lone, K.P. Symposium: Contributed Papers Presentation: Oral Contact: S. Fatima, Shafaq Fatima, Department of Zoology, Lahore College for Women University, Lahore, Pakistan Lahore Pakistan E-Mail: [email protected] Abstract: Changes in testosterone (T), 11-ketotestosterone (11-KT), estradiol (E2), progesterone (P), 17α-hydroxyprogesterone (17α-HP) and cortisol were correlated to changes in GSI, ovarian and testicular histological variations during annual reproductive cycle of Catla catla, an important commercial carp. In female, E2 level shows direct changes with 8 ovarian developmental stages and GSI, with peak (0.14 ± 0.02) at maturation. Maximum values of T and 11-KT, one month before spawning indicate the important role of androgens in late stages of maturity. In male catla, both T and 11-KT are involved in initiating recrudescence. Roles of T and 11-KT are more pronounced in spermeiogenesis and spermiation respectively. High levels of T at maturity coincide with maximum GSI values during these months. Peak of E2 (0.13 ± 0.00) in march reveals its role in spermatogonial proliferation. Serum values of both P and 17α-HP remain low during period of sexual activity and show high profiles at immature stages. Cortisol concentrations are correlated with k factor with minimum values during spawning season. The annual reproductive cycle is greatly influenced by temperature and photoperiod, while rainfall acts as a spawning cue. This study describes the reproductive biology of Catla catla for the first time in temperate region.

AN E3 UBIQUITIN LIGASE HOMOLOGOUS TO HUMAN GRAIL IS INVOLVED IN THE SEA WATER ACCLIMATION RESPONSE IN TILAPIA (OREOCHROMIS MOSSAMBICUS). Fiol, D.F., Lim, A., Sanmarti, E.. and Kültz, D. Symposium: Stress Effects and Response Presentation: Poster Contact: Diego Fiol, Physiological Genomics Group, Animal Science, UC Davis, Davis, CA 95616 Davis CA 95616 USA E-Mail: [email protected] Abstract: A novel E3 ubiquitin ligase protein, OmGRAIL, was identified and cloned in the euryhaline fish Oreochromis mossambicus using the suppression subtractive hybridization approach based on its response to hyperosmotic stress. OmGRAIL showed highest homology (40% identity) with the human ring finger protein 128 or GRAIL (gene related to anergy in lymphocytes) and belongs to the same group of E3 ubiquitin ligases as human Goliath and Xenopus GREUL1. OmGRAIL contains the characteristic PA (Protein associate) domain and a C3H2C3 RING finger indicating functional E3 ubiquitin ligase activity. Real time PCR quantification of OmGRAIL mRNA showed higher expression levels in gills. Protein abundance and tissue distribution of OmGRAIL were quantified using homologous antibodies, revealing an almost exclusive expression in gills. In addition OmGRAIL expression was higher in fish acclimated to salt water as compared with the fresh water controls. Stably transfected cell lines overexpressing Tet-inducible OmGRAIL were established from HEK-293 cells. Cell lines expressing OmGRAIL showed an increased tolerance to hyperosmotic media. Taken together, our results suggest that OmGRAIL is a novel E3 ubiquitin ligase protein involved in the response to hyperosmotic stress in fish.

Fish Biology Congress Abstracts 32

EVOLUTION OF GLYCOPROTEIN HORMONE/GLYCOPROTEIN EVOLUTION OF GLYCOPROTEIN HORMONE/GLYCOPROTEIN HORMONE RECEPTOR SYSTEMS IN VERTEBRATES FROM A SEA LAMPREY PERSPECTIVE Freamat, M., and Sower, S.A. Symposium: Biology of Lampreys Presentation: Oral Contact: Mihael Freamat, Department of Biochemistry and Molecular Biology, 46 College Road Durham NH 3824 USA E-Mail: [email protected] Abstract: The endocrine control of the gonadal and thyroid physiology in vertebrates depends ultimately on the interaction between the pituitary glycoprotein hormones (GpH) and their cognate receptors (GpH-R). Both ligands and receptors are closely related proteins, with similar overall structural organization. Subtle differences between their structures result in a high degree of selectivity in the activating interactions hormone/receptor which ensures a minimal cross interaction between these pathways in later evolved vertebrates. This selectivity however was found to be less stringent in earlier evolved vertebrates. Investigation of the GpH/GpH-R system in the sea lamprey resulted in identification of one GpH and of two GpH-R homologs. We present the structural and functional properties of these molecules and their implications for understanding of the origins of pituitary-gonadal/thyroid axes, of the mechanisms of functional divergence of related receptors and evolution of complex chemical signaling networks. Supported by NSF IBN-0421923.

CHALLENGING THE OXYGEN HYPOTHESIS: PARTITIONING OF OXYGEN AND SODIUM UPTAKE BETWEEN THE GILLS AND SKIN DURING LARVAL DEVELOPMENT OF THE RAINBOW TROUT, ONCORHYNCHUS MYKISS Fu, C.H.L., Rombough, P., Brauner, C.J. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Clarice Fu, University of British Columbia, Department of Zoology, #2370-6270 University Blvd. Vancouver BC V6T 1Z4 Canada E-Mail: [email protected] Abstract: This study is one of the first to provide physiological evidence challenging the long-held assumption that early gill development is driven by respiratory demand. According to the recent “ion hypothesis”, ionoregulation, rather than gas exchange, is the first to transition from the skin to the gills during larval development. A novel method was used to quantify oxygen and unidirectional sodium uptake by the skin and gills from 0 to 18 days post-hatch in rainbow trout, Oncorhynchus mykiss. Preliminary data supports the ion hypothesis. Results are being compared to a model that estimates the relative importance of gas and ion exchange as competing driving forces on gill development. This study not only stresses the need to challenge current assumptions on gill development, but also urges a re-examination of the belief that oxygen delivery was the major selective pressure behind the evolution of the vertebrate gill.

THE X-RAY MICROTOMOGRAPHIC STUDIES OF GALEASPID ENDOCRANIUM SHED NEW LIGHT ON THE ORIGIN OF PAIRED NOSTRILS AND JAW Gai, Z.K., Philip C.J. D., Zhu, M., Philippe, J. Symposium: Contributed Papers Presentation: Oral Contact: Zhikun Gai, Department of Earth Sciences, University of Bristol, Wills Memorial Building, Queen's Road Bristol Bristol BS8 1RJ UK E-Mail: [email protected] Abstract: The origin of jaws has been one of the longest-standing and least tractable problems in the study of vertebrate evolution. Progress has been made in comparing the development of the equivalent populations of neural crest cells in living jawless and jawed vertebrates. These studies have shown that a gnathostome fate is precluded in lampreys because the rudiment of the nasohypophysial system precludes rostral migration of mandibular neural crest. Thus, the dissociation of the olfactory system from the NHP has been interpreted as a key evolutionary event for the origin of jaw, facilitating the rostral migration of premandibular mesenchyme, but an event for which there has been no palaeontological evidence. Through detailed X-ray microtomographic analysis of the galeaspid endocranium, here, we provide the first definitive fossil evidence for this scenario. Galeaspids possess paired and laterally situated nasal sacs, as in gnathostomes, but a single median nostril, as in lampreys.

GILL NA+, K+ ATPASE ALPHA SUBUNIT ISOFORM EXPRESSION AND MIGRATION TENDENCY IN JUVENILE STEELHEAD Gale, W., Simpson, W., Kennedy, B. and Ostrand, K.

Fish Biology Congress Abstracts 33 Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: William Gale, Abernathy Fish Technology Center, 1440 Abernathy Creek Rd. Longview WA 98632 USA E-Mail: [email protected] Abstract: We examined the relationships between gonadal development, gill Na+, K+ ATPase (NKA) activity, and gill NKA alpha subunit expression in migratory and residual steelhead juveniles. The sex ratio of residuals favored males 2:1, whereas the sex ratio of migrants was 1:1. Examination of GSI data from residual males revealed a distribution that was highly positively skewed, suggesting the presence of a significant number of early maturing males. Both NKA activity and NKA alpha subunit mRNA expression were significantly higher in migrants than in residuals. A positive relationship between NKA activity and expression of the NKA alpha-1b subunit occurred in both groups. Conversely, the expression of the alpha-1a isoform was unrelated to NKA activity. NKA alpha subunit expression may provide a useful indicator of migratory readiness. However, more research is needed before the roles of different isoforms of the NKA alpha subunit are fully understood.

TEMPERATURE DEPENDENT PROPERTIES OF ACTION POTENTIALS AND SARCOLEMMAL K+ CURRENTS FROM BLUEFIN TUNA MYOCYTES Galli, GLJ and Block, BA Symposium: Fish Living on the Edge Presentation: Oral Contact: Gina Galli, Hopkins Marine Station of Stanford University, 120 Oceanview Boulevard Pacific Grove CA 93950 USA E-Mail: [email protected] Abstract: Pacific bluefin tuna, Thunnus orientalis, make extensive horizontal and vertical migrations where they inhabit a broad range of temperatures. Temperature has a profound effect on the excitability and contractility of the ectothermic heart. Thermal plasticity of proteins associated with excitation-contraction coupling is essential for the maintenance of cardiac function. The present study examines the effect of acute thermal stress and thermal acclimation on sarcolemmal K+ currents and their role in action potential generation. In atrial and ventricular cells, an acute reduction in temperature led to a prolongation of the action potential, which was mirrored by a decrease in the density of the delayed rectifier current (IKr). The density of the background inward rectifier current (IK1) was unchanged with acute thermal stress. Acclimation to cold (14oC vs. 24oC) had no significant effects on action potential shape/duration, or on IK1 density, while IKr density was markedly reduced.

THE INFLUENCE OF NATURAL ORGANIC MATTER ON CADMIUM AND CALCIUM UPTAKE IN ZEBRAFISH DURING EARLY-LIFE STAGES Galvez, F., Cox, A., and Svoboda, K.R. Symposium: Environmental Pollution Presentation: Poster Contact: Fernando Galvez, Department of Biological Sciences, 216 Life Sciences Building Baton Rouge LA 70803 USA E-Mail: [email protected] Abstract: This study utilized a radioisotopic flux technique to measure the unidirectional uptake of cadmium (Cd2+) and calcium (Ca2+) in zebrafish from 24 to 96 hours post fertilization (hpf). Time-course experiments showed high Cd2+ contents in embryos at 24 and 96 hpf, compared to the low levels measured at 72 hpf. Embryos at 24 hpf showed linear, non-saturable Cd2+ kinetics, up to a waterborne Cd2+ of 100 nM. However, by 72 hpf, zebrafish demonstrated saturable, carrier- mediated Cd2+ transport. Michaelis-Menten analyses of these kinetic relationships suggested, larval fish at 96 hpf were similar to adult zebrafish with regards to Cd2+ transport. Similarly, Ca2+ burdens were highest in embryos at 24 hpf, although most of this ion accumulation was associated with the chorion. By 72 hpf, zebrafish hatchlings had significantly lower Ca2+ burdens compared to embryos at 24 hpf, although levels were again increased in animals by 96 hpf. Current studies are investigating the influence of natural organic matter on Cd2+ and Ca2+ uptake during zebrafish development. Acknowledgements: This work was funded by a Powe Junior Faculty Enhancement Award (Oak Ridge Associated Universities) to FG, LEQSF(2005-08)-RD-A-11 (Louisiana Board of Regents) to KRS, and a NIH Grant Number P20 RR16456 ( INBRE Program of the National Center for Research Resources) to AC.

INTERSPECIFIC AND INTRASPECIFIC VARIATIONS IN THE PHYSIOLOGICAL PLASTICITY OF FUNDULUS TO OSMOTIC STRESS Galvez, F., Zhang, S., Hobson, J., and Whitehead, A. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Fernando Galvez, Department of Biological Sciences, Louisiana State University, Baton Rouge Louisiana 70803 USA

Fish Biology Congress Abstracts 34 E-Mail: [email protected] Abstract: Although Fundulus are typically resilient to osmotic challenges, considerable interspecific, and some intraspecific variations in salinity tolerances exist. Studies in our laboratories are investigating the physiological responses and functional genomics of hyperosmotic and hypoosmotic exposures in several taxa of Fundulus collected along the Atlantic and Gulf coasts of the United States. This presentation will describe temporal changes in the ionoregulatory status of Fundulus following abrupt salinity transfers. More specifically, we will present results from recent studies showing differences in the capacity of gills to remodel during salinity stress. Future work is trying to address the adaptive role of claudins, a constitutive component of epithelial tight junctions, in regulating the paracellular movement of solutes during osmotic stress. Acknowledgements: This work was funded by Louisiana State University and an NSF grant (Award no. 0723771).

CHRONIC HYPOXIC EFFECTS ON THE PHYSIOLOGY ATLANTIC COD (GADUS MORHUA): ADAPTATION OR COMPROMISED FUNCTION? Gamperl, A.K. and Petersen, L.H. Symposium: Fish Living on the Edge Presentation: Oral Contact: Kurt Gamperl, Ocean Sciences Centre, Memorial University of Newfoundland St. John's NL A1C 5S7 Canada E-Mail: [email protected] Abstract: The physiological effects of chronic hypoxia on fishes, and their capacity to adapt to such environmental challenges are poorly understood. Thus, we acclimated adult Atlantic cod to either 40-45% or 100% oxygen saturation (normoxia) for 6- 12 weeks at 10oC, and subsequently measured: 1) metabolic parameters [routine oxygen consumption (MO2), Max. MO2, metabolic scope] and cardiac function (Q, ƒH and VS) during Ucrit tests at both O2 levels; 2) maximum and resting in situ cardiac function in normoxia, and anoxia; 3) the in situ and in vivo responsiveness of cardiac function to adrenergic stimulation; and finally; 4) their acute hypoxia tolerance (at rest), tissue O2 extraction efficiency, blood Hb-O2 binding characteristics, and resting and maximal circulating catecholamine levels. These results provide important insights into how fish cardiorespiratory physiology is impacted by short-term and prolonged exposure to hypoxia, and further highlight the tremendous capacity of some aspects of the fish cardiorespiratory system to deal with environmental challenges.

Symposium: Presentation: Contact: , E-Mail: Abstract:

THE IMPACT OF ELEVATED WATER TEMPERATURE AND ZATEBRADINE-INDUCED BRADYCARDIA ON CARDIOVASCULAR FUNCTION IN MALE AND FEMALE RAINBOW TROUT (ONCORHYNCHUS MYKISS) Gamprerl, A.K., Swafford, B.L., and Rodnick, K.J. Symposium: Contributed Papers Presentation: Poster Contact: K.J. Rodnick, Dept. of Biological Sciences, 650 Memorial Drive, Idaho State University Pocatello ID 83209-8007 USA E-Mail: [email protected] Abstract: Questions remain about the regulation of fish cardiovascular function, including the role played by sex differences. This study examined the relationship between heart rate (FH), stroke volume (Vs), cardiac output (Q) blood pressure (DAP) and systemic vascular resistence (RS) in sexually-immature trout: 1) when challenged with an increase in temperature from 14- 24 C; and 2) following a 50% reduction in FH at 24 C, achieved through zatebradine administration. FH increased in a linear fashion with temperature; from ~ 60 bpm at 14 C to ~ 130 bpm at 24 C), and overall there were no significant sex differences. At 24 C, zatebradine halved FH, and yet Q remained within 10% of pre-treatment values. These results: 1) indicate that the cardiovascular response of sedentary, immature, male and female trout to elevated temperatures is similar; 2) support recent in vivo data for Atlantic cod challenging whether elevated temperature and heart rate negatively influence teleost stroke volume / myocardial contractility. Supported by NSF grant IOB-517669.

ISOTOPIC COMPARISON BETWEEN GEODUCK SHELLS AND ROCKFISH OTOLITHS: IMPLICATIONS FOR LIFE HISTORY AND BEHAVIOR Gao, Y.W. Symposium: Contributed Papers Presentation: Oral Contact: Yongwen Gao, Makah Fisheries Management, PO Box 115+J331 Neah Bay WA 98357-0115 USA

Fish Biology Congress Abstracts 35 E-Mail: [email protected] Abstract: The hard part of fishes (e.g., otoliths, shells, bones) can be used as a proxy for ocean environmental studies. In this study, we analyzed geoduck (Panopea abrupta) shells and yelloweye rockfish (Sebastes ruberrimus) otoliths from Puget Sound and the western coast of Washington, and examined the isotopic signatures and differences. The lifetime d18O variations of geoduck shells were similar to those of rockfish otoliths, but their d13C variations were significantly different from each other. From age-1 to age-5, the averaged d13C values of rockfish otoliths were increased from about -5.0‰ to -1.0‰, indicating that rockfish may change their diet for higher trophic level food. In contrast, smaller d18O and d13C variations from shells suggest that geoducks are sedentary animals with lower trophic level changes during the same growth period. Overall these comparisons suggest that stable isotope signatures of fish hard parts are a powerful tool in fisheries research.

THE BEAVERS OF THE FISH WORLD: CAN WOOD-EATING DIGEST WOOD? German, D.P. Symposium: Tropical Fish Adaptations Presentation: Oral Contact: Donovan German, Department of Zoology, University of Florida, 321 Bartram Hall Gainesville FL 32611-8525 USA E-Mail: [email protected] Abstract: I performed three feeding trials with wood-eating and detrivorous catfishes (family ) to determine whether they can assimilate wood cellulose. First, I performed a stable isotope turnover study with Pterygoplichthys disjunctivus. These fish were raised in the laboratory for 150 days on pellets consisting of 80% ground, sterilized wood (cellulose fraction delta13C = -25 ppt), and 20% corn meal and corn gluten meal (delta13C = -10 ppt) as soluble polysaccharides. After 150 days, the fish lost weight and the delta13C of their tissues indicated that they assimilated the corn products, not the cellulose. The second trial confirmed that Panaque nigrolineatus could assimilate only 6% of cellulose from wood, and the third trial showed that both species pass wood through their guts in less than four hours. My results therefore suggest that wood-eating loricariid catfishes are specialized detritivores and do not actively digest cellulose in their digestive tracts.

EVOLUTION OF HERBIVORY IN A CARNIVOROUS CLADE OF MINNOWS (TELEOSTEI: CYPRINIDAE): EFFECTS ON GUT STRUCTURE AND FUNCTION German, D.P., Nagle, B., and Evans, D.H. Symposium: Contributed Papers Presentation: Poster Contact: Donovan German, Department of Zoology, University of Florida, 321 Bartram Hall Gainesville FL 32611-8525 USA E-Mail: [email protected] Abstract: To investigate how evolutionary history affects digestion, we examined gut structure and function in eleven taxa composing sister clades of minnows with different dietary affinities: Campostoma are herbivorous whereas Nocomis are carnivorous. Additionally, Nocomis leptocephalus is carnivorous in the Chatahoochee drainage, and herbivorous in the Altamaha drainage. Thus, we were able to examine the effects of diet and evolutionary history on digestion among the clades and within a single species. The Campostoma had longer guts, higher amylase and laminarinase activities, and lower chitinase activities than the carnivorous Nocomis. This same pattern was observed among the populations of N. leptocephalus. Independent contrasts indicate that the evolution of diet and gut function are indeed correlated in these fishes. Overall, our results indicate that a longer gut and high amylase and laminarinase activities are associated with the evolution of herbivory, whereas a shorter gut and higher chitinase activities are associated with carnivory.

THE ANAESTHATIC EFFICIENCY OF CLOVE OIL IN CASPIAN SALMON SALMO TRUTTA CASPIUS RELATED TO WATER SALINITY AND PH Ghaziloua A,Saeedi H Symposium: Stress Effects and Response Presentation: Poster Contact: Amir Ghazilou, khorramshahr uni of marine science and technology tabriz eastazarbijan 5166886911 Iran E-Mail: [email protected] Abstract: The anaesthatic efficiency of clove oil in Caspian salmon Salmo trutta caspius K. related to water salinity and pH was evaluated. All fish exposed to experimental concentrations of clove oil (20, 40,60and80mg.L -1) and salinity-pH trials were anaesthetized reaching the stage 4 in ≤ 3 min and recovered within 14 min when placed in fresh aerated water. General linear model analysis indicated a significant dependence (p<0.05) of induction time on pH and salinity level and decreased induction time along with decreased salinity observed .In the case of pH, induction time was significantly different and could be divided into two groups of low & higher concentrations of clove oil. Decreased induction time along

Fish Biology Congress Abstracts 36 with decreased pH observed at higher concentrations but neutrality led to lowest induction time for fish anaesthesized at 100 mg.L-1 concentration of clove oil. Recovery time decreased to either side of neutrality significantly but the significant differences in the recovery times were independent of water salinity.

BULLIES IN THE FISH TANK: PHYSIOLOGICAL EXTREMES ELICITED BY SOCIAL STRESS Gilmour, KM Symposium: Fish Living on the Edge Presentation: Oral Contact: Katie Gilmour, Department of Biology, University of Ottawa, 30 Marie Curie Ottawa ON K1N 6N5 Canada E-Mail: [email protected] Abstract: Juvenile salmonid fish form linear dominance hierarchies in which the dominant fish enjoys the most favourable position in the environment, gains the lion’s share of available food and is aggressive towards more subordinate fish. Subordinate fish, by contrast, exhibit marked behavioural inhibition including reduced activity and feeding. These behavioural extremes are accompanied by distinctive physiologies. Most obviously, subordinate fish do not grow as rapidly as dominant fish, even when fed the same ration. The lower growth rates of subordinate fish reflect a variety of factors, including impaired digestive function and altered metabolism. For example, subordinate fish are forced to rely upon on-board energy stores and consequently exhibit low hepatic glycogen levels and pyruvate kinase activity together with elevated hepatic phosphoenolpyruvate carboxykinase activity. Subordinate fish also exhibit chronic cortisol elevation and distinctive patterns of corticosteroid receptor and heat shock protein expression. Taken together, these physiological responses reflect an extreme social environment.

MECHANISMS OF RELAXATION IN DORSAL AND VENTRAL AORTA IN NORMAL AND BENZO[A]PYRENE-EXPOSED TROUT Goertzen, M.M., and Weber, L.P. Symposium: Environmental Pollution Presentation: Poster Contact: Meghan Goertzen, Department of Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, 52 Campus Drive Saskatoon SK S7N 5B4 Canada E-Mail: [email protected] Abstract: Compared to mammals, vasorelaxation in fish is poorly characterized. We hypothesized that prostaglandin E2 (PGE2), sodium nitroprusside (SNP), and sodium hydrogen sulfide (NaSH) would mediate vasorelaxation in fish and that toxicant- exposure would alter their effects. Rainbow trout (Oncorhynchus mykiss) dorsal and ventral aorta ring responses to vasoactive agents were characterized in an organ bath. In untreated fish, carbachol contracted ventral aorta and calcium ionophore (A23187) contracted dorsal aorta, while SNP and PGE2 relaxed both vessels. NaSH contracted ventral aorta, but relaxed dorsal aorta. After benzo[a]pyrene (BaP) injections, PGE2 responses in dorsal aorta were unchanged. However, SNP caused a significantly larger relaxation in dorsal aorta from BaP compared to vehicle-injected fish, while NaSH- induced relaxation was eliminated in BaP-exposed fish. BaP exposure increased hepatic, but not aortic, CYP1A enzyme activity in BaP compared to vehicle-exposed fish. In conclusion, exposure to the toxicant, BaP, appears to alter vasorelaxation in trout dorsal aorta.

INTRASPECIFIC BRAIN SIZE EVOLUTION IN THE WILD: A CASE STUDY IN NINESPINE STICKLEBACK Gonda, A., Herczeg, G., Merilä, J. Symposium: Morphometrics Presentation: Poster Contact: Abigel Gonda, University of Helsinki, Department of Biological and Environmental Sciences, Ecological Genetics Research Unit, PO Box 65 (Biocenter 3 Viikinkaari 1) Helsinki FIN-00014 Finland E-Mail: [email protected] Abstract: Size of the total brain and different parts of the brain strongly vary among species. Yet, the vast majority of studies in brain size evolution have so far relied on interspecific comparisons which have uncovered a number of interesting correlations between brain size and putative selective factors. However, studies on local adaptation in brain size at the intraspecific level are rare. We investigated variation in brain size and size of different brain parts in first generation laboratory-bred ninespine sticklebacks (Pungitius pungitius) populations from contrasting habitats. We found significant genetically based habitat independent differences in relative brain and cerebellum size among the populations. However, relative sizes of both telencephalon and bulbus olfactorius were significantly larger in marine than in pond fish. Our results suggest genetically based brain size divergence among populations in the wild, and that at least part of this divergence can be attributable to habitat specific natural selection.

Fish Biology Congress Abstracts 37 THEORETICAL CONSIDERATIONS UNDERLYING NA+ UPTAKE MECHANISMS IN FRESHWATER FISHES Goss, G.G., Tresguerres, M., and Parks S.K. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Greg Goss, Dept of Biological Sciences, University of Alberta Edmonton AB T6H 5N7 Canada E-Mail: [email protected] Abstract: Ion and acid–base regulating mechanisms have been studied at the fish gill for almost a century. Original models proposed for Na+ and Cl− uptake, and their linkage with H+ and HCO3 − secretion have changed substantially with the development of more sophisticated physiological techniques. At the freshwater fish gill, two dominant mechanisms for Na+ uptake from been proposed, an apical Na+/H+ exchanger versus an apical Na+ channel electrogenically coupled to an apical H+-ATPase. Advances in molecular biology have greatly enhanced our understanding of the basic ion transport mechanisms at the fish gill. This talk will focus on the recent molecular advances for Na+ uptake in freshwater fish and the relative roles of NHEs and Na+ channels in Na+ transport and acid-base regulation in freshwater fishes. Emphasis will be placed on thermodynamic constraints that prevent electroneutral apical NHE function in most freshwater environments.

RADIO WAVES TAINTED WITH BLOOD: THE LATEST IN FISH TELEMETRY Gräns, A., Axelsson, M., Pitsillides, K., Olsson, C., Höjesjö, J., Kaufman, R.C., Cech J.J., Jr. Symposium: Physiological BioTelemetry Presentation: Oral Contact: Albin Gräns, Dept. of Zoology, Göteborg University, Medicinaregatan 18, Box 463 Göteborg 405 30 Sweden E-Mail: [email protected] Abstract: A fully implantable dual channel radio-based blood flow biotelemetry system was used to measure postprandial effects on cardiac output and gut blood flow in fish. A comparison was made between a continuous feeder (green sturgeon Acipenser medirostris) and an intermittent feeder (northern pike Esox lucius). The two feeding strategies constitute two of the extremes found among fish. The phasic blood flow traces allow calculation of heart rate and stroke volume which gives further information about different strategies for how cardiac output is regulated after feeding. We show that blood flow measurements are possible to conduct with animals swimming freely in large holding tanks instead of using traditional hardwired measurement techniques were the animals have to be more confined. The use of biotelemetry in comparative physiology and applied animal ecology will help scientists to collect information previously impossible to obtain and will open the possibility for new types of ecophysiological studies.

ONTOGENY OF ION REGULATION AND GROWTH IN WILD AND LABORATORY- REARED JUVENILE PINK SALMON (ONCORHYNCHUS GORBUSCHA) Grant, A.M., Gardner, M.N., Nendick, L., Sackville, M., Farrell, A.P. and Brauner, C.J. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Manuela Gardner, Department of Zoology, University of British Columbia, 6270 University Blvd. Vancouver BC V6T 1Z4 Canada E-Mail: [email protected] Abstract: A critical aspect of evaluating the consequences of sea lice infestations on the physiology and survival of juvenile pink salmon is to understand the remarkable developmental trajectory of pink salmon after emergence, since they migrate to sea almost immediately and reportedly grow at rates of 3.5-7.6% per day. Here we present preliminary data for juvenile fish sampled in the Broughton Archipelago (either directly or after transfer to our laboratory) and for hatchery-reared fish. Differences in growth trajectories, whole body ion concentrations and gill Na+/K+-ATPase (NKA) activity were very similar among the three groups. Juveniles tolerated laboratory conditions of 30 ppt salinity at 28 weeks post-hatch. In general, gill NKA activity peaked at 37.5 weeks post-hatch (held in 100% seawater) with a concurrent decline in whole body Na+ and Cl- levels. Following 37.5 weeks, gill NKA activity remained constant while whole body ion levels continued to decline.

COMPARATIVE ENERGETICS AND PHYSIOLOGY OF PARENTAL CARE FOR SMALLMOUTH BASS (MICROPTERUS DOLOMIEU) ACROSS A LATITUDINAL GRADIENT Gravel, M.-A., P. Couture and S.J. Cooke Symposium: Social Behavior Models Presentation: Oral Contact: Marie-Ange Gravel, Department of Biology, 209 Nesbitt building, Carleton University , 1125 Colonel By Drive Ottawa ON K1S 5B6 Canada E-Mail: [email protected] Abstract:

Fish Biology Congress Abstracts 38 Parental care is a costly activity, particularly for temperate species of fish that provide care at the onset of spring, where annual energy stores are depleted after winter. Because latitude and associated climate conditions are known to influence a number of ecological and physiological processes in animals, we wished to test for latitudinal effects during a challenging time, sole paternal care of smallmouth bass. We used physiological indicators such as nucleoside diphosphate kinase (NDPK), plasma indicators of nutritional status, and tissue lipid stores to test for differences at the onset of care and for differential patterns across the care period. We found that fish from southern and mid latitudes had greater potential for biosynthesis, while northern fish had higher muscle energy stores and higher incidences of recent feeding. Our study shows that physiological indicators provide useful information to help elucidate the costs and consequences of reproduction and complex behaviours such as parental care in wild fish.

MOLECULAR AND FUNCTIONAL ANALYSIS OF GOLDFISH TUMOR NECROSIS FACTOR-ALPHA. Grayfer, L., and Belosevic, M. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Leon Grayfer, University of Alberta, Biological Sciences Building Edmonton Alberta T6G 2E9 Canada E-Mail: [email protected] Abstract: In higher vertebrates the cytokine tumor necrosis factor-alpha (TNF-α) is a central molecule in the regulation and execution of pro-inflammatory responses. We recently cloned two isoforms of goldfish TNF-α (TNF-α-1& TNF-α-2), characterized their expression using quantitative PCR and functionally analyzed a recombinant protein form of TNFα-2 (rTNFα-2). The goldfish TNF-α isoforms displayed constitutive expression in goldfish tissues and were up-regulated in primary goldfish macrophages subpopulations in response to activation. The recombinant TNFα-2 enhanced phagocytosis, chemotaxis and the production of reactive nitrogen and oxygen intermediates of goldfish macrophages. The induction of these pro-inflammatory and antimicrobial responses of macrophages was dose- dependent. Unlike the phagocytic response and the induction of the reactive oxygen and nitrogen intermediates in macrophages, which increased after addition of increasing amounts of the rTNFα−2, the chemotactic response in macrophages was induced by lower concentrations (0.07-70ng/ml), and abrogated at higher doses (700-7000ng/ml) of rTNFα-2. Our findings indicate that TNFα is a central pro-inflammatory cytokine of teleosts. [Funded by NSERC]

GHRELIN-INDUCED GROWTH HORMONE AND GONADOTROPIN RELEASE FROM GOLDFISH PITUITARIES UTILIZE BOTH CALCIUM ENTRY AND THE PKC PATHWAY. Grey, C.L., and Chang, J.P. Symposium: Fish Growth Presentation: Oral Contact: Caleb Grey, University of Alberta, Biological Sciences, CW 405 Edmonton Alberta T6G2E9 Canada E-Mail: [email protected] Abstract: Ghrelin (GRL), the endogenous ligand for the growth hormone secretagogue receptor (GHSR), has been identified in goldfish and shown to increase growth hormone (GH) and maturational gonadotropin (GtH-II) release from goldfish pituitary cells. Consistent with a role of GRL in regulating pituitary functions, recent molecular studies have identified and sequenced GHSR fragments from various goldfish tissues, including brain and pituitary. Ca2+ and PKC have both been implicated in GH and GtH-II responses to neuroendocrine regulators. Here we tested the hypothesis that Ca2+ and PKC also mediate GRL actions in the goldfish pituitary. Use of PKC inhibitors bisindolylmaleimide-II and Go-6976 resulted in significant reduction of GRL-induced GH and GtH-II release from perifused goldfish pituitary cells. Similarly, inhibition of voltage gated Ca2+ channels with nifedipine or verapamil also abolished GRL action in goldfish pituitary cells. Furthermore, FURA-II Ca2+ imaging studies show that gGRL19 treatments significantly increase [Ca2+]i in goldfish somatotropes and gonadotropes.

CHARACTERIZATION OF TRANSFERRIN-DERIVED SYNTHETIC PEPTIDE THAT INDUCES PRO- INFLAMMATORY RESPONSES OF MACROPHAGES Haddad, G. and Belosevic, M. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: George Haddad, CW-405 Biological Sciences, University of Alberta Edmonton Alberta T6G2E9 Canada E-Mail: [email protected] Abstract: Transferrin is an iron transporting protein that plays a role in innate immunity of vertebrates. During an inflammatory response, transferrin is cleaved by host enzymes found in immune cells granules as well as invading pathogens, and these cleaved transferrin fragments were shown to induce antimicrobial responses of activated macrophages of teleosts and mammals. We report on the molecular and functional characterization of a macrophage-activating peptide from elastase-

Fish Biology Congress Abstracts 39 digested transferrin. The mass spectrometry-identified transferrin domain had 31 amino acids and was located in the N2 sub-domain of the transferrin N-lobe. The peptide was synthetically produced and shown to induce potent inflammatory responses in murine and goldfish macrophages suggesting that this is a highly conserved mechanism of macrophage activation. [Funded by NSERC, Canada].

DISRUPTION OF THE STRESS ENDOCRINE SYSTEM OF INSHORE WILD FISH OF THE SOUTHERN CALIFORNIA BIGHT Hamilton A.W., Reyes, J.A., Kalman, J.E., Armstrong, J.L. and Kelley, K.M. Symposium: Stress Effects and Response Presentation: Oral Contact: Andrew Hamilton, Environmental Endocrinology Laboratory, Department of Biological Sciences, California State University, Long Beach Long Beach CA 90840 USA E-Mail: [email protected] Abstract: Our studies have consistently demonstrated impaired stress-induced cortisol responses in wild fish sampled from inshore locations in the Southern California Bight (SCB) marine environment, including at sites impacted by wastewater treatment plant (WWTP) outfalls. This phenotypic effect is observed across different fish species, such as English sole (Parophrys vetulus), hornyhead turbot (Pleuronichthys verticalis), California scorpionfish (Scorpaena guttata), and others. The impaired cortisol response may be due, at least in part, to altered interrenal expression of steroidogenic enzymes, including StAR, P450-11beta hydroxylase and 11beta-HSD type-2. In addition, the same animals exhibit significant physiological impacts, such as increased rates of parasitic infestations or alterations in the growth endocrine system, suggesting associated effects on ecologically relevant functions (defense and anabolism). (Support in part by Pacific Coast Environmental Conservancy, San Francisco Estuary Institute and NOAA-USC Sea Grant)

THE COSTS OF ABDOMINAL DISTENTION TO THE PREDATOR ESCAPE RESPONSE OF GRAVID TRINIDADIAN GUPPIES (POECILIA RETICULATA) Handelsman, C., Torres Dowdall, J., Ghalambor, C.K. Symposium: Morphometrics Presentation: Oral Contact: Corey Handelsman, Colorado State University, Department of Biology Fort Collins CO 80523 USA E-Mail: [email protected] Abstract: Pregnancy in live-bearing fish may increase vulnerability to predation because abdominal distention, increases in non- muscle body mass, and altered body shape may reduce swimming performance. However, the relationship between predator escape performance and different stages of gestation in gravid females is poorly understood despite the potential for negative fitness consequences. Thus, in populations experiencing high predation selection should favor reproductive strategies that limit costs to the predator escape response. To test whether predation intensity favors morphologies that limit the costs of gestation to locomotor performance, we compared morphological characteristics and fast start performance at 5 intervals during gestation and following parturition in populations of Trinidadian guppies (Poecilia reticulata) from high and low predation streams. High predation fish showed elevated escape responses and the morphological correlates of these performance differences were compared between individuals. Variation among individuals and populations will be discussed in the context of fitness consequences in the wild.

US VETERINARY IMMUNE REAGENT NETWORK: AN UPDATE Hansen, J.D., Baldwin, C., Lunney, J., Black, S.J., Lillehoj, H., Labresh, J., Horohov, D., Miller, N., Bengten, E., Chinchar, G., Wilson, M., and Wagner, B. Symposium: Immunity and Disease Resistance Presentation: Poster Contact: John Hansen, Western Fisheries Research Center, 6505 NE 65th Street Seattle WA 98115 USA E-Mail: [email protected] Abstract: One major obstacle that limits advances in veterinary immunology and disease control is the lack of sufficient immunological reagents and tools. The US veterinary immune reagent network represents a community-based approach to develop tools and reagents in the form of biologically active proteins and monoclonal antibodies for researcher working on models of animal health including ruminants, swine, poultry, equine and two fish species; catfish and rainbow trout. Monoclonal antibodies are currently being developed to identify the major leukocyte subsets in these species, for reactivity with cytokines/chemokines and their receptors and to target specific receptors that modulate immune responses. These reagents will be useful to research scientists for assessing vaccine design and efficacy and for those evaluating pathology of important veterinary diseases. The project directors working within this network coordinate their efforts with international researchers and continually revise the prioritization of reagents by seeking input from scientists working with these species.

Fish Biology Congress Abstracts 40 COSTIMULATORY RECEPTORS IN FISH: GENOMIC ANALYSIS OF THE CD28 AND B7 FAMILIES Hansen, J.D., Du Pasquier, L., Lefranc, M.P., Sunyer, O., and Boudinot, P. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: John Hansen, Western Fisheries Research Center, 6505 NE 65th Street Seattle WA 98115 USA E-Mail: [email protected] Abstract: Vertebrate T-cells require two signals to be come fully activated. The first signal is specific to a given pathogen as it is provided by the interaction of antigenic peptide-MHC complex with the T-cell receptor. The second signal is delivered to T-cells (in an antigen independent manner) by antigen presenting cells to promote T-cell expansion, effector function and the release of secreted cytokines. Signal two involves a balance of both activating and inhibitory signals via the interaction of CD28 family members on T-cells and B7 family members on antigen presenting cells to regulate the response. To learn more about the diversity of these genes in teleost fish, we performed a comprehensive survey of CD28 and B7 family members using available genomic and EST databases. In a second effort, we assessed the transcriptional regulation of these genes in trout during the immune response to IHNV to learn about their roles in immunity.

THE MORPHOLOGICAL CORRELATES OF MATE CHOICE AND PARENTAL CARE PERFORMANCE IN BLACK BASS Hanson, K.C., Hasler, C.T., Suski, C.D., and Cooke, S.J. Symposium: Morphometrics Presentation: Oral Contact: Kyle Hanson, Department of Biology, Carleton University, 1125 Colonel By Drive Ottawa ON K1S 5B6 Canada E-Mail: [email protected] Abstract: Female mate choice represents a situation whereby a female chooses a male reproductive partner based on direct or indirect fitness benefits to the female. In separate studies, we assessed the role of individual variation in body size and shape on mate choice and parental care performance in black bass (Micropterus spp.). Previous studies have shown that larger males are preferred by females, though no mechanisms for this choice have been tested. If body size is an honest signal of the energetic and nutritional condition of the male, female bass should choose to mate with males whose body shape is indicative of sufficient energy stores to enable the male to care for the brood until it is independent. Similarly, as parental care performance is a whole organism trait that varies between individuals, certain morphological and physiological variables should predispose individuals to be better able to provide quality parental care.

INTRINSIC MECHANICAL PROPERTIES AND HYPERCAPNIC TOLERANCE OF THE PERFUSED ARMOURED CATFISH HEART Hanson, L.M., Baker, D.W., Brauner, C.J., Farrell, A.P. and Val, A.L. Symposium: Tropical Fish Adaptations Presentation: Oral Contact: Linda Hanson, University of British Columbia, c/o Aquaculture 2357 Main Mall Vancouver BC V6T 1Z4 Canada E-Mail: [email protected] Abstract: Armoured catfish (Liposarcus pardalis) possess an unparalleled ability to tolerate severe hypercapnia. Moreover, during hypercapnic exposure L. pardalis leaves extracellular pH largely uncompensated. This creates an extreme challenge for organs, such as the heart, which rely on venous blood and must continue to function optimally under these adverse conditions. In this study, maximum cardiac performance of in situ perfused hearts was not affected at levels of hypercapnia as low as 5% CO2. Conversely, exposure to 7.5% CO2 resulted in a 35% decrease in performance. Nevertheless, the effects of hypercapnia were transitory as cardiac performance was restored following the return to control conditions. Moreover, during extreme hypercapnia L. pardalis was shown to preferentially regulate intracellular pH. We also used the in situ perfused heart preparation to characterize baseline cardiac parameters and cardiac function of L. pardalis as these variables have not been previously reported. Research support provided by NSERC.

EXPRESSION AND LOCALIZATION OF AQUAPORIN WATER CHANNELS IN THE DOGFISH SHARK (SQUALUS ACANTHIAS) Harmon, S., Burch, K., Murray, D. and Cutler C.P. Symposium: Ion and Acid-base Regulation Presentation: Poster Contact: Christopher Cutler, Georgia Southern University, Department of Biology, 202 Georgia Avenue Statesboro GA 30460-8042 USA E-Mail: [email protected] Abstract: Despite publication of a considerable amount of data concerning the role of aquaporin water and small solute channels in teleost fish and in agnathans, to date little or no data has been published from elasmobranches. In the last few years this

Fish Biology Congress Abstracts 41 study has cloned four aquaporin isoforms from the dogfish shark (Squalus acanthias). These include orthologues of human aquaporin 1, 3 and 4, as well as a novel aquaporin, AQP1e. Antibodies have subsequently been raised to each of these proteins. These antibodies have been used in Western blots to determine the level of proteins in various shark tissues. Antibodies have also been used to localize the expression of these aquaporins in the cells or sub-cellular domains of various shark tissues.

