Number 18 De~ember 1,1996

pH and Temperature in Ectothermic Vertebrates

Life Histories of Noturus baileyi and N. flavipinnis (Pisces: Ictaluridae), Two Rare Madtom Catfishes in Citico Creek, Monroe County, Tennessee BULLETIN ALABAMA MUSEUM OF NATURAL HISTORY

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Copyright 1996 by The Alabama Museum of Natural History ))~((

.~ ...:-.~ »0((w ALABAMA MUSEUM of Natural History

Number 18 December 1, 1996

pH and Temperature in Ectothermic Vertebrates

by Gordon R. Ultsch and Donald C. Jackson

Life Histories of Noturus baileyi and N. flavipinnis (Pisces: Ictaluridae), Two Rare Madtom Catfishes in Citico Creek, Monroe County, Tennessee

by Gerald R. Dinkins and Peggy W. Shute

THE UNIVERSITY OF ALABAMA TUSCALOOSA, ALABAMA

December 1, 1996

pH and Temperature in Ectothermic Vertebrates

Gordon R. Ultsch

Departmen t of Biological Sciences University of Alabama

Tuscaloosa, AL ~5487

(FAX 205-~4R-178(j, e-mail Guitsch@biolob,)'.as.ua.edu)

Donald C. Jackson

Departmen t of Physiolob,)' Brown University Providence, RI 02912

ABSTRACT: Ultsch, Gordon R. and Jackson, Donald C. 1996. pH and Temperature in Ecothermic Verte­ brates. Bulletin Alabama Museum of Natural History, Number 18: 1-42, 13 tables. We have compiled data from a large number of studies of ectothermic vertebrates relating to temperature-acid-base relations. We have documented and analyzed the inverse relationship between both blood pH (pHb) and tissue pH (pHi) with temperature and have evaluated the agreement between these data, both ell masse and in selected subgroupings of species, with the theoretical constructs of Reeves (alphas tat regulation) and Rabn (constant relative alkalinity). In general the experimental data agree with these hypotheses, especially alphas tat regulation, but the changes in pHb (arterial blood pH) with temperature are consistently less than predicted by theory. The slope of the curve for 207 pHb-temperature points (each point an average of up to 36 studies), representing 81 species, is -0.016. When large data sets are considered, significant intergroup differences in the plasma pH-temperature relationship emerge related to the elevation (or y-intercept) of the linear regression relating pH to temperature, but not to its slope. Marine fishes, for example, have lower pHb values than do freshwater fishes, and reptiles have a lower pHb than do freshwater fishes and amphibians. The underlying bases for these differences are unknown. The observed fall in pHb with temperature of air-breathers is achieved principally by increases in PC02, similar to the behavior of blood equilibrated ill vitro, as has been previously noted. The regulatory mechanisms associated with temperature-dependent pHb changes in fishes are less certain, but may involve adjustments in both PC02 and [HCOs·]' We caution that "deviations" from the overall ectotherm slope of certain species or smaller taxonomic groups are in many cases based on relatively few studies, and that more extensive data are needed, particularly within the normal temperature range in which a species operates, before such deviations are accepted as real and therefore requiring an explanation.

Bull. Alabama Mus. Nat. Hist. 18:1- 41

Decem ber 1, 1996 2 BULLETIN 18 December 1,1996

Introduction however, demonstrated that the pattern of pH" change HISTORICAL ASPECTS-I t is well established that the pH of was similar among ectothermic species but that the values blood (pHb) and other body fluids of ectothermic verte­ of related acid-base variables were often quite different brates decreases with increasing body temperature. Al­ (Howell et aI., 1970; Rahn and Garey, 1973). For PCO" though this phenomenon was reported several times early and [HC03-], prominent interspecific differences can b~ in this cen tury (Austin et aI., 1927; Henderson, 1928; Dill readily explained by habitat and respiratory mode (Rahn, et aI., 1935; Dill and Edwards, 1935), it only gained wide­ 1966). Water-breathers achieve a particular pH" with low spread attention in more recent times after the study of -], values of both PC02 and [HCOg whereas air-breathers Robin on turtles (1962), followed by detailed investiga­ at the same pH" and temperature have proportionately tions by Rahn, Reeves, and their co-workers (e.g., Rahn, -] . elevated values of both PC02 and [HCOg Systematic 1966; Howell et aI., 1970; Reeves, 1972). The latter work­ differences among species in pHb at a particular tempera­ ers not only added to the growing body of data on the ture, on the other hand, have not been generally recog­ subject, but they also attempted to deduce general laws nized and have not been examined in a comprehensive governing the collected data. manner. The integrative hypotheses of constant relative alkalin­ THEORETICAL CONCEPTS-Rahn (1966) in troduced the con­ ity and alphastat regulation, in particular the latter, have cept of "constant relative alkalinity," based on the similar­ become the operative paradigms in the comparative acid­ ity benveen the temperature dependence of pH" and of base field, and much subsequent work has been carried the neutral pH of pure water (pN). He noted that pH" out to test their validity. The central issues of alphastat

and pNw both varied with temperature with a slope of regulation concern the satisfaction of two key properties: about -0.017 U 1°C, but that pHb was 0.4-0.6 U on the first, that pHb decreases with an overall slope of about alkaline side of neutrality. From a regulatory point of view, -0.018 U;oC bet\Veen 0 and 40°C (Reeves, 1976) and, Rahn proposed that the animals were maintaining a con­ second, that total plasma or extracellular CO" concentra­ stant [OH-] / [H+] ratio. An alternative interpretation of tion remains constant over this range. These-conditions, the pH,,-temperature relationship, developed by Reeves in short, require that blood in vivo behave like blood in (1972, 1977), is that the observed pHb changes conform vitro. The failure of collected data to satisfy these ideal to the temperature-dependence of the pK values of pro­ conditions set forth in the Rahn or Reeves hypotheses has tein buffers in the blood and intracellular fluid and that led some workers to question their validity (Heisler, 1986; these changes serve to maintain a constant net electrical Cameron, 1989). Heisler (1986), for example, assembled charge, and thereby a constant degree of dissociation and data from a variety of vertebrate ectotherms in which the a consistent tertiary structure, of proteins within these pH,,-temperature slope was consistently less than -0.018, fluids. The principal buffer group involved is the imida­ leading him to challenge the alphastat hypothesis. In zole moiety of the amino acid histidine. Reeves (1972) addition, Heisler presented intracellular data from his suggested that the animals were regulating the imidazole own and other laboratories that departed in both direc­ dissociation state, or alpha-imidazole (aIm)' and coined tions from the ideal pH,,-temperature slope. Cameron the term alphastat to describe the defended variable asso­ (1989) also criticized the alphastat idea because of the ciated with the inverse pH" change with temperature. The uncertainty of assigning a single value, or of confidently potential importance of alphastat regulation for intracel­ ascribing any value, to the pK of intracellular imidazole lular protein functions has been analyzed by White and groups, because of the very different rnicroenvironments Somero (1982). these groups experience in their host proteins. The issue A striking related observation, originally made on Im­ of intracellular alphastat regulation is complicated fur­ man and laboratory animal blood by Rosenthal (1948), is ther by the pH heterogeneity ofthe intracellular compart­ that the pH of blood subjected to temperature change in ment (White and Somero, 1982), by the diversity of cell vitro at constant gas content exhibits a temperature de­ and tissue types, and by acclimatory changes following pendence similar to ectothermic blood in vivo. This in a temperature change that involve acid-base-relevant vitro behavior can be explained by the effect of tempera­ ionic exchanges bet\Veen intra- and extracellular com­ ture on CO" solubility and on the equilibrium constants partments and between the organism and its environment and enthalpy of ionization of resident weak acids, notably (Heisler, 1986). The further requirement of constant CO" imidazole (Reeves, 1976). Ectothermic animals, however, content in each compartment has also been challengeci are open systems, producing metabolic products and ex­ because of uncertainty regarding this aspect in both wa­ changing respiratory gases with their environments, so ter-breathers (Heisler, 1986) and air-breathers (Stinner et the apparent agreement of their in vivo acid-base state aI., 1994; Stinner and Wardle, 1988) . with the intrinsic behavior of their blood in vitro suggests that physiological control is matched to the underlying OBJECTIVEs-The intense interest within the comparative chemistry (Jackson, 1982). Early comparative studies, physiology community in the pH,,-ternperature relation- Ultsch & Jackson pH AND TEMPERATURE IN ECTOTHERMS 3

ship has subsided in recent years, but the fundamental values. In addition, further heterogeneity results from the issues outlined above remain unresolved. We therefore common measurement techniques employed for pHi in believe that now is an opportune time to examine care­ comparative studies that give average values for large fully the wealth of primary data that has accumulated. We populations of sampled cells. Finally, pHi is generally do this with the following specific goals in mind: first, to measured on specific tissues or organs, for example skel­ document and analyze differences in the pH,,-tempera­ etal muscle, and these can exhibit tissue-specific values of ture relationship of both blood and intracellular fluids pHi or responses of pHi to temperature change that are among species or groups of species; second, to reassess related to their particular properties. The common the fit of these data with the alphastat and constant rela­ method for measuring pHi in comparative physiological tive alkalinity hypotheses; and third, to evaluate tempera­ studies is by distribution of the weak acid DMO

ture-dependent changes in body fluid C09 content in the (5,5-dimethyl-2,4-oxazolidinedione) (Waddell and But­ same body of data. A broader goal is to stimulate renewed ler, 1959), but data included in this review also include interest in this important topic. To this end, we have pHi measurements of red blood cells using the freeze­ compiled, on the basis of criteria defined below, a large thaw method and of skeletal muscle using in vivo 31 P­ body of data relevant to this issue for the ectothermic NMR spectroscopy. Data from comparative studies on vertebrates. pHrtemperature relations that we have assembled here are restricted to red blood cells, skeletal muscle, cardiac Data Criteria muscle, liver, and brain, as they are the cell types for which SPECIES-We have limited the scope of our survey to ecto­ pHi is most commonly determined. thermic vertebrates. We do not include data from heterothermic or hibernating mammals and birds, chiefly Blood pH vs. Temperature because ofthe fundamental difference in regulatory state ALL SPECIES-Data from 81 species, on blood acid-base that exists in these animals when they are not normother­ status as a function of temperature including multiple mic. studies of many species, at temperatures ranging from 3 to 42°C, are presented in Table 1. Values of pH" from more BLOOD DATA-Strict criteria have been used in selecting than one study at a given temperature for a single species data for this review. Blood data cited were, with very few are listed as a single point that is the grand mean of the exceptions, determined from systemic arterial samples collected individual mean values for each study (e.g., collected from chronically catheterized, spontaneously 7.869 for 36 studies of Oncorhynchus myhiss at 15°C). This ventilating, unanesthetized, and relatively undisturbed approach compresses multiple measurements into a animals. In addition, all cited data were from normoxic single value but prevents the skewing of the data that

animals at or close to sea level atmospheric levels of P09 would result from including large numbers of values for a

and PC09 , with the caveat that atmospheric levels are single, well-studied species. When pH" values from Table often approximated in respiration chambers. Finally, 1 (with the exception of Synbranchus manILoralus as ex­ cited blood measurements were made using electrodes plained in the legend) are plotted versus temperature equilibrated at the acclimation temperatures of the ani­ (Fig. 1), the slope (dpHI dT) is -0.016 U/DC. This slope is mals. The only exceptions with respect to the source of not significantly different from the predicted alphastat blood were heart puncture data that agreed with data slope (-0.018 U/DC) for this temperature range based on from catheterized animals of the same or related species; the thermal behavior of canine plasma proteins (Reeves, even then, such data were used only if they provided 1976) . information on a species or at a temperature for which Although the comprehensive data in Fig. 1 demon­ data from catheterized animals were unavailable. The strate a strong correlation between pH" and temperature condition that animals were minimally disturbed in col­ (1' = 0.813), considerable scatter is apparent that may lecting blood samples is important because it is well obscure consistent differences among species or groups known that handling and other stress-inducing interven­ of related species making up this larger population. These tions can lead to large and persistent changes in acid-base differences could either be in the slopes (dpH/dT) or in state, particularly in fishes. the elevations (i.e., the y-intercepts) of the curves. Table 2 therefore provides information on linear regression INTRACELLULAR DATA-Intracellular pH (pHi) data we cite analyses of pH" vs. temperature for individual species and were likewise taken from studies on animals subjected to for assemblages of species grouped on the basis of habitat minimal stress. In contrast to blood, a rather simple single or taxon. With few exceptions, these analyses reveal slopes compartment (i.e., plasma) in terms of pH measurement, that, like the overall ectotherm slope, are significantly the intracellular fluid of even a single cell is heteroge­ different from zero (and negative). Slopes (dpHI dT) of neous, consisting of cytoplasm and organelles, for ex­ subgroups, however, cover a broad range, from -0.004 to ample mitochondria, that can have distinctly different pH -0.022 U!"C. 4 BULLETIN 18 December 1, 1996

Table I, section A. Blood acid-base variables of freshwater and marine fishes, amphibians, and reptiles. Studies that did not use animals with chronically implanted arterial catheters are not included unless the results were comparable to those that did, or unless such studies were the only ones reported at a given temperature and the results were within the range of those found for related species (i.e., almost all of the data cited here are from studies using catheterized animals). Aquatic forms were in water reported (or assumed to be) with an 02 tension ~I 00 mmHg. When more than one study was involved, data are presented as the mean of means reported, as presented by authors in tables or graphs. When a study involved more than one group of animals, data from each group were used as if they were separate studies (e.g., two studies, one with one group and the second with three groups, is counted as total of four studies). When only two of the three acid-base variables (pH, PC02, [HC03-] ) were reported, the third was calculated using constants from Reeves (1976); (pH - pN) and a-imidazole were also calculated as in Reeves (1976).