USING STABLE ISOTOPES TO ASSESS POTENTIAL SEA LAMPREY DAMAGE TO LAKE SUPERIOR FISHES Harvey, C.J., Ebener, M.P., White, C.K., Fox, J.M., Madenjian, C.P., Essington, T.E., Kitchell, J.F., and Levin, P.S. Symposium: Biology of Lampreys Presentation: Oral Contact: Chris Harvey, Northwest Fisheries Science Center, NOAA Fisheries, 2725 Montlake Blvd. E Seattle WA 98112 USA E-Mail: [email protected] Abstract: Accurate diet information has been unavailable for invasive sea lamprey (Petromyzon marinus) in the Great Lakes, which leads to oversimplified impact assessment. We examined stable isotope ratios of sea lamprey in six regions of Lake Superior in order to generate quantitative diet and host mortality estimates. Top predators such as lake trout (Salvelinus namaycush) comprised 60-90% of sea lamprey diet in all areas except Black Bay, Ontario, where lake whitefish (Coregonus clupeaformis) and catostomids were the major hosts. We integrated stable isotope data and sea lamprey growth data into an individual-based sea lamprey feeding model and generated host-specific estimates of blood loss and mortality. Coregonines accounted for the majority of host deaths. Fewer lake trout deaths occurred, relative to a traditional impact model in which lake trout were the only hosts. In this diverse host assemblage, sea lamprey have more complex community-level effects than portrayed in current impact models.

CONDITION, MIGRATION BEHAVIOUR AND ENERGY USE OF AN IMPERILED STOCK OF CHINOOK SALMON IN A REGULATED RIVER SYSTEM Hasler, C.T., Sunder R., Donaldson, M.R., Guimond, E., Mossop, B., Patterson D., Hinch, S.G., and Cooke, S.J. Symposium: Physiological BioTelemetry Presentation: Oral Contact: Caleb Hasler, c/o Cooke Lab, Department of Biology, Carleton University, 1125 Colonel By Drive Ottawa ON K1S 5B6 Canada E-Mail: [email protected] Abstract: In 2007 an imperiled stock of Chinook salmon were monitored in the Puntledge River, BC. This river system is characterized by a variety of natural and artificial barriers. Male adult Chinook salmon were tagged using either electromyogram (EMG) or radio transmitters and tracked to observe energy use and movement. Each transmitter also had a thermologger attached to it so that thermal histories could be recorded. At the time of tagging, blood was sampled to evaluate physiological status. Fish were then tracked as they moved upstream to their spawning grounds. The physiological data indicated that calcium and magnesium at the time of tagging were negatively correlated with distance traveled, while gross somatic energy density was positively correlated with movement. Other key findings include variable energy use in response to pulse flows and possible utilization of intermediate flows by salmon to facilitate movement over difficult barriers.

PREDATION RELATED VARIATION IN THE SOCIAL COST OF SHOALING IN NINESPINE STICKLEBACK Herczeg, G., Gonda, A., and Merila, J. Symposium: Fish Growth Presentation: Poster Contact: Gábor Herczeg, University of Helsinki, Department of Biological and Environmental Sciences, Ecological Genetics Research Unit, PO box 65 (Biocenter 3, Viikinkaari 1) Helsinki FIN-00014 Finland E-Mail: [email protected] Abstract: The main benefit of grouping is the reduced predatory risk, while the main costs are thought to derive from competition and increased frequency of social interactions. However, as opposed to the benefits of grouping, its costs have rarely been studied. We studied growth of ninespine sticklebacks (Pungitius pungitius) from two marine (high predation) and two pond (low predation) populations by rearing them either individually or in groups. We found that living in groups had a strong (up to 14 %) negative effect on growth in fish from low predation populations, but not on the fish from high predation populations. Our results suggest that the social cost of shoaling can be high, but individuals from high predation risk populations seem to have adapted to minimize these costs better than individuals from low predation risk populations.

Fish Biology Congress Abstracts 42 EVIDENCE FOR AN APICAL NA-CL COTRANSPORTER INVOLVED IN ION UPTAKE IN TILAPIA EMBRYOS Hiroi, J., Yasumasu, S., McCormick, S.D., Hwang, P.P., and Kaneko, T. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Junya HIROI, St. Marianna University School of Medicine, 2-16-1 Sugao, Miyamae-ku Kawasaki Kanagawa 216- 8511 Japan E-Mail: [email protected] Abstract: The objectives of this study were to clone and identify ‘freshwater-type’ and ‘seawater-type’ cation-chloride cotransporters of Mozambique tilapia and to determine their localization patterns within mitochondria-rich cells (MRCs). We first cloned four cation-chloride cotransporter homologs (NKCC1a, NKCC1b, NKCC2 and NCC) from tilapia gills. The NKCC1a and NCC mRNAs were highly expressed in seawater and fresh water, respectively. We then conducted quintuple-color immunofluorescence staining for NKCC1a and NCC, together with Na+/K+-ATPase, CFTR and NHE3 with tilapia embryos acclimated to fresh water or seawater. The NKCC1a immunoreactivity was localized to the basolateral membrane of seawater-specific MRCs, whereas the NCC immunoreactivity was restricted to the apical membrane of freshwater- specific MRCs. We propose a novel ion uptake model by MRCs in fresh water, which incorporates apical NCC, and also revaluate a traditional ion uptake model incorporating apical NHE3.

FUNCTIONAL CHARACTERIZATION OF GOLDFISH CYTOKINE M17 Hitchen, S.J., Hanington, P.C., Grayfer, L., and Belosevic M. Symposium: Immunity and Disease Resistance Presentation: Poster Contact: Steve Hitchen, Biological Sciences Building, University of Alberta Edmonton AB T6G 2E9 Canada E-Mail: [email protected] Abstract: The IL-6 family of cytokines is well characterized in mammals. Members of this family play roles in neurological development, hematopoiesis, and are central to inflammatory processes. Recently we cloned and characterized the goldfish M17, a member of IL-6 cytokine family. Quantitative analysis revealed the highest expression of M17 in the brain followed by the kidney and peripheral blood leukocytes. Functional assessment of a recombinant goldfish M17 demonstrated this cytokine’s ability to induce macrophage proliferation and differentiation from monocytes to macrophages. Additionally, this cytokine enhanced the production of reactive nitrogen and oxygen intermediates, as well as chemotaxis of goldfish macrophages in a concentration dependent manner. [Funded byNSERC]

AUTONOMOUS RECORDING AND TELEMETRY OF FEEDING EVENTS IN FREE SWIMMING SHARKS Holland, K.N., Papastamatiou, Y., Meyer, C.G and Whitney, N Symposium: Physiological BioTelemetry Presentation: Oral Contact: Kim Holland, Hawaii Institute of Marine Biology, University of Hawaii Kaneohe Hi 96744 USA E-Mail: [email protected] Abstract: When fish feed and how much they ingest are critical factors in understanding their ecology. However, these phenomena are difficult to observe in mobile, wide ranging species. We tested ambient sound, stomach motility and pH logging and telemerty devices for their ability to detect and quantify feeding in free ranging blacktip and whitetip reef sharks. All three methods detected feeding events but the most promising data were obtained from the pH (stomach acidity) tags. Results from these devices indicated that changes in stomach pH are related to food type and meal size and these data can be successfully acoustically transmitted from free swimming animals to remote data collectors. Future possible directions for this approach will be discussed.

DIVERGENT CORTISOL STRESS RESPONSIVENESS IN FAMILIES OF ATLANTIC COD (GADUS MORHUA) SUBJECTED TO HANDLING STRESS Hori, T.S., Johnson, S.C., Gamperl, A.K. and Afonso, L.O.B Symposium: Stress Effects and Response Presentation: Oral Contact: Tiago Hori, Ocean Sciences Center - Memorial University of Newfoundland St John's NL A1C 5S7 Canada E-Mail: [email protected] Abstract: In this study, we examined intra-specific variation in the cod’s cortisol response to stress using ten families subjected to repeated handling stress (30 sec out of water) over 5 months, and determined for each family the proportion of high (HR) and low (LR) cortisol responders, and their growth and survival. The final mass of families with a high proportion (~70%) of LR (345g) was significantly greater than for families with a high proportion (~70%) of HR (220g). Further, free-cortisol

Fish Biology Congress Abstracts 43 (FC) levels were higher in HR than in LR fish (an important finding given that only FC interacts with cortisol receptors in target-cells). After the main experiment was completed, the remaining LR and HR fish were subjected to another stressor (heat shock), and differences in the magnitude of the cortisol response between groups was maintained. Taken together, these results suggest that low cortisol responsiveness may be a desirable trait to select for in broodstock development programs.

HEAT SHOCK-RESPONSIVE GENES IDENTIFIED AND VALIDATED IN ATLANTIC COD (GADUS MORHUA) USING GENOMIC TECHNIQUES. Hori, T.S., Kimball, J., Johnson, S.C., Afonso, L.O.B., Bowman, S., Hubert, S., Gamperl, A.K. and Rise, M.L. Symposium: Genomics of Adaptation Presentation: Poster Contact: Tiago Hori, Ocean Sciences Center, Memorial University of Newfoundland St John's NL A1C 5S7 Canada E-Mail: [email protected] Abstract: To identify genes and molecular pathways involved in the cod’s responses to acute heat-stress, we built and characterized Suppression Subtractive Hybridization (SSH) cDNA libraries enriched for genes up-regulated by heat-shock in liver, head kidney, and skeletal muscle. We generated a total of 3677 expressed sequence tags (ESTs) from the three SSH libraries, and contiguous sequences (contigs) had functional annotations including: RNA and protein metabolism, amino acid and carbohydrate metabolism and transport, protein folding, and signal transduction pathways. Further, QPCR revealed that some chaperone genes were significantly up-regulated by heat shock, including GRP78, Gp96, HSP47, HSP90α and HSP71. This latter result contradicting earlier reports suggesting that cod do not express HSP71 following heat-stress. These chaperones and other heat-stress responsive genes may be candidates for marker-assisted selection, and thus used to develop broodstock with improved temperature tolerance. This would of significant benefit to the industry given that sea- cage temperatures can approach this species’ thermal limits.

FUNCTIONAL REGULATION OF H+-ATPASE-RICH CELLS IN ZEBRAFISH EMBRYO ACCLIMATED TO ACIDIC ENVIRONMENT Horng, J.L.1, Chang, W.J.2, Lin, L.Y.3, Yan, J.J.1, Hsiao, C.D.4, Hwang, P.P.1 Symposium: Ion and Acid-base Regulation Presentation: Poster Contact: P.P. Hwang, 1. Institute of Cellular and Organismic Biology, Academia Sinica, Taipei, Taiwan; 2. Institute of bioscience and biotechnology, National Taiwan Ocean University, Keeling, Taiwan; 3. Department of Life Science, National Taiwan Normal University, Taipei, T Taipei 11529 Taiwan E-Mail: [email protected]. Abstract: H+-ATPase-rich (HR) cells, which have been identified in zebrafish skin and gills, are responsible for secreting metabolic acid. The present study was aimed to examine the regulations in function and differentiation of HR cells in zebrafish during environmental acid challenges. After acclimated to acidic (pH 4) freshwater for 3 d, HR cells showed significant increasing in cell number and the area of apical openings, which were accompanied with an enhanced H+ secretion. Zebrafish gcm2 (zgcm2), a transcription factor, was demonstrated to be associated with the differentiation of HR cells based on knockdown experiments, and its mRNA expression in zebrafish gills was found to increase after acclimation to pH4 environment. For conclusion, functional regulation of HR cells is probably achieved by enhancing the cell differentiation via zgcm2 activation.

VISUAL FUNCTION IN CHESAPEAKE BAY'S PREDATORY FISHES Horodysky, A.Z. Brill, R.W., Warrant E.J., Musick, J.A, and Latour, R.J. Symposium: Social Behavior Models Presentation: Oral Contact: Andrij Horodysky, Virginia Institute of Marine Science, Rt 1208 Greate Rd Gloucester Point VA 23062 USA E-Mail: [email protected] Abstract: Little is known about how differences in visual function reflect the lifestyles and feeding strategies of estuarine fishes. We therefore assessed day and night spectral sensitivities, light sensitivities, and flicker fusion frequencies of seven Chesapeake Bay fishes: striped bass (Morone saxatilis), bluefish (Pomatomus saltatrix), weakfish (Cynoscion regalis), spotted seatrout (Cynoscion nebulosus), red drum (Sciaenops ocellatus), summer flounder (Paralichthys dentatus), and cobia (Rachycentron canadum) using electroretinography (ERG). Subjects were presented approximately isoquantal light stimuli covering the spectral range from UV (300 nm) to the near infrared (800 nm) and six orders of magnitude of intensity via a custom- designed computer-controlled system. Peak sensitivities ranged between 450-575 nm, though retinograms showed strong interspecific differences. Weakfish responded to shorter and striped bass to longer wavelengths. Intensity-response and FFF experiments revealed interspecific differences in light sensitivity and FFF. The functional characteristics of fish visual systems appear reflective of their specific niches within the estuarine environment.

Fish Biology Congress Abstracts 44

SOME BIOLOGICAL ASPECTS OF THE GANGES RIVER SPRAT CORICA SOBORNA (PISCES: CLUPEIDAE) IN THE MATHABHANGA RIVER (SW BANGLADESH) Hossain, M.Y., and Ahmed, Z.F. Symposium: Morphometrics Presentation: Poster Contact: Md. Yeamin Hossain, United Graduate School of Agricultural Sciences, Faculty of Fisheries, Kagoshima University, 1-21-24 Korimoto Japan Kagoshima 890-0056 Japan E-Mail: [email protected] Abstract: The present study describes some biological parameters, including sex ratio, length-frequency distributions, length-weight relationship and total length-fork length relationship of the Ganges River sprat Corica soborna (Pisces: Clupeidae), an important fisheries resource in the Mathabhanga River in Bangladesh. A total of 135 specimens ranging from 30.6-48.9 mm total length were analysed in the present study. Sampling was done using traditional bamboo fabricated basket traps, between January and December 2004. The sex ratios showed no significant differences from expected value of 1:1 (P > 0.05). The allometric coefficient b values of the length-weight relationship indicated isometric growth (~3.0) of both males and females (2.946 and 2.967 respectively). The relationship analysis between total length and fork length showed a highly significant correlation in both sexes (r2 > 0.911, P < 0.001).

COMPARATIVE GENOME ANALYSIS OF THE SEX CHROMOSOME OF ATLANTIC SALMON (SALMO SALAR) WITH OTHER TELEOSTS Huang, T-K., Fujiki, K., Li, J., Boroevich, K. A., Koop, B. F., and Davidson, W. S. Symposium: Genomics of Adaptation Presentation: Poster Contact: Teng-Kai (Kevin) Huang, Department of Molecular Biology and Biochemistry, Simon Fraser University, 8888 University Drive Burnaby B.C. V5A 1S6 Canada E-Mail: [email protected] Abstract: In Atlantic salmon, Linkage Group (LG) 1 contains the sex-determining locus, and it was found that LG 1 corresponds to chromosome 2. We have completed a screen of the BAC libraries for all known microsatellite markers on LG 1, and were able to integrate much of the physical and linkage maps in this genomic region. We have also constructed BAC minimum tiling pathways that will eventually cover the entire chromosome 2 of the Atlantic salmon. The sex-determining locus, SEX, is believed to be located near the telomeric end of the q arm on chromosome 2. All the BAC-end sequences assigned to Atlantic salmon LG 1 were subjected to reciprocal-best BLAST searches with the genomes of medaka, stickleback and zebrafish. We found 52 medaka orthologues, 51 stickleback orthologues and 58 zebrafish orthologues. These were used to identify the regions of these fish genomes that are syntenic to Atlantic salmon LG 1.

A MODEL FOR ACID SECRETION/SODIUM UPTAKE IN ZEBRAFISH Hwang, P. P. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: P. P. Hwang, Institute of Cellular and Organismic Biology, Academia Sinica Taipei Taiwan E-Mail: [email protected] Abstract: Current models for acid secretion/Na+ uptake in fresh water (FW) have been proposed based on studies in traditionally used model species, but many inconsistencies and unclear points are still in debate. A new model is proposed based on the recent functional genomic and molecular physiological studies on zebrafish. Specific co-localizations of Na+/H+ exchanger (zNHE3b), carbonic anhydrases (zCA2-like a and zCA15a) in the H+-ATPase rich (HR) cells in zebrafish skin/gills reveal a similarity between zebrafish HR cells and human proximal tubular cells. Acclimation to low-Na+ or acidic FW caused changes in the mRNA expressions of H+-ATPase (zHA), zNHE3b, and zCA15a in zebrafish HR cells. Translational knockdown of zCA2-like a or zCA15a was found to affect Na+ influx and the H+ level at the apical surface of HR cells, as well as the mRNA expressions of zHA and zNHE3b in the morphants. These results support our proposed model.

A MODEL OF ENDOTHELIN SIGNALING IN THE FISH GILL Hyndman, K.A., and Evans, D.H. Symposium: Ion and Acid-base Regulation Presentation: Poster Contact: Kelly Hyndman, University of Florida, Department of Zoology, 321 Bartram Hall Gainesville Fl 326118525 USA E-Mail: [email protected] Abstract:

Fish Biology Congress Abstracts 45 The peptide endothelin-1 (EDN1) is involved in diverse processes such as the regulation of vascular tone and sodium excretion in mammals. In fishes, evidence suggests that EDN1 is also involved in these processes. Our objective was to characterize EDN1 signaling from the euryhaline killifish (Fundulus heteroclitus) gill, and determine if rapid changes in environmental salinity regulates the EDN1 system. By using molecular biology and immunohistochemistry we have modeled EDN1 signaling in the gill, and our model predicts that EDN1 acts as an autocrine and regulates lamellar pillar cell tone. In addition, we predict that EDN1 acts as a paracrine to regulate mitochondrion-rich cell physiology (the main site of ion balance). Finally, we determined that components of this cascade are affected by rapid changes in environmental salinity lending support to the hypothesis that EDN1 is a regulator of ion transport in the gill.

TESTING THE WINTER STRESS SYNDROME HYPOTHESIS IN JUVENILE FISH EXPOSED TO METAL MINING DISCHARGES Janz, D.M., Bennett, P.M., Driedger, K., Kelly, J.M. and Muscatello, J.M. Symposium: Environmental Pollution Presentation: Oral Contact: David Janz, Toxicology Centre, University of Saskatchewan, 44 Campus Drive Saskatoon SK S7N 5B3 Canada E-Mail: [email protected] Abstract: The winter stress syndrome hypothesis states that exposure of fish to contaminants can compromise their ability to allocate sufficient energy to growth and lipid storage, thus decreasing overwinter survival. To test this hypothesis in the field, we collected young-of-the-year fish immediately prior to and following the overwinter period, and measured indices of growth (length, weight, muscle RNA:DNA ratio, muscle proteins) and energy storage (whole body lipids and triglycerides, liver and muscle triglycerides and glycogen). Six species of fish were collected downstream of three metal mining operations in northern Saskatchewan and Ontario, Canada. In contrast to our predictions, most fish species collected in spring exhibited similar or greater lipid stores compared to the previous autumn. Only slimy sculpin (Cottus cognatus) supported the winter stress syndrome hypothesis. Our further research in this area is investigating influences of mining discharges on nutrient enrichment, prey quality/quantity, parasitism rates, size-dependent mortality, and alteration of intermediary metabolism in juvenile fish inhabiting north temperate aquatic ecosystems.

A NOVEL PATTERN RECOGNITION RECEPTOR ON NCC WITH BACTERICIDAL ACTIVITY: ROLE IN THE INNATE IMMUNE RESPONSE OF CATFISH. Jaso-Friedmann, L., Connor M. A., Leary, J. H. III, and Evans, D. L Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Liliana Jaso-Friedmann, Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia Athens GA 30602 USA E-Mail: [email protected] Abstract: Nonspecific cytotoxic cells (NCC) were the first described population of cytotoxic cells in lower vertebrates. Through molecular and functional analysis of the proteins expressed by NCC, it has become apparent that these potent effectors of innate immunity are armed with multiple receptors that respond to diverse stimuli. We have recently identified a new pattern recognition receptor (Ncamp-1) in NCC that preferentially binds bacterial DNA. Secreted Ncamp-1 has bactericidal activity; and Ncamp-1 appears to belong to a family of cationic anti-bacterial peptides of the histone family. WE have compared the spectrum of bactericidal activity of recombinant Ncamp-1 as well as the natural Ncamp-1 expressed in NCC granular extracts. Our results show that recombinant and naturally isolated NCAMP-1 has bactericidal against gram positibe and negative bacteria. The in vivo role of NCAMP-1 was investigated in an infectious disease model using Edwardsiella ictaluri to probe the innate immune response of channel catfish. NCAMP-1 expression on cell membrane of NCC was determined over a time course of infection and spleen and anterior kidney cell counts were done to determine cell trafficking patterns during infection. Gene expression of NCAMP-1, NCCRP-1 and TNF-α was examined by QPCR analysis to investigate transcriptional activation in response to ESC. We hypothesize that NCAMP-1 found in the spleen and anterior kidney plays a role in innate immunity during infection with Edwardsiella ictaluri. These results provide insight into the in vivo expression of Ncamp-1 and suggest that NCC elicit pleiotropic effector functions. This work was funded by grants from the USDA and BARD.

THE ADVANTAGE OF BEING SHY: DO PERSONALITY TRAITS PREDICT FITNESS OF BROWN TROUT (SALMO TRUTTA) IN THE WILD? Johnsson, J. I., and Adriaenssens, B. Symposium: Social Behavior Models Presentation: Poster Contact: Bart Adriaenssens, Gothenburg University, Department of Zoology, Section of Animal Ecology, Box 463 Gothenburg 40530 Sweden E-Mail: [email protected]

Fish Biology Congress Abstracts 46 Abstract: In many animals individual differences in behaviour show remarkable consistency across situations and contexts (i.e. animal personality or behavioural syndromes). Here, we tested for correlation between five behavioural traits in brown trout (Salmo trutta) parr: boldness, activity, aggressiveness, initial (cryptic prey) foraging performance and learning speed. Next we examined the significance of these behaviours for dominance status in a small group and performance under natural conditions (growth, survival and movement). We found support for behavioural syndromes with three pairs of behaviours showing consistent individual variation: boldness-activity, initial foraging performance-aggressiveness and initial foraging performance-activity. Laboratory measures of aggressiveness were a poor indicator of dominance status and neither of these traits affected performance in the wild. In contrast, boldness was negatively correlated with growth under wild conditions. These results emphasize that fitness interpretations based on laboratory measures of behaviour should be made with great caution, and ideally tested in nature.

EARLY HATCHERY REARING DENSITY INFLUENCE BEHAVIOUR AND POST-RELEASE PERFORMANCE IN JUVENILE BROWN TROUT (SALMO TRUTTA) Johnsson, J.I., Brockmark, S., and Adriaenssens, B. Symposium: Contributed Papers Presentation: Oral Contact: Jörgen Johnsson, Department of Zoology, University of Gothenburg, Box 463 Gothenburg Västra Götaland SE 405 30 Sweden E-Mail: [email protected] Abstract: An individual’s phenotype is determined by interactions between genes and the environment in which it develops. Fish reared for conservation or supplementation purposes are often confined at unnatural high densities, which creates a social environment substantially different from the wild. In a series of experiments on juvenile brown trout (Salmo trutta), we investigated how early rearing density influences behavioural development and performance after release in a semi-natural stream. Fish were reared from the egg stage at three densities: conventional hatchery density, intermediate density and natural density. A few months after hatching several individual behavioural traits as well as social dominance were estimated for fish from the different treatments. In addition, fish (40 from each treatment) were released in an enclosed stream and recaptured after a month. The results indicate that a reduced, more nature-like, rearing density enhances development of behaviour, social dominance and post-release growth and survival in hatchery-reared salmonids.

IDENTIFICATION AND CHARACTERIZATION OF VOMERONASAL-LIKE RECEPTOR GENES IN ATLANTIC SALMON Johnstone, K.A., Lubieniecki, K.P., Chow, W., Phillips, R.B., Koop, B.F., and Davidson, W.S. Symposium: Genomics of Adaptation Presentation: Oral Contact: Kimberley Johnstone, Simon Fraser University, 8888 University Drive Burnaby BC V5A 1S6 Canada E-Mail: [email protected] Abstract: Olfactory senses are essential for food selection, mate choice, defence against predators and migration in salmonids. To understand olfaction in Atlantic salmon, we are isolating and characterizing the vomeronasal receptor family genes. Oligonucleotide probes were designed from conserved sequence from both classes of odorant receptors to screen the Atlantic salmon BAC library. Hybridization positive BACs were matched to fingerprint contigs and representative BACs were shotgun cloned and sequenced, identifying the ora1 and ora2 genes as well as 33 putative olfC genes. mRNA was isolated from the olfactory rosette of a smolt to examine the expression of odorant genes. Several of the putative genes that have been identified are expressed in this tissue, confirming that these genes encode for odorant receptors. Future research will focus on the expression of odorant receptors during various developmental and life-history stages, which will help in understanding how discrete populations of salmonids are established and maintained.

VARIATION IN PERIPRANDIAL REGULATION OF NEUROPEPTIDE GENES BETWEEN BEHAVIORALLY DISTINCT POPULATIONS OF ZEBRAFISH (DANIO RERIO) Jones, B.S., Powell, M.S., Drew, R.E., Murdoch, G.K., Oswald, M., Benner, M.J. and Robison, B.D. Symposium: Social Behavior Models Presentation: Poster Contact: Brandon Jones, University of Idaho Department of Biological Sciences Moscow Idaho 83844 USA E-Mail: [email protected] Abstract: Adaptation to captivity in fish commonly causes decreased fear related behaviors and increased foraging. However, the genetic basis of these behavioral changes remains largely unknown. In this study we used one wild strain and one domesticated strain of zebrafish to test the hypothesis that interstrain variation in latency to feed is mediated by differential expression of the neuropeptides npy, agrp1, and cart1. Significant strain and sex effects were observed for place preference

Fish Biology Congress Abstracts 47 and foraging behaviors, with the wild strain exhibiting prolonged feeding latency and patterns of place preference indicative of higher fearfulness. In the diencephalon of the wild strain we observed significantly higher expression of agrp1, an orexigenic factor which stimulates increased appetite. The elevated level of agrp1 expression in the wild strain appears to reflect a reduced foraging rate, which may in turn be related to inhibition of feeding behavior resulting from increased fearfulness.

USE OF A SOLID PHASE IMMUNOPARTITIONING TECHNIQUE TO MONITOR THE DYNAMICS OF THE TROUT ANTIBODY RESPONSE Kaattari, I.M. and Kaattari, S. L. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Ilsa Kaattari, Dept. of Environmental and Aquatic Animal Health, Virginia Institute of Marine Science, College of William and Mary Gloucester Point VA 23062 USA E-Mail: [email protected] Abstract: Past serum affinity maturation studies in both fish and mammals have heavily relied upon classical methodologies for the assessment of antibody affinities. These methods include such techniques as equilibrium dialysis, fluorescence quenching, and more recently, solid phase analytics (e.g. Biacore). Unfortunately, such analyses reduce the usable data to a single, average affinity estimate. Thus, a plethora of potential data is lost as these techniques do not permit the resolution of the constituent affinity subpopulations that yield these estimates. The data presented herein demonstrate that the dynamics of affinity maturational processes occurring in trout to various antigens can be more precisely analyzed by immunopartitioning techniques. Such analyses reveal essential information regarding the expansion and persistence of antigen-specific B cell subpopulations of varying affinities.

UNIQUE TELEOST ANTIBODY ASSEMBLY PROCESSES MAY LEAD TO ENHANCED AFFINITY MATURATION Kaattari, S. l.., Ye, J., and Bromage, E. A. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Stephen Kaattari, Dept. of Environmental and Aquatic Animal Health, Virginia Institute of Marine Science, College of William and Mary Gloucester Point VA 23062 USA E-Mail: [email protected] Abstract: The degree of disulfide polymerization of trout antibodies is conferred in direct proportion to the affinity of the BCR interaction. Thus, low antigen concentrations, which selectively induce the highest affinity B cells, also yield antibodies that are more highly cross-linked. Also, differential antibody immunoadsorption revealed a direct correlation between affinity and disulfide polymerization. Finally, examination of antibodies generated over a 27 week period revealed that, as the process of affinity maturation progressed, so did the relative degree of disulfide polymerization. Passive transfer of antibodies to naïve hosts revealed those antibodies with a lower degree of cross-linking were more quickly removed from the circulation, thus resulting in an increased affinity of the remaining antibodies as well as more highly cross-linked antibodies. In summary, high affinity BCR interactions result in increased antibody disulfide polymerization, with the latter process effectively enabling the animal to distinguish and selectively retain high affinity and / or remove low affinity antibodies from the circulation.

THE EFFECT OF SALINITY STRESS ON APOPTOSIS IN TILAPIA GILL CELLS, ASSESSED USING LASER SCANNING CYTOMETRY (LSC) Kammerer, B., and Kültz, D. Symposium: Ion and Acid-base Regulation Presentation: Poster Contact: Brittany Kammerer, Department of Animal Science, One Shields Ave., University of California, Davis, Davis CA 95616 USA E-Mail: [email protected] Abstract: We have developed a technique for immunocytochemistry of gill cells, used here to assess apoptosis in tilapia gill cells. Epithelial cells were isolated from tilapia acclimated to 25g/l seawater (SW) over a time course: 0 (FW-control), 3, 6, 9, 12, 24, 72, and 120 h. Multivariate LSC analysis based on propidium iodide, Na+/K+ ATPase (NKA) antibody, caspase3&7 and TUNEL apoptotic staining were used to quantify changes in cells over time. We have found that apoptosis increases significantly at 6 hours and 72 hours in tilapia mitochondria-rich cells (MRCs) (NKA-positive) using both apoptotic stains. However, in cells other than MRC (NKA-negative), a peak in apoptosis identified by TUNEL assay at 72 h is missing activity. The reasons for the different kinetics of apoptosis in MRC compared to other gill cells are currently unclear, but taken together, these results indicate extensive cellular turnover in the tilapia gill epithelium during SW acclimation.

Fish Biology Congress Abstracts 48

FISH BIOMECHANICS DATA BASE AND FISH SWIMMING PERFORMANCE Katopodis, C. and Gervais, R. Symposium: Swimming Performance Presentation: Oral Contact: Chris Katopodis, Freshwater Institute, 501 University Crescent Winnipeg MB R3T 2N6 Canada E-Mail: [email protected] Abstract: This study focused on updating a fish biomechanics data base from raceway, fixed velocity (endurance) and increased velocity (critical velocity) tests and re-analysing the results. New or increased numbers of data sets for several species were added to a 1990 data base. Analyses indicate that for some fish species sufficient data exist to estimate fish swimming distance or time to fatigue at various water velocities. Analyses of data sets for groups of species, including fish with small samples, point to broader generalizations. Such generalized relationships are helpful when data for specific species are lacking. Reasonable estimates of swimming performance may be obtained from species groups such as Genera and Families. Estimates of fish speeds and endurance are key components for upstream and downstream fish movements through stream-like or more uniform fish passage systems; they also assist in designing systems to avoid unsuitable habitat conditions for fish, impingement or entrapment, and in possibly hindering the movements of invasive species.

ISOLATION AND FUNCTIONAL CHARACTERIZATION OF GOLDFISH (CARASSIUS AURATUS L.) NEUTROPHILS Katzenback, B.A., and Belosevic, M. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Barbara Katzenback, CW405 Biological Sciences Building, University of Alberta, Edmonton Alberta T6G 2E9 Canada E-Mail: [email protected] Abstract: Neutrophils are one of the first cells at the site of inflammation and central cells of innate immunity of vertebrates. Upon activation, neutrophils are capable of producing an intense respiratory burst response, they rapidly degranulate in the presence of pathogens, and deploy neutrophil extracellular traps that efficiently remove infectious agents. We report on the isolation, cultivation and characterization of neutrophils isolated from goldfish kidney. Neutrophils stained positive for acid phosphatase and myeloperoxidase, and produced a dose-dependent respiratory burst response after stimulation with phorbol esters (PMA). Furthermore, treatment of neutrophil cultures with either PMA, calcium ionophore A23187 (CaI) or zymosan induced a dose-dependent non-reversible degranulation of goldfish neutrophils. The development of methods for isolation and primary cultivation of goldfish neutrophils will permit further examination of their role in host defense against fish pathogens. [Funded by NSERC]

ENVIRONMENTAL DISRUPTION OF GROWTH ENDOCRINE FACTORS AND PROTEOME BIOMARKERS IN WILD FISH FROM IMPACTED MARINE AND ESTUARINE ENVIRONMENTS IN CALIFORNIA Kelley, K.M., Reyes, J.A., Armstrong, J.L., Brar, N.K., Adams, S.A., Waggoner, C.M., Hall, W.A., Cianfrani, D.M. and Fairey, R. Symposium: Fish Growth Presentation: Oral Contact: Kevin M. Kelley, Environmental Endocrinology Laboratory, Biological Sciences Department, California State University, Long Beach Long Beach CA 90840 USA E-Mail: [email protected] Abstract: Many locations within San Francisco Bay and the Southern California Bight are known to be polluted by a variety of human-derived contaminants. However, the extent to which these contaminants impact the biota are not understood. Our studies of several different fish species residing in contaminated environments (of different characteristics) from both northern and southern California indicate that endocrine factors important in somatic growth are exhibiting environment- associated disruptions. These factors include IGF-I, IGFBP-1, thyroxine, triiodothyronine, and cortisol. The overall pattern of alterations in these endocrine factors suggests reduced somatic growth and anabolism, which is supported in part by data indicating that cartilage proteoglycan synthetic (“growth”) activity is also reduced in impacted fish. Using proteome screening, we have identified additional phenotypic biomarkers in the impacted fish, indicative of exposure to environmental contaminants and of additional physiological effects. (Support by NOAA-USC Sea Grant, Pacific Coast Environmental Conservancy, and San Francisco Estuary Institute)

BRIDGING THE GAP BETWEEN HYDRAULICS AND FISH BEHAVIOUR: THE APPLICATION TO FISH PASSAGE

Fish Biology Congress Abstracts 49 Kemp, P. S., and Russon, I. J. Symposium: Swimming Performance Presentation: Oral Contact: Paul Kemp, International Centre for Ecohydraulic Research, School of Civil Engineering and the Environment, University of Southampton Southampton Hampshire SO17 1BJ UK E-Mail: [email protected] Abstract: The ability of fish to negotiate either natural or anthropogenic impediments to migration significantly influences individual fitness and population status. Barriers to migration are not only related to physical structures per se, but to the hydraulic conditions created. Traditionally, the development of criteria that describes the ability of fish to pass obstructions has been based on the swimming capability of economically significant species such as salmonids that exhibit subcarangiform locomotion. Fish passage criteria for non-commercial species have been less often considered, especially those that exhibit anguilliform swimming. Further, the behavioural response of fish to hydraulic conditions created are usually ignored, despite evidence that fish exhibit avoidance at impediments that they would otherwise be expected to pass based on swimming capability alone. In Europe, species of lamprey are provided legislative protection. Further, the EU Water Framework Directive requires that member states ensure water bodies are maintained under conditions of good ecological status, or potential if heavily modified. Thus, research is required to better define fish passage criteria for species of lamprey, based on both swimming capabilities and behaviour. This presentation will highlight interdisciplinary research conducted at the International Centre for Ecohydraulics Research (ICER) at the University of Southampton. Fish biologists work closely with hydraulic engineers to quantify the response of lamprey to artificial structures placed in flumes that mimic semi-natural conditions in an effort to develop generic models that may help water resource manager's better provision fish passage strategies. The results of these studies will be discussed.

ANGUILLIFORM LOCOMOTION AND CLIMBING IN LAMPREY Kemp, P. S., Tsuzaki, T., and Moser, M, J. Symposium: Biology of Lampreys Presentation: Oral Contact: Paul Kemp, International Centre for Ecohydraulic Research, School of Civil Engineering and the Environment, University of Southampton, SO17 1BJ. UK Southampton Hampshire SO17 1BJ UK E-Mail: [email protected] Abstract: Lamprey exhibit anguilliform locomotion. Some species appear to employ a modified form of anguilliform locomotion in order to climb vertical structures. The Pacific lamprey (Lampetra tridentata) is capable of climbing waterfalls during it natural migration. How climbing performance is influenced by prior experience and behaviour is of interest, however, because it will directly influence assessments of fish passage criteria. To enhance understanding of this relationship, the kinematics, behaviour, and performance of Pacific lamprey that climbed a vertical 1.4 m high experimental weir were assessed. Propulsion was achieved during powerful cycles of axial undulation initiated from a stationary position of attachment of the oral disk. Most ascents employed intermittent locomotion when climbing (representing on average approximately one-fifth of total ascent time), interspersed with periods of inactive attachment. However, two of the most rapid ascents (57 and 85s) occurred during one continuous bout, at a low cycle frequency. Probability of success and velocity of ascent were positively related to experience of climbing the weir. The duration of bouts of climbing relative to time spent stationary was negatively related to vertical distance travelled, indicative of fatigue. Ascents with long periods of activity had correspondingly high durations of recovery, while the duration of ascents that took less than 300s, was positively related to the proportion of time spent stationary. The findings suggest that modification of intermittent climbing locomotion will compensate for variation in climbing performance and extend distance travelled before exhaustion.

FUNCTIONAL AND ENERGETIC CHARACTERIZATION OF P-GLYCOPROTEIN-MEDIATED EFFLUX OF XENOBIOTICS IN TELEOSTS Kennedy, C.J., Hildebrand, J.L., Bains, O., and Lee, D.H.S. Symposium: Environmental Pollution Presentation: Oral Contact: Chris Kennedy, Department of Biological Sciences, Simon Fraser University, 8888 University Drive Burnaby BC V5A 1S6 Canada E-Mail: [email protected] Abstract: Numerous mechanisms exist to defend teleosts against contaminants in the environment, all of which have a common strategy: to reduce cellular accumulation. Here, the functional characterization and energetic costs associated with two model substrates, rhodamine 123 (R123) and doxorubicin (DOX) were examined in isolated hepatocytes of rainbow trout. Accumulation and efflux rates of both compounds were concentration dependent. Costs of DOX efflux were measured through cell adenylate concentrations and R123 by cell respiration rates. The efflux of DOX resulted in significant decreases in intracellular ATP concentrations, adenylate energy charge, and phosphorylation potential and significant

Fish Biology Congress Abstracts 50 increases in concentrations of ADP, AMP and inorganic phosphate. Respiration rates were significantly higher in cells exposed R123. Costs were attributed specifically to P-gp mediated efflux by use of the non-competitive P-gp inhibitor, XR9576. This study demonstrates that increased P-gp transport of xenobiotics may result in higher energy costs for organisms living in contaminated environments.

RESISTANCE AND RECOVERY FROM MYXOZOAN AND MICROSPORIDIAN INFECTIONS IN SALMONID FISHES Kent, M.L., Bartholomew, J.L., Hedrick, R.P. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Michael Kent, Department of Microbiology, 220 Nash Hall, Oregon State University Corvallis OR 97330 USA E-Mail: [email protected] Abstract: Parasitic diseases represent some of the most important infectious diseases of wild and captive fishes. Compared to bacteria and viruses, little research has been conducted on understanding mechanisms of resistance and recovery, or on vaccine development. Indeed, Ichthyphothirius multifiliis represents the only fish parasite for which vaccines have been developed. The following are some examples of myxozoan or microsporidian parasites of salmonid fishes for which immunity or resistance exists. Examples of innate resistance at species level include Atlantic salmon resistance to Loma salmonae (Microsporidia) and brown trout resistance to Myxobolus cerebralis (Myxozoa). Significant differences in resistance to Ceratomyxa shasta (Myxozoa) and M. cerebralis exist between strains of rainbow trout from endemic waters versus those from watersheds where the parasite is absent or newly introduced. Close to sterile acquired immunity occurs following natural exposure and recovery from infections by both Tetracapsaloides bryosalmonae (the PKX myxozoan) and L. salmonae with Oncorhynchus species.

ETHOLOGICAL EVALUATION OF CHROMATOPHORE DISTRIBUTION IN TILAPIA- OREOCHROMIS MOSSAMBICUS Kesavan, K., Padmakumar, K.B., and Menon, N.R Symposium: Stress Effects and Response Presentation: Oral Contact: K. Kesavan, Dept. Marine Biology, School of Marine Science, Fine Arts Ave, Cochin University of Science and Technology Cochin Kerala 682016 INDIA E-Mail: [email protected] Abstract: The relationship between changes in colour pattern and activity in fishes is a subject of speculation among researchers. Behavioural studies in various fishes are almost always linked with changes in the pattern of body colour even though there is no unanimity of opinion about the biological processes involved. The present work is made to identify the colour pattern in Oreochromis mossambicus, which resulted during breeding activity under captivity in glass aquaria, and perceivable alterations in body colour as an effect of stress. Variations in chromatophore concentration were observed when the specimens were exposed to the dye malachite green. Altogether seven shade patterns observed were (1) inactive (pale or dirty yellowish white), (2) activated juvenile (barred with conspicuous ‘Tilapia mark’), (3) activated adult (darkening of the dorsal half of the body- common to both sexes), (4) aroused adult (darkening of caudal, anal and pelvic fins- common to both sexes), (5) courting female (whole body turns black, margins of pectoral, dorsal and caudal fins reddish), (6) brooding female (striped with a distinct mid lateral stripe, blackening of eyes, jaw and operculum) and (7) aggressive male (intensified darkening of the body with whitish jaw and operculum, margins of dorsal and caudal reddish). The colour changes were also found to be influenced by background shades and/ or light intensity. Spurting movements with flushing of water evidenced by increased opercular rate were considered as the index of activated state of the animal. Some of the patterns were associated with behavioural responses in accordance with the changes in the situation while certain others were related to sexual courting behaviour.

IS THE BIRTH CONTROL PILL AN EFFECTIVE FORM OF CONTRACEPTION FOR WILD FISHES? Kidd, K. Symposium: Plenary Presentation: Oral Contact: Karen Kidd, Canadian Rivers Institute & Biology Department, University of New Brunswick Saint John New Brunswick Canada E-Mail: [email protected] Abstract: It is well known that municipal effluents contain substances that affect reproductive endocrine function in wild fishes. However, it is not well understood whether the responses observed at the organism level, such as the production of the egg protein precursor vitellogenin in male fish downstream of these discharges, are indicative of impacts at the population level. To investigate this, a whole lake experiment was done at the Experimental Lakes Area (ELA) in northwestern Ontario,

Fish Biology Congress Abstracts 51 Canada from 1999-2006 which followed the effects of the synthetic estrogen ethynylestradiol (EE2) used in birth control pills on the fish populations and their supporting food web. Continuous additions of EE2 (5-6 ng/L) occurred in the summers of 2001-2003; biochemical- and tissue-level endpoints were examined in several species of fish and population data were collected for all trophic levels before, during and after EE2 additions and contrasted to reference lake data. The experiment was successful at reproducing (no pun intended) the impacts observed downstream of wastewater discharges. Male fishes from the treated lake produced high concentrations of vitellogenin and exhibited gonadal abnormalities. In the second and third summer of additions, we observed reproductive failures for the fathead minnow, with losses of the smallest size classes and a subsequent collapse in the population. These results indicate that chronic exposure to low concentrations of the estrogen used in birth control pills can impact the sustainability of fish populations.

PLASMA LEPTIN LEVELS IN SALMONIDS DURING FOOD RESTRICTION AND EFFECTS OF HOMOLOGOUS LEPTIN TREATMENTS ON FEED INTAKE Kling, P., Rønnestad, I., Moen, A-G. G, Murashita, K., Kurokawa, T., Stefansson, S. O., Jönsson, E. and Björnsson, B. Th. Symposium: Fish Growth Presentation: Poster Contact: Peter Kling, Department of Zoology/Zoophysiology, Medicinaregatan 18, Gothenburg University Gothenburg Sweden E-Mail: [email protected] Abstract: In mammals, leptin (Lep) is an anorexigenic hormone regulating appetite and energy homeostasis. To gain insight into the function of Lep in fish, a salmon Lep radioimmunoassay (sLep-RIA) was established. Plasma Lep levels were assessed in rainbow trout during 1- 3 week starvation, and in Atlantic salmon during months of low (60%) ration. Both studies indicate that feed restriction results in increased plasma Lep levels. No clear correlation between condition factor and Lep levels were observed suggesting that Lep does not act as an adiposity signal in salmonids. In rainbow trout treatment studies, full- length, homologous Lep was used to examine its short-term effects on feed intake. While central (icv) injections were without effect, ip injections suppressed feed intake. Taken together these results suggest that the function of Lep in salmonids may differ in many aspects from that in mammals, possibly pointing to functional divergence between ectothermic and endothermic vertebrates.

CAN ENERGETICS EXPLAIN INTRASPECIFIC HOME RANGE ALLOMETRY IN AN APEX PREDATOR, THE NORTHERN PIKE? Knight, C. M., Gozlan, R. E., Lucas, M. C. Symposium: Physiological BioTelemetry Presentation: Oral Contact: Carolyn Knight, Norwegian Institute for Water Research (NIVA), Gaustadalléen 21 Oslo NO-0349 Norway E-Mail: [email protected] Abstract: Numerous studies have investigated cross species allometric relationships in home range size, with mammals, birds and fishes all subjects of consideration. However, despite a plethora of studies recording single species movements and home range activity through telemetry, none have considered allometric scaling of individuals within a species or the behavioural or energetic basis for such relationships. Using data collected with radio telemetry on a wide size range (14-101 cm) of riverine northern pike (Esox lucius) we found a strong allometric relationship with home range size increasing as pike age and size increased (R2 ≥ 0. 47, P ≤ 0.001 in both cases). Moreover, individual growth rate was found to be negatively correlated with home range size (R2 ≥ 0.3, P < 0.001). Using the large amount of available information on pike energetics and standard metabolic rate we consider the energetic basis of home range allometric scaling in pike.

COLLECTION OF JUVENILE SALMONIDS AT COWLITZ FALLS DAM: PHYSICAL AND HYDRAULIC CONDITIONS THAT RESULT IN DELAY OR REJECTION OF VOLITIIONAL ENTRY INTO A SURFACE COLLECTION SYSTEM Kock, T.J., Liedtke, T.L., Rondorf, D.W., Serl, J.D., and Kohn, M. Symposium: Swimming Performance Presentation: Oral Contact: Tobias Kock, U.S. Geological Survey, Columbia River Research Laboratory, 5501A Cook-Underwood Road Cook WA 98605 USA E-Mail: [email protected] Abstract: Efforts to collect juvenile salmonids at Cowlitz Falls Dam began in 1997 with the construction of a surface collection system. This system has been only marginally successful at collecting juveniles; annual mark-recapture studies suggest that the collection system successfully captures only about 20-50% of the juvenile steelhead, coho salmon, and Chinook salmon that enter the dam forebay each year. Numerous efforts have been made to improve the system including the use of directed flow, altering entrance configurations, and the installation of a prototype fish screen. Using telemetry, acoustic

Fish Biology Congress Abstracts 52 camera, and hydraulic monitoring research techniques we were able to describe spatially limited zones upstream of collection flumes and inside the fish screen where fish either delayed or rejected volitional entry into the collection system. Our presentation will focus on physical and hydraulic factors that may serve to limit the collection of juvenile salmonids at surface collection facilities.

EVALUATING THE BARRIER POTENTIAL OF INSTREAM OBSTACLES TO UPSTREAM BROOK TROUT MOVEMENT Kondratieff, M.C. Symposium: Swimming Performance Presentation: Oral Contact: Matt Kondratieff, 317 West Prospect Road Fort Collins CO 80526 USA E-Mail: [email protected] Abstract: Instream obstacles such as culverts and waterfalls have the potential to serve as barriers to the upstream movement of brook trout (Salvelinus fontinalis). The barrier potential of these obstacles can be evaluated through long-term fish mark- displacement-recapture studies or by more rapid theoretical studies that rely on model-driven analyses of the physical dimensions and characteristics of the obstacles. It is unclear whether these two approaches are comparable. The barrier potential of instream obstacles to upstream brook trout movement was evaluated using both approaches. A comparison of these approaches suggests that field evaluations, while time-consuming, were useful as a decision tool for identifying which instream obstacles functioned as barriers, monitoring barrier performance over time, and identifying strategies for increasing the barrier potential of existing obstacles. Theoretical approaches were useful for categorizing the relative vulnerabilities of instream obstacles to upstream fish passage and for guiding barrier designs.

ROLE OF HISTIDINE RESIDUE 151 IN THE THIRD TRANSMEMBRANE DOMAIN OF AN ANCESTRAL GONADOTROPIN-RELEASING HORMONE RECEPTOR FROM THE SEA LAMPREY, PETROMYZON MARINUS Kosugi, T., Fremat, M., Kavanaugh, S.I. and S.A. Sower. Symposium: Biology of Lampreys Presentation: Poster Contact: Takayoshi Kosugi, Department of Biochemistry and Molecular Biology, University of New Hampshire Rudman Hall, 46 College Road, Durham NH 3824 USA E-Mail: [email protected] Abstract: Gonadotropin-releasing hormone (GnRH) is a central regulator of reproductive function in vertebrate, and its function is mediated through a GnRH receptor (GnRHR), a class A 7-transmembrane G-protein coupled receptor (GPCR). To date, multiple forms of GnRHR, seemingly concomitant with multiple forms of its ligand, GnRH, have been cloned in different class of vertebrates. These known GnRHRs in vertebrates have a variation of DRY motif located in the third transmembrane domain, which is a conserved motif in the GPCR superfamily. In the GnRHR of the sea lamprey, one of the oldest lineage of vertebrates, the Tyr residue of the DRY motif is replaced by His but its functional property is unclear. In this study, the functional significance of the His residue of the DRH motif in the ancestral form of GnRHR has been evaluated by site-directed mutagenesis. Supported by NIH 5R21RR024477-02

DIVERSITY OF MORMYRID FISH IN SOUTHERN AFRICA: A MULTIDISCIPLINARY APPROACH Kramer, B. Symposium: Tropical Fish Adaptations Presentation: Oral Contact: Bernd Kramer, University of Regensburg, Zoological Institute Regensburg Bavaria D-93040 Germany E-Mail: [email protected] Abstract: The Okavango delta and an adjacent paleo-lake in southern Africa have been identified as an evolutionary hotspot for haplochromine cichlids, resulting in a multitude of species that have seeded adjacent river systems. This could well turn out to be true also for snoutfishes (Mormyridae). The Upper Zambezi-Okavango system and its historically related rivers often hold species complexes for what was thought to represent single, wide-spread species. The genera Marcusenius, Petrocephalus, Pollimyrus and Hippopotamyrus provide examples of allopatric, parapatric, and sympatric differentiation on the species level. Critical comparisons among allopatric populations have greatly profited from combining morphometrics, DNA sequence analyses, electric organ discharge, and mating call comparisons. Playback experiments and detailed behavioural analyses of courtship and spawning behaviour have provided additional valuable information that can assist in resolving the researcher’s dilemma “cline vs differentiation”, in addition providing insight into the evolution of a communication system.

FEMALE CHOICE BY ELECTRIC ORGAN DISCHARGE IN A MORMYRID FISH

Fish Biology Congress Abstracts 53 Kramer, B., and Machnik, P. Symposium: Social Behavior Models Presentation: Poster Contact: Bernd Kramer, University of Regensburg, Zoological Institute Regensburg Bavaria D-93040 Germany E-Mail: [email protected] Abstract: In adult males of the South African weakly electric Bulldog fish, Marcusenius pongolensis, the duration of the Electric Organ Discharge (EOD) increases with body size over lifetime. While there is experimental support for intrasexual selection (male-male competition) having shaped the males’ EOD pulse duration in evolution, nothing is known about intersexual selection, such as female choice. We conducted two series of playback tests, one using 25 natural male EODs of pulse duration varying from 320 µs (average female value) to 716 µs, for playback via a single dipole model of an electric fish, the second by simultaneously pairing the shortest pulse with one of four gradually longer ones, to 8 or 10 female experimental subjects (1st and 2nd series, respectively). In all subjects but one (1st series), response scores in behaviours such as observed in reproducing females increased significantly with pulse duration. We suggest female choice as an explanation.

A NEW SPECIES OF SLENDER STONEBASHER FROM SOUTHERN AFRICA (TELEOSTEI: MORMYRIFORMES) Kramer, B., and Swartz, E. Symposium: Morphometrics Presentation: Poster Contact: Bernd Kramer, University of Regensburg, Zoological Institute Regensburg Bavaria D-93040 Germany E-Mail: [email protected] Abstract: Previous work has revealed specimens referable to Hippopotamyrus ansorgii (Boulenger, 1905) sampled from Namibia’s part of the Upper Zambezi system, to represent a species complex the members of which are well differentiated from the Angolan Type specimens. In the present study, we sampled an Angolan River, the Cunene, one of several possible type rivers for H. ansorgii, on its lower section where it borders Namibia. The specimens are morphologically differentiated from the six other morphs within the H. ansorgii species complex we are aware of, including the Types. The pulse waveform of the Cunene specimens’ Electric Organ Discharge, as well as the associated amplitude spectra, is categorically different from all others known. Based on mitochondrial DNA analysis, the Cunene samples belong to a distinct historically isolated lineage, further supporting their status as a new species We recognise the Lower Cunene morph as the new species, Hippopotamyrus longilateralis.

TRADEOFF BETWEEN MALE ATTRACTIVENESS AND PREDATOR AVOIDANCE IN AN ELECTRIC FISH? Kramer, B., Hanika, S. Symposium: Social Behavior Models Presentation: Poster Contact: Bernd Kramer, University of Regensburg, Zoological Institute Regensburg Bavaria D-93040 Germany E-Mail: [email protected] Abstract: Unlike females, male Marcusenius pongolensis generate Electric Organ Discharges the duration of which increases over lifetime. Experimental support suggests that in evolution long male pulses have been selected for both intra- and intersexually. However, long-duration EODs not only confer fitness advantages but also alert predatory catfish. Here, we demonstrate short-term plasticity in male EOD waveform, with an increase limited to the reproductive season, as a likely solution to the dilemma "longevity versus fitness”. In 10 captive M. pongolensis, an increase in EOD duration was evoked by introducing stranger males in adjacent compartments that were separated by plastic mesh, thus increasing territorial competition and aggression. EOD duration values exceeded those found in the non-reproductive season in nature, and pulse duration receded as soon as solid plastic walls separated the fish. Similar changes occurred in males reproducing in the laboratory that receded to a short EOD as soon as reproduction was over.

PHYSIOLOGICAL RESPONSES TO WATER QUALITY IN LARGE SCALE COMMERCIAL ATLANTIC SALMON SMOLT PRODUCTION Kristensen, T., Rosten, T., Urke, H., Åtland, Å., Kroglund, F. and Rosseland, B.O. Symposium: Fish Living on the Edge Presentation: Oral Contact: Torstein Kristensen, Norwegian Institute for Water Research, Gaustadalléen 21, NO-0349 Oslo NO-0349 Norway E-Mail: [email protected] Abstract:

Fish Biology Congress Abstracts 54 Commercial production of Atlantic salmon smolts in Norway has gone through a large degree of upscaling, with the largest production tank units reaching 1200 m3. Production intensity in terms of specific water use (liters of water/kg fish/minute) is variable, but generally quite high, requiring efficient oxygenation systems and in many cases CO2 aeration. We have tested and evaluated the use of a "point of care" analyzer (I-Stat) for physiological parameters describing ionoregulatory and acid base status of the fish in large scale systems, as well as the capacity of such parameters in predicting critical water quality parameters. Despite the "crude" sampling methodology, involving netting from tanks and a time lag of up to 10 minutes before sampling, the results from several sampling sites consistently show differences between groups of fish reared at various intensity levels in both ionoregulatory parameters and acid-base balance. The robustness of these parameters were demonstrated in a power analysis by low least significant number for e.g. sodium and bicarbonate. Strong correlations were found between blood PCO2/HCO3- and water CO2 levels, The use of the I-stat point of care analyzer gave a robust analysis on relevant parameters for fish health, and have a strong predictive value on production intensity in large scale production.

THE INTRASPECIFIC STRUCTURE OF THE ARCTIC LAMPREY LETHENTERON CAMTSCHATICUM, AND ITS FORMATION IN THE RIVERS OF WESTERN KAMCHATKA (BY THE EXAMPLE OF THE UTKHOLOK RIVER) Kucheryavyy, A.V.(a,b), Savvaitova, K.A.(b), Pavlov, D.S.(a,b), Stanford, J.A.(c), Gruzdeva, M.A.(b), and Kuzishchin, K.V.(b) Symposium: Biology of Lampreys Presentation: Oral Contact: Alexandr Kucheryavyy, a) Icthyology Department, Biological Science Faculty, Moscow State University, GSP-2, 119992; b) Institution of Ecology and Evolution, Russian Academy of Science, Leninsky Prospect, 33, Moscow, Russia; c) Flathead Lake Biological Station, The University o Moscow Russian Federation E-Mail: [email protected] Abstract: In the meandering, brown-water Utkholok River (57V 372323E 63827N) 1000 or more amoecetes per m2 were commonly observed in depositional areas. Five life forms of this species were documented using morphometric and behavioral metrics: ammocoetes (4 years in river, based on statoliths), smolts (two distinct types: spring migrant, larger summer type, based on catches in fyke nets), resident adults , anadromous early maturing adults (forma praecox – 95% males, one year in ocean), typically anadromous adults form (2 years in ocean). The three adult forms spawned together in entwined masses in riffles. The tendency to residualize (mature in fresh water) was linked to pre-smolt ammoecete nutrition as interpreted by concentration of 15Sr in tissues indicating marine food source: individuals completing juvenile life history in salmon carcasses (high nutrition, high marine 15Sr signal) residualized, whereas those reared in the benthos (lower nutrition, low marine signal) migrated to marine environments.

A METABOLOMICS APPROACH TO STUDY FISH METABOLISM AND GROWTH: EFFECTS OF FOOD DEPRIVATION IN RAINBOW TROUT (ONCORHYNCHUS MYKISS) Kullgren, A., Samuelsson, L., Larsson, D.G.J., Björnsson, B.Th., and Jönsson Bergman, E. Symposium: Fish Growth Presentation: Poster Contact: Andreas Kullgren, Department of Zoology, University of Gothenburg, Box 463 Gothenburg SE-405 30 Sweden E-Mail: [email protected] Abstract: This study was undertaken as a first step in evaluating the suitability of NMR metabolomics for research on metabolism and its endocrine regulation in salmonids. Rainbow trout (Oncorhynchus mykiss) weighing 43-115 g were either fed ad libitum or deprived of food for 28 days at 10 C to investigate catabolic effects on energy reserves and metabolite patterns. Body weight and length, liver weight, mesenteric fat mass and muscle lipid content were all reduced by food deprivation compared with the control group. NMR metabolomics analysis of blood plasma showed that food deprivation affects plasma levels of several endogenous plasma metabolites. Identification and characterization of such complex profile changes in the metabolome will be used as a research tool in future studies in order to provide a comprehensive picture of metabolic flows under various environmental conditions as well as to elucidate the endocrine control of metabolism in fish.

THE MOVEMENT, LENGTH, AND CONDITION OF BROOK TROUT (SALVELINUS FONTINALIS) IN THE HURRICANE RIVER, PICTURED ROCKS NATIONAL LAKESHORE, MICHIGAN Kusnierz, P.C. and Leonard, J.B.K. Symposium: Morphometrics Presentation: Poster Contact: Paul Kusnierz, Northern Michigan University, 1401 Presque Isle Ave. Marquette MI 49855 USA E-Mail: [email protected] Abstract:

Fish Biology Congress Abstracts 55 Coaster brook trout (Salvelinus fontinalis) were once found throughout Lake Superior, Lake Nipigon, and their tributaries. These fish were popular with sport fishermen and were known to grow larger and mature later than their stream resident counterparts. The impetus for this study was to compare the movement patterns, age, length, and condition of wild coaster and resident brook trout from the Hurricane River, Pictured Rocks National Lakeshore, Michigan. Mean length was not significantly different between coasters and residents, but K was for all tagged fish at ages 1 and 3 as well as for all ages combined. Most brook trout movement took place in the fall with October being the peak month of coasting behavior. The data collected suggest that all brook trout in the Hurricane River may potentially be coasters and that coasters in this stream are smaller and mature younger than fish from other coaster populations.

EFFECTS OF CROWDING ON ENZYME ACTIVITIES AND GENE EXPRESSION OF ORNITHINE- UREA CYCLE ENZYMES IN THE GULF TOADFISH (OPSANUS BETA) Laberge, T.L., Walsh, P.J., and McDonald, M.D. Symposium: Fish Living on the Edge Presentation: Oral Contact: Tammy Laberge MacDonald, Marine Biology and Fisheries, University of Miami, RSMAS, 4600 Rickenbacker Causeway Miami Fl 33149 USA E-Mail: [email protected] Abstract: The gulf toadfish (Opsanus beta) is a facultatively ureotelic fish that excretes primarily urea when crowded. To examine the relationship between ammonia and urea production, ornithine-urea cycle (O-UC) enzyme activities and O-UC mRNA expression, we crowded toadfish for two days and one week, respectively, to activate the O-UC. Ammonia and urea levels were assayed in liver and plasma cortisol levels were measured. Liver enzyme activities were measured for glutamine synthetase (GS), carbamoyl phosphate synthetase (CPS III), ornithine transcarbamylase (OTC), and arginase (ARG). Messenger RNA expression was determined for GS, CPS III, OTC, ARG, argininosuccinate synthetase (ASS) and argininosuccinate lyase (ASL). Crowded fish had lower ammonia levels, increased urea production and elevated cortisol levels. Crowded fish also had elevated O-UC enzyme activities, but only CPS and ASS had significant changes in mRNA expression. These results suggest that enzyme activities of O-UC genes are better predictors of liver urea levels.

EXAMINING THE FATE OF DUPLICATED REGIONS IN THE ATLANTIC SALMON (SALMO SALAR) GENOME USING FABP2 GENES Lai, Y., Lubieniecki, K., Phillips, R., Koop, B.F. and Davidson, W.S. Symposium: Genomics of Adaptation Presentation: Poster Contact: Yvonne Lai, Dept of Molecular Biology and Biochemistry, 8888 University Drive, Burnaby B.C. V5A 1S6 Canada E-Mail: [email protected] Abstract: Salmonids are thought to have undergone two additional genome duplications compared to mammals. In the present study, the structure and organization of the intestinal fatty acid binding protein (FABP2) genes in Atlantic salmon were examined to understand how salmonid genomes have become reorgainized as part of the re-diploidization process. The cGRASP EST database revealed three unique FABP2 transcripts in salmon compared to one fabp2 transcript in zebrafish, suggesting there is a loss of a duplicated gene both in the zebrafish and salmon lineages. A preliminary tissue expression showed different expression pattern of the three FABP2 genes. Screening from the Atlantic salmon BAC library, representative BACs for the three FABP2 genes were identified and chosen. They fall into discrete contigs, corresponding to independent loci. We are in the process of sequencing these BACs, localizing their positions in the Atlantic salmon karyotype by FISH analysis and mapping them on the linkage map.

STURGEON, SWIMMING, STRESS, AND METABOLIC PRODUCTS - I.E., ALL THINGS JOE CECH JR. Lankford, S.E., Miller, R.A., Adams, T.E., and Cech, J.J., Jr. Symposium: Tribute to Joe Cech, Jr. Presentation: Oral Contact: Scott Lankford, University of Central Missouri, 306 W.C. Morris Warrensburg MO 64093 USA E-Mail: [email protected] Abstract: Temporal patterns of plasma cortisol, lactate and glucose concentrations were studied in cannulated green sturgeon (Acipenser medirostris) during increasing swimming speeds. Blood samples were collected before swimming initiation and during each subsequent velocity interval until exercise induced fatigue ensued. Interestingly, swimming initiation elicited a transient increase in cortisol and lactate (18.9±1.9 ng/ml and 0.27 mM, repectively) that returned to resting levels during the subsequent interval, suggesting an acute stress response. Cortisol levels then steadily increased with increasing swimming velocity, reaching a peak (56.5±11.7 ng/ml) at exhaustion. Plasma lactate patterns were similar, but the largest increase by

Fish Biology Congress Abstracts 56 far occurred between Ucrit and exhaustion velocities (0.384 and 1.87 mM, respectively). Throughout the protocol there were only small changes in glucose levels, suggesting tight homeostatic control. These data suggest that exercise induced elevation of cortisol is not due to excessive plasma lactate accumulation and support a role for cortisol in enhancing lactate clearance.

PRECOCIOUS MATURATION IN MALE SALMONIDS: A MODEL FOR UNDERSTANDING INTERACTIONS BETWEEN THE GROWTH AND REPRODUCTIVE AXES. Larsen, D.A., Beckman, B.R., and Swanson, P. Symposium: Fish Growth Presentation: Oral Contact: Don Larsen, NOAA Fisheries, Northwest Fisheries Science Center, 2725 Montlake Blvd. E. Seattle WA 98112 USA E-Mail: [email protected] Abstract: The physiological regulation of growth and age of reproductive maturation in fish is inextricably linked. In Chinook salmon, age of maturation ranges from 1-6 years post-fertilization; influenced by genetic and environmental factors. Male salmon that precociously mature at age-1 and -2 have provided a useful model for a series of studies focused on environmental regulation of growth and reproduction. These studies have shown that maturation for each age class is physiologically initiated as much as 8-12 months prior to spermiation and growth rate during this period significantly influences the physiological “decision” to mature in a given year. Thus, maturation at age-1 is influenced by conditions at the time of first feeding. The androgen, 11-ketotestosterne (11-KT), provides the first endocrine marker of the initiation of spermatogenesis in each age class and interacts with the growth-regulating hormone, insulin-like growth factor-I (IGF-I), to stimulate testicular growth and maturation.

EXPRESSION OF GROWTH RELATED GENES DURING THE TRANSITION FROM LATE STAGE EMBRYO TO EARLY STAGE JUVENILE IN RAINBOW TROUT Leatherland, J.F., Li, M., and Raine, J.C. Symposium: Fish Growth Presentation: Oral Contact: John Leatherland, Department of Biomedical Sciences, University of Guelph Guelph Ontario N1G 2W1 Canada E-Mail: [email protected] Abstract: In teleost fish, the transition from late stage embryo when the animal is reliant on yolk as the source of nutrients, to early stage juvenile when the animal is reliant on exogenous food sources is a critical developmental stage and a period of changing physiology. Very little is known about the regulation of this developmental period. Both ambient temperature and the ration size fed the swim-up animals influence the growth pattern of the early juveniles; moreover, the pattern of expression of genes in the liver and intestinal tract that encode for growth hormone (GH), insulin-like growth factor (IGF) and their receptors, and thyroid hormone (TH) receptors is also affected by the rearing temperature and ration size suggesting that GH, IGF and TH play a central role in aspects of the development of the late stage embryos as they are transformed into juveniles.

SWIMMING PERFORMANCE AND AEROBIC METABOLIC SCOPE IN DOMESTICATED STRAINS OF EUROPEAN SEA BASS, DICENTRARCHUS LABRAX. Lefrançois C., Luna Acosta A., Millot S., Péan S., Bégout M.L., Domenici P. Symposium: Swimming Performance Presentation: Oral Contact: C. Lefrançois, LIENSS (Université de La Rochelle- CNRS)- Bat. ILE. 2 rue Olympe de Gouges La Rochelle 17000 France E-Mail: [email protected] Abstract: The aim of this study was to compare the functional integrity of farmed seabass from strains characterized by different levels of domestication. Functional integrity was assessed through whole-animal performance traits i.e. aerobic metabolic scope (AMS) and swimming performance (i.e. critical swimming speed Ucrit, escape performance). Neither AMS nor Ucrit differed among different strains. Strains differed in the strength of the escape response, i.e. seabass responded to startling stimuli using either fast (FER) or slow escape responses (SER) as found in other species. The highly domesticated strain showed the highest proportion of SER. SER are characterized by low locomotor performance (low cumulative distance, velocity and turning rate) and long latency (i.e. the time interval between the stimulus onset and the first detectable movement of the escape response). SER were all ‘single bend’ type, i.e. they consisted of a single body bend (i.e. stage 1) as opposed to two (double bend type).

THE EFFECT OF PREDATION ON BODY SHAPE EVOLUTION OF THREESPINE STICKLEBACKS

Fish Biology Congress Abstracts 57 Leinonen, T.E., Cano, J.M., and Merilä, J. Symposium: Contributed Papers Presentation: Poster Contact: Tuomas Leinonen, Dept. Biological and Environmental Sciences, Biocenter 3, Viikinkaari 1, University of Helsinki Helsinki 14 Finland E-Mail: [email protected] Abstract: Measurements of quantitative genetic variation can be used for predicting outcomes of adaptive trait evolution. When predicting direction of change of correlated characters, it is useful to have measures of genetic covariances between traits. The information on covariances can be used to estimate the multivariate direction of greatest additive genetic variance, the direction along which evolutionary change is most likely. We measured quantitative genetic variation in body shape and armor of threespine sticklebacks to find out the most likely directions of change of these characters. Since predators are thought to act as the main selective force thriving the divergence in body armor, and to some degree in body shape, we performed a survival experiment to find out the multivariate selection gradient imposed by a gape-limited predator. We will present the combined results of these two experiments with discussion of how well they reflect the patterns observed in wild populations.

USING COGNITIVE TRAFFIC RULES TO IDENTIFY RELEVANT INFORMATION TOPOLOGIES WITHIN A FISH SCHOOL. Lemasson, B.H., Anderson, J.J., and Goodwin, R.A. Symposium: Social Behavior Models Presentation: Oral Contact: Bertrand Lemasson, Columbia Basin Research, School of Aquatic and Fishery Sciences, University of Washington, Box 358218 Seattle WA 98195-8218 USA E-Mail: [email protected] Abstract: Fish schools can demonstrate collective cognitive abilities in terms of social learning, or the rapid exchange of information. Although schooling dynamics can be mathematically reproduced using traffic rules, these rules have generally been based on molecular mechanics, social forces, or arbitrary distance-based behavioral phases. Here we present cognitive rules developed from simplified visual and neural mechanics. Our purpose is to explain behavioral tendencies from an information processing perspective. Environmental stimuli (i.e., neighbors) are first translated into their retinal representations and scaled by the degree of redundant neurological stimulation. A decision function then filters the information, which in turn identifies the network of influential fish within the school’s global network. The model demonstrates that these information networks display temporally dynamic topologies, which are related to both individual and group level properties. In contrast to other models, group level order results in less individual processing of stimuli and information topologies can be independent of absolute distance.

LAKE SUPERIOR’S COASTER BROOK TROUT: A COMPARISON WITH THE STREAM RESIDENT FORM Leonard, J.B.K. Symposium: Social Behavior Models Presentation: Oral Contact: Jill Leonard, Biology Department, Northern Michigan University, 1401 Presque Isle Ave. Marquette MI 49855 USA E-Mail: [email protected] Abstract: Lake Superior is home to an endemic form of brook trout termed a coaster for its nearshore, lake dwelling habit. Controversy surrounds the definition of coasters beyond their lacustrine lifestyle and so we have compared the growth, physiology and morphology of coasters to stream residents. Differences in growth and condition occur, but tend to be greater between populations than among life history strategies under common rearing conditions. There are minor differences in fin morphology between the forms. Some differences in metabolic indicators between populations were also detected. In a single partially migrant population (Hurricane R.), coasters had slightly lower condition at ages 1 and 2. Growth of the population overall was within the range expected for coaster populations. We suggest that the potential for growth and behavior similar to traditionally defined coasters may be prevalent in Lake Superior drainages and that other factors limit its modern expression.

MITOCHONDRIAL PROTEIN SYNTHESIS IS DEFENDED IN GILL BUT NOT HEART OF THE CUNNER (TAUTOGOLABRUS ADSPERSUS) IN RESPONSE TO ACUTE HYPOXIA AND HYPOTHERMIA Lewis, J.M., and Driedzic, W.R. Symposium: Fish Living on the Edge Presentation: Oral

Fish Biology Congress Abstracts 58 Contact: Johanne Lewis, 30 Marie Curie, Department of Biology, University of Ottawa, Ottawa, ON Ottawa ON K1N 6N5 Canada E-Mail: [email protected] Abstract: The cunner is a north temperate, labrid species that utilizes metabolic depression to survive the extreme low temperatures of its inshore habitat. Protein synthesis was measured in the whole tissue and mitochondrial protein pools of heart and gill in cunner exposed to either acute hypothermia or hypoxia to determine if the adaptation of decreased protein synthesis is conserved across levels of cellular organization and in response to varying environmental stressors. An active depression in whole tissue protein synthesis occurs in heart and gill in response to both acute hypothermia (80%) and hypoxia (40-50%). The synthesis of proteins slated for the mitochondrial protein pool was also depressed by 60% in heart in response to both hypothermia and hypoxia, but no depression occurred gill. The defense in gill mitochondrial protein synthesis is thought to be linked to the integral role of gill tissue in ion and gas exchange with the environment.

THE EFFECT OF HEAT STRESS ON RED BLOOD CELL GENE EXPRESSION IN RAINBOW TROUT (ONCORHYNCHUS MYKISS) Lewis, J.M., Walsh, P.J., and Currie, S. Symposium: Genomics of Adaptation Presentation: Poster Contact: Johanne Lewis, 31 Marie Curie, Department of Biology, University of Ottawa, Ottawa, ON Ottawa ON K1N 6N5 Canada E-Mail: [email protected] Abstract: Fish possess nucleated red blood cells (RBCs). Blood is also easily collected and manipulated in non-terminal experiments, making it an ideal tissue on which to study the stress response in fish. Through use of the cGRASP 16K microarray, we are currently investigating differences in RBC gene expression in fish held under control conditions (12C) and exposed to heat stress (one hour at 25C followed by recovery at 12C). Repeated blood sampling (via a dorsal aorta cannula) enables us to examine the stress response over time, as well as how much gene expression varies within an individual, with samples taken pre-heat stress (representing individual control) and at 4 and 20 hours post heat stress (representing early and late transcriptional regulation). This study will enable a broader understanding of the mechanisms underpinning the stress response in fish and the discovery of novel genes that are regulated in a stress specific manner.

ELEVATED EGG CORTISOL LEVEL EFFECTS ON EGG QUALITY AND EMBRYO GROWTH PERFORMANCE Li, M., Leatherland, J.F. Symposium: Contributed Papers Presentation: Poster Contact: Mao Li, University of Guelph Guelph On N1G2W1 Canada E-Mail: [email protected] Abstract: Oocytes were incubated in two level of cortisol rich ovarian fluid for 3 hours to examine the affects of early cortisol exposure on gene expression in embryos and growth of juveniles. Egg cortisol content was initially elevated, but was metabolized within 24 hours post-fertilization; however, some of the effects of cortisol on the early embryo were still evident in 4-month old juveniles. The expression of growth-related genes (IGF-1 and IGF-2) was affected from 7 days post- fertilization (dpf), and the expression of innate immune-related genes (TLR-5M/5S) and the glucocorticoid receptor 1 (GR 1)gene [but not the GR 2 gene] was affected by 13-dpf. Lysozyme activity levels were stimulated in a dose-dependent manner from 1-hour post-fertilization and for all collection times up to 21-dpf. Growth of the juveniles from 102 dpf to 135-dpf was also related to oocyte cortisol exposure, with the lower cortisol exposure resulting in significantly enhanced growth.

ATLANTIC SALMON PRIMARY CELL CULTURE BIOASSAYS OF TOXICITY & EFFICACY OF CHEMOTHERAPEUTANTS Lyndon, A.R. and Toovey, J.P.G. Symposium: Environmental Pollution Presentation: Oral Contact: Alastair Lyndon, School of Life Sciences, Heriot-Watt University, John Muir Building Edinburgh EH14 4AS UK E-Mail: [email protected] Abstract: Salmon cell culture was investigated as a basis for rapid screening of toxic effects and efficacy of chemotherapeutants targetted at salmon lice. Various cell types were examined and epidermis found to have the best performance in terms of cell-area expansion over time. Dose-response plots indicated a negative relationship of expansion with dose for all

Fish Biology Congress Abstracts 59 chemicals studied. A modification of the same assay system allowed assessment of chemotherapeutant efficacy versus salmon louse settlement. Ratios of EC50 from cell and louse assays gave consistent approximations of published therapeutic margins for the chemicals assayed. This study suggests that primary cell culture assays may have value in assessment of environmental toxic effects.

PROBIOTICS AND INNATE IMMUNITY TO EPIDERMAL PATHOGENS OF FISH Lyndon, A.R., Pieters, N., Brunt, J., McMenemy S., and Austin, B. Symposium: Immunity and Disease Resistance Presentation: Poster Contact: Alastair Lyndon, School of Life Sciences, Heriot-Watt University, John Muir Building Edinburgh EH14 4AS UK E-Mail: [email protected] Abstract: Probiotics can protect fish against bacterial pathogens accessing the gastro-intestinal tract. However, there is also evidence that probiotics can influence the immune system beyond the confines of the gut. We present evidence that selected probiotic bacteria can stimulate parts of the innate immune system of rainbow trout in such a way as to improve their resistance to pathogens interacting with the epidermis and blood systems. The effective mechanisms appear to vary between pathogens, while not all probiotics tested (even different strains of the same nominal species) induce the same effects. This implies that probiotics need to be carefully characterised before use, but further suggests that specific probiotic strains might be used to target particular pathogens.

ATLANTIC SALMON LACTATE DEHYDROGENASE (LDH) ISOZYMES: A MODEL FOR STUDYING A WHOLE GENOME DUPLICATION SUPERIMPOSED ON AN ANCIENT SERIES OF GENE DUPLICATIONS Ma, X., Lubieniecki, K.P., Loeve, C.A.T., Philips, R.B., Koop, B.F., and Davidson, W.S. Symposium: Genomics of Adaptation Presentation: Poster Contact: Xuezheng Ma, Molecular Biology and Biochemistry, Simon Fraser University, 8888 University Drive Burnaby B.C. V5A 1S6 Canada E-Mail: [email protected] Abstract: Lactate dehydrogenase (LDH) isozymes represent the classical example of a multi-gene system that is derived by successive gene duplications. By investigating the genes encoding the LDH isozymes in Atlantic salmon we will be able to gain insight into the effect of a whole genome duplication superimposed upon more ancient gene duplications. Five contigs coding for LDH were found in the cGRASP EST database. These correspond to two LDH-Bs, two LDH-As and one LDH- C. Exon-specific primers for each transcript were used with RT-PCR to examine their expression patterns. BAC clones containing each of the five salmon LDHs have been identified, and these have been sequenced. This has allowed us to map the LDH genes on the Atlantic salmon linkage map. FISH analysis has also been used to place the LDH-containing BACs on to chromosomes.

PROTEOMIC PROFILING FOR BIOMARKER DISCOVERY IN THE ZEBRAFISH GILL MacCormack, T.J., De Souza, A.G., Li, L., and Goss, G.G. Symposium: Environmental Pollution Presentation: Oral Contact: Tyson MacCormack, CW405 Biological Sciences Bldg., University of Alberta Edmonton Alberta T5G2E9 Canada E-Mail: [email protected] Abstract: Using high-throughput mass spectrometer-based methods (2D LC-ESI MS/MS) we have identified approximately 6172 proteins in the gill of the zebrafish, Danio rerio. Proteins were characterized according to their cellular process, molecular function, and subcellular location. Those relevant to expected physiological and metabolic processes in the gill were identified and provided excellent coverage of important biochemical pathways. Greater than 14% (2573) of the peptides in the analysis were classified as hydrophobic (GRAVY index >0.5) and more than 250 integral membrane proteins were found without using specific enrichment strategies. Approximately 40 proteins involved in xenobiotic metabolism and a number of established toxicological biomarkers were present. The goal of this project was to establish a baseline proteome for ongoing physiological and toxicological studies. Quantifying specific changes in the zebrafish proteome will be a powerful tool in identifying new biomarkers for emerging toxicants and novel engineered compounds.

ASSESSING THE POTENTIAL IMMUNOTOXICITY OF NANOPARTICLES IN CHANNEL CATFISH USING HIGH-THROUGHPUT TECHNIQUES MacCormack, T.J., Kelly, J., Dang, M., Veinot, J.G.C., and Goss, G.G. Symposium: Environmental Pollution Presentation: Poster

Fish Biology Congress Abstracts 60 Contact: Tyson MacCormack, CW405 Biological Sciences Bldg., University of Alberta Edmonton Alberta T5G2E9 Canada E-Mail: [email protected] Abstract: The potential toxicity of manufactured nanoparticles (NP) is of growing concern since they will undoubtedly find their way into the environment. We examined the effects of water soluble NP exposure on immune function in channel catfish (Ictalurus punctatus) cell lines using high-throughput microplate and flow cytometric based assays. The use of cell lines allows for the examination of interactions between specific immune cell types, including B and T cells, natural killer cells, and monocyte/macrophages. We assessed NP effects on cellular viability, proliferation, apoptosis, necrosis, antioxidant pathways, and other immune cell functions. These studies are designed to determine the potential applicability of using immune cells as a high throughput indicator of the general and specific mechanisms of NP toxicity. The goal of the study is to provide feedback to NP engineers to facilitate the development of safer NPs while preserving the unique and beneficial properties of these materials.

EFFECTS OF ETHINYLESTRADIOL IN THE ESTUARINE KILLIFISH FUNDULUS HETEROCLITUS DURING DIFFERENT REPRODUCTIVE PERIODS MacLatchy, D., Hogan, N., Cushnie, W., Lippert, K., Hanson-Lee, M. and Currie, S. Symposium: Environmental Pollution Presentation: Oral Contact: Deb MacLatchy, Faculty of Science, Wilfrid Laurier University, 75 University Avenue West+J206+J207 Waterloo Ontario N2L 4L5 Canada E-Mail: [email protected] Abstract: To determine the mechanism(s) by which endocrine status is affected in mummichog (Fundulus heteroclitus) exposed to estrogenic endocrine disruptors, adults were exposed to 0-270 ng/L ethinylestradiol (EE2) for 10-20 days during regression and recrudescence. Following exposure, gonadal tissue was incubated with steroidogenesis stimulators, including chorionic gonadotropin/forskolin, 25-OH cholesterol, or pregnenolone. No differences in testosterone production were found in regressing fish. Production was decreased in EE2-exposed recrudescing males, both at and downstream of cholesterol mobilization to P450 side-chain cleavage (P450scc) and/or P450scc conversion of cholesterol to pregnenolone. In females, effects were more predominant downstream of pregnenolone production. Hepatic transcript levels of estrogen receptor (ER)α were increased in EE2-treated recrudescing males while heat shock protein (hsp) 90α transcription showed no change and hsp90β was significantly decreased. In regressing fish, no changes in hepatic hsp 90 protein were found, although males had significantly greater overall levels due to EE2 treatment. We are presently focusing on determining how estrogenic mechanisms are affected under additional environmental stressors such as temperature and salinity change.

CELLULAR ASPECTS OF METABOLIC DEPRESSION IN CUNNER (TAUTOGOLABRUS ADSPERSUS) MacSween, C.E. (1), Staples, J.F. (2), and Gamperl, A.K.(1). Symposium: Fish Living on the Edge Presentation: Oral Contact: Courtney MacSween, (1) Ocean Science Center, Memorial University of Newfoundland, St. John’s, NL Canada. (2) Dept. of Biology, University of Western Ontario, London, ON Canada. St. John's NL A1C 5S7 Canada E-Mail: [email protected] Abstract: The cunner, a temperate member of the Labridae (wrasses), actively depresses its metabolism in winter. However, the cellular processes mediating this metabolic downregulation are largely unknown. In this experiment, adult cunners and rainbow trout (used as a comparative species) were acclimated to 1 and 10oC, and state 3 and 4 respiration rates were determined on isolated liver mitochondria at 1, 5, 10, and 20oC. Acclimation temperature did not affect either parameter, however, assay temperature (p<0.001) and species (p<0.001) both had significant effects. For example, high Q10 values (>10) for state 3 and 4 respiration were recorded in cunner from 5-10oC in both acclimation groups indicating the capacity to rapidly depress metabolism (values for trout < 2.5). Cunner proton leak was also measured at 20 and 5oC, and showed that mitochondria assayed at 20oC were much ‘leakier’ than when assayed at 5oC. CS and CCO activities will also be reported for both species.

OVERWINTERING OF SEA TROUT (SALMO TRUTTA) IN FRESHWATER: ESCAPING SALT AND LOW TEMPERATURE OR AN ALTERNATE LIFE STRATEGY? Madsen, S.S., Thomsen, D.K, Nielsen, C. and Koed, A. Symposium: Ion and Acid-base Regulation Presentation: Oral

Fish Biology Congress Abstracts 61 Contact: Steffen Madsen, Institute of Biology, University of Southern Denmark, Campusvej 55 Odense DK-5230 Denmark E-Mail: [email protected] Abstract: Salmo trutta show large phenotypic plasticity. Juveniles may reside in their native stream until maturation or migrate into the ocean as 1-3 year old smolts. Sea-going fish reside at sea for 2-3 years until return to their native stream for reproduction. However, immature fish may leave the ocean during their first or second winter and overwinter in freshwater. The question is why? We tested the hypothesis that hypoosmoregulatory capacity is compromised by low temperature in trout representing two populations: a high- and low-salinity coastal population. Sea-run, immature trout were caught in lower parts of two rivers during winter and acclimated to laboratory conditions. Subgroups were challenged with high salinity, low water temperature or both and their osmoregulatory performance was investigated. Low temperature compromised hypoosmoregulatory ability, as indicated by insufficient compensatory adjustments of ion-transport mechanisms. Thus FW-overwintering is likely driven by compromised osmoregulatory physiology but since not all fish in a population do so, the phenomenon may also reflect diverging life-strategies.