Number of Tem perature PCO~ [HCO~-l Species Studies ("C) pH (mmHg) (111M) pH-pN" a-i m iclal.ole

Freshwater Fishes-Obligate Water Breathers

Anguilla anguilla 3 15 7.96 4.69 15.3 0.79 0.87

CY/Jl7nus emIlio 5 8.09 2.6 16.3 0.72 0.89 2 10 7.99 3.4 13.4 0.72 0.85 6 15 7.907 4.18 12.52 0.74 0.85 3 20 7.845 4.25 1 0.46 0.76 0.86 5 25 7.908 3.86 9.81 0.91 0.90

CalosloulUS rommm:mni 12.5 7.96 2.5 8.4 0.64 0.83

HO/llial' mala/}(lrirus 30 7.78 5.44 8.73 0.86 0.89 lrtaiurln /nmrtalus 15 7.929 1.80 6.20 0.76 0.86 18 7.883 3.7 9.52 0.66 0.86 20 7.868 3.1 8.1 0.79 0.87 4 22 7.831 2.49 5.02 0.78 0.87 1 23 7.83 2.7 6 0.80 0.87 26 7.75 2.5 7.34 0.77 0.87 31 7.704 3.40 4.63 0.80 0.88

Ol1rorh)lnrhu~ lII)'kiss 5 8.\0 1.95 9.78 0.73 0.88 2 6-8 7.925 2.02 6.17 0.60 0.82 5 7-10 7.93 1.66 4.92 0.62 0.82 4 9 7.903 2.68 8.10 0.61 0.82 10 10 7.901 2.53 7.80 0.63 0.82 5 II 7.874 2.50 8.05 0.62 0.82 4 13 7.930 2.17 6.50 0.72 0.85 3 14 7.833 2.34 7.34 0.64 0.83 34 15 7.869 2.61 7.28 0.70 0.84 4 16 7.8\0 3.09 7.38 0.66 0.83 I 18 7.96 1.80 6.98 0.84 0.88 20 7.76 2.03 3.78 0.68 0.84

Pi'trom),zon I/Ulrinus 2 10 7.98 1.76 6.84 0.71 0.85

Saimo tnltla 5 8.01 1.5 5.3 0.64 0.83 2 15 7.85 1.75 4.03 0.68 0.84

Saivl'linus jontinaiis 11 7.957 2.28 9.74 0.71 0.85 Ultsch & Jackson pH AND TEMPERATURE IN ECTOTHERMS 5

Table 1. (rol1tinul'd)

Number [HC0 -] of Temperature PC02 3 Species Studies (OC) pH (mmHg) (mM) pH-pN", a-imidazole

1'h)'lIIaUus arrtirus 10 8.02 1.8 8.4 0.75 0.86 17 7.89 2.3 8.4 0.75 0.86

1'il1(([ linea 2 13 8.164 3.30 18.05 0.98 0.91 14 7.985 2.90 10.94 0.80 0.87 2 15 7.996 3.2 11.3 0.82 0.88 16 8.10 2.5 11.4 0.95 0.91

Marine Fishes-Obligate Water Breathers

Congn rollgl'l' 17 7.86 1.95 5.15 0.72 0.85

G),nosriol1 aH'lwrius 18 7.83 2.13 4.36 0.71 0.85 28 7.70 2.69 3.60 0.75 0.86

Dirl'l1tmrrhu~ labmx 2 10 8.005 4.58 19.2 0.74 0.86

Hl'lllilri/Jlerul alll1'l1mnUS 10-11 7.90 1.81 5.41 0.64 0.83

HijJf)()glossoides l'lassor/ol1 3 11.5 7.828 2.04 5.20 0.59 0.80

O/Jsal1m liI'la 24 7.821 2.11 5.92 0.81 0.88

"Oncorhynchus lII)'liiss 7 10 7.947 2.38 9.55 0.68 0.84 12 7.78 2.65 7.0 0.55 0.79 15 7.85 2.7 8.5 0.68 0.84

Ol1corliYllflius lii.llltch 3 9-11 7.836 2.31 5.99 0.56 0.80 I 10-12 7.83 3.49 9.81 0.58 0.81 13 7.77 2.5 6.06 0.56 0.80

Paro/J!t'j'l' VI'lullll' 2 II 7.75 2.02 4.82 0.50 0.78

Platichthys sit'llalus 4 9 7.847 2.45 5.95 0.56 0.80 3 9-11 7.808 2.74 6.43 0.54 0.79 II 7.82 3.3 9.6 0.57 0.80

Rajll Ofella I a 2 12 7.842 0.82 3.12 0.61 0.82

SllllIIO satar 12 7.94 2.21 6.96 0.71 0.85

SI),liorhi 1111.1' milieu/a 15 7.841 0.55 1.39 0.67 0.83

SI)'liorhinlls lit'llaris 2 Hi 7.86 2.0() 7.12 0.71 0.85 4 17 7.788 2.04 4.81 0.65 0.83

Squa/us afllllihit/l" 16-19 7.87 2.07 6.19 0.74 0.86

Freshwater Fishes-Facultative Air-breathers

Alllia mlva " I 24.5 7.733 5.45 8.43 0.61 0.85 " I 30 7.53 7.73 g.40 0.61 0.82 6 BULLETIN 18 December 1,1996

Table 1. (continued)

Number -] of Temperature PC02 [HCOg Species Studies (0C) pH (mmHg) (mM) pH-pN" a-imidazole

Clanas batrachus 28 7.60 3.5 4 0.65 0.83

Hoplerythnnus unitaeniatus 27.5 7.807 11.09 19.53 0.84 0.88 30 7.87 6.07 11.97 0.95 0.91

Neoceratodus fosten 18 7.64 3.6 4.89 0.52 0.78

Lepisosteus oClllatus 2 20 7.762 5.6 8.95 0.68 0.84 2 26 7.64 7.4 7.94 0.66 0.84 30 7.59 10.5 9.5 0.67 0.84

Synbranchus marmoratus 30 8.17 5.6 24.2 1.25 0.95

Freshwater Fishes-Obligate Air-breathers

Arapaima gigas 28-30 7.576 26.64 26.94 0.65 0.83

Channa argus 2 25 7.540 15.8 17.4 0.54 0.79

C ElectrojJ/wrus electncus 2 28 7.56 28.2 28.5 0.62 0.82

Protopterus aethiopicus 3 25 7.607 27.8 32.3 0.61 0.81

Aquatic Salamanders-Facultative Air-breathers

Ambystoma tigrinum-Iarvae 3 5 8.06 4.93 21.87 0.69 0.83 6 15 7.850 10.47 24.57 0.68 0.83 1 20 7.818 8.40 24.58 0.74 0.86 21 25 7.785 9.17 16.86 0.78 0.87

Cryptobranchus alleganiensis 5 8.09 2.03 9.5 0.72 0.85 1 15 7.90 4.20 10.7 0.73 0.85 3 25 7.795 6.43 10.50 0.80 0.87

Necturus maculosus 5 8.05 4.9 19.5 0.68 0.84 15 7.90 4.5 12 0.73 0.85 1 20 7.661 4.42 6.38 0.58 0.80 2 25 7.700 6.25 9.10 0.70 0.85

Aquatic Salamanders-Obligate Air-breathers

Amphiuma means 5 8.15 5.8 30.4 0.78 0.87 2 25 7.765 14.90 25.55 0.76 0.86

Sh·en lacertina 25 7.79 11.7 22.0 0.79 0.87

Terrestrial Salamanders

Amb),stoma tigrinum- adults 5 8.06 9.1 35.8 0.69 0.84 Ultsch & Jackson pH AND TEMPERATURE IN ECTOTHERMS 7

Table 1. (continued)

Number

of Temperature PC02 [HCO~-] Species Studies (0C) pH (mmHg) (mM) pH-pN" o:-imidazole

15 7.92 10.2 27 0.75 0.86 2 25 7.736 12.35 19.50 0.74 0.86

Anurans

Bufo marinus 10 8.05 4.8 22.3 0.78 0.87 13 20 7.913 8.76 24.99 0.83 0.88 1 23 7.84 11.7 23.9 0.81 0.88 11 25 7.814 11.34 20.67 0.81 0.88 30 7.75 11.6 23.5 0.83 0.88

Bufo jmmcnemis 15 7.92 9.32 24.99 0.75 0.86 32 7.70 14.6 18.97 0.81 0.88

Bufo viridis 2 21 7.720 10.8 18.6 0.65 0.83 4 23 7.675 10.9 15.8 0.64 0.82 3 26 7.661 12.6 16.7 0.68 0.84

Ph)lllomedusa sauvagl'i 24 7.705 13.4 19.3 0.70 0.84

Rana ralesbeian(/. 3 5 8.161 4.70 24.06 0.80 0.87 3 10 8.020 7.67 27.08 0.75 0.86 3 15 7.923 10.52 30.16 0.75 0.86 7 20 7.880 12.14 27.81 0.80 0.86 2 21 7.892 10.9 27.96 0.84 0.88 6 25 7.873 11.98 25.73 0.87 0.89 2 30 7.755 16.85 26.66 0.84 0.88 34 7.67 23.1 24.05 0.81 0.88

Xenojitts tarois 1 10 7.95 6.4 23.5 0.68 0.84 2 20 7.759 13.75 30.34 0.68 0.84 3 25 7.741 15.43 27.03 0.74 0.86 30 7.61 19 21.4 0.69 0.84

Turtles

A/mtone sjJinifem 10 7.945 8.11 24.14 0.67 0.84

Carl'lI a carella 25 7.63 17.2 24.8 0.63 0.82

Chelonia lIlydns 15 7.55 25.3 27.2 0.38 0.72 2 25 7.56 30.0 32.0 0.56 0.80 28 7.47 45.7 36.3 0.52 0.79 35 7.36 46.0 27.1 0.52 0.79

Chl'l),dm sellJcntiu(/. 10 7.876 15.69 39.74 0.61 0.82 ell 22-24 7.692 36.0 51.6 0.63 0.83

Chr)'sell!)'s jJirla bl'llii 3 3 8.014 9.82 39.12 0.60 0.82 1 5 7.88 13 39.6 0.51 0.78 6 10 7.865 15.22 39.12 0.60 0.81 8 BULLETIN 18 Decem ber 1, 1996

Table 1. (rolllill'lted)

Number -] of Temperature PCO~ [HCO:1 Species Studies (DC) pH (mmHg) (mM) pH-pN" a-imidazole