GERMINAL CENTRE-LIKE TISSUE ARCHITECTURE EXISTS IN FISH LYMPHOID ORGANS. Magor, B.G., Saunders, H.L., Oko, A.L., and Scott, A.N. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Brad Magor, Dept. of Biological Sciences, University of Alberta Edmonton AB T6H 2E5 Canada E-Mail: [email protected] Abstract: In fishes the development of high affinity antibodies to antigens (affinity maturation) is less pronounced than that seen in homeothermic vertebrates. One explanation for the limited affinity maturation is that fishes appear not to develop the germinal centre cell masses where affinity maturation and cell selection takes place in lymphoid organs of mammals. We are using in situ hybridization and gene expression analyses on laser-capture microdissected cells of channel catfish spleen to reconstruct the cellular organization at sites of affinity maturation. As in mammalian germinal centres, catfish cells expressing the immunoglobulin mutator protein, AID, are clustered in close association with TcR expressing cells. These cell masses appear to develop in pre-existing aggregates of melano-macrophage cells, which have been previously demonstrated to be sites of antigen trapping. This evidence is consistent with fishes having transient tissue structures that are analogous, if not homologous to the germinal centres of homeothermic vertebrates.

PERIPHERAL REGULATORS OF THYROID STATUS IN LAMPREY: PHYSIOLOGICAL, DEVELOPMENTAL AND EVOLUTIONARY CONSIDERATIONS. Manzon, R.G., Gross, T.N., Stilborn, S.S.M, Manzon, L.A. Symposium: Biology of Lampreys Presentation: Oral Contact: Richard Manzon, Department of Biology, University of Regina, 3737 Wascana Parkway Regina SK S4S 0A2 Canada E-Mail: [email protected] Abstract: Unlike other vertebrates, larval lamprey lack follicular thyroid tissue and maintain relatively high serum thyroid hormone (TH) levels. Moreover, the onset of metamorphosis in lampreys coincides with and appears to require a dramatic decline in TH levels. To further elucidate the functioning of the lamprey thyroid system and its role in metamorphosis we sought to investigate peripheral regulators of thyroid status, namely the serum thyroid hormone binding proteins, TH deiodinases and the TH and retinoid-X- nuclear receptors. We have recently identified and characterized 3 novel, developmentally regulated TH binding proteins in lamprey serum. However, despite this developmental regulation the total TH binding capacity of lamprey serum does not appear to vary and remains high throughout the lifecyle. These data will be discussed in conjunction with deiodinase cloning and expression data in the context of the evolution of the vertebrate thyroid system, the maintenance of thyroid status in lamprey, and the role of TH in lamprey metamorphosis.

ANTIOXIDANTS, LIPID PEROXIDATION AND DNA DAMAGE IN A NEOTROPICAL FISH AFTER 24H EXPOSURE TO LEAD Martinez, C.B.R.,Viléla, M.B.F.A., Monteiro, V., Sofia, S.H. Symposium: Tropical Fish Adaptations Presentation: Oral Contact: Claudia B R Martinez, Laboratório de Ecofisiologia Animal, Departamento de Ciências Fisiológicas, Universidade Estadual de Londrina. C.P. 6001 - CEP: 86051-990 Londrina Paraná 86051990 Brazil E-Mail: [email protected] Abstract:

Fish Biology Congress Abstracts 62 Inorganic lead has been measured in high concentrations in some streams in Brazil; thus, this study was designed to evaluate oxidative stress parameters of Prochilodus lineatus (Characiformes) exposed to 0.5 ppm of dissolved lead (Pb) for 24h. The results showed a significant increase in liver Glutathione Peroxidase (GPx) and a significant decrease in Glutathione S Transferase in fish exposed to Pb in relation to control (CTR). Liver catalase did not change. Metallothionein (MT) content was significantly higher in Pb group. Lipid peroxides levels did not vary, but comet assay showed significant increases in the mean score of erythrocytes DNA damage in fish exposed to Pb in relation to CTR. Taken together these results indicate that acute exposure to Pb produced the activation of an antioxidant enzyme (GPx) together with the increase in MT content, a non-enzymatic antioxidant. However these defenses were not enough to inhibit oxidative damage to DNA.

REPRODUCTION CAPACITY OF FATHEAD MINNOW (PIMEPHALES PROMELAS) IN TWO POPULATIONS FROM NORTHERN ONTARIO, CANADA Martinez, M. and Schulte-Hostedde, A. Symposium: Stress Effects and Response Presentation: Oral Contact: Mery Martinez, 935 Ramsey Lake Road, Laurentian University, Biology Department Sudbury Ontario P3E 2C6 Canada E-Mail: [email protected] Abstract: During reproduction period, fathead minnows are able to mate multiple times. Hence they need to carefully control the amount of energy allotted for sperm production. We evaluate the variation in reproduction investment (gonad size and body condition) in two populations of fathead minnow from northern Ontario, submitted to different levels of stress during two consecutive years. There was no significant year*site interaction, nor was there a significant year effect. However, males from Silver Lake had significantly higher gonado-somatic index (GSI) than males from Espanola site (P< 0.001). Body condition (fulton index) was positively correlated with GSI only for the Espanola population (P= 0.027). Furthermore, fish form Espanola also show a positive relationship between fulton index and testis asymmetry (P< 0.001), whereas in Silver Lake, there was no significant relationship (P= 0.30). These results suggest that fish from Silver Lake are heavily investing in gonad development despite their lower body condition.

THE USE OF ELECTROMYOGRAM TELEMETRY TO ASSESS THE SWIMMING BEHAVIOUR OF AN IBERIAN CYPRINID IN A POOL-AND-WEIR TYPE FISHWAY PROTOTYPE. Mateus, C.S., Quintella, B.R., Silva, A.T., Ferreira, M.T. & Almeida, P.R. Symposium: Swimming Performance Presentation: Poster Contact: Catarina Mateus, Institute of Oceanography, Faculty of Sciences of the University of Lisbon, Campo Grande Lisbon 1749-016 Portugal E-Mail: [email protected] Abstract: Dams and weirs block fish migration and contribute to species and habitat loss. Fishways can minimize part of the negative impacts associated with these obstructions but their effectiveness must be guaranteed by adjusting the design of the facilities to the potential users. One of the most common designs of fish passage is the pool-and-weir type. In this study we used an experimental fishway with a 10 m long flume (1 m x 1.20 m cross section), on a slope of 8.7%, with 6 pools (2 m x 1 m) to study the swimming behaviour of the Iberian barbel Barbus bocagei (Steindachner 1865). This cyprinid is a potamodromous species widely spread in Portugal and Spain. The barbels used in this experiment were tagged with EMG radio transmitters equipped with electrodes that register the muscle activity. A relationship was developed in a swimming tunnel for all tagged specimens whereby swim speed could be estimated from the EMG telemetry signals. The behaviour of the tagged barbels during the fish passage was video recorded and compared with the EMG signal (i.e., swim speed). To analyse the impact of different hydraulic conditions on fish while overtaking the fishway, the hydraulic behaviour of this structure using a three-dimensional Accoustic Doppler Velocimeter was studied. The information gathered with this study suggests that EMG telemetry can be used to study remotely the behaviour of barbels in fish passages.

GENE DUPLICATION AND THE EVOLUTION OF CHONDROGENIC NEURAL CREST McCauley, D.W. Symposium: Biology of Lampreys Presentation: Oral Contact: David McCauley, Department of Zoology, University of Oklahoma, 730 Van Vleet Oval Norman OK 73019 USA E-Mail: [email protected] Abstract: Neural crest cells contribute to the increased complexity of vertebrates relative to protochordates. It is widely accepted that an increase in vertebrate complexity is also related to gen(om)e duplication that occurred at the base of vertebrates and may be related to the origin of the neural crest. Study of the development of one neural crest derivative, pharyngeal cartilage, in the lamprey and comparison to chondrogenesis in other may provide clues regarding acquisition of a

Fish Biology Congress Abstracts 63 chondrogenic fate by the vertebrate neural crest. The transcription factor Sox9 is a SoxE gene that regulates expression of Type II collagen in development of vertebrate cartilage. Duplication of the ancestral SoxE transcription factor into the three SoxE genes present in vertebrates might have been important in partitioning the chondrogenic neural crest role of Sox9. Presently we are investigating the role of duplicated SoxE genes in lamprey to understand evolution of the neural crest.

SALINITY DEPENDENT ISOFORMS OF GILL NA+,K+-ATPASE IN ATLANTIC SALMON: PROTEIN CHARACTERIZATION, ABUNDANCE AND LOCALIZATION. McCormick, S.D. and Regish, A. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Stephen McCormick, USGS, Conte Anadromous Fish Research Center, PO Box 796, One Migratory Way Turners Falls MA 1376 USA E-Mail: [email protected] Abstract: Gill Na+,K+-ATPase (NKA) in teleost fish is involved in ion regulation in both fresh water seawater. We have developed and validated antibodies specific to the α1a NKA and α1b NKA protein isoforms of Atlantic salmon, and used Western blots and immunohistochemistry to characterize their size, abundance and localization. The apparent molecular weight of NKA α1a is slightly less than that for NKA α1b, and the overall abundance of NKA α1a is less than NKA α1b. The abundance of gill NKA α1a was higher in fresh water than in seawater, whereas NKA α1b was more abundant after seawater acclimation. In fresh water, NKA α1a was preferentially located in gill chloride cells on the lamellae, whereas NKA α1b was primarily found in chloride cells on the filament. Evidence for changes in abundance and localization of NKA α1a and α1b during the parr-smolt transformation and after treatment with cortisol will also be presented.

ALL STRESSED OUT AND NO WAY TO GO: THE RELATIONSHIP BETWEEN STRESS AND NITROGEN WASTE EXCRETION IN TOADFISH. McDonald, M.D. Symposium: Fish Living on the Edge Presentation: Oral Contact: Danielle McDonald, University of Miami, Rosenstiel School of Marine and Atmospheric Science, 4600 Rickenbacker Causeway Miami Fl 33149 USA E-Mail: [email protected] Abstract: When in its natural environment, it is believed that the gulf toadfish, Opsanus beta, excretes a combination of urea and ammonia. However, when exposed to environmental stressors, toadfish shut down ammonia excretion at the gill, likely by activating a gill-specific isoform of glutamine synthetase, and become ureotelic. Certain environmental stressors, such as waterborne exposure to low levels of the antidepressant, fluoxetine, seem to result in a dramatic increase in urea excretion. The switch to ureotely, urea production and the toadfish’s unique pulsatile urea excretion mechanism are all affected when circulating levels of the stress hormone, cortisol, are elevated. However, the role of stress in the regulation of toadfish nitrogen excretion is complex; natural elevations in cortisol as well as exogenous cortisol loading appear to stimulate mRNA expression of the toadfish urea transporter (tUT) but inhibit urea excretion. The relationship between stress, cortisol and nitrogen waste excretion in toadfish will be discussed.

EFFECTS OF AGE AND SALINITY INCREASE ON GILL EPITHELIA OF JUVENILE GREEN STURGEON (ACIPENSER MEDIROSTRIS). McEnroe, M., Forostyan, T., Perumal G., Allen, P.J, and Cech, J.J., Jr Symposium: Tribute to Joe Cech, Jr. Presentation: Oral Contact: Maryann McEnroe, School of Natural and Social Sciences, Purchase College, SUNY, 735 Anderson Hill Rd. Purchase NY 10577 USA E-Mail: [email protected] Abstract: We used SEM and TEM to study the gill epithelia of juvenile (<10 mo.) green sturgeon (Acipenser medirostris). The epithelium consists of pavement cells, mucous cells (MCs), chloride cells (CCs), mitochondria-rich cells (MRCs), and undifferentiated cells. FW fish had abundant MRCs and MCs on filaments. MRC apical surface was flat, and had dense microplicae. SW fish had CCs on the filaments with slightly invaginated apical surfaces; MCs were also abundant on filaments. Apical surface area of CCs was less than that of MRCs. Viewed with TEM the SW-acclimated fish had abundant CCs on filaments, few on lamellae. SW CCs were larger, with numerous, larger, mitochondria and a more developed tubular system than FW MRCs. We demonstrate early development of CCs in green sturgeon: SW-acclimated fish (7-10 mo) had abundant CCs and fish > 5 mo developed CCs when transferred to SW.

Fish Biology Congress Abstracts 64

VALIDATING THE BASIS OF DIVERGENT MUSCLE AND LOCOMOTOR DESIGNS FOR SWIMMING USING THRESHER SHARKS McGillivray, D., Syme, D.A., Bernal, D., Aalbers, S.A., Donley, J.M., and Sepulveda, C.A. Symposium: Swimming Performance Presentation: Poster Contact: Doug Syme, Dept of Biological Sciences, University of Calgary, 2500 University Drive NW Calgary AB T2N 1N4 Canada E-Mail: [email protected] Abstract: Threshers (Alopiidae) are large, pelagic sharks and the only having species with laterally and medially positioned red, aerobic, locomotor muscle (RM). This allows testing of the hypothesis that medial RM (as in Common threshers Alopias vulpines, lamnid sharks and tuna) provides a propulsion system different from sharks with lateral RM. Sonomicrometry was used to quantify RM and white muscle (WM) strain during steady swimming of Common threshers off the coast of California. RM strain was greater than WM while swimming, but decreased significantly during passive body bending (when RM was not activated). Shortening of RM during swimming showed instances of both leading and trailing the surrounding WM, with no phase difference during passive bending. Thus, the medial RM of Common threshers sheers relative to WM, allowing uncoupling of RM shortening from local body bending as in the similarly designed lamnid sharks/tuna, but not in fish with lateral RM.

SPAWNING SITE SELECTION AND INCUBATION ENVIRONMENT OF INTERIOR FRASER COHO SALMON: MCKINLEY CREEK, BRITISH COLUMBIA McRae, C.J., Shrimpton, J.M., Warren, K.D. Symposium: Contributed Papers Presentation: Poster Contact: Crystal McRae, University of Northern British Columbia, 3333 University Way Prince George BC V2N 4Z9 Canada E-Mail: [email protected] Abstract: Interior Fraser coho (IFC) (Oncorhynchus kisutch) have been designated as an endangered species by the Committee on the Status of Endangered Wildlife in Canada (COSEWIC). Despite this designation, little is known regarding the spawning and incubation habitat requirements of IFC. Radio telemetry was used to determine IFC spawning areas within McKinley Creek watershed, an indicator stream for this population. Once spawning areas were identified physical/chemical characteristics of the stream and inter-gravel environment were measured at selected and non-selected spawning sites. Significant differences between selected sites and non-selected sites were stream width (p=0.03) and velocity (p<0.01). Significant differences between inter-gravel environments of selected and non-selected sites included conductivity (p=0.001), pH (p=0.04) and dissolved oxygen (p<0.001) at 60cm, and dissolved oxygen at 30cm (p<0.001). Additionally, survival and growth of larvae is being compared between selected and non-selected sites.

MIGRATORY GENOMICS AND THE FATE OF FRASER RIVER SOCKEYE SALMON Miller, K.M., Li, S., Kaukinen, K., Schulze, A., Raap, M, Ginther, N., Farrell, A.P., Hinch, S.G., Cooke, S.J., Patterson, D.A., Crossin, G.T., English, K.K., and Lapointe, M. Symposium: Genomics of Adaptation Presentation: Oral Contact: Kristi Miller, Pacific Biological Station, 3190 Hammond Bay Rd. Nanaimo BC V9T 6N7 Canada E-Mail: [email protected] Abstract:

Fish Biology Congress Abstracts 65 The transition between FW and SW for migrating adult salmon is a period of variable and significant mortality. The physiological condition of fish and environmental conditions they encounter likely influences survivorship, but our mechanistic understanding of what differentiates fish in “good” and “poor” condition remains largely unexplored. These relationships are now being explored through functional genomics approaches. “Wild” ecological genomics experiments on adult sockeye salmon covered a 1,300 km migratory trajectory from the Queen Charlotte Islands to spawning sites in the Fraser River and included individual experiments on brain, gill, liver, white muscle and hypothalamus tissues. These studies uncovered distinctive physiologies associated with ocean migratory routes, navigational plasticity, osmotic preparation, white muscle tissue remodelling, and weak genetic signals in the ocean. By combining biotelemetry, individual stock ID, non-destructive sampling, and microarrays, we tracked the success of river migration to spawning grounds 1000’s of Fraser sockeye salmon, and uncovered powerful physiological signals associated with fate in river.

SWIMMING UNDER THE INFLUENCE: ETHANOL PRODUCTION DURING EXERCISE IN GOLDFISH Milligan, C. L, Butts, K.I. and Bartolini, M. Symposium: Fish Living on the Edge Presentation: Oral Contact: C. Louise Milligan, Department of Biology, University of Western Ontario London ON N6A 5B7 Canada E-Mail: [email protected] Abstract: Goldfish are known to produce ethanol as an alternate glycolytic endproduct under hypoxic/anoxic conditions as a strategy to extend survival. We asked if, during exercise to exhaustion, which also invokes anaerobic glycolysis, ethanol is produced and if so, what are the consequences to muscle metabolic recovery. Following exercise to exhaustion, muscle glycogen declined from ~ 40 to 5 mmol/kg, lactate increased to ~ 7 mmol/kg with ethanol as the major endproduct (~ 50 mmol/kg). The majority of the ethanol was excreted and muscle glycogen was not completely resynthesized, even after 24 hour, suggesting the loss of carbon is limiting to muscle glycogenesis. Feeding after exercised enhanced glycogen synthesis; in fed goldfish, muscle glycogen was completely restored by 24 hours, suggesting muscle glycogenesis is supported mainly by extramuscular substrates. Thus, while ethanol production reduces negative consequences associated with endproduct accumulation, it also eliminates carbon, compromising subsequent glycogenic capacity.

IMPACTS OF SHORT-TERM ACID AND ALUMINUM ON THE ION REGULATORY PHYSIOLOGY OF ATLANTIC SALMON SMOLTS: MECHANISMS OF IMPAIRED SEAWATER TOLERANCE Monette, M.Y., Yada, T., Matey, V. and McCormick S.D. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Michelle Monette, Yale School of Medicine, Department of Cellular and Molecular Physiology, 333 Cedar Street New Haven CT 6520 USA E-Mail: [email protected] Abstract: To investigate impacts of short-term acid and aluminum (Al) on the seawater tolerance of Atlantic salmon, we exposed smolts for 2 and 6 days to control and acid + three Al concentrations. Exposure to acid and moderate-high Al impaired seawater tolerance, decreased gill Na+,K+-ATPase activity and CFTR I mRNA, increased gill apoptosis and cell proliferation, and decreased chloride cell abundance. In contrast, exposure to acid and low Al impaired seawater tolerance in the absence of impacts on gill Na+,K+-ATPase, CFTR and chloride cell abundance, decreased gill cell proliferation, and altered chloride cell size, staining intensity and morphology. Thus, the mechanisms underlying impaired seawater tolerance in smolts depend on the severity of Al exposure: when Al is moderate-high chloride cell number and function are reduced whereas when Al is low, impaired seawater tolerance may result instead from the upregulation of ion uptake mechanisms leading to a shift in chloride cell phenotype.

GROWTH REGULATION BY INSULIN-LIKE GROWTH FACTOR IN EEL Moriyama, S. Symposium: Fish Growth Presentation: Oral Contact: S. Moriyama, School of Marine Bioscience, Kitasato University Ofunato Iwate 022-0101 Japan E-Mail: [email protected] Abstract: Insulin-like growth factors (IGF-I and II) play essential roles in regulating development, somatic growth, and metabolism. To better understand the role of IGFs in eel, Anguilla japonica, we cloned IGF-I and II cDNAs from the liver, and examined their tissue distribution and expression after treatment with growth hormone (GH) and seawater acclimation. Two eel IGF-Is originate by alternative splicing, but eventually yield a single form of mature peptide, while two eel IGF-IIs are transcribed from different genes, suggesting that paralogous genes may have evolved in this species. Both eel IGF-I mRNA levels in liver were positively correlated to the body size of glass eels, and the hepatic IGF mRNA levels increased after

Fish Biology Congress Abstracts 66 GH treatment in vivo and in vitro. Higher levels of IGF-I mRNAs were observed in the gills and liver of seawater-reared eels than freshwater-reared fish, while no changes of IGF-II mRNAs were observed.

CHLORIDE CELL DENSITY CHANGES OF GILL EPITHELIUM IN SPOTTED SCAT SCATSCATOPHAGUS ARGUS K. DURING SALT WATER ACCILIMATION Morovati H, Zolgharnain H, Noori mogehi M,Ghazilou A Symposium: Stress Effects and Response Presentation: Poster Contact: Amir Ghazilou, Khorramshahr Uni of Marine Science and Technology khorramshahr khoozestan 669 Iran E-Mail: [email protected] Abstract: Patterns for lamellar and filament Chloride cell density changes of gill epithelium in spotted scat investigated, during 30 day acclimation period to different salinities. For fish transferred from fresh water to low salinities, significant higher number of chloride cells during 10 day post transfer was revealed. After ward the mean number of lamellar chloride cells remained constant. The mean number of filament chloride cells decreased significantly during 10 day post transfer. After ward the mean increased cell numbers observed. For fish transferred from fresh water to higher salinities, the densities of the cells increased significantly and were 3-fold more than their initial number by 10 day post transfer. Unlike lower salinities, mean number of lamellar chloride cells decreased to its near initial from day15 until day 30 after transfer. Density fluctuations of filament chloride cells in higher salinities followed the same pattern as low salinities. General factorial design analysis concluded significant effect of all time and salinity on chloride cell density. When Regression analysis was used, chloride cell density fluctuations modeled to be following on: Lamellar chloride cell density= 12.7 + 0.492 time + 0.243salinity; Filament chloride cell density= 4.34 - 0.102 time - 0.0771salinity

DEVELOPMENT OF LAMPREY-SPECIFIC FISHWAYS Moser, M.L., Pennington, H.T., and Peery, C.A. Symposium: Swimming Performance Presentation: Oral Contact: Mary Moser, National Marine Fisheries Service, 2725 Montlake Boulevard East Seattle WA 98112 USA E-Mail: [email protected] Abstract: Hydropower dams, culverts, irrigation diversion dams, and weirs can delay or obstruct pre-spawning migration of anadromous lampreys. During the past five years, we have worked to develop lamprey-specific fishways that facilitate passage of adult Pacific lamprey (Lampetra tridentata). These prototype fishways provide a passage route over obstacles to lamprey movement in fishways at Bonneville Dam, the first dam these fish encounter on their spawning migration in the main stem Columbia River. We have exploited the climbing ability of lamprey to develop structures that can be operated in a small space and afford maximal flexibility. Improvements to lamprey fishway design require attention to attractive flows, provision of adequate attachment surfaces, recognition of swimming and climbing performance limits, and proper site selection.

RESPONSES OF JUVENILE PACIFIC LAMPREY TO LIGHT AND WATER VELOCITY DURING SEAWARD MIGRATION Moser, M.L., Russon, I.J., and Kemp, P.S. Symposium: Biology of Lampreys Presentation: Oral Contact: Mary Moser, National Marine Fisheries Service, 2725 Montlake Boulevard East Seattle WA 98112 USA E-Mail: [email protected] Abstract: Anadromous lampreys undergo profound physiological changes as they transform from relatively sedentary ammocoetes to highly migratory macropthalmia. We investigated the responses of juvenile lamprey to varying intensities of lighting and current velocity. Our experiments were conducted during the night in a large flume supplied with ambient Columbia River water. Activity of lamprey was scored during a series of exposures to both bright lighting and changing water velocity. In addition, the propensity for vertical (vs horizontal) movement was also tested in a separate chamber. We found that both bright light (>1500 Lux/m2) and current velocities < 25 cm/s elicited short term (< 5 min) responses by the fish, but that they quickly acclimated to these conditions. In addition, macropthalmia appeared to exhibit negative rheotaxis and were reluctant to move vertically. Along with physiological adaptations for marine parasitism, changes in rheotaxis and swimming orientation probably prepare these fish to orient during seaward migration.

DR. JOSEPH J. CECH – A SALUTE TO A FORTY-YEAR CAREER OF EXCELLENCE AS A FISH PHYSIOLOGIST Myrick, C.A. Symposium: Tribute to Joe Cech, Jr. Presentation: Oral

Fish Biology Congress Abstracts 67 Contact: Chris Myrick, Department of Fish, Wildlife, and Conservation Biology, Colorado State University, Campus Delivery 1474 Fort Collins CO 80523-1474 USA E-Mail: [email protected] Abstract: Dr. Joseph J. Cech, Jr. has an impressive forty-year career as a fish physiologist, educator, and colleague. During those forty years, he has conducted research on at least 58 different species of fish, published over 140 peer-reviewed journal articles, collaborated with at least 125 other scientists, and trained more than 30 graduate students. As one of the many students and colleagues who have been influenced by Dr. Cech, I will provide a brief, somewhat authorized, and sometimes humorous review of his career and contributions to the field of fish physiology.

“LITTLE FISH” PASSAGE – MEASURING SWIMMING AND JUMPING PERFORMANCE OF NON- GAME FISHES Myrick, C.A., and Ficke, A.D. Symposium: Swimming Performance Presentation: Oral Contact: Chris Myrick, Department of Fish, Wildlife, and Conservation Biology, Colorado State University, Campus Deliver 1474 Fort Collins CO 80523-1474 USA E-Mail: [email protected] Abstract: Research on the swimming and jumping performance of fishes related to the design of fish passage structures has historically focused on large migratory species, particularly salmonids. With the increased regulatory attention being given to smaller non-game species, there is now a need for more studies on the swimming and jumping performance of smaller fishes. A study on the swimming and jumping performance of brassy minnow (Cyprinidae: Hybognathus hankinsoni), common shiner (Cyprinidae: Luxilus cornutus) and Arkansas darter (Percidae: Etheostoma cragini) is used to illustrate some of challenges of developing passage requirements for smaller fish species.

PLASTICITY OF DIFFERENCES IN PERFORMANCE AMONG BLACKNOSE DACE (RHINICHTHYS ATRATULUS) POPULATIONS ACROSS A GRADIENT OF URBANIZATION; THE ROLE OF MORPHOLOGY Nelson, J.A., Gastrich K., and Handelsmann, C. Symposium: Morphometrics Presentation: Oral Contact: Jay Nelson, dept. of Biol. Sci., Towson University Towson MD 21252 USA E-Mail: [email protected] Abstract: Urbanization alters stream and watershed hydrology so that fish from urban stream systems experience extreme flows during storms. To test whether residence in urban streams requires altered swimming ability, we compared sprint and endurance (Ucrit) swimming performances of eight populations of blacknose dace along an urban/rural gradient. Impervious surface cover (ISC), a measure of urbanization, was significantly correlated with both sprint and endurance swimming performance. Ucrit was repeatable up to six months, but dace populations with exceptional Ucrit 's lost ability under no-flow conditions. After 10 weeks of no-flow conditions, dace with high sprint performance lost ability whereas those with initial poor performance gained. Both swimming performances significantly increased after training to constant flow (16 cm/s and 7 cm/s) for 50 days. Morphology (geometric morphometrics) correlated significantly and oppositely with the two swimming performance types, suggesting a potential tradeoff between the optimal body shape for each performance test.

ULTRAVIOLET RADIATION AS A LIMITING FACTOR IN THE ECOLOGY OF SHALLOW WATER FISHES Nelson, P.A. Symposium: Fish Living on the Edge Presentation: Oral Contact: Peter Nelson, H.T. Harvey and Associates, 2202 I Street Eureka CA 95501 USA E-Mail: [email protected] Abstract: Studies of disparate systems, ranging from the rocky intertidal to coral reefs and high altitude lakes, suggest that solar ultraviolet radiation (UVR) is a potent factor in the ecology of fishes, contributing to adaptations manifest in their biochemistry, physiology, behavior, and distribution. I review the literature on how fishes respond to UVR, and the evidence that deleterious aspects of UVR are pertinent to conservation and are likely to change with global warming. Direct evidence for population-scale effects attributable to UVR are scarce, but at least one recent study marshals strong indirect evidence that solar radiation has influenced salmonid populations. While UVR is a stressor in many aquatic systems, it is also source of visual information. I conclude with an overview of ultraviolet vision and the evidence that some species or life history stages may be balancing the positive aspects of ultraviolet photosensitivity with the deleterious effects.

Fish Biology Congress Abstracts 68

COMPARISON OF JUVENILE PINK SALMON UMAX AND UCRIT SWIMMING PERFORMANCE , AND THE EFFECT OF SEA LICE ON UMAX Nendick, L.J., Gardner, M., Grant, A., Sackville, M.A., Brauner, C.J. and Farrell, A.P. Symposium: Swimming Performance Presentation: Oral Contact: Laura Nendick, 1816 McNicoll Ave Vancouver BC V6J1A4 Canada E-Mail: [email protected] Abstract: Swimming capacity is an important contributor to the fitness level of a fish. The parasitic sea louse, Lepeoptheirus salmonis, has been shown to have negative effects on the swimming performance of salmonids in the adult life stage, but effects of lice infection at salmonid juvenile life stage have not been reported. The objective of this study was two fold: 1) to estimate Ucrit and Umax in juvenile pink salmon (Oncorhychus gorbuscha) of size < 5g and, 2), to quantify the effect on swimming performance by the parasitic sea louse, Lepeoptheirus salmonis, using a Umax test. In juvenile Pink salmon, Umax and Ucrit protocols were found to produce the same final swimming speed. When swum with natural sea lice loads, 1 louse per fish had no effect on Umax swimming performance when compared with uninfected or previously infected fish.

DEVELOPMENT OF A HIGH DENSITY MARKER MAP IN ZEBRAFISH USING RAD MARKERS AND MASSIVELY PARALLEL SEQUENCING TECHNOLOGY Nipper, R.W., Atwood, T., and Johnson, E. Symposium: Genomics of Adaptation Presentation: Poster Contact: Rick Nipper, Floragenex, Inc., 1900 Millrace Drive Eugene OR 97403 USA E-Mail: [email protected] Abstract: SNP markers have become the de facto standard for linkage mapping of traits in forward genetic screens. The vertebrate model organism Danio rerio (Zebrafish) has thousands of SNPs available for mapping purposes, however implementation of available SNPs for use in recombination studies is relatively expensive on a per individual basis. Here we report the fusion of restriction site associated DNA (RAD) technology with Illumina massively parallel sequencing to produce a rapid, inexpensive, high density marker map in zebrafish. Using genomic complexity reduction, we created RAD libraries from single AB and WIK individuals, sequenced each library using the Illumina Genome Analyzer, and aligned the recovered RAD sequences to the zebrafish Ensembl reference assembly. Our dataset provided a panel of several thousand polymorphic AB and WIK markers over the entire 1.7 Gb zebrafish genome, at an average of 1-3 markers per megabase. Utilization of RAD markers found through high-throughput sequencing is an attractive, cost-effective option for researchers seeking to perform genetic mapping studies.

THE OREGON HATCHERY RESEARCH CENTER: A RESEARCH LABORATORY IN THE NATURAL ENVIRONMENT Noakes, D.L.G. Symposium: Plenary Presentation: Oral Contact: David Noakes, Oregon Hatchery Research Center, Fisheries & Wildlife Oregon State University Corvallis OR 97331-3803 USA E-Mail: [email protected] Abstract: Pacific Northwest hatcheries have changed over the past 100 years in reaction to numerous societal and environmental demands. Now hatcheries are not only responding to changing management needs they are also conducting basic and applied research that will determine their future roles. The Oregon Hatchery Research Center is an innovative facility designed to address both practical management issues and fundamental research questions. It is owned by the Oregon Department of Fish and Wildlife and is operated jointly by ODFW and Oregon State University. We investigate the differences between hatchery and wild origin fishes, in relation to the management and conservation of native species. Research includes tests of conventional hatchery procedures as well as innovative experimental analyses under controlled environmental conditions. We are conducting genetic pedigree analyses of mate choice and spawning success in steelhead, coho and Chinook salmon, assessing the role of jack males in mating, and relating genetic parentage to subsequent survival and growth of juveniles. We are using stable isotopes to determine food web structure in coastal coho, and to assess the impacts of carcass placement programs. We are developing and testing techniques for production of sterile steelhead to minimize genetic and behavioral impacts of hatchery fish on sympatric wild populations. We are tagging and tracking individual juveniles to determine the factors that regulate smolting success in wild and hatchery steelhead and coho. All these activities include collaborations with colleagues across Oregon, the rest of the Pacific Northwest, and from as far afield as Iceland, Japan and Korea. Our research is integrated with education and outreach to encourage the public

Fish Biology Congress Abstracts 69 awareness and participation, with internship programs at universities and colleges, and with electronic and print publication of information.

QUANTITATIVE GENE EXPRESSION IN THE AMAZONIAN TAMBAQUI (COLOSSOMA MACROPOMUM) EXPOSED TO COPPER AND CADMIUM. Nozawa, S.R., Casanova, F.M., Ferreira-Nozawa, M.S., Honda, R.T., Aride, P.H.R., Val, A.L., and Almeida-Val, V.M.F. Symposium: Tropical Fish Adaptations Presentation: Poster Contact: Sergio Ricardo Nozawa, UniNiltionLins, Av. Prof. Nilton Lins, 3259 Manaus Amazonas 69058040 Brazil E-Mail: [email protected] Abstract: Fish exposed to transition metals exhibit disturbances of ionoregulation, olfaction and swimming performance. Some of these metals are essential for many physiological processes. However, they are toxic at high levels. Many fish species have been chronically exposed to copper, arsenic, and cadmium, even in pristine environments as the Amazon waters. Gonzales et al. (2006, Aquatic Toxicology, 77:43-52) described up-regulation of myosin-2 (4.2-fold) and tropomyosin (3.1-fold) in juveniles of mummichogs exposed to arsenic. We exposed juveniles of C. macropumum to copper (20 mg/L) and cadmium (10 mg/L) and observed, using. qPCR, a significant up-regulation of tropomyosin in white muscle: 3.3-fold in copper and 3.7-fold in cadmium exposed animals. Similar changes were found in liver. This gene is important during embryogenesis and their differential expression may be linked to the morphological changes observed in the fingerlings. (CNPq, FAPEAM).

IMPOSING ADDITIONAL STRESS ON WILD FISH DURING ALREADY CHALLENGING PERIODS: PHYSIOLOGICAL AND BEHAVIOURAL CONSEQUENCES OF CHRONIC STRESS DURING PARENTAL CARE O’Connor, C.M., Gilmour, K.M., Arlinghaus, R., Van der Kraak, G., and Cooke, S.J. Symposium: Fish Living on the Edge Presentation: Oral Contact: Constance O'Connor, Ottawa-Carleton Institute of Biology, Carleton University, 1125 Colonel By Drive Ottawa Ontario K1S 5B6 Canada E-Mail: [email protected] Abstract: Stressors during parental care have the potential to negatively affect both parent and offspring. However, the consequences of stress during parental care have never been examined in fish. We used intraperitoneal cortisol implants to elevate plasma cortisol during the first 120 hrs of parental care in wild Micropterus salmoides. We then compared the physiology, behaviour, and nest success of stressed animals to unstressed conspecifics. Individuals were blood sampled, and analysis of plasma physiological variables focused on reproductive function, catabolic activity, and ionic balance. Behaviour was assessed every other day, and included measures of brood defense and maintenance. Monitoring continued until the parental fish could be considered either reproductively successful (fry were motile) or unsuccessful. Throughout, fish were assessed for immunocompetence (resistance to Saprolegnia sp. infection). Understanding the consequences of stress during this critical period is necessary to predict how fish with diverse life histories will respond to anthropogenic activities.

IDENTIFICATION OF GERMINAL CENTRE-LIKE STRUCTURES IN THE CATFISH SPLEEN Oko, A.L., Saunders, H.L., Scott, A.N. and Magor, B.G. Symposium: Immunity and Disease Resistance Presentation: Poster Contact: Brad Magor, Dept. of Biological Sciences, University of Alberta Edmonton AB T6H 2E5 Canada E-Mail: [email protected] Abstract: Germinal centers are transient tissue structures formed in secondary lymphoid organs during antibody affinity maturation. The 'maturation' process is characterized by hypermutation of immunoglobulin genes and subsequent selection of B- lymphocytes producing antibodies with higher binding affinity for pathogen antigens. The selection process requires additional antigen trapping cells and a subset of T-lymphocytes. Though easily distinguishable by histochemistry and immunocytochemistry in mammalian lymphoid organs, germinal centre-like structures have previously not been identified in poikilothermic vertebrates. Using in situ hybridization, immunocytochemistry, and laser capture microdissection coupled with RT-PCR, we have determined that channel catfish have germinal centre-like structures in their spleens. Cells expressing the immunoglobulin gene mutator, AID, co-localize with melano-macrophages, which have previously been implicated in trapping antigens. T-cell receptor-expressing cells are also found in these seemingly transient cell masses. Together these observations suggest that affinity maturation developed in the context of germinal centres early in vertebrate evolution.

CORRELATED RESPONSES TO SELECTION ON BOLDNESS BEHAVIOR IN ZEBRAFISH

Fish Biology Congress Abstracts 70 Oswald, M., and Robison, B.D. Symposium: Social Behavior Models Presentation: Oral Contact: Mary Oswald, University of Idaho, Dept. of Biological Sciences Moscow Idaho 83844-3051 USA E-Mail: [email protected] Abstract: Wild and laboratory strains of zebrafish often display patterns of behavioral variation consistent with the effects of domestication. We performed a selection experiment to create behaviorally divergent lines of zebrafish, testing for additive genetic variation and correlated responses to selection on a single behavioral trait. Within one generation of selection in a domesticated strain, we observed a significant response in our selected trait (proportion of time near the observer). Within two generations, we observed correlated responses in other behaviors (depth preference and latency to begin foraging from the water’s surface). These three behaviors were genetically correlated, and the genetic architecture of these behaviors is consistent with the patterns of behavioral adaptation to captivity observed in zebrafish and other fishes, where populations maintained in captivity tend to evolve increased boldness, increased orientation towards the surface of the water, and decreased latency to feed.

UPSTREAM MIGRATION OF FISH AT NATURAL OBSTRUCTIONS AND FISH PASSES EFFICIENCY IN SOUTHERN BELGIUM Ovidio, M., Orban, P., Lambot, F. and Philippart, J.C. Symposium: Swimming Performance Presentation: Oral Contact: Michael Ovidio, Biology of Behaviour Unit. Laboratory of Fish Demography and Hydroecology Tihange 4500 Belgium E-Mail: [email protected] Abstract: Fragmentation of rivers by physical obstacles has resulted in the drastic range reduction or extinction of numerous diadromous and potadromous species of fish worldwide. In order to implement to partially or completely re-establish the free movements of fish in the entire watercourse, the Walloon regional government (Belgium) initiated an integrated restoration project whose objectives are 1) To perform a complete inventory of all obstacles that may potentially interfere with the longitudinal connectivity and the free movements of fish. 2) To test, using radio telemetry, the ability of representative fish species (Salmonidae, Thymallidae, Cyprinidae, Esocidae and Cottidae) to clear different typologies of physical obstacles. 3) To determine the problematic sites that should be improved priority. 4) To construct fish passage facilities and to examine their biological efficiencies. During our talk we will synthesize the results of ten years of research and define the different options for the future.

INTRACELLULAR PH IMAGING REVEALS CELLULAR MECHANISMS FOR NA+ AND CL- TRANSPORT IN GILLS FROM FRESHWATER FISH Parks, S.K., Tresguerres, M.T., and Goss, G.G. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Scott Parks, Department of Biological Sciences, CW 405, Biological Sciences Centre, University of Alberta Edmonton Alberta T6G 2E9 Canada E-Mail: [email protected] Abstract: We have optimized a technique to monitor intracellular pH (pHi) in real time in isolated mitochondria-rich (MR) cells. Our primary focus lies in ion and acid-base regulation. Previously we demonstrated that only one type of MR cell displays phenamil, acetazolamide, and DIDS-sensitive transcellular Na+ transport, and hypothesized it is mediated by apical Na+ channels, intracellular carbonic anhydrase and basolateral Na+-HCO3- cotransporters. We now present functional evidence for the presence of a Cl-/HCO3- exchanger in two MR cells subtypes. Our data suggests that the Cl-/HCO3- exchanger would function in transepithelial Cl- uptake in one cell type and for pHi regulation in the other. Cl- removal from the bath caused a peculiar cellular acidification which will be further explored. Finally, recent advances in the specific mechanisms of Na+ uptake will be discussed to provide an overall model of Na+ and Cl- uptake and H+ and HCO3- secretion at the freshwater fish gill.

VARIATIONS OF METABOLITES IN RELATION TO THE OVARIAN CYCLE IN TILAPIA OREOCHROMIS MOSSAMBICUS (PETERS). Pathan, J.G.K., and Baile, V.V. Symposium: Fish Growth Presentation: Oral Contact: J.G.K. Pathan, College of Fishery Science, Maharashtra Animal & Fishery Sciences University, Nagpur-440 001 Nagpur Maharashtra 440 001 India E-Mail: [email protected]

Fish Biology Congress Abstracts 71 Abstract: In brain (telencephalon, diencephalon) and pituitary, highest protein content in preparatory phase and lowest was in spawning. Highest carbohydrate and cholesterol was in prespawning, carbohydrate was lowest in postspawning and cholesterol was lowest in the spawning. Among somatic, liver and muscles showed highest protein, carbohydrates and cholesterol in preparatory; lowest in the spawning. Ovary showed highest proteins and carbohydrates in the spawning; lowest in preparatory and lowest carbohydrates in the postspawning. Cholesterol was highest in prespawning and lowest in spawning. Telencephalon, 60KDa protein band was more prominent in all the phases. Whereas, in diencephalon, 66KDa protein,in all phases.Pituitary had 27KDa protein band in all phases except in the preparatory phase. In ovary, in spawning, both lower and higher molecular weight bands ranging from 2.5 to 235.7KDa were present. In liver, higher molecular weight proteins dominated in preparatory and prespawning. In muscles, 60KDa band, all phases.