1 15 7.862 15.5 ~9.0 0.69 0.84 8 20 7.748 2~.19 40.05 0.67 0.83 3 30 7.626 33.23 40.07 0.70 0.85

Chr),sem)'s jJicla dm:wiis 3 7.975 10.88 ~7.4 0.57 0.80

CojJ/wrus jJOi)'jJhemIH 25 7.608 22.87 27.84 0.61 0.82

CmjJlem)'s gl!ogmjJ!tim 3 7.960 7.83 29.49 0.55 0.80

Maiacochenus lomin'i 20 7.64 15.1 21.4 0.56 0.80 35 7.47 30.9 22.9 0.63 0.83

Podorl1l!lIlis l!xjJrlU \(/ 28 7.57 31 31.8 0.62 0.82

P~l!lld(,//I)'S jloritillll(l 22.5 7.69 20.82 29.56 0.65 0.83 30.7 7.59 29.70 31.05 0.68 0.84 36.2 7.51 34.78 28.20 0.68 0.85

Slemol/tI'l"lts odomlus 2 3 7.925 8.44 28.08 0.52 0.78 2 10 7.868 10.9 28.9 0.60 0.81 lI!studo II/bulllill 28 7.55 25 25 0.60 0.81

7)"(/c1wlII)'-~ srrijJIIl I!/I'gll /IS 2 10 7.80 14.05 33.66 0.54 0.79 4 20 7.69 22'<38 35.46 0.61 0.81 24 7.604 31.8 ~7.58 0.59 0.81 2 ~o 7.57 ~2.95 ~4.00 0.65 0.84

Crocodilians

Alligillor mississijJjJil'1I \'i ~ 2 15 7.658 11.8 16.5 0.48 0.77 2 25 7.491 17.7 15.~ 0.49 0.77 2 ~5 7.~82 2~.6 14.2 0.54 0.80

Crorotlyills j)()roSlls 30 7.431 32.8 2~.5 0.51 0.78

Lilards

A III bl)'llty II rhus rrislalus 15-17 7.67 13.71 16.68 0.52 0.78 2~-25 7.60 15.16 18.23 0.59 0.81 34-36 7.44 23.84 16,76 0.60 0.82

DijJsOSallrus dOI:~alis 17.5 7.795 32 18.7 0.66 0.84 2 25 7.778 21 20.4 0.78 0.86 32 7.585 19 21.3 0.70 0.85 35 7.58 19.5 18.5 0.74 0.86 37 7.512 14 18.2 0.70 (J.8S 41.5 7.447 9 19.2 0.70 (J.85 42 7.46 32 22.4 (J.n (J .86 Ultsch & Jackson pH AND TEMPERATURE IN ECTOTHERMS 9

Table 1. (continued)

Number [HCO -] of Temperature PC02 g (0C) Species Studies pH (mmHg) (mM) pH-pNw a-imidazole

Iguana iguana 2 35 7.48 31.4 23.8 0.64 0.83

Physignathus lesueurii 1 19 7.78 10.2 24.4 0.68 0.84 1 22 7.76 9.4 28.4 0.71 0.85 1 23 7.68 12.9 20.9 0.65 0.83 1 24 7.71 12.1 24.8 0.70 0.84 29 7.62 10.8 21.2 0.69 0.84

Tupinambis nigromaculatus 2 25 7.59 20.2 21.3 0.59 0.81

Vamnus nitoticus 2 25 7.56 27.6 34 0.56 0.80 35 7.52 31 35 0.68 0.84

Vamnus exanthemalicus 1 20 7.56 19.84 22.31 0.48 0.76 2 25 7.545 19.34 19.78 0.54 0.80 1 30 7.51 23.34 20.91 0.59 0.81 4 35 7.488 28.07 22.62 0.65 0.83

Vamnus salvator 1 25 7.57 22.6 23.9 0.57 0.80 2 35 7.53 29.4 28.6 0.69 0.85

Snakes

Acrochonlus amJume 25 7.517 37.0 34.6 0.52 0.78

Coluber constrictor 3 15 7.626 12.3 19.8 0.45 0.76 2 20 7.608 13.0 17.8 0.53 0.78 3 25 7.577 15.5 17.9 0.58 0.81 2 30 7.530 19.4 18.1 0.61 0.82 3 35 7.471 21.2 16.2 0.63 0.82 2 40 7.365 28.2 15.5 0.60 0.82

Constrictor ronstrictor 28 7.52 15 14 0.57 0.80

LajJelnis hm'dwicllii 25 7.41 25.2 18.4 0.41 0.74

Pituo/lhis melanoleucus 20 7.693 13.0 19.40 0.61 0.82 30 7.536 20.6 18.87 0.62 0.82

.1 Sea-water adapted rainbow trout h Water PCO~-4 mm Hg c Water P02 hypoxic (ca. 48 mm Hg) in study by Garey & Rahn (1970), bm acid-base values are included here as they are similar to those found by.lohansen et al. (1968) and because the VO~ of the fish is primarily aerial. Ii Values shown here are averages of those at the ends ofventilatory and nonventilatory periods 10 BULLETIN 18 December 1,1996

']. Table 1, section B. Additional data from papers that reported arterial plasma pH but did not report PC02 or [HC03 These data meet the same criteria set for those in section A of this table, and are incorporated as appropriate into all analyses of relation­ ships between pH and temperature.

Species Temperature (0C) pH Source

Freshwater Fishes-Obligate Water-breathers

Aripellser tmnS11l0ntanllS 15 7.84 Burggren, 1978

Nfgaj)rion breuil'Ostris 24 7.71 Bushnell et aI., 1982

Oncorhynchus lnykiss 5 8.094 Holeton, 1977 5-7.5 8.115 Primmett et aI., 1986 9 8.02 Wright, Wood & Randall,198B 10 8.011 Tufts et aI., 1988 10 8.048 Tufts et aI., 1988 10 7.934 Tufts et aI., 1988 10 8.068 Tufts et aI., 1988 10 7.91 Ye et aI., 1991 10 7.948 Boutilier et aI., 1986 10 7.909 Tang & Boutilier, 1988b 12 7.99 Perry et aI., 1989 12 7.99 Perry et aI., 1989 15 7.90 Wilson et aI., 1994 15 7.729 Cameron & Heisler, 1983 15-16 7.90 Perry et aI., 1989 15-16 8.07 Perry et aI., 1989 15-16 7.83 Perry et aI., 1989 15-16 7.90 Perry et aI., 1989 20 7.8 Randall & Cameron, 1973

Marine Fishes-Obligate Water-breathers

Oncorhynchus 'IIIykiss 10 7.92 Tang & Boutilier, 1988b (Seawater-adapted)

Pialichlh),s stella/us 7.5-10.5 7.872 Wood et aI., 1979

P.5fudoj)ifltl'Onl!ctes amel'icanuJ 5 7.90 Cech et aI., 1976 10 7.86 Cech et aI., 1976 10 7.86 Cech et aI., 1977 15 7.83 Cech et aI., 1976

Sallllo safar 10 8.113 Ferguson & Boutilier, 1988 10 8.045 Ferguson & Boutilier, 1988

Sryiiorhinlls mnicuia 7 7.885 Butler & Taylor, 1975 12 7.813 Butler & Taylor, 1975 15 7.76 Short et aI., 1979 17 7.743 Butler & Taylor, 1975

TOIj)('ilomanllol'(//a 16 7.9 Hughes &Johnston, 1978

Anurans

BuJo 'mari711tS 22 7.89 Tufts et aI., 1987

BuJo /Htmml'lItis 15 7.95 Branco et aI., 1993 25 7.82 Branco et aI., 1993 27 7.83 Wang. Branco & Glass,1994 Ultsch & Jackson pH AND TEMPERATURE IN ECTOTHERMS 11

Table lb. (rnnlinued)

Species Temperature (0C) pH Source

35 7.79 Branco et aI., 1993

XfI10jms lal'Vis 20 7.601 Boutilier & Lantz, 1989

Lizards

Iguana iguana 20 7.64 Wood & Moberly, 1970 30 7.50 Wood & Moberly, 1970 35 7.47 Wood & Moberly, 1970

Snakes

Crotalus r/uriss1ts 25 7.63 Wang et aI., 1993

Neror/m IJijJPf{ol1 15 7.73 Dean & Gratz, 1983 25 7.59 Dean & Gratz, 1983 30 7.55 Dean & Gratz, 1983 15 7.67 Gratz, 1984 25 7.54 Gratz, 1984 30 7.48 Gratz, 1984