TOPMOUTH GUDGEON PSEUDORASBORA PARVA, A NEW NON-INDIGENOUS SPECIES IN THE INTERNATIONAL WETLAND OF ALMA-GOL (NORTHERN IRAN) Patimar, R., Abdoli, A., and Baensaf, S. Symposium: Fish Growth Presentation: Poster Contact: Rahman Patimar, Department of Natural Science, Higher Institutes of Gonbad, Shahid Fallahy Street Gonbad Golestan 4971799151 Iran E-Mail: [email protected] Abstract: Observation of P.parva was actually the first record of its being in this wetland, indicating increasing of its distribution in new habitats. Its biological characteristics were studied in Alma-Gol wetland from September 2000 to August 2002. There were significant differences in 3 morphometric and 1 meristic traits between sexes. The maximum age observed were 4+ for males and 3+ for females. The largest specimen was a male weighing 4.03gr with a total length 82mm. Both sexes grow negative allometrically (b=2.74 males, b=2.94 females). The overall ration of females to males was 1:1.6. The smallest mature specimen was 2+ in both sexes. The GSI examination showed that the peak of spawning activity of this species in the wetland is April with an average 11.93 and 6.98 for females and males respectively.

INVESTIGATIONS INTO GHRELIN’S ROLE IN REGULATING FISH GROWTH AND METABOLISM Peddu, S.C., Nilmeier, S.E., Fox, B.K., Breves, J. P., Kaiya, H., Picha, M.E., Borski, R.J., Hirano, T., Grau, E.G., Riley, L.G. Symposium: Fish Growth Presentation: Oral Contact: Larry Riley, California State University at Fresno, Department of Biology Fresno CA 93740 USA E-Mail: [email protected] Abstract: Discovery of ghrelin has broadened our understanding of the regulation of energy homeostasis. Ghrelin is an important endocrine peptide that links gastrointestinal system and brain in regulating food intake and energy expenditure. Ghrelin and its receptor (GHS-R) have been identified in several fish species. To date, biological roles of ghrelin in fish are still unclear. Ghrelin stimulates the short-term release of growth hormone (GH) in vitro and in vivo. In contrast, chronic exposure to ghrelin in tilapia does not alter plasma GH levels, but significantly increases food intake, total fat content, and weight gain. Interestingly, plasma ghrelin levels are not elevated preprandially in tilapia. Further, short-term fasting does not alter plasma ghrelin levels, whereas chronic fasting significantly elevates plasma ghrelin levels in the tilapia and hybrid striped bass. Our findings suggest that ghrelin does not play a significant role in daily feeding but may play a role in preventing catabolism.

THE IMMUNE AND STRESS RESPONSES OF ATLANTIC COD TO LONG-TERM INCREASES IN WATER TEMPERATURE Pérez-Casanova, J.C.(a), Rise, M.L.(a), Dixon, B.(b), Afonso, L.O.B.(c), Hall, J.R.(a), Johnson, S.C.(c), and Gamperl, A.K.(a) Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Juan Carlos Pérez-Casanova, (a) Ocean Sciences Centre - Memorial University of Newfoundland, St. John’s, NL; (b) Biol. Dept. Univ. of Waterloo, Waterloo, Ontario; (c) IMB-NRC, Halifax, NS. St. John's NL A1C5S7 Canada E-Mail: [email protected] Abstract: To define the upper thermal tolerance of juvenile cod and to investigate their physiological response to summer-like temperatures, we exposed 10 C acclimated juvenile cod to a 1 C increase in water temperature every 5 days and measured: plasma cortisol and glucose levels, the respiratory burst activity (RB) of blood leukocytes and the expression of specific immune-related genes [MHC Class I, Interleukin-1beta (I1B), beta 2-microglobulin (B2M), Immunoglobulin M (IgM) – light (L) and heavy (H) chains] in blood using qRT-PCR. The maximum temperature tolerated was 19.1 C. Although RB

Fish Biology Congress Abstracts 72 did not change, plasma glucose increased slightly with temperature. Cortisol levels increased significantly at 14 and 16 C, but returned to control levels thereafter. IgM-H expression remained stable during the experiment. In contrast, IL-1β expression increased significantly at 19 C, and changes in β2-M, MHC and IgM-L expression followed a pattern similar to that of cortisol. These results suggest that immune function is influenced by complex interactions between thermal effects and temperature-induced stress.

THE INVOLVEMENT OF NA+/H+ EXCHANGERS IN BRANCHIAL AND RENAL ACID-BASE REGULATION IN RAINBOW TROUT Perry, S.F, Esbaugh, A., and Ivanis, G. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Steve Perry, Department of Biology, University of Ottawa, 30 Marie Curie Ottawa Ontario K1N 6N5 Canada E-Mail: [email protected] Abstract: Current models of branchial ionic and acid-base regulation incorporate Na+/H+ exchangers (NHE) on the apical membrane of a subset of mitochondria-rich cells (MRC). Using a combination of approaches including molecular cloning, real-time PCR, immunocytochemistry and in situ hybridization, we demonstrated that NHE2 and NHE3 are localized to MRCs exhibiting positive binding of peanut lectin agglutinin (PNA+ MRCs). NHE2, the predominant gill isoform was transcriptionally activated during hypercapnic acidosis owing to elevated levels of the stress hormone, cortisol. Within the kidney, NHE3 (the predominant renal isoform) was confined to the apical membrane of proximal tubules where it was co- localized with the vacuolar type H+-ATPase (V-ATPase). NHE3 mRNA and protein levels were increased during hypercapnia and by cortisol treatment. Thus, we speculate that the elevation of plasma cortisol during hypercapnia contributes to transcriptional activation of NHE3 that ultimately promotes acid-base regulation by stimulating H+ secretion and HCO3- reabsorption.

NUTRITIONAL REGULATION OF IGFS IN CHANNEL CATFISH Peterson, B.C., Waldbieser, G.C., and Small, B.C. Symposium: Fish Growth Presentation: Oral Contact: Brian Peterson, 141 Experiment Station Rd Stoneville MS 38776 USA E-Mail: [email protected] Abstract: We examined changes in hepatic IGF-I and IGF-II mRNA, insulin like growth factor binding proteins (IGFBP-1 and IGFBP-2) mRNA, muscle IGF-I and IGF-II mRNA in fed (fed daily for 45 days) and restricted (not fed for 30 days followed by feeding for 15 days) channel catfish. By day 30, liver IGF-I mRNA decreased 14.3 fold compared to fed controls (P < 0.01). Abundance of IGFBP-1 mRNA increased (P < 0.05) 2.4 fold in restricted fish while IGFBP-2 mRNA was similar between treatments. Muscle IGF-II mRNA decreased (P < 0.05) 3.4 fold while levels of IGF-I mRNA were similar between treatments. After 15 days of refeeding, liver IGF-II mRNA was higher (P < 0.05) in restricted fish while all other genes were similar between treatments. A change in IGF-II mRNA due to restricted food intake suggests a physiological role for IGF-II in growth regulation of channel catfish.

COMPARATIVE CHROMOSOME MAPPING IN RAINBOW TROUT AND ATLANTIC SALMON Phillips, R.B., Ventura, A.B ., Lubienlecki, K.P., Danzmann, R.G., and Davidson, W.S. Symposium: Genomics of Adaptation Presentation: Poster Contact: Ruth Phillips, Washington State University Vancouver, 14204 NE Salmon Creek Ave. Vancouver WA 98686 USA E-Mail: [email protected] Abstract: We present comparative data on the chromosome maps of rainbow trout and Atlantic salmon that shows conservation of large blocks up to whole chromosome arms with the same gene order. The results are consistent with the hypothesis that there was a whole genome duplication in an ancestor of both species, and that the resulting 50 chromosome arms were subject to different fusions to produce the current karyotypes. All of the chromosome fusions in rainbow trout were centric fusions, while both tandem and centric fusions occurred to produce the current karyotype of Atlantic salmon. A small block of repetitive DNA marks the site of the tandem fusions. Each chromosomal segment in Atlantic salmon corresponds to a specific chromosome arm in rainbow trout. Most of the fusions appear to have been centromere to telomere or centromere to centromere followed by pericentric inversions. The integration of the two maps will be very useful for identification of candidate genes for QTLs for important traits that have been mapped in the two species.

REGULATION OF GROWTH HORMONE (GH) FAMILY RECEPTORS IN PRIMARY CULTURED TILAPIA HEPATOCYES: EFFECTS OF PITUITARY HORMONES (GH AND SOMATOLACTIN: SL) AND

Fish Biology Congress Abstracts 73 METABOLIC HORMONES (INSULIN, GLUCAGON, CORTISOL, AND TRIIODOTHYRONINE) ON EXPRESSION OF THE GH RECEPTOR Pierce A.L., Moriyama S., Uchida K., Hirano T., and Grau E.G. Symposium: Fish Growth Presentation: Oral Contact: Andrew Pierce, Hawaii Institute of Marine Biology, University of Hawaii, 46-007 Lilipuna Rd, P.O. Box 1346 Kaneohe HI 96744 USA E-Mail: [email protected] Abstract: The vertebrate GH/insulin-like growth factor (IGF1 and IGF2) axis is the principal endocrine axis that regulates growth in mammals. In mammals, GH stimulates liver production of IGF1, which stimulates growth. Recent evidence suggests that the teleost GH/IGF axis differs substantially from mammals. In fish, unlike adult mammals, IGF2 is strongly regulated by GH and metabolic hormones. Further, the fish pituitary produces SL, a GH family peptide hormone not found in tetrapods. The function of SL is currently unknown; however, the putative SL receptor (GHR1) is highly expressed in the teleost liver. We established a system for primary culture of tilapia (Oreochromis mossambicus) hepatocytes, and tested the effects of pituitary and metabolic hormones on the expression of GHR1 and GHR2 in vitro. GH strongly stimulated GHR2 (0.5 – 50 nM; biphasic). However, GHR1 did not respond to GH over physiological concentrations, increasing only at 500 nM GH. This confirms our hypothesis that GHR2 is the receptor for GH in tilapia. Native tilapia SL slightly but significantly increased GHR1 at 50 nM. There was a significant trend toward increased GHR1 with increasing SL concentration (0 – 50 nM SL). These findings are consistent with the hypothesis that GHR1 is a receptor for SL, and also suggest that SL may bind GHR2 at higher concentrations. Insulin (1 µM) suppressed stimulation of GHR2 by GH. Insulin, glucagon, and cortisol (1 µM) increased GHR1. This strong regulation of GHR1 by metabolic hormones suggests that this receptor plays a role in the regulation of metabolism. This study shows that GH, SL, insulin, glucagon, and cortisol differentially regulate hepatocyte expression of GHR1 and GHR2 (Supported by NRI Competitive Grants 2006-35206-16447, 2005-35206- 15285, and 2008-35206-18785 from the USDA CSREES).

HOLDING-DURATION-ASSOCIATED STRESS OF GREEN STURGEON (ACIPENSER MEDIROSTRIS) AT SOUTH DELTA FISH COLLECTION FACILITIES, CALIFORNIA Portz, D.E., and Crocker, C.E. Symposium: Tribute to Joe Cech, Jr. Presentation: Poster Contact: Don Portz, US Bureau of Reclamation, Denver Federal Center, P.O. Box 25007 (86-68290) Denver CO 80225- 0007 USA E-Mail: [email protected] Abstract: Information about the effects of capturing and holding processes of threatened and endangered species is needed to improve their survival during the salvage of fishes at large water diversions. Exposure of fishes to environmental stressors, such as capture and handling, can be a great concern, in that extreme or prolonged stressors may plague fish performance and overall health, adversely affecting population size and sustainability. Monitoring holding-associated stress during the salvage process was accomplished by comparing changes in plasma constituents (i.e., cortisol, lactate, glucose, hematocrit) in green sturgeon (Acipenser medirostris) that were exposed to simulated salvage-tank-related stresses over a 24-hour holding duration. Indwelling catheters were used to minimize disturbances during sampling and gain insight to the physiological changes throughout the holding period. Preliminary results show the value of using this approach in identifying stress-related effects of the fish salvage process. Research funded by USBR Mid-Pacific Region, and Science and Technology Program.

EXPRESSION OF GLYCOLYTIC AND GLUCONEOGENIC ENZYMES SHOW INTRASPECIFIC VARIATION AMONG SELECTED AND NON-SELECTED STRAINS OF RAINBOW TROUT FED HIGH LEVELS OF DIETARY CARBOHYDRATE Powell, M.S., Wacyk J., Plant, K., Stone, D.A.J., Rodnick K.J., Robison, B.D., Drew, R.E., Murdoch G.K., Hill, R.A., and Hardy, R.W. Symposium: Fish Growth Presentation: Poster Contact: Madison Powell, University of Idaho / Hagerman Fish Culture Experiment Station, 3059F National Fish Hatchery Road Hagerman ID 83332 USA E-Mail: [email protected] Abstract: Plant protein-based feeds for carnivorous fish inevitably contain carbohydrates (CHO) of starch origin, and carnivorous fish do not utilize starch as an energy source very efficiently. Intra-specific variation in CHO utilization has been reported but not thoroughly studied. Three strains of rainbow trout (RBT) were used in a factorial design to examine variation in tolerance of CHO considering performance parameters and gene mRNA levels at regulatory points related with CHO

Fish Biology Congress Abstracts 74 utilization. We were successful in identifying differences in growth performance, HSI and plasma glucose levels in three strains of RBT fed varying levels of dietary starch. Increasing dietary starch level also led to elevated levels of plasma glucose and HSI. The responses of these classic growth parameters were expected and are characteristic of RBT fed diets high in CHO. With increasing inclusion of dietary starch, levels of glucokinase and glycogen synthase mRNA expression increased correspondingly but varied significantly among strains.

CHARACTERIZATION OF PATHOGENESIS AND HOST IMMUNE RESPONSES IN TWO CHINOOK SALMON POPULATIONS THAT DIFFER IN THEIR ABILITY TO SURVIVE RENIBACTERIUM SALMONINARUM CHALLENGE Purcell, M.K., Metzger, D., Elliott, D.G., and Park, L.P. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Maureen Purcell, 6505 NE 65th ST Seattle WA 98115 USA E-Mail: [email protected] Abstract: In the late 1960s, Chinook salmon from the Green River in Washington State (WA) were successfully introduced into Lake Michigan. Beginning in 1988, large disease outbreaks occurred among Chinook salmon in the lake due to Renibacterium salmoninarum, the causative agent of bacterial kidney disease (BKD). In laboratory challenges, the contemporary Lake Michigan stock injected with R. salmoninarum had significantly greater survival relative to WA. Histological evaluation following challenge revealed delayed onset of severe granulomatous lesions and delayed bacterial replication in the Lake Michigan stock relative to the WA stock. The host responses to R. salmoninarum in the two stocks were profiled using a salmon microarray and revealed significant regulation of a number of immune-related genes. Using quantitative PCR, we observed differential regulation of interferon- and inflammatory-related genes following challenge in both stocks. In summary, higher survival was associated with delayed pathogenesis but the protective immune mechanisms remain uncertain.

MAKING SENSE OF NON-SCENTS IN METAL-CONTAMINATED WATERS Pyle, G.G. Symposium: Environmental Pollution Presentation: Oral Contact: Greg Pyle, Dept. of Biology, Nipissing University North Bay ON P1B 8L7 Canada E-Mail: [email protected] Abstract: Olfaction is one of the most important sensory modalities utilized by aquatic animals to convey information about predation risk and food availability, reproductive status of mates, kin recognition, etc. Impairment of chemosensation by toxicants at the molecular or cellular level can potentially lead to perturbations at higher levels of organisation. We have examined the behavioural, neurophysiological, and ecological consequences of metal-impaired chemosensory function in a range of aquatic animals that represents several trophic levels of a typical aquatic ecosystem. In each case, metal concentrations that are insufficient for causing overt toxicity were sufficient to cause chemosensory dysfunction. Because chemosensory systems demonstrate a high degree of phylogenetic conservation, metal-impaired chemosensation among phylogenetically disparate animal groups probably results from a common mechanism. It may be possible to exploit the general nature of these effects to develop a predictive model which has the potential to significantly improve the current environmental risk assessment paradigm.

INTERGENERIC AND INTERFAMILIAL RELATIONSHIP IN AS SUGGESTED BASED ON THE SPERMATIC CELLS Quagio-Grassiotto, I., Ortiz, R.J. and Oliveria, C. Symposium: Contributed Papers Presentation: Poster Contact: I. Quagio-Grassiotto, Departamento de Morfologia, Instituto de Biociências de Botucatu, UNESP, Rubião Júnior S/N. Botucatu São Paulo 18618000 Brazil E-Mail: [email protected] Abstract: Doradidae intergeneric relationships are still not well definite, and also its relationships within the remaining Siluriformes. Considering that sexual characteristics can have phylogenetic significance and be useful in cladistic analyses, spermatogenesis, spermiogenesis, and spermatozoal ultrastructure of elected Doradidae members were studied. The data were analyzed considering the previous relationship hypotheses within Doradidae, between Doradidae and Auchenipteridae, between Doradoidea (Doradidae+Auchenipteridae) and Aspredinidae, and between Doradoidea and Ariidae. The diversity of spermatogenesis, spermiogenesis and spermatozoa of Doradidae are according to the intergeneric groups recognized for the family. Doradidae and Auchenipteridae share a few characteristics, apparently due to the sperm modifications of the inseminating Auchenipteridae. The high similarity of spermatogenesis, spermiogenesis and

Fish Biology Congress Abstracts 75 spermatozoa agree with the proposal that Aspredinidae and Doradoidea as sister-groups, and may also indicate a possible basal position for Astrodoradinae within Doradidae. The characteristics of spermatogenesis and spermatozoa shared by Doradidae and Ariidae suggest some level of relationship between these families.

USING GENOMICS TO IDENTIFY GENES ASSOCIATED WITH UPPER TEMPERATURE TOLERANCE IN SALMONIDS WITH APPLICATIONS TOWARD AQUACULTURE AND CLIMATE CHANGE Quinn, N.L., Lubieniecki K.P., Koop, B.F., Davidson, W.S. Symposium: Genomics of Adaptation Presentation: Oral Contact: Nicole Quinn, SSB 6150-8888 University Drive Burnaby BC V5A1S6 Canada E-Mail: [email protected] Abstract: It is predicted that climate change will severely affect the northern regions of Canada, with devastating ecological and economical impacts. Indeed, the aquaculture industry in the Yukon, Canada has reported massive losses in Arctic charr stocks over recent summers due to elevated ambient water temperatures. It is therefore of interest to develop temperature tolerant strains of salmonids using genomics-assisted selective breeding. We are using the well-studied Atlantic salmon genome to identify upper temperature tolerance (UTT) genes in salmonids. Specifically, we are using a unique approach that combines full region sequencing, comparative genomics and expression analysis to refine previously-identified salmonid UTT quantitative trait loci (QTL). The information produced from this research will directly feed into a gene- and marker-assisted breeding program for Artic charr. This research also stands to benefit aquaculture and fisheries breeding programs for other salmonid species, and can be used to develop population-specific conservation strategies for wild salmonids.

REFERENCE LEVEL OF SOME SERUM ENZYMES IN HUSO HUSO LINNAEUS, 1758 Rajabipour, F. ; Shahsavani, D.; Moghimi A. ; Jamili, S. ; Mashaii, N. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Farhad Rajabipour, PoBoX; 89715-1123, Brackish water Fisheries Reasearch Center, Bafq, Yazd, Iran Bafqh Yazd PoBoX; 89715-1123 Iran E-Mail: [email protected] Abstract: Hematological analyses of serum enzymes proved useful in the detection of metabolic disturbances, especially in cultured fishes. Blood samples were collected from 60 five-years cultured great sturgeons, Huso huso Linnaeus, 1758, to establish activities of the serum enzymes: AST, ALT, LDH, CK, ALP and ACP. Fishes were caught from two sites of brackishwater earthpond of Bafgh (n=30) and freshwater of Gorgan (n=30) at the center and northeast of Iran, at May 2006. Values of the serum enzymes activities of H. huso samples from Bafgh and Gorgan were: AST, 502.9±258.2 and 436.1±186.8; ALT, 104.4±35.1 and 53.1±38.7; LDH, 3094.2±1277.5 and 2486.3±1393.3; CK, 3632.9±2618.7 and 3967±5054.9; ALP, 281.2±112.7 and 762.2±600.2; ACP, 13.3±2.5 and 33±6.8 IU/L, respectively. Mean values of ALT, ALP and ACP were significantly different in fishes of two sites (p<0.05), but not any significant difference between AST, LDH and CK (p>0.05) by t-test.

MOLECULAR ECOLOGY AND EVOLUTION OF INTERTIDAL FISH Ramon, M. L. Symposium: Contributed Papers Presentation: Oral Contact: Marina L. Ramon, University of Southern California, 3616 Trousdale Parkway Los Angeles California 90089 USA E-Mail: [email protected] Abstract: Some of the well-known studies done on fish that address species formation and adaptive radiation have been done on freshwater fishes. However, patterns from freshwater systems seem to be of limited applicability to marine organisms. Specifically the degree to which genetic drift, selection, historical biogeography, gene flow and local adaptation each contributes to the speciation between closely related species. This study examined aspects of some of these evolutionary forces. I focused on a group of sculpins (Family: Cottidae) off the Pacific west coast. Sculpins serve as good candidates to study the speciation process. Approximately twenty genera are found in the north Pacific, they are found intertidally and subtidally, and have several closely related genera. A phylogeny was constructed to examine historical biogeographic factors that have contributed to the speciation of cottids. Second, the evolution of a mitochondrial gene rearrangement discovery in several genera was used to evaluate the speciation of sculpins. Lastly, a phylogeographic study explored the process of gene flow and population structure for Clinocottus analis.

SEARCHING FOR ADAPTIVE GENES USING A GLOBAL APPROACH

Fish Biology Congress Abstracts 76 Ramon, M.L. and Gracey, A.Y. Symposium: Genomics of Adaptation Presentation: Poster Contact: Marina L. Ramon, University of Southern California, 3616 Trousdale Parkway Los Angeles California 90089 USA E-Mail: [email protected] Abstract: Molecular evolution has had a difficult time making the connection between natural selection at the molecular level and an organism's phenotype, its ecology or how it effects populations with ongoing gene flow. Traditionally studies attempting to detect natural selection at the molecular level (i.e. nucleotide) have focused on the effects of one gene with one trait instead of multiple genes and this is due to the complexity of gene interactions. The primary goal of this research is to understand the relationship between an organism's genotype in conjunction with the environment it lives in and how these two factors contribute to its phenotype. I will utilize modern genomic techniques that are capable of monitoring the interactions of several hundred genes at once and identify genes being expressed under different environmental conditions for Clinocottus analis that might be ecologically and evolutionarily important. Genomic approaches using fish have already provided insights into the mechanisms that underlie short-term and long-term environmental adaptations and this study will contribute to this body of work. It will also contribute to the growing recognition of the need to move beyond model organisms and explore the mechanisms by which natural populations operate.

INCIDENCE OF SCOLIOSIS IN THE INLAND SILVERSIDE (MENIDIA MENIDIA) IN AN URBAN LAKE Ramroop, D., and Ma, P.M. Symposium: Environmental Pollution Presentation: Poster Contact: PoKay M. Ma, Department of Biology, Queens College, City University of New York, 65-30 Kissena Boulevard Flushing NY 11367 USA E-Mail: [email protected] Abstract: Flushing Meadow Lake is located in an urban park in the Borough of Queens in New York City. The lake is connected to Flushing Bay through an underground channel. The inland silverside (Menidia menidia) is one of the most predominant species in this lake. This fish appears to be annual and semelparous. Spawning occurs between May and August. Young- of-the-year grows up rapidly and over-winters as medium-sized juveniles. In early spring, rapid feeding, growth and maturation occur. Adults die off after spawning. Between July and August, approximately 10 – 15% of the young fish between 1 and 2 months old exhibit various degrees of vertebral column deformity. Scoliosis is the most common abnormality. The number of scoliotic fish dwindles rapidly towards the end of September, probably the result of attrition due to compromised ability to grow and to avoid predation. The deformity may be due to genetic, environmental, or parasitic factors.

HUSBANDRY STRESS AND DISEASES OF ZEBRAFISH Ramsay, J.M., Schreck, C.B., Kent, M.L. Symposium: Stress Effects and Response Presentation: Oral Contact: Jennifer Ramsay, Dept. Fisheries and Wildlife, 104 Nash Hall, Oregon State University Corvallis OR 97331 USA E-Mail: [email protected] Abstract: Zebrafish are widely used as vertebrate research organisms in the biomedical and environmental sciences. Husbandry stress has been implicated in disease outbreaks in zebrafish colonies, yet little is known about how stress affects the susceptibility of zebrafish to pathogens. Zebrafish are susceptible to numerous Mycobacterium spp., many causing severe outbreaks. Pseudoloma neurophilia is a common parasite of the central nervous system of zebrafish, associated with emaciation, although some fish appear clinically normal. We chronically stressed zebrafish infected with Mycobacterium marinum and M. chelonae. Stress significantly increased mortality and prevalence of clinical disease in the M. marinum- but not the M. chelonae-infected fish. We stressed zebrafish that had chronic P. neurophilia infections, which resulted in greater mortality and inflammation of the muscle and spinal tissues. Stress also increased the area of parasite occupying spinal tissue. There appears to be a role of stress in exacerbating some diseases common to zebrafish husbandry.

OXYGEN TRANSFER IN FISH AND GENTLEMAN JOE Randall, D.J. Symposium: Tribute to Joe Cech, Jr. Presentation: Oral Contact: David Randall, Zoology, University of British Columbia, 6270 University Boulevard Vancouver BC V6T 1Z4 Canada E-Mail: [email protected]

Fish Biology Congress Abstracts 77 Abstract: Joe and I helped organize a symposium in Sacremento in 1990. Joe presented a paper on some Californian freshwater fish showing that temperature and hypoxia are major determinants of fish species distribution. Joe showed that the species with the lowest metabolic rate were best able to survive temperature change and hypoxia. Like other vertebrates, fish increase ventilation and gill diffusing capacity to maintain oxygen transfer during hypoxia. Teleosts, however, are the only vertebrates to reduce organic phosphates and increase hemoglobin oxygen affinity during hypoxia. This will enhance oxygen uptake across the gills but may be detrimental to oxygen delivery to the tissues. Hypoxic trout maintain high muscle oxygen levels, probably above venous levels, during hypoxia. It is postulated that teleosts decrease organic phosphates during hypoxia to maintain arterial oxygen levels at the expense of PaO2 and deliver oxygen to tissues through a Root off shift.

LIFE-HISTORY TRAITS AND ENERGETICS OF MOUTH BROODING IN DIVERGENT OXYGEN ENVIRONMENTS Reardon, E.E., and Chapman, L.J. Symposium: Tropical Fish Adaptations Presentation: Oral Contact: Erin Reardon, Biology Department, McGill University, 1205 Dr. Penfield Ave. Montreal QC H3A1B1 Canada E-Mail: [email protected] Abstract: Although hypoxia occurs naturally in many tropical aquatic systems, this environmental stressor is becoming increasingly widespread globally as influx of municipal wastes and fertilizers accelerates eutrophication and pollution. Thus, it has become extremely important to understand consequences of hypoxia in fishes. We explore the relationship between DO, life-history traits, and energetics in the African cichlid, Pseudocrenilabrus multicolor using field surveys and lab experiments. This fish reproduces under a range of conditions from extreme hypoxia to normoxia. Fish from hypoxia were characterized by smaller size-at-maturity than fish from normoxia. The egg size/number relationship differed between divergent DO field sites, and between fish reared under alternative oxygen environments, though not in the same manner. Costs of mouth brooding under hypoxia were evident in metabolic rates measured under alternative DO levels. The apparent costs of reproduction under hypoxia may be offset by the exploitation of hypoxia habitats as physiological refugia under field conditions.

PHYSIOLOGICAL CONSEQUENCES OF SELECTION FOR ANGLING VULNERABILITY IN A RECREATIONAL SPORTFISH Redpath, T.D., Wahl, D.H., Philipp, D.P., Suski, C.D., Couture, P., Arlinghaus, R., and Cooke, S.J. Symposium: Stress Effects and Response Presentation: Oral Contact: Tara Redpath, Carleton University Biology Department, 1125 Colonel By Drive Ottawa Ontario K1S 5B6 Canada E-Mail: [email protected] Abstract: Commercial marine fisheries can exert selective pressures on the life history traits of fish, although the consequences for recreational sportfish are less known. Beyond documenting selectivity associated with recreational fishing, there is an interest in understanding the mechanistic basis for differential angling vulnerability, a trait shown to be genetically heritable. We evaluated the physiological correlates of angling vulnerability using the popular sportfish, the largemouth bass (Micropterus salmoides). Over 30 years, fish were subjected to four generations of artificial truncation selection for vulnerability to angling by selecting for high (HVF) and low (LVF) vulnerability fish. A series of studies evaluated their metabolic characteristics and physiological response to exercise as a stressor. By quantifying oxygen consumption, it was revealed that HVF have higher standard metabolic rates and maximal metabolic rates than LVF. Trends in the exercise recovery profiles revealed that HVF experienced more severe physiological disturbance than LVF immediately following exercise.

HEMOGLOBIN-PROTON SENSITIVITY IN FISHES OCCUPYING A TRANSITIONAL PHASE IN ROOT EFFECT EVOLUTION Regan, M.D., and Brauner, C.J. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Matthew Regan, Department of Zoology, University of British Columbia, 6270 University Blvd Vancouver BC V6T 1Z4 Canada E-Mail: [email protected] Abstract: The Root effect, in conjunction with a gas gland and rete, represents a complex physiological system to maximize oxygen (O2) delivery to the swimbladder and eye of fishes. The Root effect is defined by a reduction in blood O2 carrying capacity with a reduction in pH and could therefore compromise O2 uptake during a generalized acidosis. To compensate, most

Fish Biology Congress Abstracts 78 teleost fishes have the ability to regulate red blood cell (rbc) pH adrenergically (β-NHE). Basal actinopterygian fishes, however, lack β-NHE but possess a Root effect and thus potentially jeopardize O2 uptake. The present study analyzed the magnitude of the Root effect (% Hb-O2 desaturation) along a pH spectrum (pH 8.5 – 5.5) in seven basal species within the actinopterygian order. The results demonstrate that although some of these species exhibit a relatively large Root effect, the pH of onset is too low to be operational in vivo. Consequently, O2 uptake is not jeopardized in these species during generalized acidoses despite the lack of rbc β-NHE.

HIGH-SPEED VIDEO ANALYSIS OF A UNIQUE PACIFIC LAMPREY CLIMBING BEHAVIOR Reinhardt, U.G. ,Eidietis, L.E. , Friedl, S.E. and Moser, M.L. Symposium: Biology of Lampreys Presentation: Poster Contact: Ulrich Reinhardt, Biology Department, Eastern Michigan University, 316 Mark Jefferson Hall, Ypsilanti MI 48197 USA E-Mail: [email protected] Abstract: New lamprey-friendly fishways involve inclined ramps that allow passage of Pacific lampreys over Bonneville Dam. We observed the lampreys moving against water at two flow volumes and on two ramps of 45˚ and 18˚ angle. We documented climbing movements using high speed video (125 frames per second). Lampreys advanced on the ramps by cycles of attaching to the ramps, pulling their bodies into a W-shape, and, then, rapidly flexing the body to propel themselves up, with simultaneous brief (20 to 140 ms) release of suction. Lampreys were using a burst swimming method to climb, because we observed inflection points in the body curvature traveling toward the posterior of the body and the center of mass moving up. This behavior has not been described for any other fish species. Vertical motion during each cycle of movement was greatest on the 45 ramp with low water flow (mean of 0.066 L/ cycle). These findings may be useful for developing a ramp design that allows the most efficient lamprey passage.

INVESTIGATION OF THE ROLE OF MYOSTATIN IN SMOLTIFICATION OF ATLANTIC SALMON Reitsma, J. and Bradley, T. Symposium: Fish Growth Presentation: Poster Contact: Joshua Reitsma, Dept. of Fisheries, Animal and Veterinary Science, University of Rhode Island, Bldg. 14, East Farm Route 108 Kingston RI 2881 USA E-Mail: [email protected] Abstract: For successful adaptation to the marine environment, Atlantic salmon (Salmo salar) must go through a critical parr-smolt transformation (PST) associated with a marked increase in growth. To investigate the potential role of muscle regulatory factors, levels of myostatin-1a, -1b, MyoD1b and myogenin mRNA, as well as myostatin protein, were measured in white muscle tissue during PST. Limited differences were observed in MyoD1b or myogenin mRNA levels, while myostatin mRNA levels were generally higher in smolts than parr. A short term reduction in levels of myostatin mature peptide in smolt was observed at the one week time point in both seawater and freshwater. A negative correlation was observed between levels of myostatin mature peptide and fish length and weight, consistent with the hypothesis that myostatin may have a negative regulatory role in fish muscle growth. This project was supported by National Research Initiative Competitive Grant no. 2003-35206-13506 from the USDA CSREES.

TOWARDS A PHYLOGENY OF LAMPREYS OF THE WORLD Renaud, C.B., and Lang, N.J. Symposium: Biology of Lampreys Presentation: Oral Contact: Claude Renaud, Research Services Division, Canadian Museum of Nature, P.O. Box 3443, Station D Ottawa Ontario K1P 6P4 Canada E-Mail: [email protected] Abstract: There are 38 lamprey species distributed in temperate and cold waters of the two hemispheres. In 2003 a first cladogram based on morphological characters was constructed for the 18 species that feed as adults. A global effort involving 16 collaborators has recently produced a cladogram for over 35 species, parasitic and nonparasitic, using a mitochondrial gene, cytochrome b. While a number of the relationships proposed by the morphologically-based cladogram were also recovered by this new study; others are novel. For example, the monotypic genus Okkelbergia is resurrected for the least brook lamprey and the western North American Lampetra, L. ayresii and L. richardsoni, form a monophyletic group quite distinct from Eurasian members of the genus. Future work will utilize nuclear genes and integrate the morphological and molecular data in order to produce a total evidence cladogram.

Fish Biology Congress Abstracts 79 RESPONSES OF HEAT SHOCK PROTEINS TO THERMAL STRESS IN POPULATIONS OF GAMBUSIA Reneau, P.C. and Broughton, R.E. Symposium: Genomics of Adaptation Presentation: Oral Contact: Paulette Reneau, University of Oklahoma, Oklahoma Biological Survey and Department of Zoology, 111 E. Chesapeake St. Norman OK 73019 USA E-Mail: [email protected] Abstract: Abiotic stressors play a fundamental role in defining the limits of adaptation. One of the mechanisms by which fishes adapt to stressful environments involves differential expression of genes that aid in stabilization and recovery. We investigated the relationship between gene expression and adaptation in thermally variable habitats in natural populations of Gambusia, fishes known for their thermotolerance and success in colonizing and persisting in ecologically harsh environments. We utilized three species (Gambusia affinis, Gambusia geiseri, and Gambusia speciosa) naturally occurring in different thermal environments in order to compare the relative expressions of hsp 90, hsp70 and hsc 70. Quantitative real time PCR of cDNA from liver tissues revealed a direct relationship between an increase in temperature and the expression of specific heat shock genes in several cases. In contrast, one obligate cool-spring dwelling population showed a decrease in relative expression of heat shock protein transcripts with increased temperature. These results provide direct evidence that species can respond to variable and stressful environments via differential expression of the same suite of genes. This mechanism may allow more rapid adaptation than evolutionary modification of primary gene coding sequences and confer enhanced ability to colonize environments with novel stresses.

SHORT TERM VENTILATORY DEPRESSION AND PHENOTYPIC PLASTICITY MAY PROTECT THE ARMOURED CATFISH (LIPOSARCUS PARDALIS) FROM REOXYGENATION INJURY AND OXIDATIVE STRESS. Renshaw G.M.C., Paula-Silva M.N. and Almeida-Val V.M. Symposium: Tropical Fish Adaptations Presentation: Oral Contact: Gillian Renshaw, 1Hypoxia & Ischemia Research Unit, Heart Foundation Research Centre & School of Physiotherapy & Exercise Science. Griffith University, Gold Coast Campus. Parklands Queensland 4222 Australia E-Mail: [email protected] Abstract: Early vertebrates evolved in hypoxic environments and some still retain the ability to reversibly reprogram their metabolism in order to prolong their endurance in extremely “oxygen impoverished” tropical environments in the black water systems of the Amazon. The respiratory and metabolic defence strategies used in response to sudden anoxia followed by 24 hours of reoxygenation was examined in the Amazonian armoured catfish (Liposarcus pardalis), a facultative air breather. We quantified: respiration rate, lactate dehydrogenase (LDH) isoenzyme profiles, the level of citrate synthase (CS) and glutathione S-transferase (GST). During anoxic exposure, there was a rapid onset of ventilatory depression and after 24 hours of reoxygenation there were significant tissue specific changes in LDH and GST but not CS. The up- regulation of anaerobic isoforms of LDH coupled with the increased GST may confer resistance to oxidative stress associated with reoxygenation and provide a means of extending the time between air breathing intervals.

CHANGES IN THE EXPRESSION OF HYPOXIA-INDUCIBLE FACTOR-1 AND HEAT- SHOCK COGNATE 70 IN THE LIVER AND HEART OF THE TROPICAL ANOXIA-TOLERANT EPAULETTE SHARK (HEMISCYLLIUM OCELLATUM), IN RESPONSE TO AN ANOXIC CHALLENGE. Renshaw, G.M.C., Castelli A.V., Pritchard G.A., Ashton K.J., Symposium: Fish Living on the Edge Presentation: Poster Contact: Gillian Renshaw, Hypoxia & Ischemia Research Unit, School of Physiotherapy & Exercise Science. Griffith University, Gold Coast Campus. QLD 4222. Southport Queensland 4222 Australia E-Mail: [email protected] Abstract: While most vertebrates are anoxia intolerant, a few fish and amphibian species have evolved both protective and reparative mechanisms that enable them to survive anoxia at temperatures close to freezing. However the epaulette shark (Hemiscyllium ocellatum) can survive extended periods of anoxia at tropical temperatures and maintains constant blood glucose levels during these periods, indicating that some metabolic pathways may remain intact during anoxia. Both long- and short-term protection and repair strategies may involve translational as well as transcriptional modification of a suite of protective genes. Expression levels of a master regulator of over 200 oxygen-sensitive genes, Hypoxia inducible factor 1 alpha (HIF-1 α) and one of its downstream target genes Heat shock cognate 70 (HSC70) were measured using real- time PCR in order to determine the effect of and anoxic challenge on stress-activated repair mechanisms (HSC70) and transcriptional modification (HIF-1 α).

Fish Biology Congress Abstracts 80

ELEVATED ESTROGEN IN MALE FLATFISH IN SOUTHERN CALIFORNIA'S URBAN OCEAN: BIOACCUMULATION OR TESTICULAR SYNTHESIS? Reyes, J.A., Hagstrom, K.R.E., Petschauer, D.M., Koons, K.S. and Kelley, K.M. Symposium: Environmental Pollution Presentation: Oral Contact: Jesus Reyes, Environmental Endocrinology Laboratory, Department of Biological Sciences, California State University, Long Beach Long Beach CA 90840 USA E-Mail: [email protected] Abstract: The indigenous flatfish, hornyhead turbot (Pleuronichthys verticalis), collected within the Southern California Bight (SCB) marine environment has been found to exhibit extraordinarily high circulating concentrations of 17beta-estradiol (E2), particularly in locations impacted by municipal wastewater treatment plant outfalls. Male turbot with this phenotype show up to >1000 pg/ml plasma E2, as high or higher than that measured in females; in contrast, males of other flatfish species have low E2 levels (<50 pg/ml) irrespective of location. Further work suggests that bioaccumulation of environmental E2 is possible, since sediment invertebrates that make up the specific diet of turbot have high tissue E2 concentrations. An additional, likely possibility is that endogenous E2 synthesis may be activated in the impacted turbot, since testicular mRNA expression of StAR, P450arom and 17beta-HSD-1 are altered in a pattern indicative of enhanced steroid synthesis. (Support by NOAA-USC Sea Grant #CE-17 and Pacific Coast Environmental Conservancy)

EARLY RESPONSE GENES OF CHINOOK SALMON MAY BE IMPORTANT IN INFECTION BY RENIBACTERIUM SALMONINARUM Rhodes, L.D., Wallis, S., and Demlow, S.E Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Linda Rhodes, Northwest Fisheries Science Center, 2725 Montlake Blvd. East Seattle WA 98112 USA E-Mail: [email protected] Abstract: The early response to bacterial infection can be a crucial decision point in the host’s efforts to contain a pathogen. We exploited differences in the response of juvenile Chinook salmon to infection by a virulent or attenuated strain of Renibacterium salmoninarum, the etiologic agent of bacterial kidney disease, to identify potentially important early genes. Pronephros from fish exposed to either a virulent or attenuated strain was subjected to suppression subtractive hybridization at 72 hours post-infection, and over 130 library clones of expressed sequence tags (ESTs) were sequenced. Twenty ESTs with high quality GenBank matches were selected for stable transcript analysis at 24 and 72 hours post-infection. ESTs matching genes in interferon-inducible pathways, apoptosis, monocyte/macrophage function, and transmembrane receptors were expressed at significantly higher levels among fish exposed to the attenuated strain. These ESTs provide insight into an effective host response, and they may be useful tools in vaccine development.

GENOMIC RESEARCH FOR IMPROVING FISH HEALTH Rise, M.L. Symposium: Genomics of Adaptation Presentation: Oral Contact: Matthew Rise, Ocean Sciences Centre, Memorial University of Newfoundland, 1 Marine Lab Road St. John's NL A1C 5S7 Canada E-Mail: [email protected] Abstract: In my research program, experiments involving aquatic animal models and genomic techniques (e.g. suppression subtractive hybridization cDNA library construction and sequencing, expressed sequence tag database creation and mining, DNA microarray hybridization, and quantitative reverse transcription – polymerase chain reaction) are used to study the genes and molecular pathways involved in biological processes such as embryonic development, organogenesis, growth, and responses to pathogens (e.g. viral, bacterial, protozoal) and environmental toxicants (e.g. heavy metals, pesticides, dioxins). In addition to providing valuable insight into the molecular basis of these processes, information acquired in this largely collaborative, functional genomics research will lead to applied projects such as the development of new molecular tools and techniques for detecting and combating emerging fish diseases, evaluating the impact of environmental stressors on wild fish populations, and selecting fish with superior production traits.

CHARACTERIZATON OF MYOSTATIN INTERACTING PROTEINS IN TROUT Roberts, S.B., and Goetz, F.W. Symposium: Fish Growth Presentation: Oral Contact: Steven Roberts, University of Washington, 1122 NE Boat St Seattle WA 98105 USA E-Mail: [email protected]

Fish Biology Congress Abstracts 81 Abstract: Myostatin is a member of the transforming growth factor-beta superfamily that has been shown to be a significant regulator of skeletal muscle growth in mammals. To date, clear evidence that myostatin is a potent regulator of muscle growth in fish is lacking. In order to characterize the role myostatin has in salmonid muscle growth, a yeast two-hybrid system is being used to identify proteins that interact with that active form of myostatin. Recombinant proteins have been generated based on these results and used to treat primary cultures of trout muscle cells. Muscle cell cultures were routinely imaged and sampled eight days after plating. Gene and protein expression levels across treatments were analyzed. Significant muscle hypertrophy was not observed, however cell morphology and expression patterns suggest addition of certain molecules can influences muscle cell physiology. This research is supported by US Department of Agriculture grant no. 2005-35206- 1526.

GENOMIC ANALYSIS OF BEHAVIORAL ADAPTATION TO CAPTIVITY IN THE ZEBRAFISH Robison, B.D. Symposium: Genomics of Adaptation Presentation: Oral Contact: Barrie Robison, University of Idaho, PO Box 443051 Moscow ID 83844 USA E-Mail: [email protected] Abstract: Captive rearing of endangered fishes is often used to mitigate population declines. However, captive rearing can cause significant changes in fear related behaviors. The less fearful phenotypes of domesticated populations are often deleterious in the natural environment. We are using the zebrafish to study the genetic changes associated with domestication. We compared place preference, foraging, and aggression behaviors of zebrafish strains with different domestication histories, and observed reduced fearfulness in domesticated strains. Aggression also varied among strains, but was not associated with domestication history. Microarray analysis of whole brain RNA from two wild and two domesticated strains indicated over 1,500 transcripts as differentially expressed among strains. Categories of genes that differentiated strains included steroid hormone biosynthesis, response to stress, appetite regulation, and sequestration of selenium. We are now using these data to nominate candidate genes that may underlie behavioral variation arising from domestication.

SEXUAL DIMORPHISM IN THE HEPATIC TRANCIPTOME OF ZEBRAFISH AND THE EFFECT OF DIETARY CARBOHYDRATES Robison, B.D., Drew, R.E., Murdoch, G.K., Powell, M., Rodnick, K.J., Settles, M., Stone, D., Churchill, E., Hill, R.A., Papasani, M.R., Lewis, S.S., and Hardy, R.W. Symposium: Fish Growth Presentation: Oral Contact: Robert Drew, Department of Biological Sciences, University of Idaho Moscow ID 83844 USA E-Mail: [email protected] Abstract: We tested for sexual dimorphism in the hepatic transcriptome of adult zebrafish and examined the effect of long term manipulation of dietary carbohydrate levels. Zebrafish were fed diets with 0%, 15%, 25%, and 35% carbohydrate levels from the larval stage to maturity. Using microarrays and qRTPCR, we detected substantial sexual dimorphism in the hepatic transcriptome. Males up-regulated genes involved in oxidative metabolism, carbohydrate metabolism, energy production, and response to oxidative stress, while females up-regulated genes involved in translation. Carbohydrate deprivation (0%) significantly affected gene expression, growth rate, and retention of protein and energy. For some genes, the response to nutritional manipulation differed between the sexes. With increased carbohydrate levels, males up- regulated genes involved in oxidative metabolism while females up-regulated genes involved in glucose phosphorylation. This is the first report of sexual dimorphism in the hepatic transcriptome of a fish species and highlights the importance of this phenomenon in nutrition studies.

CORTISOL-SENSITIVE BASOLATERAL UREA TRANSPORT ACROSS ACROSS THE GILLS OF THE GULF TOADFISH, OPSANUS BETA Rodela, T.M, Gilmour, K.M., Walsh, P.J., and M.D. McDonald Symposium: Fish Living on the Edge Presentation: Oral Contact: Tammy Rodela, Dept of Biology, University of Ottawa, 30 Marie Curie Ottawa ON K1N 6N5 Canada E-Mail: [email protected] Abstract: Gulf toadfish, Opsanus beta, employ a unique pulsatile urea excretion mechanism allowing the fish to void urea from the body in large, irregular pulses via the periodic insertion or activation of a branchial urea transporter. The precise cellular and sub-cellular location of the facilitated diffusion mechanism(s) still remains uncertain. This study examined the cortisol- sensitive contribution of basolateral membrane urea transport in ureotelic toadfish. Toadfish demonstrated two components of the rate urea uptake, a linear constituent at high urea concentrations and saturation kinetics at low (<1 mM) urea

Fish Biology Congress Abstracts 82 concentrations. Saturable urea uptake was inhibited by phloretin and reduced in the presence of urea analogues. In vivo manipulations demonstrated a reduction in the rate of urea transport in response to cortisol treatment, which suggest that cortisol inhibits the recruitment of urea transporters to the basolateral membrane which may ultimately affect the size of the urea pulse event in gulf toadfish.

MATING SYSTEMS IN LAMPREYS: THE ROLE OF GENETIC COMPATIBILITY Rodríguez-Muñoz, R. Symposium: Biology of Lampreys Presentation: Oral Contact: R. Rodríguez-Muñoz, Centre for Ecology and Conservation, University of Exeter, Cornwall Campus, Tremough Penryn Cornwall TR10 9EZ UK E-Mail: [email protected] Abstract: Lampreys exhibit a wide variety of social mating systems, ranging from monogamy-polygyny for some big parasitic species, to promiscuity in brook lampreys. As in many other vertebrate species, social behaviour is not always closely related to genetic mating system. Species like the sea lamprey, lacking social polyandry, have been recently reported to have a polygynandrous mating system, derived from female polygamous behaviour as opposed to male territoriality. Male polygamy is easier to explain due to the positive correlation between the number of partners and male fitness. Polyandry, in turn, is more controversial, as in most species a single male has the potential to fertilize all the progeny of several females. Many of the hypotheses proposed to explain polyandrous behaviour focus on the existence of genetic benefits for multiply mated females. I discuss the potential genetic benefits of multiple mating in lampreys and show some evidence for the existence of genetic compatibility issues in the sea lamprey.

USE OF ARCTIC ROCKLING (ONOGADUS ARGENTATUS) FOR DEEP-WATER ENVIRONMENTAL MONITORING, BIOMARKER BASELINE LEVELS IN MATERIAL COLLECTED OFF NORWAY. Rolf C Sundt, Nadia Aarab and Jan Fredrik Børseth Symposium: Environmental Pollution Presentation: Poster Contact: Nadia Aarab, International Research Institute of Stavanger (IRIS), Marine Environment, Mekjarvik 12, Randaberg Randaberg N 4070 Norway E-Mail: [email protected] Abstract: The increased extent of deep sea oil and gas exploitation increases the demand for environmental research and monitoring at great depths. To investigate the suitability of deep-water fish for biomonitoring, Arctic rockling, (Onogadus argentatus) was collected at two deep-water sites where oil exploration is planned. The sites are situated in the Vøring area on the continental slope off Norway and are characterized by clay bottom and depths of approximately 1.100 m. Fish were analyzed for a suite of chemical (PCB, PFOS, PAH metabolites) and biological markers of contamination (EROD, histology) in order to define baseline levels of the different parameters. Levels in Arctic rockling were similar to levels obtained in fish species where these biomarkers are already established, indicating that this species may be used as a sentinel organism for environmental investigations in arctic deep-sea waters.

GETTING TO THE ROOT OF TISSUE OXYGENATION IN TELEOST FISH: A MORE UBIQUITOUS ROLE FOR THE ROOT EFFECT IN OXYGEN DELIVERY Rummer, J.L., and Brauner, C.J. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Jodie L. Rummer, University of British Columbia, Department of Zoology, 6270 University Blvd. Vancouver BC V6T 1Z4 Canada E-Mail: [email protected] Abstract: Teleost fish possess Root effect hemoglobins (Hb) that, during an acidosis, exhibit low affinity and carrying capacity for O2. This mechanism benefits O2 delivery to swim bladder (SB) and retinal tissue, but during a general acidosis may compromise O2 uptake at the respiratory surface. Most teleosts adrenergically safeguard O2 uptake during an acidosis; catecholamines are released which stimulates erythrocyte Na+/H+ exchanger (beta-NHE) that increases pHi. In rainbow trout blood, we simulated the Root effect, catecholamine release, and beta-NHE short-circuiting via plasma-accessible carbonic anhydrase (CA), and monitored changes in real-time. CA short-circuits adrenergic pH regulation and facilitates an increase in PO2 over 30-fold that which would be accomplished with the Bohr effect alone. If the Root effect can also facilitate general O2 delivery, which our data currently support, this could clarify selection for the Root effect prior to the evolution of the beta-NHE and specialized retinal and SB tissues.

Fish Biology Congress Abstracts 83 THE EFFECT OF SEA LICE (LEPEOPHTHEIRUS SALMONIS) ON IONOREGULATION IN JUVENILE PINK SALMON (ONCORHYNCHUS GORBUSCHA) Sackville, M.A., Gardner, M., Grant, A., Nendick, L.J., Farrell, A.P., and Brauner, C.J. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Michael Sackville, Department of Zoology, University of British Columbia, 6270 University Blvd. Vancouver BC V6T 1Z4 Canada E-Mail: [email protected] Abstract: Sea lice are endemic to British Columbia’s Broughton Archipelago, but elevated levels associated with fish farms have been implicated in reducing local pink salmon populations. Despite these allegations, little is known about the relationship between sea lice infection levels and associated sub-lethal disturbances in juvenile pink salmon. Due to the physically disruptive nature of louse attachment to the fish’s skin and the challenge of hypo-osmoregulation in a seawater environment, we hypothesize that negative impacts of sea lice on fish performance are ionoregulatory in origin. Combining field and laboratory studies, we examined the relationship between sea lice infection levels and sublethal indicators of ionoregulatory status (gill Na+/K+-ATPase activity, drinking rate, whole-body ion concentrations) to establish a threshold infection level above which significant physiological impacts arise. In conjunction with other measures of whole animal performance, these data will be used to model the effect of sea lice infection on pink salmon fitness.

GENETIC STRUCTURE OF A SPAWNING SCHOOL: A TRACK TO KNOWLEDGE ON THE POPULATION STRUCTURE OF THE FRESHWATER MIGRATORY FISH, BRYCON HILARII Sanches, A., and Galetti Jr., P.M. Symposium: Tropical Fish Adaptations Presentation: Oral Contact: Pedro Manoel Galetti Junior, Departamento de Genética e Evolução, Universidade Federal de São Carlos, Via Washington Luis Km 235 São Carlos SP 13565-905 Brazil E-Mail: [email protected] Abstract: Two or more genetic populations of migratory fish can co-exist in a single hydrographic system, but they must segregate during spawning to maintain their genetic integrity. The genetic variation of a Brycon hilarii spawning school and samples from different collection sites in the Miranda River basin (larger Paraguay River basin, Brazil) were surveyed using seven microsatellites. FST values, gene and genotype frequencies indicate a population structuring revealed by a significant differentiation of the spawning school from the supposed local populations. However, no structuring was observed regarding the local populations. Bayesian clustering analysis revealed that the genetic composition of spawning school was significantly homogenous in comparison to the heterogeneity of the local populations. These results may be supported by a behavior during the reproductive season that promotes the maintenance of the genetic integrity of different subpopulations, such as spawning waves or homing instinct, and are very important for conservation.

EFFECTS OF DIFFERENT STRESSORS IN BLOOD CHEMISTRY IN CULTURED NILE TILAPIA OREOCHROMIS NILOTICUS Sandoval-Gío, J., J., Quintero-Mármol, E., S., Rosado-Vallado, M., E., and Rodríguez-Canul, R. Symposium: Stress Effects and Response Presentation: Poster Contact: Juan José Sandoval Gío, Laboratorio de Inmunología y Biología Molecular, Centro de Investigación y de Estudios Avanzados del IPN (CINVESTAV-IPN) Unidad Mérida, Carretera Antigua a Progreso Km, 6 AP 73 Cordemex. Mérida Yucatán 97310 México E-Mail: [email protected] Abstract: The objective of this study was to establish baseline in the selected blood chemistry values from 36 cultured Nile Tilapia Oreochromis niloticus and to compare them with values from tilapia exposed to several stressors. Three groups with ten fishes each were inoculated respectively, with saline extracts of Cichlidogyrus spp., with Streptococus spp.extracts, and were exposed a therapeutic treatment with formalin (1ml/L) for 60 min. Mean (minimum-maximum) values, obtained by colorimetric methods using ELISA microplate (96 well) were as follows: glucose, 100.83 (45-183) mg/dL; cholesterol, 210.10 (119-287) mg/dL; total protein, 3.69 (2-5) g/dL; albumin, 1.73 (1-3) g/dL; globulins, 1.95 (1-3) g/dL; uric acid, 4.93 (3-12)mg/L; calcium, 17.40 (6-32) mg/dL; chloride, 263.49 (130-445) meq/L, and magnesium, 2.59 (1-4) mg/dL. Our results establish that for this species, the induced parasitic and bacterial infection, and the therapeutic treatment caused changes in several blood chemistry values that can serve as index of fish health status.

SALINITY STRESS IN GREEN STURGEON: THE EFFECTS OF GLOBAL CLIMATE CHANGE ON SAN FRANCISCO BAY-DELTA POPULATIONS. Sardella, B.A. and Kültz, D.

Fish Biology Congress Abstracts 84 Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Brian Sardella, Dept. of Animal Science, UC Davis, One Shields Ave Davis CA 95616 USA E-Mail: [email protected] Abstract: Abiotic factors within the San Francisco Bay-Delta are forecasted to become less stable due to global climate change. This region is inhabited by a wide range of fish species including the threatened green sturgeon (Acipenser medirostris). Forecasted changes to will result due to a precipitation shift from snow to rain at higher altitudes, and thus less snow pack, causing large flushes of freshwater through the estuary during winter, and greater tidal influence during summer when run- off is low. We simulated these effects by exposing estuary- or bay acclimated sturgeon to a rapid (3 h) freshwater pulse. To simulate summer conditions, freshwater-acclimated sturgeon were exposed to an oscillation of salinity to simulate a tidal cycle, with salinity peaking at 24 g/l at 12 and 18 h. At each sampling time we assessed plasma osmolality, muscle hydration, Na+/K+-ATPase and H+-ATPase activities/abundance, heat shock protein expression and effects on apoptosis.

CHANGES IN THE WHITE STURGEON PROTEOME FOLLOWING TEMPERATURE ACCLIMATION. Sardella, B.A. and Kültz, D. Symposium: Stress Effects and Response Presentation: Poster Contact: Brian Sardella, Dept. of Animal Science, UC Davis, One Shields Ave. Davis CA 95616 USA E-Mail: [email protected] Abstract: White sturgeon (Acipenser transmontanus) were transferred from 18 to 27 ºC (50% of CTMax for 18ºC-acclimated fish) and acclimated for two weeks. Physiological responses to temperature stress were measured at the level of the proteome using two-dimensional gel electrophoresis, followed by mass spectrometry and T3 sequencing. Proteins were extracted from gill tissue samples and separated using immobilized pH gradient focusing for the first dimension and sodiumdodecylsulphate-polyacrylamide gels (SDS-PAGE) for the second dimension. To generate a list of proteins that were specifically up regulated or down regulated during acclimation, gels were scanned, digitized, and analyzed using Delta 2D software which provided an index of any differences in the proteome between control and experimental conditions. Ten proteins were up regulated at least 100% and nine were down regulated by at least 50%. These individual proteins were extracted and identified using mass spectronomy.

A COMPARATIVE GENOMICS APPROACH FOR DISCOVERY OF CYTOKINES AND THEIR REGULATORY REGIONS Savan, R Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Ram Savan, National Cancer Institute, Laboratory of Experimental Immunology, MD , USA Frederick MD 21701 USA E-Mail: [email protected] Abstract: A comparative study of immune system is critical in understanding the mechanism of pathogenesis and disease resistance. As fish forms a major evolutionary link between the invertebrates and the mammals, it is a good model organism to understand the evolution of immune processes. Recent releases of the draft sequence of fish genomes have lead to a rapid acceleration in the discovery of cytokine genes. The major cytokines isolated and characterized are IL-2, IL-4, IL-6, IL-7, IL-12, IL-15, IL-17 family, IL-10 family (IL-10, IL-19, IL-22 and IL-26), IFNg and TNF superfamily ligands and receptors. Interestingly, novel genes related to IL-2, IL-17, IFNg and TNF families have also been discovered in fish. Using comparative genomic approaches, we have identified several important conserved non-coding sequences which may have important role in regulation of cytokines. By comparative sequence analyses of these genes will gain insights on evolution of the immune system and also help understand the regulation of these molecules during disease.

FISH IMMUNE SYSTEM AND INTERACTIONS WITH ENVIRONMENT Scapigliati, G., Randelli E., Casani D., Boschi I., Buonocore F. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Giuseppe Scapigliati, Department of Environmental Sciences, University Of Tuscia, Largo, dell'Universita' Viterbo 1100 Italy E-Mail: [email protected] Abstract: Animal species live in homeostasis with the environmental and body microbiome, constituted by a huge number of microorganisms. A dynamic equilibrium exists between aggression strategies developed by many microorganisms and defence strategies invented by animal eukaryotes, with the immune system that must guarantee full immune defence in every environmental condition. Employing as main investigated species the sea bass Dicentrarchus labrax, we defined the

Fish Biology Congress Abstracts 85 distribution and the ontogenesis of lymphocytes, their involvement during antigenic stimulation “in vivo” and “in vitro”, the cloning of genes coding for immunoregulatory molecules, and expression of these genes during immune responses. Current research focus on transcriptomic analysis through PCR-arrays of gene groups coding for the sea bass immunome. Of great interest is the gut-associated immune system, rich in T lymphocytes, whose functional organisation could be related to food habits, and work is in progress to study non-predatory fish species. European Union funded Research (6FP, 000173)

ENERGETIC COSTS OF OVERWINTERING SUMMER STEELHEAD, ONCORHYNCHUS MYKISS Schemmel, Eva; Clements, Shaun; Schreck, Carl and Noakes, David Symposium: Physiological BioTelemetry Presentation: Poster Contact: David Noakes, Fisheries & Wildlife Department and Oregon Hatchery Research Center, 104 Nash Hall, Oregon State University Corvallis Oregon 97331-3803 USA E-Mail: [email protected] Abstract: We evaluated energetic costs of summer steelhead in the Clackamas River, Oregon with coded electromyographic (EMG) radio tags. We implanted EMG tags internally with electrodes placed in the red muscle to measure the muscle activity. The EMG code is related to activity levels and swimming speed of tagged steelhead. We calibrated the EMG tag code to swimming speed using a Blazka swim tube at the Oregon Hatchery Research Center. We tagged returning hatchery summer steelhead with EMG radio tags at the Clackamas Hatchery and released them downstream. We tracked these tagged steelhead from July to December to evaluate diel activity patterns and energetic cost of overwintering in the river. We recorded positions and movements of tagged fish for 24 hour cycles, as well as from fixed receivers over the total time period. We used measures of locomotor activity, water temperature and habitat quality to estimate the energy budget of steelhead.

EFFECTS OF INJURY AND STRESS ASSOCIATED WITH TRANMSPORT OF LIVE TILAPIA: DO THEY INOCULATE EACH OTHER WITH PATHOGENS? Schreck, C., Momoda, T., Chitwood , R., Feist, G., and Colt, J. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Carl Schreck, Oregon Cooperative Fish and Wildlife Research Unit, USGS, 104 Nash Hall, Oregon State University Corvallis Oregon 97330 USA E-Mail: [email protected] Abstract: Tilapia are transported long distances from fish culture facilities to urban centers for the live fish market where a variable amount of mortality is experienced. We are studying the transportation and distribution processes to identify stressful and injurious practices. Importantly, we are testing the hypothesis that stress and trauma, particularly physical injury, experienced during the transport process predisposes the fish to infection. This is supported by data on fish from both laboratory simulated transport and actual transport from the growers to the market. Physical injury was assessed by using fluorescein dye on fish from various discrete aspects of the hauling process including crowding, truck loading, unloading, and subsequent holding. Our work indicates that such things as the type of net used may affect the injury incurred by the fish. Furthermore, the addition of salts to the hauling medium or a pre-transport salt dip reduces mortality despite the injury to fish.

MINUTE TEMPERATURE DECREASE TRIGGERS HORMONE MEDIATED DOWNSTREAM MOVEMENT IN PACIFIC SALMON Schreck, C., Munakata, A., Miura, G. Kitamura, S., Aida, K, and Hansen, B. Symposium: Stress Effects and Response Presentation: Oral Contact: Carl Schreck, Oregon Cooperative Fish and Wildlife Research Unit, USGS, 104 Nash Hall, Oregon State University Corvallis Oregon 97331 USA E-Mail: [email protected] Abstract: Downstream orientation in juvenile salmon (Oncorhynchus spp.) appears triggered by slight decreases in temperature independent from changes in flow and mediated by hormones of smoltification. Small decreases in water temperature (< 2 C) were associated with elevations in plasma cortisol levels in smolts but not resident masu salmon in a small river. Cortisol implantation increased downstream orientation in masu smolts, and a decrease in temperature (< 1C) increased plasma cortisol to levels of natural stream migrants. Chinook exposed to a similar decrease in temperature exhibited an increase of cortisol levels. Coho salmon parr in an artificial stream exhibited downstream orientation when exposed to a 1.2C drop in temperature. Coho salmon parr given thyroxine or cortisol implant showed constant downstream behavior

Fish Biology Congress Abstracts 86 independent of water temperature, similar to that of un-hormone implanted fish exposed to a temperature decrease. Steelhead exposed to temperature increases and decreases will also be discussed.

EXPLORING THE EVOLUTION OF PHYSIOLOGICAL TRAITS USING COMPARATIVE GENOMICS Schulte, P.M. Symposium: Genomics of Adaptation Presentation: Oral Contact: Trish Schulte, Department of Zoology, University of British Columbia, 6270 University Blvd Vancouver BC V6T 1Z4 Canada E-Mail: [email protected] Abstract: Research in my laboratory is focused on understanding the physiological adaptations that allow fish to live in particular environments. In this presentation I will discuss work that uses genomics approaches to explore two different systems: the evolution of variation in exercise performance in populations of stream-resident and anadromous three-spine stickleback (Gasterosteus aculeatus), and variation in thermal tolerance in populations of Atlantic killifish (Fundulus heteroclitus) from different latitudes. In both cases, we have utilized microarray analyses of gene expression in fish held under common conditions in the laboratory to provide clues as to the biological processes that differ between closely related groups of fish that differ in whole-organism performance. This work, which has taken advantage of hybridization to heterologous microarrays, demonstrates that comparative genomics approaches are accessible even in species for which few molecular tools are available.

ENVIRONMENTAL AND GENETIC SOURCES OF VARIATION IN JUVENILE SEA LAMPREY DISPERSAL AND PHENOTYPE Scribner, K., Jones, M., Rosa, G., and Gilmore, S. Symposium: Biology of Lampreys Presentation: Oral Contact: Kim Scribner, Department of Fisheries and Wildlife and Department of Zoology, Michigan State University, 13 Natural Resources Building East Lansing MI 48824 USA E-Mail: [email protected] Abstract: Control methods that disrupt invasive sea lamprey reproduction are based on assumptions pertaining to unresolved aspects of the species’ breeding behavior, reproductive success, and patterns of larval dispersal and growth. We used replicated experimental streams stocked with small numbers of adults and microsatellite loci to genetically determine parentage, to elucidate aspects of sea lamprey behavior and larval life history. Reproductive success of male and female lamprey was highly variable. High rates of polyandry and polygyny and high variances in male and female reproductive success were observed. Full and half sibs were typically found in close proximity in the stream, though larval dispersal distances >5 km were observed. Estimates of heritability for larval body weight and length were 0.458 and 0.443, respectively across streams. We discuss the implications of additive genetic covariance between relatives and stream environmental heterogeneity to recruitment dynamics and compensatory demographic responses to sea lamprey control.

GIRLS GONE WILD: DEFEMINIZATION IN THE ELKHORN RIVER Sellin, M.K. and Kolok, A.S. Symposium: Environmental Pollution Presentation: Oral Contact: Marlo Sellin, Department of Biology, University of Nebraska at Omaha, 6001 Dodge Street Omaha NE 68182 USA E-Mail: [email protected] Abstract: Cattle feedlot runoff has been identified as a potential source of endocrine-disrupting compounds in the aquatic environment due to the use of growth-promoting implants in beef production. The Elkhorn River watershed is characterized by a high density of beef feedlots making it an ideal site to investigate the impacts of feedlot runoff on fish health. The goal of this study was to determine whether caged fathead minnows deployed in the Elkhorn River experience alterations in endocrine function. To accomplish this, the expression of estrogen- and androgen-responsive genes was measured in minnows deployed in the Elkhorn River and at two reference locations. Females deployed in the Elkhorn River experienced significant reductions in the expression of vitellogenin and estrogen receptor alpha relative to females deployed at the reference locations indicating a defeminization of females at this site. The reproductive consequences of reduced vitellogenin expression, while axiomatic, remaining to be experimentally validated.

GROWTH IN COASTER BROOK TROUT: SEASONAL AND HORMONAL EFFECTS Serfas, C., Holman, R., Watch, L., Karner, J., Leonard, J.B.K. Symposium: Fish Growth Presentation: Poster

Fish Biology Congress Abstracts 87 Contact: Carla Serfas, Northern Michigan University, Biology Department, 1401 Presque Isle Marquette MI 49855 USA E-Mail: [email protected] Abstract: Some brook trout (Salvelinus fontinalis) are termed coasters because they spend all or part of their life in a large lake. Recently, coaster populations have declined in Lake Superior. Different populations of Lake Superior brook trout (age 1) were compared over time to determine if they varied in growth rate, condition factor or thyroid hormone concentration. Three coaster strains (Nipigon, Siskiwit, Tobin Harbor) and 1 stream resident strain (Iron River) were held at constant conditions and natural photoperiod with a subset of fish sampled monthly. Differences were found in condition factor across sampling periods and strains (p<0.001). Nipigon strain had a higher condition factor than Iron River, Siskiwit, and Tobin Harbor strains (5.6%, 5.3%, and 9.6% respectively). We suggest that the differences in growth in field studies are likely due to environmental effects given the similarity in growth parameters seen under common rearing conditions.

ASSESSEMENT OF THE WATER QUALITY DURING THE TRANSPORTATION OF PIAU JUVENILES (LEPORINUS FRIDERICI). Serra, M.; Wolkers, C.P.B.; Hoshiba, M.A.; and Urbinati, E.C. Symposium: Stress Effects and Response Presentation: Poster Contact: Mônica Serra, Rua Castro Alves, 2211 Jaboticabal São Paulo 14875-270 Brazil E-Mail: [email protected] Abstract: This work evaluated the water quality and physiological parameters of juveniles of piau (Leporinus friderici) during a 4h- transport (133g.L-1). Temperature and total ammonia (NH4) concentration increased significantly during the transport (1, 2, 3 and 4 h) but the concentration of the un-ionized ammonia (NH3) did not differ, and was maintained at levels below of the level considered inappropriate for tropical fish, probably because pH values were maintained lower than 7.0 along the transport. Glucose increased during the transport especially after 3 h of the beginning. The mean corpuscular volume of erythrocytes increased during the transport but the increase of both parameters was not related to the water quality changes but to the fish handling to the transport.

REGULATION OF POSTPRANDIAL HYPEREMIA - IMPORTANCE OF MECHANICAL AND CHEMICAL STIMULI Seth, H. and Axelsson, M. Symposium: Contributed Papers Presentation: Oral Contact: Henrik Seth, Göteborg University, Department of Zoology, Zoophysiology, Medicinaregatan 18, Box 463 Göteborg 405 30 Sweden E-Mail: [email protected] Abstract: The regulation of the gastrointestinal hyperemia that occurs after feeding is complex and several factors are important, such as mechanical and chemical stimuli, gastrointestinal hormones as well as metabolic factors. This postprandial hyperemia is attributable to either an increase in cardiac output (Q) or a redistribution of blood and enables an effective absorption and redistribution of nutrients, while supplying the metabolically active tissue with oxygen. We have shown that mechanical stimuli induce a consistent pressor response in fish. This increase is most likely important in the subsequent redistribution of blood as food enters the proximal intestine and as with mammals different food components such as proteins or fats are not equally important a shown when injecting different nutrient solutions into the proximal intestine. All in all there is a complex interaction between mechanical and chemical stimuli and future studies will hopefully reveal the exact nature of this regulation.

STUDY OF MTDNA VARIATION OF STELLATE STURGEON (ACIPENSER STELLATUS)POPULATION FROM THE NORTH(VOLGA RIVER)AND SOUTH(GORGANRUD RIVER)CASPIAN SEA USING RFLP ANALYSIS OF PCR AMPLIFIED ND5/6 GENE REGIONS. Shabani, A., Pourkazemi, M. Rezvani, S. And Shabanpour, B. Symposium: Contributed Papers Presentation: Poster Contact: Ali Shabani, Gorgan Unniversity of Agricultural Sciences and Natural Resources, Faculty of Fishery and environmental Sciences P.O. Box: 49165-386-Gorgan-Iran, GORGAN Iran E-Mail: [email protected] Abstract: Genetic Variation of Stellate Sturgeon (Acipenser Stellatus) from the North (Volga river) and South (Gorganrud river)Caspian sea was investigated using NADH 5/6 gene and PCR-RFLP analysis. In this Study a total of 60 samples of Stellate Sturgeon from the North (Volga river) and 14 samples from the Southern (Gorganrud river) Caspian sea were Collected. MtDNA ND 5/6 gene was amplified by polymerase chain reaction (PCR). PCR-product digested using 31

Fish Biology Congress Abstracts 88 endonucleases restriction enzyme. Five enzymes of 31 enzymes showed polymorphism (RsaI,HinfI, HpaII, HaeIII, MboI). Totally 18 composite haplotypes among 74 samples were detected. Haplotypes AAAAA, BAAAA, ABABA from the Volga River showed 28.33%, 11.66% frequency respectively and from the Gorganrud river haplotypes AAAAA, BAAAA showed 64.28% and 14.28% frequency respectively. Average haplotype and nucleotide diversity within Populations in Volga river was 0.880.027 and 0.0122 and in Gorganrud river was 0.59340.144and 0.00398 respectively. Average nucleotide diversity and nucleotide divergence among populations of North and South Caspian sea was 0.008644and 0.000542 respectively. Monte-Carlo simulation using 1000 replication was shown nonsignificant difference between sampling site in the southern and Northern Caspian sea (X2=0.9260.0083).

ISOMERIC SPECIFIC BIOACCUMULATION AND TOXICITY OF PERFLUOROOCTANE SULFONATE (PFOS) TO ADULT AND EMBRYONIC ZEBRAFISH (DANIO RERIO) Sharpe, R.L., Laarman, A., Benskin, J.P., Macleod, S.L., Martin, J.W., Wong, C.S. and Goss, G.G. Symposium: Environmental Pollution Presentation: Oral Contact: Rainie Sharpe, University of Alberta, Department of Biology, CW 405 Biological Sciences Centre Edmonton Alberta T6G 2E9 Canada E-Mail: [email protected] Abstract: Perfluorooctane sulfonate (PFOS) is a fully fluorinated organic compound that has been found in measurable quantities in waters and animals, including humans, around the world. It is environmentally persistent, toxic and bioaccumulative. Challenges in measuring PFOS concentrations in biota and water have been great due to its unique chemistry and a lack of commercially available chemical standards. The present study examined the toxicity of PFOS to adult and embryonic zebrafish (Danio rerio). Developmental defects were evident in the embryos prior to hatch at concentrations in the low ppm range, and evidence of maternal transfer has also been detected. When specific isomer profiles were examined, a dose- response in isomeric uptake was evident in adult zebrafish exposed to PFOS; fish preferentially take up higher concentrations of non-linear PFOS when exposed to lower concentrations of linear PFOS in the water. This study establishes the mutagenicity, embryo and adult toxicity limits and isomeric preferential bioaccumulation of PFOS in zebrafish.

CHARACTERIZATION OF GENES INVOLVED WITH GROWTH AND IMMUNITY IN THE YELLOW PERCH (PERCA FLAVESCENS) Shepherd, B., Rees, C., Binkowski, F. and Goetz, R. Symposium: Fish Growth Presentation: Poster Contact: Brian Shepherd, Great Lakes WATER Institute/ARS/USDA, 600 East Greenfield Avenue Milwaukee WI 53204 USA E-Mail: [email protected] Abstract: In most vertebrates, growth hormone (GH) stimulates growth, metabolism and immunity. In yellow perch, GH does not appear to stimulate growth which suggests a condition of GH insensitivity. Furthermore, females grow faster and larger than males and estrogen preferentially stimulates this growth. The connection between estrogen-dependent growth and GH insensitivity is of particular interest given current efforts to improve growth in this species via selective breeding. A primary concern here is that animals which have been selectively bred for improved growth can experience decreased immunity. We hypothesize that the estrogen-dependent growth seen in yellow perch may be a causal factor underlying GH insensitivity in this species and that any genetic gain in growth may be offset by decreases in immunity. Data will be presented that describe the regulation of genes which encode for a family of proteins that control the signaling of hormones that affect growth and immunity in yellow perch.

EFFECT OF HYPERCAPNIC ACIDOSIS ON THE FORCE-FREQUENCY RELATIONSHIP OF VENTRICULAR MUSCLE FROM THE ARMOURED CATFISH, LIPOSARCUS PARDALIS. Shiels, HA, Santiago, DA, Quinn, DA, Galli, GLJ Symposium: Fish Living on the Edge Presentation: Poster Contact: Holly Shiels, Faculty of Life Sciences, University of Manchester, 46 Grafton Street Manchster M13 9NT UK E-Mail: [email protected] Abstract: The armoured catfish, Liposarcus pardalis, is a freshwater teleost adapted to high levels of seasonal hypercapnia which precipitates acidosis of body tissues. An isometric muscle preparation was used to investigate the mechanisms underlying the acidotic tolerance of the heart of L. pardalis. Acidotic tolerance was assessed by comparing force development before, during and after an acidotic challenge (7.5% CO2, pH = 7.14  0.03; 15% CO2, pH = 6.77  0.05). Frequency had no effect on peak force under normoxic conditions, however, acidosis caused the relationship to becomes

Fish Biology Congress Abstracts 89 biphasic. At all frequencies acidosis resulted in a significant decrease in force that did not recovery upon return of normoxic conditions. Inhibiting intracellular Ca2+ cycling in the SR did not significantly affect the response to increased stimulation frequency, nor did it significantly change the response of the myocardium to acidotic challenges. The results suggest myocardial force is sensitive to extracellular acidosis.

AMMONIA EXCRETION IN THE SKIN OF ZEBARFSIH LARVAE Shih, T. H. and Lin, L. Y. Symposium: Ion and Acid-base Regulation Presentation: Poster Contact: Li-Yih Lin, 88, Sec.4, Ting-Chou Rd. Taipei 116 Taiwan E-Mail: [email protected] Abstract: Branchial ammonia transport in freshwater teleosts is not well understood. In this study, electrophysiological approach, scanning ion-selective electrode technique ( SIET ) was applied to measure the H+ and NH4+ fluxes in specific cells of larval skin. NH4+ extrusion was found significantly in the H+-pump-rich-cell (HRC), which was identified as the H+ secreting ionocyte in zebrafish. Though the pavement cell covers the most surface of larvae, their secretion of NH4+ was minor. Coupled decrease of NH4+ and H+ extrusion was revealed in larvae with H+-pump inhibitor treatment or H+-pump gene knockdown. In addition, significant decline of surface H+ gradient was found in the larvae acclimated to 5mM ammonia, indicating that the secreted acid by HRC was buffered by secreted NH3. This study demonstrates that HRC is the major site for ammonia excretion in zebrafish, H+ pump plays critical role in generating acid gradient and consequently facilitates ammonia excretion.

IONOREGULATORY AND GROWTH DIFFERENCES IN PLOIDY MANIPULATED OCEAN-TYPE CHINOOK SALMON Shrimpton, JM, JW Heath, RH Devlin, DD Heath Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Mark Shrimpton, University of Northern British Columbia, 3333 University Way Prince George BC V2N 4Z9 Canada E-Mail: [email protected] Abstract: In this study, ocean-type chinook salmon were bred using two 3 x 3 factorial mating designs to create 18 families and test whether triploidization resulted in changes in growth performance and smolting. Eggs were pressure shocked after fertilization to create triploid fish from a subset of each family. In spring, freshwater fish were sampled for size and gill Na+,K+-ATPase activity. Plasma chloride and cortisol were measured following a 24-hr saltwater challenge. Growth performance was monitored for a further 4-mo following transfer to an ocean sea-cage. For smolts, we found differences in size due to ploidy and a significant difference in weight among family groups (paternal effect). Significant differences in gill Na+,K+-ATPase activity due to ploidy and family (maternal effect), corresponded with performance in a 24-hr saltwater challenge. Following 4-mo of ocean growth, diploid animals were consistently larger, with greater circulating levels of insulin-like growth factor-1 and higher specific growth rates. Conversely, gill Na+,K+-ATPase activity was significantly higher in triploid than diploid fish. The strong family effect indicates that genotype has a significant effect on ionoregulatory and growth performance in ploidy manipulated fish.

PUTATIVE PHEROMONES OF THE PACIFIC LAMPREY (ENTOSPHENUS TRIDENTATUS) Siefkes, M.J., Close, D., Wildbill, A., and Yun, S.-S. Symposium: Biology of Lampreys Presentation: Oral Contact: Michael Siefkes, Great Lakes Fishery Commission, 2100 Commonwealth Blvd., Suite 100 Ann Arbor Michigan 48105 USA E-Mail: [email protected] Abstract: This study was performed on Pacific lamprey (Entosphenus tridentatus) from the Pacific coast, to characterize putative migratory and sex pheromones. Mass spectrometry analysis of larval washing extract revealed the presence of petromyzonol sulfate, 3keto petromyzonol sulfate, allocholic acid, 3keto allocholic acid, petromyzonamine disulfate, and petromyzosterol disulfate. Mass spectrometry analysis of spermiating male washing extract revealed the main component as petromyzonol sulfate. Electro-olfactogram was performed on adult Pacific lamprey to determine the sensitivity and specificity of the compounds. EOG analysis revealed that the olfactory system of Pacific lamprey was able to detect identified compounds at low concentrations. This study may suggest that the identified compounds have pheromonal roles in Pacific lamprey. Further research is required to confirm the biological roles of the compounds.

Fish Biology Congress Abstracts 90 IBERIAN BARBEL (BARBUS BOCAGEI, STEINDACHNER,1865 ) RESPONSE TO DIFFERENT HYDRAULIC ENVIRONMENTS ON A EXPERIMENTAL POOL-TYPE FISHWAY Silva, A.T; Pinheiro, A.N.; Ferreira, M.T; and Katopodis,C. Symposium: Swimming Performance Presentation: Oral Contact: Ana Silva, Departamento de Engenharia Florestal, Instituto Superior de Agronomia, Tapada da Ajuda, 1349-017 Lisboa Lisboa 1349-017 Portugal E-Mail: [email protected] Abstract: This study focused on Barbus bocagei upwards movements and its hydraulic preferences in a real scale pool-weir fishway prototype. Altogether, 80 barbels were tested to evaluate the behavioural responses to discharges from 36.7 l/s to 77 l/s and to different submerged orifices dimensions (0.03m2 to 0.06m2). Experiments lasted 1.5 h each and were conducted using two adult fish of different dimensions (15

GENOMIC AND PROTEIN-LEVEL RESPONSES TO HEAT STRESS IN A COLD-ADAPTED ANTARCTIC FISH Sleadd, I.M. and Buckley, B.A. Symposium: Genomics of Adaptation Presentation: Poster Contact: Brad Buckley, Portland State University, Department of Biology, 1719 SW 10th Ave. Portland OR 97201 USA E-Mail: [email protected] Abstract: The teleost suborder Notothenioidei comprises highly stenothermal species of Antarctic fish that for millions of years have evolved in a thermally stable environment at or near the freezing point of seawater. During this time, some species have lost the heat shock response, an otherwise well-conserved cellular response to thermal stress. Using cDNA microarrays, gene expression profiles of Trematomus bernacchii were generated in order to gauge what other conserved patterns of gene expression may have been lost or gained during this species’ adaptation to the cold. Hundreds of genes responded to experimental heat stress, including the transcription factor CCAAT/enhancer binding protein delta (C/EBP delta), which was significantly upregulated at the mRNA level. Here we show that the protein concentration of C/EBP delta also increases in T. bernacchii, providing a possible link between environmental heat stress and cell cycle arrest.

EXPRESSION OF GROWTH-RELATED GENES DURING GONADAL STEROIDOGENESIS AND GAMETOGENESIS Small, B.C. Symposium: Fish Growth Presentation: Oral Contact: Brian Small, USDA, ARS, Catfish Genetics Research Unit, P.O. Box 38 Stoneville MS 38776 USA E-Mail: [email protected] Abstract: There is a well-known association between energy homeostasis and gonadal development. In mammals, several factors known to regulate energy balance and metabolism have been observed as modulators of the gondal axis, participating in gonadal steroidogenesis, gametogenesis and ovulation. Many of these factors also have regulatory roles for growth (e.g. GH, IGF-I, Ghrelin), which suggests a link between the somatropic and gonadotropic axes. As a first step in the exploration of potential involvement of growth-related genes in channel catfish reproduction, the seasonal expression of several growth-related genes was evaluated using real-time PCR. The genes evaluated were GHRH, SS-14, GH, IGF-I, and Ghrelin. Gonadal mass increased from October through May and was highly correlated to circulating concentrations of estradiol and testosterone. Of the five genes evaluated, GHRH and SS-14 were significantly correlated to gonadal steroidogenesis and gametogenesis, suggesting a tenuous link between these somatotropic genes and the gonadotropic axis in channel catfish.