REFERENCES: FISHES- tllIlia mlva (Randall et aI., 1981); tlIIg1lil/a rlllg1li/la (Bornancin et aI., 1977; Thomas et aI., 1980; Wood &Johansen, 1973); tlm/Jllillla giga.l' (Randall et aI., I 978b) ; Call1sllllllll.l' rtllIlllwt:WJtli (Hobe et aI., 1983); Chal/I/a arg1l.l' (Ishimatsu & ltaZal\~I, 1983); C/mias balmrllll.l' (Rahn & Care)" 1973); ClIlIger mllg{!/' (Toews et aI., 1983); Cy"oscirJtl rmmmills (Cameron, 1978); G),/JliIlIlS rw11ill (Claiborne & Heisler, 1984, 1986; Dt:jours, 1973; Dt:jours & Armand, 1973; Fuchs & Albers, 1988; Class et aI., 1990; Itazawa & Takeda, 1978;Jensen et aI., 1987; Takeda, 1990, 1991; Ultsch et aI., 1981, van DUk et aI., 1993); Dir""lmrrhlls /abmx (Thomas & Hughes, 1982a); E/n.·lm/I/lIIl'IIs "'I'rllims (Gare)' & Rahn, I 970;Johansen etal., 19(8); HI!lIIifli/I/I!l'IIs all/(!/imllllS (Milligan & Farrell, I 98fi); Hi/I/lIIg/OSSllidl's 1'/((.ISm/1II1 (Turner, Wood & Hobe, 1983); HO/I/I'I)'1/lIillll.l IlIIi/(((mia/II.I' (Cameron & Wood, 1978; Randall et aI., I 978a) ; I-Ill/i/ias lIIa/ablllil'lI.I' (Cameron & Wood, 1978); Ir./a/llnts /l/Il/r/a/II.1' (Burggren & Cameron, 1980; Cameron, 1980, 1985; Cameron & Iwama, 1987; Cameron & KOl'lnanik, 1982a,b); Le/li.l'os/I!IIS lim/a/liS (Smatresk & Cameron, 1982a,b); NI'llInll/lIdlls(tIS/I!I7' (Lenlill1t et aI., 1966/(7); 011 cllrh)'I/(h11.1' hi,HI/l'h (Milligan et aI., 1991; Perry, 1982; van den Thillart et aI., 1983); Olcllrhy"r!lIIs lIIyhiss (Andreasen, 1985; Booth et aI., 1982; Boutilier et aI., 1988; Cameron & Randall, 1972; Currie & Tufts, 1993; Edd), et aI., 1977; CihllOlIl' & Perry, 1994; Gilmour et aI., 1994; Goss & Wood. 1990, 1991; Hobe et al.. 1984; Holeton. Booth &Jansz, 1983; Holeton. Neumann & Heisler • 1~)83/I,Janssen & Randall, 1975; Kieffer et al.. 1994; Larsen &Jensen. 1993; Mmdme et al.. 1991; McDonald. 1983; McDonald, Hobe & \Yood. 1980; McDonald & Prior. 1988; McDonald & Wood. 1981; Milligan & Wood. I 986a,/1, Perr),. 1982; Perry & Thomas. 1991; Perry etal., 1981; Tang & Boutilier. I 988a; Tang et al.. 1988, I 989a,/1, Thomas. 1983; Thomas & Hughes. 1982/1, Thomas & Le Ruz. 1982; Thomas & Poupin. 198.~; Thomas et al.. 1980, 1983. 1986; Turner. Wood & Clark. 1983; Wilkie & Wood. 1991; Wilson & Ta),lor. 1992; Wilson et al.. 1994; Wood &Jackson, 1980; Wood & LeMoigne, 1991, Wood & Munger. 1994; Wood, Walsh. Thomas & PelT)'. 1990; Wright & Wood, 1985); O/mlllll., bl'/a (Barber & Walsh, 1993); Prml/lhl~l'S 111'/III!n (McDonald et aI., 1982; Wright, Randall & Wood, 1988); I'l'Imlll)'ZIIII IIImillll.l' (Tulb. 1991; Tufts et al.. 1992); I'/a/icillhys s/ella/IIS (Milligan & Wood. 1987 a.b; Milligan et al.. 1991; ""ood & Milligan, 1987; Wood. C.M. et aI., 1977. 1979); I'ro/O/I/I'ntS al'lhill/lims (Delane), et al.. 1974. 1977; Lahiri et al.. 1970); Rairlllo'/Ia/a (Graham. et OIL. 1990); .')a/lIIlIsa/ar (Maxime etal.. 1990); Sa/lilli/rill/a (Butler & Da)" 1993a.b); Sa/vl'iillllsjiJtl/illa/iJ (Wood et al.. 1988); SI)'/illrhilllls rrtl/imla (Tnlchot et aI., 1980); Sc:y/illlhil/IIS sM/mis (Heisler, 1982a; lIeisler et al.. 1976a,b. 1988; Piiper & Schumann, 1967; Piiper et aI., 1972); S'I'la/lls a((lll/hias (Claiborne & Evans, 1982); Sy""m"rllllol IIIlIrlllom//I.I (Hei~ler, 1982"); '1111'111111111\' IIrr/irlll' (Cameron, 197(1); '/)'II((J /illm (Eddy. 1974;.Jensen. 1987; .Jensen & 'Nebel', 1985, 1987), SALAMANDERS-tllII/~'.I/1I1II11 /iglill II III larvae (Aguinaga & Stiffler. 1993; Burggren & Wood, 1981; Crocker & Stiffler, 1991; DeRuyter & Stiffler, 1988; l'licks & Stiffler. 1980, 1984; Rohrbach & Stiffler, 1987; Stiffler, 1991; Stiffler & Bachoura, 1991; Stiffler et al.. 1983, 1987. 1990; Talbot & StillIer, 1991. 1992; Wood etal.. 1989); tllII/~'.\/IIIIIII/iglillllllladuIL~ (l'licks &Stiml'r. 1984; Stiffler ,1991); AIII/I/lllilllll III('((IIS (Heisleretal.. 1982; I-licks & Stiffler, 1984); c,),/I/II/I/(/lIrhll,I'lIl1l'gallil!ll.lis (Boutilier & Toews. 1981; Boutilier et al.. 1980; Moalli et aI., 198 I); NI'r/lIl'11\ IIIII(/t/II.\II., (l'licks & Stimer. 1984; Stimer et aI., 1983; Wood et al.. 1989); Simll /l1l'1'r/il/ll (I'leisler et aI., 1982). ANURANS-/J/I/II IIImillll\' (Boutilier, 1984; Boutilier & Heisler, 1988. BOlllilier et al.. 197911,",1'. 1980. 1987; Garland & Toews. 1992; McDonald. Boutilier & Toews, 1980; l'iirtner etal.. 1991; Snyder & Nestler, 1991; StillIer & Toews. 1992; Toews & Heisler, 1982; Toews & Kirby. I ~)85; Toews & Stiffler, 1990; Tufts & Toews. 1985. I 98(); ",,rood et al.. 1989); Bllfil/lllmwl'lIIis (Kruh0lfer et al.. 1(87); /J1I/illlilidi.l (Katz, I 980a,b. 1981); I'hy/lllllln/II,m J({1I11agl'i (Stinnel' & Shoemaker, 1987); Hrll/II m/I',III'illl/a (Gottlieb &Jackson. 1986; Howell etal., I 970;Jacksnn & Braun. 1979; Kinkead & r-'Iilsom, 1994; Lillo. 1978; Lindinger & McDonald, 1986; r-lacKelllie &Jackson, 1978; Pindel; 1987; Reeves. 1972; Tazawa et al.. 1979; Toews & Stiffler. 1990; Warburton et aI., 1989; Wasser et .11., 1993; Wood et al.. 1989); XI'IIO/I/I\' 1t1l~lis (Boutilier. 1984; Boutilier & Shelton. 1986; Boutilier et al.. 1987; Emilio & Shelton. 1980; Wond et aI., 1989). REI'TILES-tlITor/llm/II' am/llml' (Seymour et 011..1981); illliga/orllli,\.\issijlflil!llsis (Davies, 1978; D,l\'ies etal., 1982); Alllb~rdl.l' "dlllol (.'/i"/IIII1.1 (Ackerman & While. 1980); A/IfI/IIIII's/lilli/l'm (Ultsch l·t aI., 1984); Cllrt'l/a mrl'l/a (Lutcamge & Lutz. 1991); CIII'/Ollill IIIJI/I101 (Jackson & Prange. 1979; Kraus &Jackson, 1980; Wood et al.. 1984); ClII'/y/m 1'I'11"'I1/illll (UIL~ch et aI., 1984; West et OIl.. 198~j); C/II)'selllys/lir/a hl'/Iii (Glass et al.. 1983. 1985; Herbert & Jackson. 1985; Jackson & Heisler. 1983; Nicol et al.. 1983; Silver & Jackson. 1985. 1986; UIL~ch. 1989; Ultsch & Jackson. 1982; Ultsch et a!., 1984; Wasser &.Jackson. 1988; Wassel l't aI., 19~)]); OIlYI'I"I/YI' /Iir/II dona/i, (Ultsch et aI., 1985); CII/II/II'r mlls/lidor (Nolan & Frankel. 1982; Stinner. 1987; Stinner & Wardle, 1988); CIII/.\/lic/m (IJ"'/Iirlllr (Rahn & Garey, 1973); CI1Imdyhls /10111""' (Se)'lllour et a!.. 1985); /)i/ISII.\(I/I/'II.I dona/is (Bickler, 1981. 1984); GII/II",I'IIS /IIJ/Y/I/WIIIIIS (Ultsch. 1987); Gm/I/I!IIIYS gl'"gm/lhim (Ultsch &Jackson, 1995); IgUlIlIII igual/a (Gleeson & Bennett, 1982; Milchell el al.. 1981); LII/II"lIis hrm/widlii (Seymour & Webster, 1975); Ma/llmdwwls /ol'llimi (Wood et al.. 1978); I'hysigllll/I/IIIIS it',,,wlllii (Courtice. 1981); I Ji/llo/lhis IIIdlll/o/mlrllS (Slinner. 1982); 1'0dl}(;III!IIIYS I'X/IIIIISII (Rahn & Garey. 1973); PI'I'lIdl'lIIys/loridlllla (Kinney et OIl.. 1977); S/I'''''"//'''I'IIS odom/lls (Jackson et 011.,1988; Ultsch, 1988; UIL~ch & Cochran, 1994; Ultsch etal.. 1984); '/('S/IIl/II/II/I11/a/1l (Rahn & Gare),. 1973); '/1'(11'111'111),.1' sfli/I/a I~it'glllls (Jackson & Kagen. 197(j; Hitzig. 1982; Hitzig & Nattie, 1982;Jackson & Kagen, 1976;Jackson & Silverblatt. 1974;Jackson et al.. 1974); '/II/lilllllllbis lIigl1l/lIIlIcla/lIs (Glass & !'leislel; 198(1); I'tmlll/IS e,wlllllllmlll/iC/l" (Gleeson & Bennett. I ~)82; Milchell et al.. 1981; Wood. S.C. et al.. 1977, 1981); '''mlll/IS lIi/o/it:IIS (l'licks et aI., 1987; Wood & Johansen, 1974); 'lrmllllls.m/1I11/lir (Gleeson & Bennett. 1982; Mitchell & Gleeson. 1985), 12 BULLETIN 18 December 1, 1996

8 .2 II " ter teleosts also shows no significant difference, sowe have 8.1 A" II " combined these two groups. Reptiles are a diverse group, 8 .0 a" I 4 I:J.Il. and a sizable body of data is available on all major orders, A C. J'IIS: "".0. 6 "~ b including snakes, lizards, turtles, and crocodilians. As 7 .9 " A b. I:J. 1""".0. .aftll. II .o.~ " shown in Table 3, significant differences exist in the eleva­ 7 .8 A~" " " I "" " tions of the pHb-temperature curve for several of these " " A" " a. 7.7 "" groups of reptiles, but the curve for the lizards, the rep­ ",," " " 7.6 " " " tiles with the highest elevation, is still significantly below " " "f the curve for the freshwater teleosts and amphibians. We 7.5 " " tf' have therefore combined the reptiles and have compared 7.4 " " them as a single group with the freshwater teleosts and " 7.3 +1---,,---,----.-----,------,--,---.------,------, amphibians (Fig. 3) . This illustration reveals a rather o 5 10 15 20 25 30 35 40 45 striking displacement in these curves with little overlap of TEMPERATURE (C) values. Clearly, the reptiles are regulating their blood pH at a substantially lower level than the freshwater teleosts Figure 1. Arterial pH as a function of temperature in and amphibians, but again the slopes for the change in ectotherms. Each of the 207 values represents one of 81 pHb with temperature are similar. species at a given temperature (see Table 1). Species A similar separation is revealed if freshwater species, included are those acclimated to their usual environment, including fishes, amphibians, and reptiles, are compared and also those euryhaline forms that can be acclimated to with marine species, including fishes and reptiles (Fig. 4). either freshwater or seawater (e.g., Petromyzon marinus in The slopes of these two curves are not significantly differ­ freshwater and Oncorhynchus mykiss in seawater). One ent, but the elevation (intercept) of the curve for freshwa­ temperature-species point is the mean of means of study ter species is significantly higher than that for the marine groups reported for that species at that temperature, as species. detailed in Table 1. A further broad inter-group comparison that includes all the data plotted in Fig. 1 reveals a significantly lower pH-intercept for amniotes than for anamniotes (Fig. 5) . INTER-GROUP COMPARISONS-An objective in creating Table Anamniotes involved in this comparison include amphib­ 2 was to assemble species taxonomically (e.g., freshwater ians and water-breathing fishes, both freshwater and ma- teleosts), to make statistical comparisons, using analysis of covariance, between such groups, and then to create even larger groupings by combining subgroups that do not differ from each other in either slope or elevation. Com­ parisons among groups for both slope and elevation are 8 .2 shown in Table 3. Space does not allow us to discuss all the possible permutations, but several broad relationships 8.1 emerge from this analysis. A first comparison concerns o FRESHWATER TELEOSTS 8.0 fishes. The pHb-temperature curve for freshwater teleosts m=-.012 has a significantly higher elevation when compared to I either marine teleosts or elasmobranchs, but marine te­ a. 7.9 leosts and elasmobranchs do not differ from each other. We therefore have combined these latter two groups and 7 .8 show the comparison between freshwater teleosts and marine fishes in Fig. 2. As for nearly all the comparisons to 7.7 - be described, the slopes of these regression lines in Fig. 2 are not statistically different but the elevations are. This 7.6 means that a freshwater teleost will usually have a higher 0 5 10 15 20 25 30 35 pHb than a marine teleost or elasmobranch tested at the TE MP ERATURE (DC) same temperature, but if temperature changes the pHb values will change in similar fashion. Figure 2. Arterial pH as a function of temperature in In the various amphibian orders, comparison of the freshwater teleosts (excluding air-breathing species) pHb-temperature relationship reveals no significant dif­ compared to elasmobranchs plus marine teleosts (not ferences in either slope or elevation, so we have combined including seawater-adapted forms that are typically all amphibians into a single group. A comparison of the freshwater, such as seawater-adapted Ollcorhynchus mykiss). resultant amphibian curve with the curve of the freshwa- Individual species-temperature data points are as in Fig. 1. Ultsch & Jackson pH AND TEMPERATURE IN ECTOTHERMS 13

Table 2. Regression data for extracellular (plasma) pH and intracellular pH as a function of temperature for various species and groups of species of ectotherms. Data are for arterial blood of resting animals, sampled from indwelling catheters. When an analysis of data for extracellular pH includes air-breathing fishes, SY1lbra1lchus marmoratus is not included because of its uniquely high pH; analyses of intracellular pH include this species. Regression equations for individual species use all data available at all temperatures. For equations for species groups, one data point is comprised of the mean of data for one species at a given temperature, which ranges from a single study of a species to 36 study groups, where a given report may have had more than one study group. For example, 01lcorhYllchus mykiss at 15°C: 36 study groups (see Table 1, with 34 studies from section A and 2 from B) does not mean 36 separate reports; all 36 data points are used in the regression analysis for the species, only the grand mean value at 15°C is used in any analysis that includes this species in a species group, so as to not unduly weight the multispecies regression toward a single species. In addition, in the accounting of number of species, the larval and adult forms of the salamander Ambystoma tigrillllm are counted separately, as are the subspecies of Chrysemys picta (bellii and dorsalis). If P (slope) is (~0.0500, the slope is significantly different from zero. Values not reported or uncalcuable from reported data are indicated by -. The intercept is the pH predicted by the regression at O°C.

Temp- 95% rature Number Number confidence range of of data interval of P Group (0C) species points Slope slope Intercept r (slope)

Extracellular pH-individual species

Oncorhynchus my/I iss (freshwater) 5-20 94 -0.014 -0.009 to -0.019 8.079 0.531 *** Cyjirinlls ull/lio 5-25 17 -0.007 0.007 to -0.014 8.033 0.441 0.0762 I ria /uru s Inl'lu:la/Us 15-31 II -0.014 -0.012 to -0.017 8.150 0.966 *** Ir/all/rlls jmnc/a/us (Cameron & Kormanik, 1982) 14-33 51 -0.013 8.120 0.95 AlIlb)'s/oll/a ligrinum (larval) 5-25 30 -0.011 -0.007 to -0.015 8.052 0.706 *** Ambys/oll/(l IigTinulI/ (metamorphosed adult) 5-25 4 -0.016 -0.011to-0.021 8.150 0.995 0.0048 CI)'II/o/mmdms alleganiellsis 5-25 5 -0.014 -0.006 to -0.022 8.145 0.953 0.0121 Nu/urus II/(lru[osus 5-25 5 -0.019 -0.004 to -0.033 8.138 0.921 0.0262 13ufo II/arinlls 10-30 28 -0.018 -0.014 to -0.021 8.256 0.882 *** X/!/IOIIIIS [rllmis 10-30 9 -0.013 -0.002 to -0.025 8.020 0.711 0.0319 Rrm(l. ca/esbeimw 5-34 27 -0.014 -0.011 to -0.017 8.185 0.890 *** .Ina/la wtr!sbr!ial1a (Malan et a!. , 197fi) 3.5-30 52 -0.021 -0 .016 to -0.025 8.184 0.79 Alligator lIIississijJlliensis 15-35 6 -0.014 -0.0 II to -0.017 7.857 0.985 0.0003 Tmc/wlII)'.I· srrijl/a eir!gans 10-30 9 -0.012 -0.008 to -0.016 7.926 0.931 0.0003

.1 'Fmc/II!III)'.I' mill/a eiegans (Malan et aI., 197fi) 9-32 lfi -0,021 -0,019 to -0.023 8,092 0.99 Chry,wmlYs Ilir/(1 bellii 3-30 22 -0.013 -0.011 to -0.015 8.011 0.947 *** Chdollia lIIydas 15-35 5 -0.010 0.009 to -0.028 7.749 0.694 0.1933 PI'r!urh!IIlYs jloridalla 22.5-36.2 3 -0.013 -0.005 to -0.022 7.986 0.999 0.0313 \!rlra/lus e.wllI/hellla/ir:us 20-35 8 -0.005 -0.001 to -0.011 7.670 0.951 0.0318 Amb/yhynrhus r;ris/a/us 16-35 3 -0.012 0.008 to -0.033 7.876 0.991 0.0836 Di/mmulru,1 do/:Hlli,~ 17.5-42 8 -0.016 -0.009 to -0.023 8.135 0.917 0.0013 Igualla igurl/Irl 20-35 5 -0.010 -0.006 to -0.014 7.824 0.975 0.0046 Physiglla/hll.1 II'slII!lIrii 19-29 5 -0.016 -0.007 to -0.025 8.093 0.931 0.0213 Co/ub/!/' ((}/ls/rir/or 15-40 15 -0.010 -0.007 to -0.012 7.797 0.931 *** N{!/'oilia silll't/o/l 15-35 (j -0.013 -O.OOfi to -0.019 7.885 0.930 0.0071

Extracellular pH-species grouped by taxonomy and habitat

Freshwater teleosts (excluding air-breathers and marine salmonids in fre~hlVater; e.g., Suill/o su/ar) 5-31 II 40 -0.012 -0.008 to -0,015 8.097 0.714 *** Air-breathing (facultative and obligate) freshwater teleosts 18-30 9 13 -0.004 -0.015 to -0.022 7.753 0.137 0.655('; l'vlarine te1eosts (excluding seawater-adapted freshwater species) 5-28 II 19 -0.007 0.000 to -0.015 7.949 0.468 0.0433 14 BULLETIN 18 December 1,1996

Table 2. (continued)

Temp- 95% rature Number Number confidence range of of data interval of P Group (0C) species points Slope slope Intercept r (slope)

Teleosts, marine (in seawater) and freshwater (in freshwater), without air-breathers 5-31 22 59 -0.009 -0.006 to -0.013 8.036 0.560 *** Marine teleosts (excluding salmonids) 5-28 9 14 -0.006 0.000 to -0.012 7.925 0.555 0.0394 Marine salmonids (including seawater-adapted Oncorhynchus mykiss) 10-20 3 9 -0.020 0.004 to -0.044 8.111 0.593 0.0926 Elasmobranchs 7-24 6 10 -0.009 0.000 to -0.018 7.953 0.616 0.0577 Elasmobranchs and marine teleosts 5-28 17 29 -0.008 -0.003 to -0.013 7.953 0.523 0.0036 Aquatic salamanders 5-25 5 14 -0.017 -0.012 to -0.021 8.153 0.920 *** Anurans 5-35 6 27 -0.013 -0.009 to -0.018 8.117 0.778 *** Amphibians (aquatic and terrestrial) 5-35 12 44 -0.014 -0.011 to -0.017 8.126 0.849 *** Amphibians and freshwater teleosts 5-35 23 84 -0.014 -0.011 to -0.016 8.120 0.823 *** Turtles 3-36.2 14 30 -0.016 -0.013 to -0.018 8.005 0.927 *** Snakes 15-40 7 15 -0.011 -0.006 to -0.016 7.841 0.814 0.0002 Lizards 16-42 8 27 -0.011 -0.008 to -0.015 7.898 0.799 *** Lizards, snakes, and turtles 3-42 29 72 -0.014 -0.012 to -0.016 7.964 0.886 *** Crocodilians 15-35 2 4 -0.014 -0.009 to -0.019 7.857 0.993 0.0073 Reptiles (= ectothermic amniotes) 3-42 31 76 -0.014 -0.013 to -0.016 7.962 0.882 *** Reptiles (excluding marine species and crocodilians) 3-42 24 62 -0.014 -0.012 to -0.016 7.979 0.925 *** Marine reptiles 15-35 5 10 -0.011 -0.002 to .0.020 7.798 0.722 0.0183 All freshwater ectotherms, excluding air-breathing fishes (freshwater teleosts, aquatic salamanders, Alligator mississippiensis, Nerodia sipedon, freshwater turtles) 3-36.2 27 81 -0.016 -0.013 to -0.018 8.110 0.810 *** All freshwater species, including air-breathing fishes 3-36.2 36 94 -0.016 -0.014 to -0.019 8.110 0.815 *** All marine species (marine teleosts, elasmobranchs, marine reptiles including Crocodylus poroSlts) 5-35 24 40 -0.018 -0.015 to -0.022 8.064 0.822 *** All ectotherms (including air-breathing fishes, Petromyzon mal'inlts in freshwater, and Oncorhynchus m)'/liss in seawater) 3-42 81 207 -0.016 -0.015 to -0.018 8.090 0.813 *** Anamniotes (excluding air-breathing fishes) 5-35 41 118 -0.012 -0.010 to -0.014 8.063 0.722 *** All anamniotes (including air-breathing fishes) 5-35 50 131 -0.013 -0.011 to -0.015 8.077 0.741 *** Amniotes (equivalent to 'reptiles' above) 3-42 31 76 -0.014 -0.013 to -0.016 7.962 0.882 ***

Intracellular pH-individual species letaluTUs jmnrtatus (Cameron & Kormanik, 1982) white muscle 15-32 27 -0.015 -0.014LO -0.015 7.584 red muscle 15-32 26 -0.018 -0.018 to -0.019 7.683 heart muscle 15-32 27 -0.012 -0.010 to -0.013 7.778 brain 15-32 28 -0.019 -0.017 to -0.020 7.893 whole-body 15-31 28 -0.016 -0.015 to -0.018 7.707

ScyliorhinU.l stpllaris (Heisler et aI., 1976) white muscle 10-23 396 -0.018 -0.016 La -0.020 7.60 red muscle 10-23 258 -0.033 -0.030 to -0.037 7.88 heart muscle 10-23 104 -0.0 IO -0.004 to -0.016 7.50

Anguilla rostmta (Walsh & Moon, 1982) white muscle 5-20 28 -0.009 -0.004 to -0.014 7.38 red muscle 5-20 28 -0.003 0.004 to -0.0 I 0 7.52 liver 5-20 28 -0.018 -0.013 to -0.022 7.95 heart muscle 5-20 28 -0.020 -0.010 to -0.031 7.56 Ultsch & Jackson pH AND TEMPERATURE IN ECTOTHERMS 15

Table 2. (continued)

Temp- 95% rature Number Number confidence range of of data interval of P Group (0G) species points Slope slope Intercept r (slope)

Rana ralesbl'iana (Malan et aI., 1976) skeletal muscle 3.5-30 31 -0.015 -0.009 to -0.022 7.275 0.66

Trachemys scripta elegans (Malan et aI., 1976) skeletal muscle 9-31 17 -0.019 -0.013 to -0.024 7.421 0.91 heart muscle 9-30 16 -0.012 -0.009 to -0.015 7.452 0.93 liver 9-29 14 -0.023 -0.015 to -0.032 7.753 0.87 smooth muscle (esophagus) 9-30 18 -0.014 -0.008 to -0.020 7.513 0.82

DijJsosaur1ts dorsalis (Bickler, 1982) skeletal muscle 18-42 24 -0.010 -0.006 to -0.014 7.530 0.88 heart muscle 18-42 24 -0.010 -0 .006 to -0.015 7.604 0.88 whole-body 18-42 33 -0.015 -0.012 to -0.Ql8 7.654 0.91

Intracellular pH-species grouped by taxonomy

All ectotherms-brain 9-31 7 14 -0.022 -0.014 to -0.030 7.918 0.866 0.0001 All ectotherms-heart muscle 3-42 15 38 -0.007 -0.003 to -0.012 7.464 0.460 0.0037 Anamniotes-heart tissue 5-32 12 26 -0.006 0.001 to -0.011 7.476 0.330 0.0993 Amniotes (reptiles)-heart muscle 3-42 3 12 -0.005 0.001 to -0.011 7.345 0.531 0.0760 All ectotherms-liver 3-37 7 16 -0.018 -0.009 to -0.027 7.777 0.744 0.0010 Anamniotes-liver 5-25 4 8 -0.019 -0.001 to -0.037 7.880 0.726 0.0416 Amniotes (reptiles)-liver 3-37 3 8 -0.013 -0.003 to -0.022 7.578 0.794 0.0187 All ectotherms-erythrocytes 6-24 13 23 -0.015 -0.005 to -0.025 7.572 0.560 0.0067 Freshwater fishes-erythrocytes 6-20 4 13 -0.011 -0.002 to -0.020 7.556 0.641 0.0183 All ectotherms-white (or skeletal) muscle 3-42 23 63 -0.013 -0.009 to -0.018 7.441 0.625 *** Freshwater fishes-white muscle 5-31 4 18 -0.006 -0.002 to -0.011 7.376 0.580 0.0116 Freshwater ectotherms-white (or skeletal) muscle 5-31 12 37 -0.016 -O .OlD to -0.021 7.465 0.698 *** Anamniotes-white (or skeletal) muscle 5-31 19 50 -0.018 -0.013 to -0.024 7.526 0.727 *** Amniotes (reptiles)-white (or skeletal) muscle 3-42 3 12 -0.004 0.004 to -0.013 7.257 0.329 0.2966