LEGACY EFFECTS OF DIRECT EXPOSURE TO X-RAYS AND TO THE X-RAY INDUCED BYSTANDER EFFECT IN RAINBOW TROUT (ONCORHYNCHUS MYKISS) Smith, R.W., Mothersill, C.E., Moccia, R.D., and Seymour, C.B. Symposium: Environmental Pollution Presentation: Oral Contact: Richard Smith, McMaster University, Department of Biology, 1280 Main Street West Hamilton Ontario L8S 4K1 Canada

Fish Biology Congress Abstracts 91 E-Mail: [email protected] Abstract: Our group uses HPV-G reporter cells to detect the effects of direct X-ray exposure and of bystander signals, emitted from X-rayed fish, on non-irradiated fish. Using this model the legacy of X-ray exposure was investigated in rainbow trout. Tissue from eyed eggs, yolk sac larvae (YSL) and juveniles, X-rayed as eggs (24h post-fertilisation), and tissue from YSL X-rayed as eyed eggs reduced HPV-G growth. In 1 year old fish skin increased and kidney decreased HPV-G growth, irrespective of when irradiation occurred. Tissue from non-irradiated YSL and juveniles, receiving bystander signals from similar fish, and non-irradiated adults receiving signals from fish X-rayed as eyed eggs increased HPV-G growth. However tissues from non-irradiated adults receiving bystander signals from fish X-rayed as 24h post-fertilisation eggs reduced HPV-G growth. We conclude X-ray exposure of early life stages leaves a legacy in terms of the direct effects of radiation and bystander effects in non-irradiated fish.

MRNA EXPRESSION OF MULTIPLE CLAUDIN ISOFORMS IN ATLANTIC SALMON (SALMO SALAR) INTESTINE AT DIFFERENT SALINITIES AND AFTER IN VITRO HORMONE TREATMENT Sorensen, K.J., Tipsmark, C.K., and Madsen, S.S. Symposium: Ion and Acid-base Regulation Presentation: Poster Contact: Christian Tipsmark, Department of Biology, University of Southern Denmark, Campusvej 55 Odense Denmark 5230 Denmark E-Mail: [email protected] Abstract: Claudins are membrane proteins that constitute a central part of tight junctions being involved with regulating membrane ion permeability through the para-cellular space and thereby influencing ion transport. In this study on Atlantic salmon (Salmo salar), expression of three claudin isoforms (claudin 15, 25a and 25b) were investigated. The tissue specificity was examined in a series of tissues including sections of the intestinal system. All three claudin isoforms were specifically expressed in the three intestinal sections: pyloric ceca, anterior intestine and posterior intestine. Furthermore, claudin 25b showed expression in the liver. A salinity transfer experiment showed that claudin 15 expression was elevated in seawater when compared to freshwater. In another experiment the in vitro hormone effects of cortisol, prolactin and growth hormone on claudin expression were investigated. This experiment showed that cortisol has an overall inhibiting effect on claudin mRNA expression in the anterior part of the intestine.

ORIGINS OF GNRH IN VERTEBRATES: IDENTIFICATION OF A NOVEL GNRH IN A BASAL VERTEBRATE, THE SEA LAMPREY Sower, S.A., Kavanaugh, S.I. and Nozaki, M. Symposium: Biology of Lampreys Presentation: Oral Contact: Stacia Sower, Dept Biochemistry and Molecular Biology, Rudman Hall, 46 College Road Durham NH 3824 USA E-Mail: [email protected] Abstract: We cloned a cDNA encoding a novel gonadotropin-releasing hormone (GnRH) from the sea lamprey. In situ hybridization of the brain showed expression and localization of the transcript in the hypothalamus, medulla and olfactory regions; whereas immunohistochemistry only showed GnRH-II cell bodies and processes in the preoptic nucleus/hypothalamus areas. Lamprey GnRH-II was shown to stimulate the hypothalamic-pituitary axis using in vivo and in vitro studies. Lamprey GnRH-II was also shown to activate the inositol phosphate signaling system in COS-7 cells. These studies provide evidence for a novel lamprey GnRH that has a role as a third hypothalamic GnRH. In summary, the newly discovered lamprey GnRH-II offers a new paradigm of the origin of the vertebrate GnRH family. We hypothesize that due to a genome/gene duplication event, an ancestral gene gave rise to two lineages of GnRHs—the gnathostome GnRH and lamprey GnRH-II. Supported by NSF IBN-0421923 and NSF DBI-0618719

OLD FISH IN COLD STREAMS: EVIDENCE FOR EXOTIC BROOK TROUT NICHE CONTRACTION IN THE PRESENCE OF NATIVE BULL CHARR. Steimle, K.M. and Kershner, J.L. Symposium: Fish Living on the Edge Presentation: Oral Contact: Kai Steimle, 3125 SE Morrison St. Portland OR 97214 USA E-Mail: [email protected] Abstract: We used comparative observations of brook trout invasion in Idaho streams to address whether temperature was a limiting factor for invading brook trout and whether the presence of bull trout limits the ability of brook trout to colonize cold streams. Despite variation in the thermal threshold for brook trout presence among streams, all streams showed a similar pattern where brook trout colonization limits were correlated with stream temperature in the presence of bull trout at

Fish Biology Congress Abstracts 92 temperatures that have not limited brook trout colonization in allopatry. Brook trout sympatric with bull trout had much lower apparent survival, moved less during the summer, and yet maintained their numbers during our five year study. Our observations highlight that factors other than temperature and growth may generate differences in life history expression. Constrained life history expression may limit brook trout colonization of cold streams in the presence of native bull trout.

THE RANGEWIDE PACIFIC LAMPREY CONSERVATION INITIATIVE - STATUS AND FUTURE Streif, B. Symposium: Biology of Lampreys Presentation: Oral Contact: Streif, Bianca, U.S. Fish and Wildlife Service USA E-Mail: [email protected] Abstract: The Pacific Lamprey Conservation Initiative is an effort coordinated by the U.S. Fish and Wildlife Service (FWS) to facilitate communication and coordination relative to the conservation of Pacific lampreys throughout their range from Baja, Mexico, around the Pacific Rim to Japan. The goal of the initiative is to develop a Pacific Lamprey Conservation Plan (Plan) that will lead to restored Pacific lamprey populations and improvement of their habitat. Currently, a ground swell of interest in lamprey conservation has emerged, yet efforts to conserve and recover lampreys are uncoordinated and generally lack a strategic approach. The FWS hopes to utilize and build on existing efforts to increase support for Pacific lamprey conservation through partnerships with various entities, communication with the scientific community, assistance with restoration projects, and the development of the Plan. The FWS seeks to identify interested parties who can contribute to this Plan. A review of the status of this effort and near future actions will be shared.

PHYSIOLOGICAL DISTURBANCES IN LARGEMOUTH BASS: CONSEQUENCES OF TEMPERATURE AND OXYGEN SHOCK Suski, C. D., VanLandeghem, M. M. and D. H. Wahl. Symposium: Stress Effects and Response Presentation: Oral Contact: Cory Suski, University of Illinois, Department of Natural Resources and Environmental Sciences, 1102 South Goodwin Ave. Urbana IL 61801 USA E-Mail: [email protected] Abstract: The objective of this study was to quantify the physiological disturbances in largemouth bass at two levels of heat- and cold-shocks and at two levels of hypoxia and hyperoxia shock. For the temperature shock treatment, resting bass acclimated to 20 C were instantly exposed to water at either 15 C, 8 C, 25 C or 32 C. For the oxygen shock experiment, bass acclimated to 8mgO2/L were instantly exposed to water containing 4mgO2/L, 2mgO2/L, 12mgO2/L, and 18mgO2/L. Fish were held at these temperatures/oxygen concentrations for either 1- or 6-h and then sampled for blood and muscle. Results showed the physiological disturbances were most pronounced at the extreme treatments for both temperature and dissolved oxygen shocks. Plasma glucose concentrations increased at 8 C and 32 C for 1h but returned to near control concentrations at 6h. Some temperature shock treatments resulted in ionic disturbances. Muscle phosphocreatine was consumed and tissue lactate was produced following 6h exposure to 2mgO2/L.

JOURNEYS WITH JOE: RESEARCH AT THE INTERFACE OF PHYSIOLOGY AND BEHAVIOR Swanson, C. Symposium: Tribute to Joe Cech, Jr. Presentation: Oral Contact: Christina Swanson, The Bay Institute, 2128 Bueno Drive #13 Davis CA 95616 USA E-Mail: [email protected] Abstract: I think I was always a bit an oddity in the Cech lab - a physiologist who had little interest and no aptitude for inserting a cannula but who really liked to watch fish. But under Joe’s guidance, my graduate and post-doctoral research flourished, nurtured by energetic discussions and provocative questions. How can euryhaline milkfish grow faster and have lower metabolic rates in hyper-saline conditions? Why does delta smelt experience swimming failure at sub-maximal velocities? For me, what evolved was a broad understanding of how fish interact with their environment, and our manipulations of their habitat, and the conclusion that what a fish can do (i.e., its physiological capacity) may not be as important as what a fish actually does do (i.e., its behavior). I like to think that I brought a new dimension to the Cech lab with my research - but I suspect he already knew this stuff.

EMERGING HALLMARKS OF THUNNIFORM SWIMMING: MUSCLE DYNAMICS AND POWER IN YELLOWFIN TUNA (THUNNUS ALBACARES) Syme, D.A., and Shadwick, R.E. Symposium: Swimming Performance Presentation: Poster

Fish Biology Congress Abstracts 93 Contact: Doug Syme, University of Calgary, Dept of Biological Sciences, 2500 University Drive NW Calgary AB T2N 1N4 Canada E-Mail: [email protected] Abstract: Fish with internalized red muscles appear to adopt stiff-bodied (thunniform) swimming. Mechanical properties of internal red muscle of yellowfin tuna (Thunnus albacares) were studied, revealing advantages of this design for fast, sustained swimming. Muscle length changes (sonomicrometry) and activation (electromyography) were recorded as fish swam in a water tunnel. Muscle shortening lagged local body curvature by 31±4deg and onset of muscle activation progressed rapidly along the body (1.7 lengths/tailbeat) such that high power was transmitted to the caudal tendon but caused little body bending. Strain amplitude in deep red muscle was double that predicted from midline curvature, resulting in near-maximal power produced by isolated bundles of red muscle. Finally, the timing of muscle activation and tail beat frequencies in swimming fish were very similar to those resulting in maximal muscle power. Thunniform swimmers use internalized red muscles to produce near-maximal power that is directed to the tail, not local bending.

DIRECT ACTIONS OF PROLACTIN, MINERALO- AND GLUCOCORTICOIDS ON THE ESOPHAGUS OF EURYHALINE FISHES Takagi, C., Takahashi, H., Kudose, H., Kato, K., and Sakamoto, T. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Chiyo Takagi, Ushimado Marine Laboratory Faculty of Science, Okayama University, 130-17 Kashino, Ushimado, Setouchi 701-4303, Japan Setouchi Okayama 701-4303 Japan E-Mail: [email protected] Abstract: Apoptosis throughout teleost esophageal epithelium is important for the simple epithelium and high permeability/absorption in seawater, whereas cell proliferation induced for the stratified epithelium and low permeability in freshwater. In the cultured esophagus of euryhaline medaka, prolactin, the freshwater-adapting hormone, induced the cell proliferation. Together with the prolactin receptor distribution, we first demonstrate the direct effects on the osmoregulatory esophagus. Although 11-deoxycorticosterone recently identified as a potential agonist of fish mineralocorticoid receptor had little impact on the cell turnover, cortisol stimulated the apoptosis at 10 nM and the cell proliferation was induced at higher doses within physiological range. The blocks by glucocorticoid receptor (GR) antagonists suggest that these dual functions are via GRs and specific target genes dependent on cortisol doses. Using medaka oligonucleotide microarray, we have identified these genes as those related to differentiation, stress response and osmoregulation. Cortisol-GR may be more important in the osmoregulation, than mineralocorticoid signaling.

INVOLVEMENT OF MELATONIN ON ITS RECEPTOR EXPRESSION IN THE CULTURED HEPATOCYTES OF TILAPIA OREOCHROMIS MOSSAMBICUS Takeuchi, Y., Nikaido, Y., Park, Y.J., and Takemura, A. Symposium: Contributed Papers Presentation: Poster Contact: Y. Takeuchi, Sesoko Station Tropical Biosphere Research Center University of the Ryukyus 3422 Sesoko Motobu Okinawa 905-0227 Japan E-Mail: [email protected] Abstract: Melatonin receptor is considered to mediate the specific and cyclic functions of melatonin in target organs. The aim of the present study was to examine day-night variations in MT1 mRNA expression in neural and peripheral tissues, and to evaluate the involvement of melatonin in MT1 mRNA expression in the liver of the tilapia Oreochromis mossambicus. A high expression of MT1 mRNA that increased at night was observed in the retina, brain, liver, and heart. This increase coincided with fluctuations in plasma melatonin levels. When MT1 mRNA expression in tilapia hepatocytes was examined using a primary culture technique, it tended to increase in the subjective night. The addition of melatonin into the culture media resulted in the induction of MT1 mRNA expression. These results suggest that melatonin is involved in the transcription of MT1 in the liver and in the daily functions of the liver through the activity of its receptors.

DETERMINING MORPHOLOGICAL AND IMMUNOCHEMICAL PARAMETERS ASSOCIATED WITH EARLY OVARIAN FOLLICULAR ATRESIA IN WHITE STURGEON (ACIPENSER TRANSMONTANUS) FEMALES Talbott, M.J., Guy, C.S., Van Eenennaam, J.P., and Webb, M.A.H. Symposium: Contributed Papers Presentation: Poster Contact: Mariah Talbott, Montana Cooperative Fishery Research Unit, U.S. Geological Survey, Department of Ecology, Fish & Wildlife Management Program, Montana State University Bozeman MT 59717 USA E-Mail: [email protected]

Fish Biology Congress Abstracts 94 Abstract: In order for the caviar industry to maximize quality and quantity of caviar, it is essential to correctly assess stage of ovarian maturity in white sturgeon females and avoid harvesting females with atretic ovarian follicles. To detect early stages of follicular atresia blood and ovarian follicle samples have been taken repeatedly prior to and after inducing atresia by transferring females from cold water (10–13 C) to warm water (20 C). Results of morphological (follicle diameter and oocyte polarization indices) and immunochemical characteristics (blood plasma sex steroid, total protein, and calcium concentrations) associated with early onset of atresia will be presented. This research will also benefit the conservation of wild sturgeon by providing less-invasive techniques to detect ovarian atresia as an alternative to the current techniques of biopsy or urogenital catheterization.

GILL NA+/K+-ATPASE ACTIVITY OF EURYHALINE TELEOSTS INCREASES WITH OSMOTIC STRESS IN ENVIRONMENTS DISTINCT FROM THEIR PRIMARY NATURAL HABITATS Tang, C.H., and Lee, T.H. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Tsung-Han Lee, Department of Life Sciences, National Chung-Hsing University, 250 Kuo Kuang Road Taichung 402 Taiwan E-Mail: [email protected] Abstract: Since previous studies revealed that the gill Na+/K+-ATPase (NKA) activities of euryhaline teleosts exhibited different patterns upon salinity challenge, we hypothesized that the NKA activity in gills of euryhaline teleosts was elevated to facilitate acclimatization to stressful environments with salinities that were different from their primary natural habitats. To demonstrate the hypothesis, the euryhaline tilapia (Oreochromis mossambicus), spotted green pufferfish (Tetraodon nigroviridis), and milkfish (Chanos chanos), whose primary natural habitats are fresh water, estuaries, and seawater, respectively, were acclimatized to fresh water, brackish water, and seawater in this study. The levels of hepatic HSPs 60, 70, and 90, and branchial HSP 90, examined by immunoblotting, which were lowest in FW-acclimatized tilapia, BW- acclimatized pufferfish, and SW-acclimatized milkfish, positively correlated with branchial NKA activity in these three euryhaline teleosts from different primary natural habitats. The abundance of hepatic HSPs, the stress proteins, indicates that gill NKA activity of euryhaline teleosts is induced by osmotic stress to maintain osmotic homeostasis.

THE RESPONSES OF NA+/K+-ATPASE AND HEAT SHOCK PROTEIN 90 IN THE MARINE EURYHALINE GRASS PUFFER, TAKIFUGU NIPHOBLES, UPON OSMOTIC STRESS Tang, C.H., and Lee, T.H. Symposium: Stress Effects and Response Presentation: Poster Contact: Cheng-Hao Tang, Department of Life Sciences, National Chung Hsing University, 250 Kuo Kuang Road Taichung 402 Taiwan E-Mail: [email protected] Abstract: The grass puffer, Takifugu niphobles were captured from a local estuary and acclimatized to hyposmotic (i.e., fresh water; FW) or hyperosmotic (i.e., seawater; SW; 35‰) environment. In physiological responses, muscle water contents and plasma osmolality were maintained in homeostasis. Furthermore, Na+/K+-ATPase (NKA) immunoreactive (NKIR) cells in gills of grass puffer acclimatized to either FW or SW were distributed mainly in the filaments and NKIR cell numbers were similar between FW and SW groups. Relative protein abundance and specific activity of branchial NKA was higher in the FW grass puffer compared to those of the SW individuals. For stress response, relative protein abundance of liver and gill HSP 90 was higher in the long-term acclimatized FW group rather than in the SW group. Immunological detection reveled that most gill HSP 90 expressed in NKIR cells. In addition, short-term effects of hyposmotic shock resulted in elevating protein amounts of gill HSP 90 immediately in 1 h post-transfer. This study illustrates the impressive osmoregulatory ability of this euryhaline species in response to salinity challenges.

THE ABUNDANCE OF NHE2-LIKE AND NHE3-LIKE PROTEINS IN GILLS OF GOLDFISH, CARASSIUS AURATUS, ALTERED IN RESPONSE TO ION-POOR AND ACIDIC ENVIRONMENTS, RESPECTIVELY Tang, C.H.1, Lee, T.H.1, Claiborne, J.B.2 Symposium: Ion and Acid-base Regulation Presentation: Poster Contact: Tang, Cheng-Hao, 1. Department of Life Sciences, National Chung Hsing University, Taichung, 402 Taiwan 2. Department of Biology, Georgia Southern University, Statesboro, Ga. 30460-8042 USA Taiwan E-Mail: [email protected] Abstract:

Fish Biology Congress Abstracts 95 Na+/H+ exchangers (NHEs) are electroneutral antiporters. NHEs transport H+ across the plasma membrane to extracellular medium and exchange Na+ into the cell. NHE2 and NHE3 have been localized to the apical membrane of epithelial cells in previous studies. The roles of NHEs in Na+ uptake or acid-base regulation in fresh water fish gills, however, are not conclusive. To clarify the putative mechanisms of NHE2 and NHE3, the goldfish was acclimatized to reverse osmosis water as ion-poor water and acid water (pH= 4.5±0.1) and relative abundance of branchial NHE2-like and NHE3-like proteins was determined by immunoblotting. The levels of NHE2-like proteins were induced by ion-poor water while the amounts of NHE3-like proteins increased when acclimatized to acidic water. Therefore, this study presumed that in gills of goldfish NHE-2-like protein play major roles for Na+ uptake in ion-poor water and NHE3-like protein responded significantly for H+ efflux in acidic water.

THE INFLUENCE OF MATERNAL CONDITION AT SPAWNING ON OFFSPRING SWIMMING PERFORMANCE Tierney, K.B., Patterson, D.A., and Kennedy, C.J. Symposium: Stress Effects and Response Presentation: Oral Contact: Keith Tierney, Biology Department, Univeristy of Windsor, 401 Sunset Avenue Windsor ON N9B 3P4 Canada E-Mail: [email protected] Abstract: Many adult salmon complete their return migrations but are unable to complete spawning. In this study, eggs were collected from returning adult sockeye capable and incapable of spawning, and then fertilized by healthy males. Egg number, size, hatching success and offspring growth did not differ across maternal groups, suggesting that persisting physiological maternal effects were absent. However, the swimming behaviour and physiology of the offspring at parr stage differed between maternal groups. In a 30 min schooling test (groups of five), parr from females unable to naturally spawn were more likely to fatigue, have looser schools, and show a decreased startle response. When given individual swim tests, these offspring produced more lactate (used more white muscle). Offspring lactate was positively correlated to maternal lactate at spawning, suggesting heritable differences in muscle dynamics. This study shows that sockeye propagated from unfit females have swimming performance-based differences that could affect survivorship.

A REALISTIC MIXTURE OF PESTICIDES AFFECTS SALMON OLFACTORY TISSUE RESPONSES Tierney, K.B., Ross, P.S., Sekela, M.A., and Kennedy, C.J. Symposium: Environmental Pollution Presentation: Oral Contact: Keith Tierney, Biology Department, University of Windsor, 401 Sunset Avenue Windsor ON N9B 3P4 Canada E-Mail: [email protected] Abstract: Contaminants within the salmon producing waterways of the world may impair olfactory sensory neurons (OSNs) critical to important behaviours. 96-hr exposures to realistic and higher concentrations of a mixture of the ten most frequently occurring pesticides in British Columbia’s Nicomekl River reduced OSN responses of rainbow trout to a behaviourally- relevant odorant, whereas responses were not altered by a (20%) lower concentration, potentially through an upregulation in detoxification enzymes. After the initial exposures, a further 10-min exposure to a high (20×) concentration of the mixture caused greater decreases in odour-evoked responses than predicted by an additive model of the pesticide effects, which suggests the presence of synergistic interactions. This study demonstrates that an environmentally-observed pesticide mixture can injure salmon olfactory tissue, suggesting that some behaviours such as migration may be impaired in the environment.

EXPRESSION AND LOCALIZATION OF GILL ISOFORMS OF FXYD PROTEINS AND NA,K-ATPASE ALPHA-SUBUNIT DURING DEVELOPMENT OF SEAWATER TOLERANCE IN ATLANTIC SALMON Tipsmark C.K., and Madsen, S.S. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Christian Tipsmark, Institute of Biology, University of Southern Denmark, Campusvej 55 Odense 5230 Denmark E-Mail: [email protected] Abstract: The Na,K-ATPase is the driving force for ion secretion in teleost gill chloride cells. In Atlantic salmon, gill enzyme abundance increases and isoform composition changes in response to elevated salinity and during the smoltification process that occurs prior to seaward migration in the spring. FXYD proteins associate with Na,K-ATPase and may regulate the catalytic rate in these situations. During smoltification, expression of FXYD9 and FXYD11 peaked in May, coinciding with the peak in SW-tolerance and gill Na,K-ATPase in FW dwelling salmon. Abrupt SW-transfer induced increased Na,K- ATPase abundance but no change in FXYD expression. The parallel increase in FXYD9 and FXYD11 expression with Na,K-ATPase abundance during smoltification but not after SW-transfer suggests that these auxiliary subunits have a

Fish Biology Congress Abstracts 96 specific role in controlling catalytic rate when the FW smolt prepares for SW entry. Cellular localization of gill FXYD proteins and Na,K-ATPase isoforms is currently being investigated by use of in situ histochemistry.

CHANGES IN CLAUDIN ISOFORM EXPRESSION IN THE GILL DURING SALINITY SHIFT AND SMOLTIFICATION OF ATLANTIC SALMON Tipsmark, C.K., and Madsen, S.S. Symposium: Ion and Acid-base Regulation Presentation: Poster Contact: Christian Tipsmark, Department of Biology, University of Southern Denmark, Campusvej 55 Odense Denmark 5230 Denmark E-Mail: [email protected] Abstract: Permeability changes of gill epithelium of euryhaline teleosts are essential during salinity acclimation. This study examined expression patterns of claudin isoforms in the salmon gill. These membrane proteins are major determinants of selective and gross tight junction permeability. Five isoforms: 10e, 27a, 28a, 28b and 30, were identified in the gill transcriptome from Atlantic salmon, and tissue specific expression was confirmed by RT-QPCR. We examined expression profiles throughout smoltification in freshwater and during acclimation to seawater (SW). During smoltification, claudin 10e expression peaked in May, coinciding with optimal SW tolerance. SW-transfer induced a 5-fold increase in expression of claudin 10e and reduced expression of 27a and 30. The study demonstrates for the first time that SW acclimation involves differential regulation of claudin gene expression in the salmon gill, one of which is induced in preparation to SW migration during smolting. We suggest discrete functional roles of the identified claudins.

LESSONS FROM THE COLD: PROTEIN HOMEOSTASIS IN ANTARCTIC FISHES Todgham, A.E., and Hofmann, G.E. Symposium: Fish Living on the Edge Presentation: Oral Contact: Anne Todgham, Department of Ecology, Evolution and Marine Biology, University of California Santa Barbara Santa Barbara CA 93106-9610 USA E-Mail: [email protected] Abstract: Notothenioids, the dominant group in the Antarctic fish fauna, occupy the cold and ice-filled waters of the Southern Ocean. Historically, this group has been the hallmark of cold adaptation in fish with antifreeze proteins, elevated plasma osmolality and decreases in haematocrit being a few of the distinguishing adaptations to life in the cold. In addition to the danger of freezing, recent research suggests that subzero cold also impacts the cellular protein pool and drives cold-denaturation of proteins. By examining the ubiquitin proteasome pathway in a diversity of Antarctic notothenioids we have started to address the physiological constraints of maintaining protein homeostasis in polar species. Through comparative studies with temperate New Zealand notothenioids, we have shown that subzero temperature increases the cellular levels of damaged proteins in Antarctic fishes and thermal compensation of 26S proteasome activity is one strategy used to defend protein homeostasis in these cold-adapted fishes. (Supported by NSF grant ANT 0440799)

MORPHOLOGICAL CORRELATES OF INCREASING PREDATION INTENSITY IN TWO TRINIDADIAN GUPPY SPECIES (POECILIA RETICULATA AND POECILIA PICTA) Torres Dowdall, J., Handelsman, C., Bathrick, C., and Ghalambor, C.K. Symposium: Morphometrics Presentation: Oral Contact: Julian Torres Dowdall, Colorado State University, Department of Biology Fort Collins CO 80523 USA E-Mail: [email protected] Abstract: Poecilia reticulata inhabit tropical waters where they experience a gradient in predation risk; low-predation risk in headwater streams and increasing predation risk downstream in rivers where a diversity of piscivorous species coexists. These different predation regimes have been shown to drive the evolution of several life-history, behavioral, and morphological traits. Recent work shows that low-predation populations differ in morphology and exhibit decreased escape performance compared to high-predation populations, but such dichotomous comparisons do not capture the potential for continuous trait variation in response to a predation gradient. To test for the presence of a continuum in phenotypic variation in response to a predation gradient, we analyzed morphology of three guppy populations that vary in predation intensity. Further, we compared these findings to Poecilia picta, a closely related species thought to be a “hyper-predation guppy”. Relationships between morphology, escape performance, and predation intensity are discussed in the context of species distributions.

STRESS–RELATED HORMONES AFFECT CYTOKINE EXPRESSION IN SEABREAM HEAD KIDNEY

Fish Biology Congress Abstracts 97 Tort, L., and Castillo, J. Symposium: Stress Effects and Response Presentation: Oral Contact: Lluis Tort, Dpt. Cell Biologi, Physiology and Immunology. Universitat Autonoma de Barcelona. Cerdanyola Barcelona 8193 Spain E-Mail: [email protected] Abstract: We have determined whether adrenaline, cortisol and ACTH modulate the expression of several cytokines in the head kidney (TNF-α, IL-1β, TGF-β, IL-6) and the caspase-1 gene. Incubation with ACTH (150 ng/mL 1h) caused a 3-fold increase of TNF-α mRNA, whereas adrenaline (1µM) showed no significant effects. IL-1β decreased after 1h incubation with ACTH or Adrenaline (60% of control) at 1h but not at 2h. Caspase-1 was inhibited by both ACTH and adrenaline. The 1h ACTH incubation stimulated TGF-β (1,4-fold) and IL-6 (4-fold), although in the latter the mRNA levels decreased rapidly after 2h. Adrenaline was ineffective to alter IL-6 mRNA levels. In conclusion, cytokine expression in head kidney can be highly influenced by hormones involved in stress situations, indicating their relevant influences on the immune system taking place in the head kidney especially under stress episodes.

SOLUBLE ADENYLYL CYCLASE (SAC): NATURE’S HCO3-/CO2 SENSOR? Tresguerres, M., Parks, S.K., Goss, G.G., Buck, J., Levin, L.R. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Martin Tresguerres, Weill Medical College, Cornell University, 1300 York Ave, Rm E-505 New York NY 10021 USA E-Mail: [email protected] Abstract: Mammalian soluble adenylyl cyclase (sAC) is a novel source of the ubiquitous second messenger cAMP, and it is the only signaling molecule identified so far that is directly modulated by bicarbonate. We have cloned a dogfish sAC ortholog and showed it is bicarbonate sensitive and present in gill. Specific sAC inhibitors blocked the V-H+-ATPase translocation from cytoplasmic vesicles to the basolateral membrane and prevented recovery from blood alkalosis in whole-animal experiments. These data suggest that branchial sAC senses changes in [HCO3-] and triggers cellular responses that regulate blood acid/base balance. Bicarbonate-regulated sAC homologues are predicted in organisms from bacteria to mammals. Therefore, it is likely that sAC-like adenylyl cyclases play important roles in CO2/HCO3- sensing in many different physiological processes and organisms.

SWIMMING IN AN UNSTEADY WORLD: MODIFIED BODY-CAUDAL FIN KINEMATICS DUE TO ACTIVE STABILITY REQUIREMENTS Tritico, H.M. Symposium: Swimming Performance Presentation: Oral Contact: Hans Tritico, University of Michigan, 2350 Hayward, 2340 G.G. Brown Building Ann Arbor MI 48109 USA E-Mail: [email protected] Abstract: Turbulence has been shown to affect habitat selection (Smith et al. 2005, Cotel et al. 2006) and respiration (Enders et al. 2003) however the methods for describing turbulence have relied upon point measurements which are poor at describing the spatial scale of turbulence. Additionally, the effects of turbulence on fishes’ active stability control system have not been described. This talk presents data from a series of increasing velocity tests which change both the spatial and temporal scales of turbulent eddies and relates the fluid properties to swimming modes in creek chub, Semotilus atromaculatus. Various propulsive swimming modes were observed including caudal dominated (continuous and twitch and coast) along with pectoral dominated (synchronized and asynchronous rowing) in addition to a number of new stability dominated swimming modes. The effects of turbulent eddy properties on specific fin use metrics are compared both within and across swimming modes.

ROLE OF STANNIOCALCIN-1 IN REGULATION OF THE EXPRESSION AND FUNCTION OF ZEBRAFISH EPITHELIAL CALCIUM CHANNEL Tseng, D.Y., Chou, M.Y., Tseng, Y.C., Hsiao, C.D., Huang, C.J., Kaneko, T., and Hwang, P.P. Symposium: Ion and Acid-base Regulation Presentation: Poster Contact: Deng-Yu Tseng, National University of Tainan, 33, Sec. 2, Shu-Lin St., Tainan Taiwan 700 Taiwan E-Mail: [email protected] Abstract: Stanniocalcin (STC, formerly called hypocalcin or teleocalcin) is a 50-kDa disulfide-linked homodimeric glycoprotein that was originally identified in fish and secreted from the corpuscles of Stannius. The main function of STC-1, a human orthologue of STC, is Ca2+ uptake inhibition; however the mechanisms remain unknown. In the present study, we provide

Fish Biology Congress Abstracts 98 molecular evidence to elucidate how zebrafish STC-1 regulates Ca2+ uptake in zebrafish embryos. zstc-1 was expressed in a wide variety of tissues including the kidneys, brain, gills, muscles, and skin. Incubating zebrafish embryos in low-Ca2+ (0.2 mM) freshwater stimulated whole-body Ca2+ influx and zebrafish epithelial Ca2+ channel (zECaC) mRNA expression while downregulating zstc-1 expression. A morpholino microinjection approach was used to knock down the zSTC-1 protein, and the results showed that the Ca2+ content, Ca2+ influx, and zECaC mRNA expression all increased in morphants. These data suggest that zSTC-1 negatively regulates zECaC gene expression to reduce Ca2+ uptake in zebrafish embryos.

SIZE MATTERS - LONGER FLUMES INCREASE CRITICAL SWIMMING SPEEDS Tudorache, C., Viaenen, P., Blust, R. and de Boeck, G. Symposium: Swimming Performance Presentation: Oral Contact: Christian Tudorache, Canadian Rivers Institute/Dept. of Biology, University of New Brunswick Fredericton, NB E3B 6E1 Canada E-Mail: [email protected] Abstract: Carp (Cyprinus carpio, L.) alters the repertoire of swimming behaviour with increased flume length. While the transition speed from steady to burst-and-coast swimming was unaffected by flume length, fish reached higher Ucrit, consequently swimming for longer periods of time in burst-and-coast mode and hence performing more work before becoming fatigued. Analysis of swimming behaviour of burst-and-coast swimming revealed an increase in duration and a decrease in distance of forward burst movements with increasing water speeds. Frequency was unaffected by water speed. Overall longer flumes increase Ucrit by allowing for less restricted burst-and-coast swimming behaviour.

COMPARATIVE GENE EXPRESSION PATTERNS AMONG AND WITHIN ATLANTIC SALMON POPULATIONS OF THE BAY OF FUNDY Tymchuk, W.E., O'Reilly, P.T., and Schulte, P.M. Symposium: Genomics of Adaptation Presentation: Oral Contact: Wendy Tymchuk, Department of Zoology, University of British Columbia Vancouver BC V6T 1Z4 Canada E-Mail: [email protected] Abstract: Salmon populations of the Bay of Fundy in south eastern Canada show evidence for restricted gene flow that appears to be geographically structured, and populations between the inner and outer Bay of Fundy have unique life-history characteristics. Microarrays were used to detect differences in gene expression profiles among and within Bay of Fundy populations reared in a common environment. Not only were there clear differences in expression profiles between the inner and outer Bay, but populations within the inner Bay of Fundy tended to have unique gene expression patterns. Knowledge of population-specific expression profiles will assist with conservation of these endangered populations through an ability to monitor introgression of escaped aquaculture fish and to detect effects of domestication arising during captive rearing.

MICROARRAY ANALYSIS OF GENE EXPRESSION DIFFERENCES BETWEEN SLOW-GROWING WILD AND FAST-GROWING DOMESTIC RAINBOW TROUT Tymchuk, W.E., Sakhrani, D., and Devlin, R.H. Symposium: Genomics of Adaptation Presentation: Poster Contact: Wendy Tymchuk, Department of Zoology, University of British Columbia Vancouver BC V6T 1Z4 Canada E-Mail: [email protected] Abstract: Domestic and wild rainbow trout show differences in growth, behaviour and physiology that are at least partly controlled by genotype. Microarray analysis of gene expression was used to identify genes that may have been altered through domestication leading to the observed phenotypic differences between these fish. Gene expression was measured in liver, muscle and brain from fast-growing domestic and slow-growing wild fish reared within a common environment. Domestic and wild fish showed strong differences in expression profiles for all three tissues. There were also differences observed among the tissues with liver having more down-regulated genes (domestic relative to wild) while the brain tended to have more up-regulated genes. Use of potential candidate loci identified in this analysis can be used in subsequent genetic mapping studies to help identify the genes leading to specific phenotypic differences arising due to selection within a culture environment.

NEWS ON EIF2 IN THE AMAZONIAN FISH COLOSSOMA MACROPOMUM Val, A.L., Nozawa, S.R., Honda, R.T. and Almeida-Val, V.M.F. Symposium: Tropical Fish Adaptations Presentation: Oral

Fish Biology Congress Abstracts 99 Contact: Dal Val, INPA/LEEM, Ave Andre Araujo 2936 Manaus AM 69060-001 Brazil E-Mail: [email protected] Abstract: ATP and GTP are modulators of Hb-O2 affinity in fish. A reduction of GTP occurs within the first 10min of of the Amazon. Why GTP decreases faster than ATP? We propose that this is achieved when the eIF2 binds to GTP and then to the initiator. eIF2-GTP is then hydrolyzed to eIF2-GDP with Pi release. GDP is replaced by GTP in subsequent rounds. We cloned the eIF2and monitored its expression in C. macropomum exposed to deep hypoxia using qRT-PCR. The number of copies of this gene increased within the first 15 minutes in blood cells, subsequently decreasing dramatically. Similar trend was observed in gills and liver cells. This is in accordance with the changes of GTP levels in the red cell and suggests that increased eIF2 assures the synthesis of proteins needed to face hypoxia with a subsequent shut down to reduce energy demand. (PRONEX, CNPq, FAPEAM).

BIOLOGICAL SENSITIVITY OF AMAZONIAN ORNAMENTAL FISHES TO COPPER Valdez Domingos, F. X.; Duarte, R. M.; Nascimento, T. L. A. S; Vilarinho, G. C.; Brust, S. M. A.; Alemida Val, V. M. F.; Val, A. L. Symposium: Tropical Fish Adaptations Presentation: Oral Contact: Fabíola Valdez, LEEM-Laboratório de Ecofisiologia e Evolução Molecular, INPA - Instituto Nacional de Pesquisaas da Amazônia, Avenida Andre Araujo, 2936 Manaus Amazonas 69060001 Brazil E-Mail: [email protected] Abstract: Copper is an essential element required by all living organisms in several physiological functions and biochemical reactions, but in excess this metal can be toxic to fish. Anthropogenic activity represents one of the copper sources in the Amazon aquatic environments. Rio Negro, one of the major rivers of the Amazonian basin presents waters with extremely low ion content and large quantities of DOC (dissolved organic carbon). Eight fish species of Amazonian ornamental fish were evaluated: schwarts, Corydoras pigmaeus, Dicrossus maculatus, Hyphessobrycon socolofi, Carnegiella strigata, Otocinclus hasemani and Apistogramma hyppolytae. Copper sensitivity was tested under different DOC concentrations. In the lower DOC concentration the LC50 ranged from 38.06 (Hyphessobrycon socolofi) to 270.7µgCu/L (Apistogramma hippolytae) and in the highest concentration ranged from 305.17 (Carnegiella strigata) to 915.5 µgCu/L (Corydoras schwarts). Our results confirm the protective role of DOC on copper toxicity.

EVALUATION OF FISHES LIVING IN AN AMAZON AREA INFLUENCED BY PETROLEUM TRANSPORT Valdez Domingos, F. X.; V. M. Brandão; Paula-Silva M. N.; Silva, K. Y. S.; Almeida-Val, V. M. F. Symposium: Tropical Fish Adaptations Presentation: Oral Contact: Fabíola Valdez, LEEM - Laboratório de Ecofisiologia e Evolução Molecular, INPA - Instituto Nacional de Pesquisas da Amazônia, Av. André Araújo, 2936 Manaus Amazonas 69060001 Brazil E-Mail: [email protected] Abstract: Urcucu is the biggest inland petroleum province in Brazil and is located on the Amazonian forest, nearby Solimões River. Petroleum is transported by petroleum ships for 400 Km from Coari to the petroleum refinery in Manaus along the Solimões River. To evaluate possible impacts of crude oil transport, different fish species were collected along Solimões River: Astronotus ocellatus, Brycon amazonicus, Cichla monoculus, Colossoma macropomum, Liposarcus pardalis and Pygocentrus nattereri. Sampling was performed in 2007 on nine lakes along transect of transportation during four hydrological periods: rising, high, descending, and low water levels. White muscle was collected, freezed (-180) and transported to the laboratory. GST and catalase enzymes were evaluated. The activity of the enzymes varied according to the species as well as the hydrologic period of the year. Enzyme activities were higher on rising water level for C. monoculus and higher on high water level for P. nattereri.

THE USE OF FISH DRAG STRIP TO MEASURE ACCELERATION PERFORMANCE IN A COHORT OF EUROPEAN SEA BASS ((DICENTRARCHUS LABRAX) VanDamm, J.P., Nelson J. A., Claireaux, G and Handelsmann, C. Symposium: Swimming Performance Presentation: Poster Contact: Jay Nelson, Department of Biological Sciences Towson MD 21093 USA E-Mail: [email protected] Abstract: Because the traditional methods of high speed video analysis and accelerometry have a number of limitations, the potential use of a fish drag strip for measuring acceleration performance in fish was explored. A cohort of 99 European sea bass were each chased past a series of laser light "curtains" a minimum of 4 times. Breakage of the laser "curtain" was recorded to

Fish Biology Congress Abstracts 100 0.05ms accuracy by a photodector array connected to an analog/digital converter and a PC. Maximal acceleration was generally recorded in the first 2cm of the 1m long drag strip, was evenly found among the animals' successive trials and individual variation in successive trials was far less than variation amongst individuals. The average maximal acceleration of 1m*s-2 was actually higher than that recorded by high-speed filming of a subset of the same fish (0.81m*s-2). Acceleration did not depend upon fish size over a narrow size range.

PILOT STUDIES FOR THE TOXICOLOGICAL ASSESSMENT OF WHITE STURGEON (ACIPENSER TRANSMONTANA) EARLY LIFE STAGES EXPOSED TO LIQUID EFFLUENTS OF THE TECK COMINCO METALS LTD. FACILITY AT TRAIL, B.C., CANADA Vardy, D., Hecker, M., Giesy, J.P. Symposium: Environmental Pollution Presentation: Poster Contact: David Vardy, Toxicology Centre, University of Saskatchewan, 44 Campus Dr. Saskatoon SK S7N 5B3 Canada E-Mail: [email protected] Abstract: The primary purpose of this pilot study was to develop an artificial stream exposure system to specifically address the potential impact of contaminants in the liquid effluent released by Teck Cominco Metals Ltd. into the Upper Columbia River early life-stages of white sturgeon. White sturgeon eggs and 14-day old larvae were exposed to increasing concentrations of Teck Cominco liquid effluent in order to characterize potential impacts on egg hatchability and embryo development, as well as survival and growth of fry and juveniles. Based on initial results, the liquid effluent does not appear to be toxic to white sturgeons at the dilutions studied. Overall, the pilot study was a success but limitations and deficiencies were identified that promoted fish mortalities. Future needs and issues were identified to effectively conduct definitive studies, some of which include: Upper Columbia River surface water toxicity experiments and metal sensitivity characterization in white sturgeon.