*** p < 0.0001 .• Blood samples obtained by cardiac puncture rine. Air-breathing fishes were not included in the latter the scale, so that the effect of combining these data is to category, however, because they are transitional forms tilt the overall slope to a steeper value. Note also that these whose acid-base status fails to fit either pattern. Because amniote and anamniote slopes are not different statisti­ we do not include data from mammals and birds, the cally from each other. amniote data represent the reptiles. An examination of Fig. 5 will reveal that the inclusion of data from mammals and birds at their usual body temperatures or about 37 Intracellular pH vs. Temperature and 40°C, respectively, would place their values on the ALL SPECIEs-Compiled data on pH; are shown by species, amniote curve at about 7.4-7.5. An anamniote in this by tissue type, and by taxon in Table 4, together with (lIm. temperature range would be predicted to be at a pHb of calculated values ofpH;-pNw and These values reveal about 7.6. Interestingly, the slopes of both the anamniote rather consistent relationships between pH; and tempera­ and amniote data (dpH/dT = -0.012 and -0.014, respec­ ture for ectotherms as a group. The data for individual tively) are less than the slope for the combined groups (all tissues of all ectotherms are plotted in Fig. 6. The ecto­ ectotherms) as shown in Fig. 1. Note, however, that the therm-inclusive slopes of these data (dpH/dT) are all amniote (= reptiles) data tend to be concentrated toward close to the predicted alphastat slope except for heart the high end of the temperature scale whereas the muscle, which is significantly more shallow. It is important anamniote data are concentrated toward the low end of to note, however, that slopes for individual species and for 16 BULLETIN 18 December 1. 1996

B.2 B.2 ° AMPHIBIANS AND FRESHWATER TELEDSTS B.l B.l m=-.OlJ o ANAMN IO TES

0 B.O m = -.0 12 B.O ° 0 o 0 0 / 7.9 o...... a. 0 0 7.9 0°0 7.B " :r: 7.B Q. :r: 7.7 Q. 7.7 7.6

7.6 7.5 • 00 .0 7.4 7.5 • 00 00 7.3 , 7.4 0 5 10 15 20 25 30 35 40 45

7.3 TEMPERATURE ("C) 0 10 15 20 25 30 35 40 45 Figure 5. Arterial pH as a function of temperature of TEMPERATURE (oC) anamniotes (excluding air-breathing fishes) and amniotes (reptiles). Individual species-temperature data points are Figure 3. Arterial pH as a function of temperature in as in Fig. 1. freshwater teleosts (excluding air-breathing species) plus amphibians compared with all reptiles. Individual species­ temperature data points as in Fig. 1. 7.8

I.. BRAIN. m=-.022 B.2 7.6 ,/ • il 8.1

B.O 0" 7.4

00 7.9 7.2 7.B :r: • RED CELL. m--.015 0.. 7.7 :r: A Q. 7.0 I" LIVER. m=-.Ol B 7.6 c:: <: ...J :::> 0 5 10 15 20 25 30 35 40 7.5 ...J ...J W 7.4 U 7.B <: a:: 7.3 r-- Z 0 5 10 15 20 25 30 35 40 7.6 A '" m~- 007 AA TEMPERATURE (oC) .,. 7.4 Figure 4. Arterial pH offreshwater ectotherms (amphibians, turtles, a snake [Nerodia sipedoll] , fishes including air­ 7.2 breathing species and excluding freshwater-adapted marine species, and Alligatormississippiellsis) compared with that of 7.0 .. WHITE MUSCLE. m~- .013 t marine ectotherms (teleosts excluding seawater-adapted 6.B freshwater species, elasmobranchs, and the reptiles Crocodylus POroSllS, Chelollia mydas, Caretta caretta, 6.6 AmblyrhYllchllS cristatus, Acrochordlls arafurae, and Lapemis 0 5 10 15 20 25 30 35 40 45 hardwickii). Individual species-temperature data points are TEMPERATURE (C) as in Fig. 1. Figure 6. Intracellular pH of brain, red blood cells, liver, heart, and white (or skeletal) muscle of ectotherms as a function of temperature. For details of data, see Table 4. Ultsch & Jackson pH AND TEMPERATURE IN ECTOTHERMS 17

Table 3. Analysis of covariance comparisons of regression relationships of plasma pH vs. body temperature of paired groups of ectothermic vertebrates. Data from two groups are considered similar when the probabilities that the slopes and the elevations of the regression lines are equal are both >0.050; in such a case the data from the two groups are combined for further comparisons with additional groups.

P PI Can be Comparison (slope) (elevation) combined?

Marine teleosts vs freshwater teleosts 0.264 (NS) <0.001 (S) No Marine teleosts vs elasmobranchs 0.844 (NS) 0.619 (NS) Yes Freshwater teleosts vs elasmobranchs 0.592 (NS) <0.001 (S) No Freshwater teleosts vs elasmobranchs plus marine teleosts 0.239 (NS) <0.001 (S) No

Salamanders vs. anurans 0.258 (NS) 0.274 (NS) Yes Freshwater teleosts vs amphibians 0.278 (NS) 0.478 (NS) Yes Freshwater teleosts and amphibians vs. reptiles 0.604 (NS) 0.003 (S) No Freshwater teleosts and amphibians vs lizards 0.268 (NS) <0.001 (S) No

Snakes vs lizards 0.957 (NS) 0.003 (S) No Snakes vs turtles (all data) 0.129 (NS) 0.012 (S) No Turtles vs. lizards (all data) 0.038 (S) No Crocodilians vs lizards 0.510 (NS) 0.001 (S) No Crocodilians vs snakes 0.531 (NS) 0.054 (NS) Yes Crocodilians vs turtles 0.738 (NS) 0.003 (S) No Crocodilians vs all other reptiles 0.933 (NS) 0.003 (S) No

All freshwater vs. all marine 0.410 (NS) <0.001 (S) No Anamniotes vs amniotes (= reptiles) 0.061 (NS) <0.001 (S) No

taxonomic subgroupings often deviate considerably more using for comparison the ideal slope over the tempera­ from the ideal alphastat slope than do the ectotherm­ ture range for each data set. For blood, both well-studied inclusive slopes (Table 2). individual species plus subgroups are considered, whereas for pHi only ectotherm-inclusive data for the INTER-GROUP COMPARISONS-Because of the relatively small various tissues are given. The results are equivocal. If the size of the pHi-temperature data base, it is difficult to test applied is whether the ideal slope falls within the 95% make inter-group comparisons of slope and elevation with confidence interval for the data for the individual species, the same confidence as with the much larger data base for then the positive outcome in 16 of24 studies of individual blood. The most extensive data are for skeletal muscle (63 species represents a partial affirmation of the alphastat points), but comparisons between marine and freshwater hypothesis. Subgroup comparisons are less convincing species and between anamniotes and amniotes fail to since only 8 of 18 satisfY the criterion, although the slope reveal the differences seen in the blood data (Fig. 7). An of all ectotherms together does conform. A non-paramet­ even smaller data set for heart muscle (38 points) again ric sign test produces a less favorable outcome for the reveals no difference between freshwater and marine spe­ hypothesis. Here 22 of 25 studies of species and 17 of 18 cies, although the amniotes have a lower elevation of the subgroups have mean slopes that are less than the ideal pHi slope than do anamniotes, similar to the blood pic­ slope, indicating a very consistent deviation from the ture (Table 2.). predicted relationship. Intracellular data showed some­ what better agreement on these bases with 4 of 5 tissues including the ideal slope within the 95% confidence lim­ Agreement With Theoretical Concepts its and only 301'5 exhibiting a slope less than the alphastat dpHldT SLOPE-As discussed earlier, ideal conformity ideal. The large inter-tissue and intra-tissue variability, with the alphastat hypothesis requires an overall dpHI dT however, must be also taken into account here. slope ofabout-O.018 U/"C and a constant compartmental Alphastat regulation is often evaluated by calculating CO2 content. Comparisons of ectotherm blood and pHi the fractional dissociation of imidazole, the so-called data with the ideal slope are presented in Table 5 and a-imidazole (aIm)' This is calculated from the equilibrium the non-linearity of this slope is taken into account by of the imidazole (1m) moiety 18 BULLETIN 18 December 1,1996

A constant value of aIm over a range of body temperatures, 7.5 which is equivalent to conforming to the ideal dpH/ dT 00 slope discussed above, is the explicit definition of 7.4 alphastat regulation. The compiled data in this paper can o ". be analyzed in this fashion by calculating, based on the

7.2 imidazole dissociation constants of Reeves (1976), values of aIm for the blood data presented in Table 1 and for the skeletal muscle from Table 4. These results, plotted as a 7.0 - function of temperature, are shown in Fig. 8. As with the overall plot of pHb vs. temperature (Fig. 1), this presenta­ 6.B tion of aIm vs. temperature for all ectotherms appears to I D- provide strong support for the alphastat hypothesis. w Again, however, dissection of these data into subgroups of cj 6.5 ::----~----~----r-----r----,----~-----r------If) 0 5 10 15 20 25 30 35 40 45 species reveals discrepancies from the ideal, constant aIm' :::J ::. relationship (Fig. 9). Note that in 4 of 7 groups depicted W aIm varies significantly with temperature. What is not obvi­ ~ 7.5 I ous from this statistical result, however, is how meaningful :;: these temperature effects are physiologically. The value of • MARINE 7.4 FRESH WATER aIm in turtles, for example, increases significantly with a 0 0 temperature butonly from 0.80 to 0.82 over a 30 e range. o • f~""'? .., 0 o Fig. 9 shows a similar analysis for the constant relative

7.2 o --'.9 alkalinity hypothesis (pHb - pN,,.) and, although the statis­ ". o 0 tical outcome is much the same, the magnitude of change with temperature in this variable, as noted by Reeves B 0 7.0 COMBINED (1976), is greater than for aIm' In addition, the differ­ m=-.015 ences among subgroups in pHb-pNw are much greater (i.e., per cent change from one group to another) than 6.B -+1--.----.-...... --.----.---.--.----.----, for aIm' suggesting that aIm is more of a controlled vari­ o 5 10 15 20 25 30 35 40 45 ' able than is pH,,-pNw Finally, in the two largest taxo­ TEMPERATURE tC) nomic subgroups, the anamniotes and amniotes, both aIm and pHb-pNw increase significantly with temperature, but Figure 7. Intracellular pH of white (or skeletal) muscle of the rate of increase in pHb-pNw is greater than in aIm' ectothermic anamniotes and amniotes (= reptiles, upper again suggesting that that the latter is the more tightly panel) and the two groups combined. Lower panel shows controlled of the two (Fig. 10) combined data for marine (data available only on fIShes) A feature of the data point distribution in Fig. 8 is the and freshwater ectotherms (lower panel); the latter group relatively small variation of the data for plasma in contrast includes fishes, amphibians, and reptiles. Separate to the data for skeletal muscle. Two factors may account regression lines are not shown for each group because of for this difference. First is the inherently greater variabil­ the small sample for marine ectotherms. For details of ity in reported pHi values, due likely to measurement data, see Table 4. techniques and to the heterogeneity of cellular samples discussed earlier. The second possible reason for the greater spread of intracellular aIm values is revealed in Fig. 11, a plot of calculated aIm vs. pH for a series of Hlm+ H H+ + 1m temperatures. Mean ectotherm-inclusive plasma and tis­ KIm = [H+] [Im]/[Hlm+] sue aIm and pH data points are added to this figure. Note as follows: that at any given temperature, the rate that aIm changes with pH depends on the pH value; the change becomes steeper at lower pH values. In terms of the plasma vs. pHi a-imidazole = [1m] / ([Hlm+] + [1m]) comparison, the steeper change in aIm at low pH will contribute to a larger variability in intracellular aIm be­ Or, a-imidazole can be calculated at a particular tempera­ cause of the lower pH in the intracellular compartment. ture from the measured pH and the assumed pKlm accord­ Furthermore, because of this non-linearity, published val­ ing to Reeves (1976), as follows: ues of aIm stability should be interpreted cautiously with this graph in mind since apparently small deviations in a-imidazole = 10rl-l-rK / (1 + 10pl-I-rK ) aIm may represent rather large differences in pH. To be 1111 1111 Ultsch & Jackson pH AND TEMPERATURE IN ECTOTHERMS ----- 19

Table 4. Intracellular pH, pH(pNw' and (a-imidazole of ectotherms under resting, normoxic, normocapnic conditions. pHi for erythrocytes is for arterial blood. If a study reported data for more than one skeletal muscle, the values were averaged. If pHi was determined by more than one method in a report, the results were averaged. If regression equations were given for pHi and data were not grouped about specific temperatures, pHi was calculated at 5°C intervals within the domain of measurements (data of Malan et al., 1976, Heisler et al., 1976, 1980); if data were grouped about specific temperatures, means were calculated at all temperatures for which there were at least three measurements, rather than using regression equations (data of Cameron & Kormanik, 1982 and of Bickler, 1982). When pHi was determined during the day and during the night (Bickler, 1986), daytime values were used on the assumption that most other studies were done during the daytime.

Number Temperature

Species of Studies ("C) pH; pH; - pNw a-imidazole

Freshwater fishes

Anguilla rastmta (freshwater) white muscle 5 7.34 -0.03 0.51 10 7.29 0.02 0.53 15 7.26 0.09 0.57 20 7.19 0.11 0.58

red muscle 5 7.48 0.11 0.59 1 10 7.52 0.25 0.66 15 7.48 0.31 0.69 20 7.44 0.36 0.71

liver 5 7.84 0.47 0.76 10 7.79 0.52 0.78 15 7.70 0.53 0.79 20 7.57 0.49 0.77

heart 5 7.41 0.04 0.55 10 7.39 0.12 0.59 15 7.32 0.15 0.65 20 7.10 0.02 0.53

Cyprinus caT/Jio erythrocytes 20 7.32 0.24 0.65

/rtalurus punrtatus white muscle 15 7.28 0.11 0.58 19 7.30 0.20 0.63 1 20 7.38 0.30 0.68 1 21 7.34 0.27 0.67 2 25 7.22 0.22 0.65 29 7.]3 0.20 0.63 31 7.14 0.24 0.66

red muscle 17 7.33 0.19 0.63 19 7.34 0.24 0.65 20 7.28 0.20 0.63 ] 21 7.39 0.32 0.70 2 25 7.22 0.22 0.65 29 7.13 0.20 0.63 31 7.09 0.19 0.63

Ir/alums /mne/a/us heart muscle 17 7.53 0.39 0.73 20 7.56 0.48 0.76 21 7.57 0.50 0.78 20 BULLETIN 18 December 1, 1996

Table 4. (rontinllerl)

Number Temperature Species of Studies (DC) pH; pH; - pNII a-imidal.ole

Irtalurlls /mnctatlls (continued) heart muscle 2 25 7.49 0.50 0.78 32 7.44 0.55 0.80

brain 15 7.49 0.32 0.69 18 7.57 0.45 0.75 20 7.56 0.48 0.76 21 7.47 0.40 0.73 2 25 7.40 0.40 0.73 29 7.35 0.42 0.74 31 7.29 0.39 0.75 32 7.24 0.35 0.71

skull bone 20 7.42 0.34 0.70

vertebral bone 20 7.44 0.36 0.71

whole-body 19 7.41 0.31 0.69 19-22 7.35 0.26 0.68 21 7.35 0.28 0.69 2 9-_!J 7.32 0.32 0.70 29 7.23 0.30 0.69 31 7.23 0.33 0.70

Onrorhynrhus lIlylliss white muscle 5 7.48 0.11 0.64 10 7.29 0.02 0.53 12-14 7.30 0.09 0.57 6 15 7.26 0.09 0.57 I 18 7.28 0.16 0.61

red muscle 15 7.25 0.08 0.56

liver 13 7.58 0.37 0.72 15 7.38 0.21 0.64 heart muscle 15 7.35 0.18 0.62

brain 3 15 7.59 0.42 0.44

gill 15 7.43 0.26 0.66

erythrocytes 5-7.5 7.49 0.15 0.61 2 9 7.51 0.22 0.64 9 10 7.38 0.14 0.60 10.5 7.43 0.17 0.62 2 12 7.38 0.16 0.61 3 15 7.32 0.15 0.60 4 15-16 7.43 0.27 0.67

whole-body 12-14 7.30 0.09 0.57 2 15 7.28 0.10 0.58 PI'trolllyzon I/U//'inus (freshwater) muscle 10 7.21 -0.06 0.49

eryth rocytes 5 10 7,45 0.18 0.62 Ultsch & Jackson pH AND TEMPERATURE IN ECTOTHERMS 21

Table 4. (continued)

Number Temperature Species of Studies (OC) pHi pHi - pN", a-imidazole

Sal1llo trutta erythrocytes 15 7.42 0.25 0.66

red muscle 5 7.18 -0.19 0.42

white muscle 5 7.22 -0.15 0.44 15 7.19 0.02 0.53

heart muscle 5 7.36 -0.01 0.52 15 7.38 0.21 0.64

Synbranchlls 1IlaT11l0ratus white muscle 30 6.88 -0.04 0.50

heart muscle 30 7.22 0.30 0.69

Tinea tinea erythrocytes 13 7.50 0.30 0.68 1 14 7.38 0.18 0.62 3 15 7.41 0.24 0.65 1 16 7.40 0.25 0.63

Marine fishes

He1llitrijJterus a1llPJ1ranus white muscle 10-11 7.51 0.25 0.66

heart muscle 10-11 7.50 0.24 0.66

Onrorh),nchlls kimtrh (seawater) erythrocytes 13 7.42 0.21 0.64

Onrorh),nchlls 1Il)'/liss (seawater) White muscle 10 7.31 0.04 0.55

erythrocytes 10 7.46 0.19 0.63

ParojlhT)'S vptulus erythrocytes 11 7.25 0.00 0.52

white muscle 11 7.29 0.04 0.54

heart muscle 11 7.32 0.07 0.56

brain 11 7.75 0.50 0.77

Platirhth),s stPllaills whole-body 2 11 7.57 0.32 0.70

white muscle 9 7.29 0.00 0.52 11 7.56 0.31 0.69

liver 11 7.60 0.35 0.71 22 BULLETIN 18 December 1,1996

Table 4. (continued)

Number Temperature Species of Studies ((Ie) pHi pHi - pN", a-imidazole

Platichthys stpliatus (continual) erythrocytes 2 9 7.38 0.10 0.58 ]] 7.16 -0.09 0.47

brain 9 7.79 0.50 0.78

Raja ocellata erythrocytes 2 12 7.39 0.16 0.61

heart muscle 12 7.62 0.39 0.73

white muscle 12 7.47 0.24 0.65

brain 12 7.56 0.33 0.70

Scyliorhinus stplimis white muscle 2 10 7.44 0.17 0.62 2 15 7.34 0.18 0.62 2 20 7.26 0.18 0.62

red muscle 2 10 7.55 0.28 0.68 2 15 7.39 0.22 0.64 2 20 7.23 0.15 0.60

heart muscle 2 10 7.40 0.13 0.60 2 15 7.36 0.18 0.62 2 20 7.32 0.24 0.65

Amphibians

A 1IIb),st01lla tigtinu1ll (larval) skeletal muscle 20 7.35 0.27 0.67

Bufo 1IlminllS skeletal muscle 10 7.36 0.09 0.58 3 20 7.26 0.18 0.62 2 25 7.16 0.16 0.62 30 6.97 0.05 0.55

heart muscle 10 7.60 0.33 0.70 2 20 7.24 0.16 0.61 2 25 7.24 0.24 0.66

skin 25 7.59 0.59 0.81

brain 25 7.16 0.16 0.61

liver 25 7.44 0.44 0.75

erythrocytes 22 7.34 0.29 0.68

kidney 20 6.99 -0.09 0.47

Npc/urus '/Il(lCU[OSU.I skeletal muscle 20 7.26 0.18 0.62 3 10 7.24 -0.03 0.51 Ultsch & Jackson pH AND TEMPERATURE IN ECTOTHERMS 23

Table 4. (rontinued)

Number Temperature (0C) Species of Studies pHi pHi - pNw a-imidazole

Nprlurlls lIluculo.ms (con tinued) skeletal muscle 2 20 7.06 -0.02 0.51 22 7.05 0.00 0.52 3 30 6.89 -0.03 0.51

NolojJhthai1nlls viririescells skeletal Illuscle 3 10 7.24 -0.03 0.51 2 20 7.06 -0.02 0.51 22 7.05 0.00 0.52 3 30 6.89 -0.03 0.51

Pll'thorlon ril1l'rl'llS skeIetalllluscle 10 7.49 0.22 0.64 22 7.10 0.05 0.55 30 6.66 -0.26 0.38

Ran{/. call'sbl'irl1Ja skeletal muscle 10 7.12 -0.15 0.44 15 7.05 -0.12 0.45 2 20 7.04 -0.04 0.50 25 6.90 -0.11 0.46 30 6.82 -0.10 0.47

XmojJUs IIll'Vis skeletal muscle 10 7.23 -0.04 0.50 2 20 7.03 -0.05 0.50 30 6.92 0.00 0.52

heart muscle 10 7.22 -0.05 0.49 2 20 7.27 0.14 0.60 30 7.08 0.16 0.61

erythrocytes 20 7.33 0.25 0.66

SiTl'1l lllfl'riina white (skeletal) Illuscle 25 6.88 -0.12 0.45

heart muscle 25 7.23 0.23 0.65

Reptiles

Chl),SI'III)'S jJir(a {)('llii skeletal Illuscle 3 7.27 -0.14 0.44 20 7.23 0.15 0.60

heart muscle 3 7.25 -0.16 0.43 20 7.17 0.09 0.57

lil'cr 3 7.48 0.07 0.56 20 7.49 0.41 0.74

erythrocytes 2 24 6.96 -0.05 0.49

brain 20 7.47 0.39 0.73

DijJsosauntl rionalil liver 37 7.21 0.40 0.74 24 BULLETIN 18 December 1,1996

Table 4. (continued)

Number Temperature Species of Studies (0C) pHi pHi - pN,\ a-imidazole

DipsosaU1'1IS dorsalis (continued) heart muscle 18 7.43 0.31 0.69 25 7.33 0.33 0.70 35 7.27 0.43 0.71 37 7.04 0.23 0.65 42 7.15 0.41 0.75

skeletal muscle 18 7.35 0.23 0.65 25 7.28 0.25 0.68 35 7.22 0.38 0.69 37 7.23 0.42 0.75 42 7.08 0.34 0.71

smooth muscle (esophagus) 37 7.04 0.24 0.66

whole-body 18 7.39 0.27 0.67 25 7.28 0.28 0.63 35 7.19 0.35 0.67 42 7.04 0.30 0.70

Tmchelll),s scri/Jta ell'grl11s skeletal muscle 10 7.24 -0.04 0.50 ]5 7.]4 -0.03 0.50 20 7.05 -0.03 0.50 25 6.96 -0.04 0.50 30 6.86 -0.06 0.49

heart muscle 10 7.33 0.06 0.56 15 7.27 0.10 0.58 20 7.21 0.13 0.59 25 7.15 0.15 0.61 30 7.09 0.17 0.62

liver 10 7.52 0.25 0.66 15 7.40 0.23 0.65 20 7.29 0.2] 0.64 25 7.] 7 0.] 7 0.62 30 7.05 O. ]3 0.60

smooth muscle (esophagus) \0 7.37 0.10 0.58 15 7.30 O. ]3 0.59 20 7.23 O. ]5 0.60 25 7.16 O. ]6 0.6\ 30 7.09 0.\7 0.62