MAPPING IMMUNE GENES IN RAINBOW TROUT INVOLVED IN THE INFLAMATORY RESPONSE Ventura, A. B., DeKoning, J.J. and Phillips, R.B. Symposium: Immunity and Disease Resistance Presentation: Poster Contact: Abbigail Ventura, Washington State University Vancouver, 14204 NE Salmon Creek Ave. Vancouver WA 98686 USA E-Mail: [email protected] Abstract: We report the genetic map location of fifteen genes involved in the inflammatory response to bacterial and viral pathogens in rainbow trout. These genes were mapped as possible candidate genes that may be involved in resistance to BKD and IHNV. Several of the genes (COX2, iNOS, CCR7, CXCR4, and COX1) are found in duplicate copies in the sequenced teleost genomes of zebrafish, fugu, stickleback or medaka. We will present results of phylogenetic analysis of the rainbow trout genes with similar genes found in Atlantic salmon, and the sequenced teleost genomes. This analysis has enabled us to determine orthologous and paralogous relationships among the genes and to identify conserved genomic blocks between rainbow trout and the sequenced teleost genomes.

REGULATION OF INSULIN-LIKE GROWTH FACTOR-1 RECEPTORS Very, N., Poudyal, D., Hanson, A., Kittilson, J., and Sheridan, M.A. Symposium: Fish Growth Presentation: Oral Contact: Mark Sheridan, Department of Biological Sciences, North Dakota State University Fargo ND 58105 USA E-Mail: [email protected] Abstract: Rainbow trout, which possess two distinct insulin-like growth factor-1 receptors, IGFR1A and IGFR1B, were used to examine the regulation of IGFRs. Fasting for 6 weeks reduced somatic growth as well as 125I-IGF-1 binding and mRNA expression of both IGFR1s in gill. Implantation with somatostatin-14 (SS-14), which is known to inhibit growth hormone, for 20 days reduced IGFR1A and IGFR1B mRNA levels and lowered 125I-IGF-I binding in gill. SS-14 inhibited the expression of IGFR1 mRNAs in gill filaments incubated in a time- and dose-dependent manner and inhibited 125I-IGF-I binding in filaments incubated in vitro. E2 reduced mRNA levels of IGFR1s and 125I-IGF-I binding in filaments in vitro; whereas, T stimulated the expression of IGFR1 mRNAs and increased 125I-IGF-I binding. These data indicate that IGFRs are important in the regulation of growth and that the mRNA and functional expression of IGFR1s are directly regulated by SS-14, E2, and T.

MELANOCORTIN 2 RECEPTOR AND ITS ROLE IN THE STRESS RESPONSE Vijayan, M.M. and Aluru, N. Symposium: Stress Effects and Response Presentation: Oral

Fish Biology Congress Abstracts 101 Contact: Matt Vijayan, Department of Biology, University of Waterloo Waterloo Ontario N2L 3G1 Canada E-Mail: [email protected] Abstract: Adrenocorticotropic hormone (ACTH) binds to melanocortin 2 receptor (MC2R) and activates the signaling cascade leading to corticosteroid biosynthesis. However, little is known about MC2R regulation and its role in ACTH signaling during stress adaptation in fish. We cloned and sequenced MC2R from rainbow trout (Oncorhynchus mykiss) interrenal tissue. Quantitative real-time PCR revealed a marked tissue-specific difference in MC2R mRNA abundance with the highest levels observed in the interrenal tissue, ovary and testis. Acute ACTH stimulation in vitro elevated MC2R mRNA abundance in a time- and dose-related fashion in the interrenal tissue. Acute stressor exposure in vivo transiently elevated plasma ACTH and cortisol levels at 1 h, followed by a significant increase in MC2R mRNA abundance at 4 h post- exposure. Overall, our results demonstrate that ACTH regulation of MC2R is highly conserved in vertebrates, underpinning a critical role for this receptor in the evolutionarily conserved cortisol response during stress adaptation.

TRANSCRIPTIONAL REGULATION OF THE ACTIVATION INDUCED CYTIDINE DEAMINASE (AID) GENE IN THE CATFISH AND ZEBRAFISH Villota, D.P., Singh, A.P., and Magor, B.G. Symposium: Immunity and Disease Resistance Presentation: Poster Contact: Brad Magor, Dept. of Biological Sciences, University of Alberta Edmonton AB T6H 2E5 Canada E-Mail: [email protected] Abstract: Activation-induced cytidine deaminase (AID) introduces point mutations and double strand breaks to the immunoglobulin gene during antibody affinity maturation in B cells. In mammals expression of the AID gene is restricted to germinal centre B-cells (centroblasts) via a centroblast specific promoter and enhancer in the first intron. We are using an established reporter assay to identify the equivalent transcriptional regulatory elements of fish AID genes in catfish B-cell lines, AID expressing cell lines, as well as other immune cell lines. Though we know from other work that catfish AID has restricted expression in vivo, we have been unable to identify any centroblast specific functional regulatory elements in the promoter or introns of either the catfish or zebrafish AID genes. We are currently testing more distal regions of the fish AID gene to see just how disparate fish AID gene regulation is from that of the mammalian AID gene.

CHOICE FOR NUMBER OF SHELTERS IN FISH Volpato, G. L., and Nomura, H. A. de K. Symposium: Stress Effects and Response Presentation: Oral Contact: Gilson Luiz Volpato, Laboratory of Animal Physiology and Behavior, Dept. of Physiology, Research Center on Animal Welfare, Instituto de Biociências, Caunesp, Unesp Botucatu São Paulo 18618-000 Brazil E-Mail: [email protected] Abstract: The role of shelters for environmental enrichment to Nile tilapia juveniles was investigated. The fish were isolated in an aquarium divided into two equal-sized compartments: one empty and the other either empty or with one, two, three or four shelters. Fourteen fish were tested in isolation for each number of shelters. Position of the compartment with shelter was balanced and the fish were introduced always into the empty compartment. After 2 days, fish position (compartment) was registered each min for 1 h. Choices of each fish and frequency of fish for each choice were determined by Goodman’s proportion test. No preference for a compartment was found when fish faced only empty compartments. Fish chose shelters and proportion of fish that chose shelters increased linearly with the number of shelters presented. These results sustain that increasingly number of shelters might be an improved condition to the Nile tilapia.

ANESTHESIA WITH PROPOFOL OR BENZOCAINE IN FISH: WHICH FITS BETTER? Volpato, G.L., Sales, N., and Giaquinto, P.C. Symposium: Stress Effects and Response Presentation: Oral Contact: Gilson Luiz Volpato, Laboratory of Animal Physiology and Behavior, Dept. of Physiology, Research Center on Animal Welfare, Instituto de Biociências, Caunesp, Unesp. Botucatu São Paulo 18618-000 Brazil E-Mail: [email protected] Abstract: Ventilation frequency (VF) and posture loss were investigated in isolated Nile tilapia juveniles under anesthesia by benzocaine or propofol. After a 2-h adjustment, basal VF was measured and then anesthesia (or control) was imposed in another aquarium (benzocaine = 12 g/L; propofol = 3.5 mL/L) (15 fish each treatment; ~23 g). VF was quantified until fish assumed a flank position for 5 min and continued until recover the posture (in another aquarium). Time (min ± sd) for posture loss was similar for the anesthetics (benzocaine = 6.33 ± 1.95; propofol = 4.80 ± 1.92), but recovery occurred earlier in benzocaine than in propofol (2.00 ± 0.81 vs 4.87 ± 1.88; p < 0.05). VF decreased during anesthesia and increased

Fish Biology Congress Abstracts 102 during recovery. This profile was much more stable for propofol (U-curve: exponentially lower over anesthesia and exponentially higher over the recovery time).

QUORUMS, CONSENSUS AND LEADERSHIP IN FISH SHOALS Ward, A.J.W., Sumpter, D., Couzin, I.D. & Krause, J. Symposium: Social Behavior Models Presentation: Oral Contact: Ashley Ward, School of Biological Sciences, University of Sydney Sydney NSW 2006 Australia E-Mail: [email protected] Abstract: Despite the growing interest in collective phenomena such as 'swarm intelligence' and ‘wisdom of the crowds’, little is known about the mechanisms underlying decision-making in vertebrate animal groups. We examined this field using a combination of experimental and modelling approaches. In our experimental work, we used two different automated approaches to test hypotheses relating to group movement decisions in fish. We found that quorum responses were of key importance to group decision-making and that the accuracy of decision-making increases with group size. In addition, we found that the ability of leaders to successfully initiate movements in a group changes over time as group members gather more information about their environment. Model simulations based on our data show that quorum responses enable fish to improve the accuracy and speed of their decision-making over that of independent decision-makers or those using a weak linear response.

INTRASPECIFIC INTERACTIONS AND INTERIOR FRASER COHO (ONCORHYNCHUS KISUTCH) HABITAT UTILIZATION Warren, K.W., Shrimpton, J.M., McRae, C.J. Symposium: Contributed Papers Presentation: Poster Contact: Kyla Warren, University of Northern British Columbia, 3333 University Way Prince George BC V2N 4Z9 Canada E-Mail: [email protected] Abstract: Territorial and schooling behaviours in juvenile Interior Fraser coho were analysed to better understand their effect on habitat use. Surveys within the Horsefly River watershed (central British Columbia, Canada) found that fish aggregated in particular habitats, sometimes with densities in excess of 100 individuals/m2 . To further examine behavioural interactions, juvenile coho were captured and introduced to a 5.5m x 2m raceway and observed for 14 days. Observations of position within the channel were used to characterize spatial distribution, while underwater footage of individual interactions examined the behaviours contributing to this pattern. Analysis of recorded interactions indicates that schooling and passive behaviours were far more common than aggressive or territorial displays. This contrasts with other populations of coho studied, which describe dominant territorial individuals surrounded by subdominants and solitary “floaters”. Our findings may indicate behavioural plasticity between subpopulations of coho, possibly as a result of differing environmental pressures.

COSTLY INFORMATION: PREDATION RISK AND SOCIAL LEARNING Webster, M.M. Symposium: Social Behavior Models Presentation: Oral Contact: Mike Webster, University of St Andrews, School of Biology, Bute Building E36 St Andrews, Fife KY16 9TS UK E-Mail: [email protected] Abstract: Private information can be accurate but expensive to acquire, while social information is cheap, but less reliable. Theory predicts that private information should be used when the risks associated with doing so are low, but that social information use should increase as the costs of gathering private information become greater. We tested this idea in two studies of social foraging behaviour. Study (1) revealed that minnows were more likely to forage in locations where they had previously seen other fish feeding only when predation risk was high, but not when it was low. Study (2) showed that populations of poorly-armoured and predator-vulnerable ninespine sticklebacks used social information to gauge the quality of prey patches, whereas sympatric populations of well-armoured threespine sticklebacks did not. Our findings suggest that fish can evolve both flexible and rigid social learning strategies in order to balance risk and accuracy when foraging in variable environments.

A TALE OF TWO BROOK LAMPREYS: ANCIENT VICARIANCE AND RECENT DISPERSAL White, M.M., and J. Kunkle Symposium: Biology of Lampreys Presentation: Oral

Fish Biology Congress Abstracts 103 Contact: M. White, Department of Biological Sciences, Ohio University Athens OH 45701 USA E-Mail: [email protected] Abstract: The Least Brook lamprey and the American Brook lamprey are non-parasitic species with widespread distributions in the eastern United States. Although both members of the genus Lampetra, they are members of two different clades within the genus. Sequence variation in the mitochondrial DNA control region was used to evaluate patterns of geographic variation. Data from the Least Brook lamprey indicated extensive sequence divergence with significant differentiation even in adjacent drainages. Divergences suggested that some populations have been isolated since the Pliocene with little evidence of recent dispersal. In contrast, the American Brook exhibited little differentiation across its distribution suggesting a recent range-wide dispersal event, likely from an Alaska/Asia sister taxon. Although both species are non-parasitic, there may be life history differences that contribute to contemporary dispersal in one species (appendix) but not in the other (aepyptera).

FUNCTIONAL GENOMICS OF ACCLIMATION AND ADAPTATION IN KILLIFISH Whitehead, A. Symposium: Genomics of Adaptation Presentation: Oral Contact: Andrew Whitehead, Biological Sciences, Louisiana State University Baton Rouge LA 70808 USA E-Mail: [email protected] Abstract: Killifish of the genus Fundulus have long been of interest to science in part because of their extraordinary physiological resilience in the face of environmental stress and as a model for studying adaptive evolutionary variation. Our research seeks to reveal the role of global gene expression regulation in compensatory acclimation responses to environmental stress, and how variation in expression patterns among populations and species may account for evolved differences in ability to tolerate those stresses. Several populations of F. heteroclitus have independently evolved dramatic resistance to pollution, and dose-response experiments are identifying what resistant fish are doing differently at the transcriptional level, compared to closely-related sensitive fish, that may offer insight into the mechanism of resistance. Fundulus species also vary in physiological plasticity; some species can compensate for wide extremes in environmental conditions such as osmotic stress, whereas others have more narrow tolerance ranges. Comparative genomics experiments seek to identify gene regulatory differences among taxa that account for differences in physiological plasticity.

ELECTROFISHING FOR LARVAL LAMPREY: THE INFLUENCE OF CAPTURE EFFICIENCY ON ESTIMATES OF DISTRIBUTION AND ABUNDANCE Whitesel, T.A., Luzier, C.W. and Silver G. Symposium: Biology of Lampreys Presentation: Oral Contact: Timothy Whitesel, Columbia River Fisheries Program Office, US Fish and Wildlife Service, 1211 SE Cardinal Ct., Suite 100 Vancouver WA 98683 USA E-Mail: [email protected] Abstract: The status of lamprey in the Columbia River Basin is unclear. Electrofishing is commonly used for determining distribution and abundance of fish; however, little work has been done to assess the effectiveness of electrofishing for larval lampreys. To evaluate the efficiency of electrofishing at capturing larval lamprey, we performed controlled trials with varying levels of larval density, larval size and sample effort. Cumulative capture efficiency increased as density increased for all sizes. Cumulative capture efficiency was highest for small ammocoetes. The probability of detection increased as density increased for all sizes of ammocoetes. At low densities, smaller ammocoetes were more easily detected than larger ones. We also conducted a mark-recapture study in a local stream and found lower capture efficiencies than those observed in the controlled trials. This study provides insights into the utility of electrofishing for estimating capture efficiency in and predicting the probability of detection for larval lampreys.

INTEGRATING SELECTIVE BREEDING WITH MICROBIAL GENOMICS TO IMPROVE RAINBOW TROUT DISEASE RESISTANCE Wiens, G. D. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Greg Weins, USDA/ARS, National Center for Cool and Cold Water Aquaculture Kearneysville WV 25430 USA E-Mail: [email protected] Abstract: Infectious disease is a substantial problem in aquaculture and improved methods are needed to diminish disease-related loss. One approach is to selectively breed fish from crosses that exhibit superior disease-resistance phenotypes thereby increasing fitness. A current limitation in breeding programs is the incomplete knowledge of the heritability, duration, and mechanisms of resistance, as well as the response to pathogen variants. In 2005, the NCCCWA initiated a selective

Fish Biology Congress Abstracts 104 breeding program designed to evaluate and improve rainbow trout survival following challenge with the bacterial cold- water disease agent Flavobacterium psychrophilum. We determined that survival is a moderately heritable trait and that spleen size is correlated with resistance. In conjunction with these studies, we have also determined the complete genome sequence of F. psychrophilum strain CSF 259-93, and I will discuss strategies of how we are combining this knowledge with selective breeding to improve fish health.

THE HAGERMAN NATIONAL FISH HATCHERY Wiese, N.J. Symposium: Contributed Papers Presentation: Poster Contact: Nathan Wiese, Hagerman National Fish Hatchery, 3059-D Nat'l Fish Hatchery Rd Hagerman ID 83332 USA E-Mail: [email protected] Abstract: The Hagerman National Fish Hatchery (Hatchery) is located along the Snake River, about 30 miles west of Twin Falls, Idaho. The Hatchery was authorized by 46 Stat, 371 on May 21, 1930 and was established in 1932. Construction of the physical facilities commenced in 1932, and fish production began in 1933. The initial goal of the Hatchery was the production of rainbow trout for stocking in Idaho, eastern Oregon, and northern Nevada. In the late 1970's, the Hatchery became part of the Lower Snake River Fish and Wildlife Compensation Plan (LSRCP) The Hatchery's primary production goal was changed to steelhead trout and the Hatchery was rebuilt and expanded, for $7.0 million, from June 1982 through April 1984. The Hatchery now produces over 1.4 million steelhead smolts annually under the LSRCP program. The Hatchery is currently undergoing a Hatchery Review under the USFWS.

EFFECTS OF INCREASING SALINITY ON SURVIVAL AND METABOLIC RATE OF THREE EURYHALINE FISHES, CYPRINODON VARIEGATUS, GAMBUSIA AFFINIS AND POECILIA LATIPINNA. Wilkes, A.A. and Bennett, W.A. Symposium: Tribute to Joe Cech, Jr. Presentation: Oral Contact: Allison Wilkes, University of West Florida, Department of Biology, 11000 University Parkway Pensacola Fl 32514 USA E-Mail: [email protected] Abstract: We evaluated chronic upper salinity tolerance, weight specific routine oxygen consumption and maximum survivable salinity of Sheepshead Minnow, Cyprinodon variegatus, Sailfin Molly, Poecilia latipinna and Western Mosquitofish, Gambusia affinis. Salinity tolerance of Sheepshead Minnow, Sailfin Molly and Western Mosquitofish were 120, 92, and 48‰, respectively. More efficient renal excretion of divalent ions may account for higher euryhalinity in Sheepshead Minnow and Sailfin Molly. Metabolic rates of all three fishes were unchanged at salinities above 5‰, despite increasing salinity, suggesting that these fishes conserve energy by rearranging their energetic budgets, rather than increasing metabolism to meet higher ion-osmoregulatory costs. Maximum survivable salinity estimates for Sheepshead Minnow, Sailfin Molly and Western Mosquitofish of 168, 113, and 72‰, respectively, were the highest reported for these species. Salinity adaptations of fishes in our study allow them to exploit euryhaline habitats along the Gulf of Mexico so extreme that all other fishes are excluded.

METAMORPHOSIS, METABOLIC PLASTICITY, & FEEDING IN THE SEA LAMPREY Wilkie, M.P. Symposium: Biology of Lampreys Presentation: Oral Contact: Michael Wilkie, Department of Biology, 75 University Avenue West, Wilfrid Laurier University, Waterloo, ON, N2L 3C5. CANADA Waterloo Ontario N2L 3C5 Canada E-Mail: [email protected] Abstract: The sea lamprey (Petromyzon marinus) spends its first 3-7 years as a filter-feeding larvae before metamorphosing into a parasitic juvenile which feeds on the blood of fishes. In larvae, basal rates of amino acid catabolism are low due to the poor nutritional quality of their diet, and low metabolic rate. Basal rates of amino acid catabolism increase following metamorphosis, as demonstrated by 6- to 20-fold greater basal nitrogenous waste (N-waste) excretion rates, and greater amounts and activities of glutamate dehydrogenase, and other amino acid catabolizing enzymes in liver. This metabolic reorganization allows parasitic lampreys to catabolize intermittently large amino acid loads following the ingestion of protein rich fish blood. Indeed, post-feeding N-waste excretion may increase by 25-fold. The energy-rich C-skeletons generated following feeding can be burned, or retained for glycogen and fatty acid synthesis, which are essential fuels for the subsequent upstream-migratory and spawning phases of the lamprey’s life cycle.

EVIDENCE FOR A NOVEL MECHANISM OF CL- UPTAKE IN FRESHWATER SEA LAMPREYS

Fish Biology Congress Abstracts 105 Wilkie, M.P., and Tufts, B.L. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Michael Wilkie, Department of Biology, Wilfrid Laurier University, Department of Biology, Queen's University Waterloo Ontario N2L 3C5 Canada E-Mail: [email protected] Abstract: Like other freshwater fishes, sea lampreys (Petromyzon marinus) use branchial Cl- transport to help maintain internal ion balance, but the mechanism(s) of Cl-uptake remains unresolved. We provide evidence of a saturable (Jmax=190 nmol/g.h), moderate affinity (Km=72 nmol/L) chloride uptake system in the gills of freshwater larval lampreys. Acclimation to Cl- free water resulted in a 3-fold increase in the Jmax and a 70% reduction in the Km of the lamprey Cl- transport system. Unlike many teleosts, however, Cl- uptake by lampreys was insensitive to the Cl-/HC03- exchange inhibitors DIDS and thiocyanate. Moreover, inhibition of branchial carbonic anhydrase, through exposure to acetazolamide, did not alter Cl- uptake. These findings suggest that larval lampreys may not use branchial Cl-/HCO3- exchange. The K+:Cl- co-transport inhibitor, DIOA, was also without effect. However, furosemide markedly inhibited Cl- uptake by 75%, suggesting that larval lampreys may use Na+:K+:2Cl- co-transport to translocate Cl- across the gill.

MECHANISMS OF HIGH AMMONIA TOLERANCE IN THE GOLDFISH Wilkie, M.P., Dhiyebi, H., Sangha, N., and Skelton, G. Symposium: Fish Living on the Edge Presentation: Oral Contact: Michael Wilkie, Department of Biology, Wilfrid Laurier University, 75 University Avenue West Waterloo Ontario N2L 3C5 Canada E-Mail: [email protected] Abstract: We demonstrate that goldfish (Carassius auratus) have unusually high ammonia tolerance. Acute toxicity studies revealed that this fish had a NH3 96h LC50 of 250-290μmol/L, which was 4-5 fold greater than that of most other freshwater fishes. Chronic exposure (>5d) to 5 mmol/L total ammonia did not stimulate urea production in goldfish as a means of detoxifying ammonia. Rather, goldfish tolerated internal ammonia concentrations approaching 2 mmol/L in plasma, and 8 mmol/L in brain. In other vertebrates, brain ammonia concentrations in this range overactivate N-Methyl-D-Aspartate receptors (NMDAr), leading to excitotoxicity and death. Attempts to block the NMDAr with ethanol and MK801, which protect mammals against ammonia toxicity, had no protective effect in goldfish challenged at lethal external ammonia concentrations. However, these drugs did protect ammonia-sensitive rainbow trout from ammonia toxicity. We conclude that a lower sensitivity of the NMDAr to ammonia contributes to the unusually high ammonia tolerance of the goldfish.

EFFECTIVE FISH PASSAGE SYSTEMS MATCH HYDRAULICS TO FISH BEHAVIOR Williams, J.G. Symposium: Swimming Performance Presentation: Oral Contact: John Williams, Northwest Fisheries Science Center, 2725 Montlake Blvd. E. Seattle WA 98112-6-2097 USA E-Mail: [email protected] Abstract: The design of effective fish passage facilities requires understanding migrant fish behavioral responses to varying hydraulic conditions. Fish passage facilities that do not take these evolutionarily derived behaviors into account will either fail or work much more poorly than hoped. Developing facilities for downstream migrants is harder than for upstream migrants as the former have much less time to make decisions and react to hydraulic conditions that they do not prefer. The results of Columbia River basin juvenile and adult salmonid (Oncorhynchus sp.) behavioral research over the last 50 years suggest that migrant salmonids have evolved to use changes in water velocity as cues to most effectively complete their migration either to the ocean or return to spawning grounds. However, what works for salmonids, may not work for all species. The talk will illustrate how lack of adherence to behavioral principles may lead to less than effective passage conditions.

REMODELING OF THE GILL IN THE CLIMBING PERCH Wilson, J.M. Choe, K.P. and Ip, Y.K. Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Jonathan Wilson, Ecofisiologia-CIMAR, Rua dos Bragas 289 Porto 4050-123 Portugal E-Mail: [email protected] Abstract: The climbing perch (Anabas testudineus) is an air-breathing, euryhaline freshwater fish. It is capable of acclimating to full strength seawater following gradual but not direct transfer, as well as prolonged periods of emmersion and ammonia exposure. In order to understand the mechanisms of ionoregulation at play, the expression of a number of ion transport proteins (Na+/K+-ATPase, Na+:K+:2Cl- cotransporter, V-ATPase, Na+/H+ exchanger 3 and Rhesus C glycoprotein 1

Fish Biology Congress Abstracts 106 ammonia transporter) was determined in a time course salinity acclimation experiment and following exposure to high ammonia, and emmersion using immunofluorescence microscopy and immunoblotting techniques. The gill, which is the dominate ionoregulatory organ, undergoes a high degree of remodeling at the cellular and protein level following salinity acclimation but only subtle changes occur during HEA and emmersion. Of note is the apical colocalization of NHE3 and RhCG1 to Na+/K+-ATPase-IR cells in seawater acclimated climbing perch and the rapid elimination of V-ATPase following salinity transfer.

DOES THE SEAHORSE GILL POSSESS ACCESSORY CELLS? Wilson, J.M., Laurent, P., Chevalier, C., Ip, Y.K., Lam, T.J., Sivalonganathan, S., and Reddy, K. Symposium: Ion and Acid-base Regulation Presentation: Poster Contact: Jonathan Wilson, Ecofisiologia-CIMAR, Rua dos Bragas 289 Porto 4050-123 Portugal E-Mail: [email protected] Abstract: In marine teleost fishes, Na+ and Cl- are eliminated by branchial paracellular and transcellular mechanisms, respectively, associated with the chloride-accessory cell complex. Chloride cell Na+/K+-ATPase drives intracellular Cl accumulation via a Na:K:2Cl cotransporter and an apical CFTR-like Cl channel facilitates the efflux of Cl down its electrochemical gradient. Na on the other hand accumulates in the extracellular space and exits via leaky tight junctions found between chloride and accessory cells down its electrochemical gradient. Transepithelial potential (TEP) measurements have been found to be close to the Nerst equilibrium potential of Na supporting this model; however, in some marine species (including seahorse), TEP measurements are not in agreement (Evans DH, Cooper K 1976. Nature 259:241-43) suggesting an alternative mechanism for Na+ elimination. In the present study, the existence of accessory cells by TEM and SEM was determined in seahorse gills as well as the presence of the ion transport proteins involved in secondary active Cl- elimination.

CHARACTERIZATION OF BRANCHIAL ATPASE ACTIVITY IN LAMPREY (PETROMYZON MARINUS L.) AMMOCOETES AND TRANSFORMERS Wilson, J.M., Reis-Santos, P.N., McCormick, S.D. Symposium: Biology of Lampreys Presentation: Poster Contact: Jonathan Wilson, Ecofisiologia-CIMAR, Rua dos Bragas 289 Porto 4050-123 Portugal E-Mail: [email protected] Abstract: The lamprey ammocoete larvae are a substrate-dwelling, filter-feeder and in an earlier study we found that their gills have a high percentage (90%) of ouabain-insensitive activity that was not present following metamorphosis. In the present study, we used a pharmalogical approach to characterize this residual activity in vitro. We found that the P-type ATPase inhibitor, orthovanadate, and F-type mitochondrial H+- ATPase inhibitors, azide and oligomycin, accounted for approximately 10- 20% of total activity each in ammocoetes but 90% and 3%, respectively, in transformers. The V-type H+-ATPase inhibitor, bafilomycinA1, and myosin ATPase inhibitor, BDM, accounted for negligible activity. The ABC-transporter inhibitors verapamil, probenecid and MK571 inhibited significant levels of activity in both ammocoetes and transformers although cyclosporinA stimulate activity. However, probenecid-sensitive activity was significantly greater in ammocoetes versus transformers accounting for ~ 20% total activity. The probenecid-sensitive ABC-transporter(s) may serve to eliminate toxicants that the ammocoetes may be exposed to in the substrate.

FUNCTIONAL CHARACTERIZATION OF THREE DISTINCT IGD BEARING CELL POPULATIONS IN CHANNEL CATFISH, ICTALURUS PUNCTATUS Wilson, M., Edholm, E.-S., Sahoo, M, Miller, N.W. and Bengten, E. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Melanie Wilson, Department of Microbiology, University of Mississippi Medical Center, 2500 North State Street Jackson MS 39216 USA E-Mail: [email protected] Abstract: Two immunoglobulin (Ig) isotypes have been identified in catfish, IgM and IgD. While IgM has been studied in detail, little is known concerning IgD function. To this end, a monoclonal (mAb) anti-IgD antibody was generated. Flow cytometry and cell separation techniques using this mAb demonstrated three different IgD+ populations in catfish PBL. Two are lymphocyte-like that differentially express IgM (IgM+/IgD+ and IgM-/IgD+) and are classified as B cells since PCR protocols amplify rearranged Ig messages from each cell population. Interestingly, the IgM-/IgD+ cell population in some catfish can represent as much as 60% of the total PBL. The third population is composed of IgM-/IgD+ granular-like cells of unknown origin that do not express Ig message, but appear to be armed with exogenously produced IgD via a putative IgD-binding receptor. Currently, the function of the IgM-/IgD+ granulocytes is unknown, however they degranulate in response to cross-linking of the surface bound IgD.

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ACID-BASE REGULATION IN A VERTEBRATE SALINITY-EXTREMOPHILE Wilson, R.W*., Cooper, C.A*., and Goren, M.^ Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Rod Wilson, *University of Exeter, School of Biosciences, Hatherly Laboratories, Exeter, UK., ^Department of Zoology, Tel Aviv University, Tel Aviv 69778, Israel. Exeter Devon EX4 4PS UK E-Mail: [email protected] Abstract: Hypersaline conditions cause up-regulation of drinking and intestinal HCO3- secretion for osmoregulatory purposes. This promotes greater CaCO3 base excretion via the intestine, requiring compensatory acid excretion via the gills - an unusual anatomical separation of acid and base excretion. Intestinal HCO3- is derived from metabolic CO2, which reduces gaseous CO2 excretion at the gills relative to O2 uptake - atypical for respiratory gas exchange processes. Aphanius dispar lives near the Dead Sea, in salinities up to 5-times, and [Ca2+] up to 17-times, greater than sea water. This is the most extreme salinity for any vertebrate and is expected to generate massive intestinal base excretion rates that require constitutive gill acid excretion similar to the mammalian kidney for acid-base balance. Furthermore, the majority of its metabolic CO2 excretion will probably be excreted as solid CaCO3 via the intestine. In vivo physiological studies are exploring these predictions in this remarkable vertebrate extremophile.

SUPPRESSION OF AGRESSIVE BEHAVIOR IN JUVENILE MATRINXÃ (BRYCON AMAZONICUS) FED WITH DIET SUPPLEMENTED WITH L-TRYPTOPHAN Wolkers, C.P.B., Serra, M., and Urbinati, E.C. Symposium: Social Behavior Models Presentation: Poster Contact: Carla Patrícia Bejo Wolkers, 651 José Bonifácio St. Jaboticabal SP 14882-035 Brazil E-Mail: [email protected] Abstract: The aggressiveness of juvenile matrinxã (Brycon amazonicus) causes substantial losses during the farming. This study investigated the effect of the diet supplementation with L-tryptophan (Trp), precursor of serotonin, known by its effect on reducing the aggressiveness in vertebrates. Fish distributed in individual aquaria were fed with experimental diets D1 - control (4.7g Trp/kg), D2 (9.6g Trp/kg), D3 (18.8g Trp/kg) and D4 (37.6g Trp/kg). After 7 days of feeding, fish were submitted to a resident-intruder test by introducing another fish into the aquarium. D2 fish showed elevated latency to first attack (p<0.0048), a lower number of bites (p<0.0080) and a trend to lower number of chases (p<0.6329) against intruder compared with D1, in addition to be more chased by the intruder than in the other treatments. The results suggest that the diet supplementation with Trp can be effective in reducing of the aggressiveness of matrinxã.

TEMPERATURE AND ONTOGENETIC EFFECTS ON THE SWIM PERFORMANCE, BEHAVIOR, AND KINEMATICS OF A DECLINING SPECIES, SACRAMENTO PERCH Woodley, C.M., and Cech, J.J. Jr. Symposium: Swimming Performance Presentation: Oral Contact: Christa Woodley, Center for Watershed Sciences, University of California- Davis Davis CA 95616 USA E-Mail: [email protected] Abstract: California’s only native centrarchid, Sacramento perch (Archoplites interruptus), is threatened due to its extirpation within its native range. Altered water flow regimes and diversions throughout the Sacramento perch’s native range puts this species at risk of entrainment and mortality. We hypothesized that this species would perform similarly to its confamilial species due to its similar body morphology, and lentic or large, slow-moving lotic habitats in which they currently reside. We measured critical swimming velocity, swimming behavior and kinematics for three life stages of Sacramento perch at 4 temperatures. There was a positive correlation between increasing temperature and final swimming velocity for the post- larval and juvenile stages. The adults’ swimming performance, however, exceeds that of the juveniles by only a small margin and plateaus or declines at temperatures greater than 18 C. Adult Sacramento perch and warm-water basses have similar final swimming performances.

ADAPTATIONS FOR AN AMPHIBIOUS LIFE IN MANGROVE KILLIFISH Wright, P.A. Symposium: Fish Living on the Edge Presentation: Oral Contact: Patricia Wright, University of Guelph, Integrative Biology Guelph ON N1G 2W1 Canada E-Mail: [email protected] Abstract:

Fish Biology Congress Abstracts 108 The mangrove killifish Kryptolebias marmoratus frequently leave water-filled burrows to take temporary refuge amongst moist leaf litter or rotting logs in the forest. In the lab, some killifish voluntarily emerse, spending approx. 50% of their time in air. Within a few days of air exposure, dramatic (reversible) gill remodeling occurs reducing gill surface area and altering the composition of gill cells. The cutaneous surface appears to be an important exchange surface with an extensive capillary network close to the surface. Changes in the mitochondrial-rich cell population in the skin during air exposure is dependent on the salinity of the moist substratum. Nitrogen excretion during air exposure is achieved through NH3 volatization at the skin surface. This may be facilitated by ammonia transporters or Rh glycoproteins. K. marmoratus have a flexible exchange system, reversibly remodeling branchial and cutaneous surfaces in response to changing environments.

CADMIUM AND COLD SHOCK INDUCE METALLOTHIONEIN-2 EXPRESSION BUT NOT SIMILAR TO METALLOTHIONEIN-B (SMT-B) IN ZEBRAFISH (DANIO RERIO) LARVAE Wu, S. M., Zheng, Y. D., and Kuo, C.H. Symposium: Stress Effects and Response Presentation: Oral Contact: Su Mei Wu, 300 University Road, Chiayi, Taiwan Chiayi Taiwan 600 Taiwan E-Mail: [email protected] Abstract: The objective of this study was to compare the roles of MT-2 and sMT-B after cadmium exposure (0, 0.08, 0.26, and 0.89 μM Cd) and cold shock (28 and 12℃) in zebrafish larvae with whole-mount in situ hybridization and RT-PCR reaction. Results were: (1) during developmental stage, MT-2 mRNA existed at 6 hpf, and the level rapidly increased to 24 hpf, then it gradually increased with further larval growth. sMT-B mRNA was found at 12 hpf, and it also rapidly increased to 24 hpf, but remained constant with further larval development. (2) The transcripts of MT-2 mRNA and Cd contents displayed dose and time dependence after Cd exposure and cold shock. But transcripts of sMT-B mRNA were not induced by either challenge. Apparently, MT-2 may be related to Cd detoxification and cold-shock adaptation in the zebrafish larvae, but sMT-B was not found to have these physiological functions.

TRIMETHYLAMINE OXIDE AS A PROTEIN STABILIZER IN ELASMOBRANCH AND DEEP-SEA FISHES Yancey, P.H. Symposium: Plenary Presentation: Oral Contact: Paul Yancey, 345 Boyer Ave, Whitman College, Biology Dept Walla Walla WA 99362 USA E-Mail: [email protected] Abstract: Marine elasmobranchs have long been called ureosmotic, due to their high levels of urea as an osmolyte. They also have TMAO (trimethylamine oxide) at about a 1:2 ratio to urea. Research starting in the 1970s has shown that TMAO is a strong protein stabilizer that can counteract the protein-destabilizing effects of urea at this ratio. TMAO is also found in shallow- living teleost fishes, but at much lower levels. However, recent work has revealed an increase in TMAO with depth in both teleost and elasmobranch fishes, with a corresponding decrease in urea in the latter. Skates at 3000m have a 2:1 TMAO:urea ratio and thus might be termed "TMAO-osmotic." Laboratory studies show that TMAO can counteract the effects of hydrostatic pressure on protein stability and kinetics. It may be the need for TMAO to offset pressure effects that sets the depth limit for fishes.

ENERGY METABOLISM FOR FISH ION REGULATION: IDENTIFICATION AND FUNCTIONAL ANALYSIS OF GLUCOSE TRANSPORTERS IN ZEBRAFISH IONOCYTES Yung-Che Tseng, Pung-Pung Hwang Symposium: Ion and Acid-base Regulation Presentation: Oral Contact: Yung-Che Tseng, R144, Institute of Cellular and Organismic Biology, Academia Sinica, Nankang, Taipei, Taiwan, ROC Taipei City 11529 Taiwan E-Mail: [email protected] Abstract: A timely and sufficient energy supply is crucial for the iono- and osmo-regulation in fish gills. Recently, glycogen-rich (GR) cells, a novel type of gill cells surrounding ionocytes, have been identified to supply emergent energy to ionocytes during acute environmental challenges. This study aimed to examine the expressions and functions of glucose transporter (GLUT) in ionocytes and GR cells. Eighteen members of GLUTs were cloned from zebrafish. Based on triple labeling and gene knockdown experiments, only zGLUT13a was demonstrated to specifically express in and to function in absorbing glucose as energy supply to H+-ATPase-rich (HR) cells, the ionocytes responsible for Na+ uptake, while zGLUT6 is responsible for transporting glucose into GR cells, which deposit glucose as glycogen. Taken together, the 2 specific zGLUTs in HR cells and GR cells absorb glucose into the respective cells with different affinities, but appear to fulfill different physiological demands in different types of epithelial cells.

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A NOVEL B CELL SUBSET OF TELEOST FISH EXPRESSING SURFACE IgT: ROLE IN PHAGOCYTOSIS Zhan, Y., Li, J., Sunyer, J.O. (Sunyer is the presenting author) Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Oriol Sunyer, School of Veterinary Medicine, University of Pennsylvania, 413 Rosenthal Building, 3800 Spruce St. Philadelphia PA 19104 US E-Mail: [email protected] Abstract: Up until recently, granulocytes and macrophages were also thought to be the main phagocytic cells in fish. Contrary to this belief, our studies have led to the exciting finding that IgM+ B cells from teleost fish have a potent phagocytic and killing ability. We have now discovered a new phagocytic population of rainbow trout B cells that uniquely express surface IgT. Recently developed monoclonal antibodies against trout IgT were instrumental for the purification of phagocytic and non- phagocytic populations of IgT+ B cells. The IgT+ B cells display the typical morphological features of lymphocytes and comprise ~5-15% of all rainbow trout peripheral blood and spleen B cells. Similar to the IgM+ B cell subset, IgT+ B cells were also found to be highly phagocytic. Our results indicate for the first time the presence of two major phagocytic B cell subsets in teleost fish, one expressing IgM, and the other expressing IgT.

GENERATION OF IMMUNOGLOBULIN DIVERSITY IN ZEBRAFISH: EVIDENCE FOR INVERSIONAL VJ-REARRANGEMENTS BOTH WITHIN AND BETWEEN IGL GENE CLUSTERS Zimmerman AM, Yeo G, Howe K, Maddox BJ, Steiner LA. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Ana Zimmerman, Department of Biology, College of Charleston, 66 George Street Charleston SC 29424 USA E-Mail: [email protected] Abstract: To elucidate IgL loci in the zebrafish genome assembly, conventional sequence similarity searches and a novel scanning approach based on recombination signal sequence (RSS) motifs were applied. RT-PCR with zebrafish cDNA was then used to confirm annotations, evaluate VJ-rearrangement possibilities, and show that each locus is expressed. In contrast to other vertebrates in which IgL exon usage has been studied, inversional rearrangement between (V(L)-J(L)-C(L)) clusters were found. Such rearrangements may convey a selective advantage for editing self-reactive receptors and poise zebrafish by virtue of their extensive numbers of V(L), J(L) and C(L) to have greater potential for immunoglobulin gene shuffling than traditionally studied mice and human models.

CATECHOLAMINE RELEASE FOLLOWING ACUTE STRESS IN JUVENILE LAKE STURGEON – ACIPENSER FULVESCENS Zubair, S.N., Allen, P.J., Peake, S.J. and Anderson, W.G. Symposium: Stress Effects and Response Presentation: Poster Contact: Sadaf Zubair, Dept. of Biological Sciences, University of Manitoba Winnipeg MB R3T 2N2 Canada E-Mail: [email protected] Abstract: The stress response, as measured by plasma cortisol concentration, has been noted to be lower in Acipenserids (sturgeons) compared to teleost fishes. While cortisol concentrations have been measured in a number of sturgeon species, little data is available on the catecholamine response to acute stress. Baseline circulating catecholamine, cortisol, osmolarity, lactate, glucose and ion concentrations were measured in cannulated lake sturgeon, Acipenser fulvescens. The same parameters were then measured in blood samples taken following a 30-second aerial exposure. Comparisons were made with the baseline and post stress values. Duration and magnitude of the stress response in regard to epinephrine, nor-epinephrine and cortisol were assessed. Plasma cortisol levels increased significantly, from 2.95 +/- 0.80 to 29.03 +/- 1.95 within 20 minutes post stress and returned to baseline levels within 240 minutes. These values will be compared to the measured values for catecholamines and discussed in the context of the “reduced” stress response reported for Acipenserids.

MOLECULAR AND CELLULAR ANALYSES ON THE ACTIVATION STATE OF B CELLS IN THE RAINBOW TROUT USING FLOW CYTOMETRY. Zwollo, P., Rosato, P., Mott, K., Haines, A., Barr, M., and Coyner, K. Symposium: Immunity and Disease Resistance Presentation: Oral Contact: Patty Zwollo, Department of Biology. The College of William and Mary Williamsburg VA 23185 USA E-Mail: [email protected] Abstract:

Fish Biology Congress Abstracts 110 The rainbow trout provides an important vertebrate model, as they possess a highly diverse B cell repertoire, yet do not possess bone marrow or lymph nodes. In an attempt to elucidate B cell activation processes in the trout, we used B cell- specific transcription factors as molecular markers. In mammalian species, Pax5 and XbpI define terminal B cell differentiation stages; Pax5 is expressed during all stages except plasma cells, while XbpI is not induced until B activation. We investigated activation stages of the B cell lineage using the following four markers: Pax5, XbpI, secreted IgM, and BrdU. Fixed and permeabilized immune cells were analyzed by flow cytometry. We then determined the cell frequency of mature B, activated B, plasmablast, and plasma cells. Our data show that unstimulated spleen contains mostly resting mature B cells, while PBLs, anterior and posterior kidney tissues contain multiple B cell subsets. Our approach provides an important and novel tool to measure the trout humoral immune response.

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