REFERENCES: FISHES-Allgllilla roslrala (Walsh & Moon, 19H2); CY/"'illl/\' fmllio (Fuchs & AJI)ers, 19H8); H"'lIillil,"'/I/\, allll'rifllllll.1 (1\lilligan & Farrell, 1986); irla/lIrlls IJllIIClallls (Cameron & Kormanik, 19H2a); Ollflllh.\'III/II11 /rillll,.h (\'an den Thillart et a!., 19H~); OltllrhY",}II11 III)'irill (Boutilier et a!., 1986, 1988; Ferguson et a!., 199~; !-lobe et a!., 1984; Kieflel et a!., 1994; Milligan & Wood, I 986a,/r, Perry et a!., 1988, 1989; Primmell et a!., 19H6; Portner et a!., 1990; Tang & Boutilier, 1991; Tang el a!., 1988; Tuft, et a!., 1988; Wang, Ileigenhauser & Wood, 1994; Wood & LeMoigne, 1991; Wood & Munger, 1994; Wood, Walsh, Thomas & Perry, 1990; Wrighl, Randall & Wood, 1988; Ye el a!., 1991), [>al'O/lIII)'1 111'1111111 (Wrighl, Wood & Randall, 1988); /'1!lrolll)WII lIIarillllS (Boulilier el a!., 199~; Tufts, 1991; Tufls el a!., 1992), /'/alichlhys sId/allis (Milligan & Wood, 1987a,b; Wood & Milligan, 1987); RajalJcd/ala (Graham et a!., 1990; Wood, Turner, Munger & Graham, 1990); 8a/lllo II'II//a (Butler & Day, 199~a,b); SI)'lilJrhillllS sld/aris (Heisler el a!., 1976, 1980); S)'Iil}f(lIIchus III a fIIwral liS (Heisler, 1982); Tillfa lillfa (Jensen, 1987; Jensen & Weber, 1985, 1987). AMPI-IIBIANS-Alllbyslollla I i.!".,.i II II III-larval (Wood el a!., 1989). BujlJ lIIarillllJ (Boulilier, et a!.. 1987; Portner Cl a!., 1990; Snyder & Nestler. 1991; Toews & Heisler. 1982; Tufts el a!., 1987; Wood el a!.. EJ89). NIJIIJ/lhl/ta/II/It., 1Iiriri,',wms (Johnson el a!.. 199~; Hitzig, et a!.. 1994). /'lt~lh(}dllll rilll'mlls (Johnson ct a!.. 199~); Raila trlll'Jb,~ialla (Malan et a!.. 1976; Wood et a!.. 1989). Sil'l~1I /(feerlilla (I-Ieisler el a!.. 1982); XI!IW/JIIS /almiJ (Bolltilier & Lantz, 1989; Wood cl a!.. 1989). REPTILES-CIII.\''''III.\'''llifl(f bellii (jackson & Heisler, 198~; Maginniss & Hitzig. 1987; Wasser el a!., 1991); /)iIISOJ(IIII'11S dorSl//is (Bickler. 1982, 1986); lim'hlwl.\'s scrilila I'!"WIIIS (Malan el a!., 1976). Ultsch & Jackson pH AND TEMPERATURE IN ECTOTHERMS 25

1.0 1.0 09 --" - " " § l"" .. "ft .. ~ I"~a"l ,. "" a" .. ",. "- - 0 .9 .. fi " ~~" "I! .. F" ""9,/:,, " eat''' I"" tf' 0.8 .. 0 A b. A 4 8: W .. .. _ ab.__ A I H 0 00 ....J i\ 0 0 0 8 N 0 .7 O'j 9 <{ t~ 0.7 . ~ 7i" ~o • .8 •• ~ • 00 . A 8 fio.,e&: · ° 0 a A AA AA A A o 0 0 0 0 0 ::::E 0.6 ~ 0 ~ A A 0 .6 I I AA * PLASMA I"~ WHITE OR 0.. 0 .5 At t ts I SKELETAL A 0. 5 t MUSCLE 0.4 0.4

0 .3 0.3 a 5 10 15 20 25 30 35 40 45 a 5 10 15 20 25 30 3 5 40 45 TEMPERATURE tC) 1.00 Figure 8. a-imidazole of plasma and muscle (white or skeletal) of ectotherms as a function of temperature. Ranges 0.95 of a-imidazole of 0.15 (plasma) and 0.30 (muscle) are shown to indicate that the variability of the data for muscle W 0.90 ....J is about twice that for plasma. For details of data, see 0 N Tables 1 and 4. <{ a 0 .85 ::::E I 00 0.80 ~ 0 0 .9 ts FT 0.75 0.8 A 0 .70 L a 5 10 15 20 25 30 35 40 45 Z " 0.7 - a. TEMPERATURE (C) Figure 10. pHb-pN and a-imidazole as functions of :c 0.6 w a. temperature in anamniotes and amniotes (reptiles). Individual species-temperature data points are as in Fig. 1. 0.5 ~C 1.0

0 .9 ./--~::. - - :~~:-:::.::; 0.4 0 10 20 30 40 0 .1\ FTafRESHWATER TELEOSTS ~';Z:~'"' 0 .7 0.95 EHIT~ELASMOBRA N CHS AND ,- = SIGNif iCANT SLOPE. PS:O.0 5 MARINE TELEOSTS .-' ~ ... .. ·•.. LlVER W r.J -----.. . .' A-AMPHIBIANS ....J 0.6 Il:: 0 ::::J ... ~-);:,----.. ERYTHROCYTES T-TURTLES N «>- L ~L1 ZARDS <{ ." ...... HEART 0 .5 "-' ~ 0 .90 C =CROCODILIANS a 0..'" ' .' MUSCLE ~ I S= SNAKES ~ r.J W 0.4 ...... J FT- I 40 0 A ts N 0.3 <{ a 0 .85 L· 30 ::::E I E+MT· 10 ts 0. 1 O· 0 .80 "120 0 ~C 6 .5 7.0 7.5 8 .0 8.5

0.75 -+I------.------r----.------,--- pH OF TISSUE OR PLASMA a 10 20 30 40 Figure 11. Plasma and intracellula a-imidazole as a function TEMPERATURE tC) of pH and temperature in ectotherms. Data points were

Figure 9. Regression curves for pHb-pNw and a-imida­ generated by calculation of pH at a given temperature from zole as functions of temperature for several groupings of the regression data in Table 2; the resultant pH was then ectotherms. See Table 6 for details of the regression used to calculate a-imidazole using the pK' for plasma analyses. proteins of Reeves (1976). 26 BULLETIN IS December 1,1996

Table 5. A comparison of slopes of regression plots of pH vs temperature from a variety of studies with that predicted for the relationship of the pK' of plasma proteins as reported by Reeves (1976).

Observed Predicted Predicted above(+) or in 95% CI Temperature Observed slope below (-) of observed Temperature range and name of species or group range (0C) slope (Reeves,1976) predicted (Yes/ No)

Oncorhynchus mykiss 5-20 -0.014 -0.019 Yes CY/Jrinu5 carpio 10-25 -0.005 -O.OlS Yes Ictalurus punctatus 15-31 -0.015 -0.016 Yes l. punctatus (Cameron & Kormanik, 19S2) 14-33 -0.013 -OJJ17 Ambystoma tigrinum, larval 5-25 -0.016 -O.OlS No A. tigrinum, adult 5-25 -0.016 -0.0 IS Yes CrY/Jto&ranchus alleganiensis 5-25 -0.014 -O.OlS Yes NertUTlIS maculosus 5-25 -0.019 -O.OlS + Yes Bulo marinus 10-30 -0.0176 -0.017S Yes Xenopus laevis 10-30 -0.0\3 -O.OlS Yes Rana cates&eiana 5-34 -0.014 -O.OIS No R. catesbeiana (Malan et aI., 1976) 3.5-30 -0.021 -O.OIS + Yes Tracltemys scripta elegans 10-30 -0.012 -O.OlS No T .s. elegans (Malan et aI., 1976) 9-32 -0.02\ -O.OlS + No Alligator mississippiensis 15-35 -0.014 -0.017 Yes Chr)'semys picta bellii 3-30 -0.013 -O.OIS No Chelonia mydas 15-35 -0.010 -0.017 Yes Pseudemys .floridana 22.5-36.2 -0.013 -0.017 Yes \l{lranlls exanthematieus 20-35 -0.005 -0.017 No Am&lyrhynrhus cristatus 16-35 -0.012 -0.017 Yes DipsosauTllS dorsalis 17.5-42 -0.016 -0.017 Yes Iguana iguana 20-35 -0.010 -0.017 No Physignathus leSlleurii 19-29 -0.016 -0.017 Yes Coluber constrictor 15-40 -O.OlD -0.017 No Nerodia sipedon 15-35 -0.013 -0.017 Yes

Freshwater teleosts 5-31 -0.012 -O.OIS No Marine teleosts 5-2S -0.007 -O.OIS No Elasmobranchs 7-24 -0.009 -0.0 IS No Aquatic salamanders 5-25 -0.017 -O.OIS Yes Anurans 5-35 -0.013 -O.OlS Yes Amphibians 5-35 -0.014 -O.OIS No Amphibians and freshwater teleosts 5-35 -0.014 -O.OlS No Turtles 3-36.2 -0.0\5 -O.OIS Yes Snakes 15-40 -0.011 -0.017 No Lizards 16-42 -0.011 -0.0\7 No Crocodilians 15-35 -0.014 -0.017 Yes Amniotes (= reptiles) 3-42 -0.014 -O.O\S No All ectotherms 3-42 -O.OHi -O.OIS Yes All anamniotes 5-35 -0.013 -O.OIS No Anamniotes (excluding air-breathing fishes) 5-35 -0.012 -0.018 No All marine species 5-35 -0.01S3 -0.0179 + Yes All freshwater species 3-36.2 -0.016 -0.018 Yes All freshwater species (excluding air-breathing fishes) 3-36.2 -0.016 -O.OlS Yes

All ectotherms-brain 9-31 -0.022 -0.018 + Yes All ectotherms-heart 3-42 -0.007 -O.oI8 No All ectotherms-muscle 3-42 -O.(Jl3 -O.OlS Yes All ectotherms-erythrocytes 6-24 -0.015 -O.OlS Yes All ectotherms-\iver 5-25 -0.0180 -0.0179 + Yes Ultsch & Jackson pH AND TEMPERATURE IN ECTOTHERMS 27 ---

specific, the same change in pH in both the intracellular decreased [HC03·]. A common assumption has been that fluid and in the plasma will result in a larger change in aim water-breathing and air-breathing ectotherms achieve in the intracellular fluid than in the plasma. this pH status in different ways (Prosser, 1991, p. 137); air­ -] It is important to emphasize that even though the breathers by maintaining [HC03 constant in their body accumulated data do not consistently match the ideal f1uids and increasing PCO') by ventilatory control; water­ alphastat slope, the data do show a quite consistent nega­ breathers, due to the dedication of their gill ventilation to tive relationship between pH and temperature. For ex­ oxygen acquisition, by actively reducing CO') concentra­ ample, the plasma data for the 25 studies of species in tion at constant PCO') (Jackson, 1982). According to these Table 5 have a mean dpH/dT (±S.E.) of -0.014±O.001 generalizations, air-breathers, but not water-breathers, compared to the predicted mean slope of -0.018. The satisfy the constant CO') concentration condition of ideal finding of a consistently lower slope may have at least alphastat control, but this assumption has also been chal­ three explanations. First, ideal conformity of blood or lenged based on conf1icting experimental evidence intracellular fluid to the alphastat hypothesis and equiva­ (Stinner et aI., 1994; Stinner and Wardle, 1988). In addi­ lence of in vivo and in vitro behavior depends on the tion, data from fishes often show a significant rise in PCO') dominance of imidazole moieties in the acid-base behav­ with temperature (e.g., Cameron and Kormanik, 1982af ior of the solution in question. As Reeves (1976) pointed To investigate this second criterion for ideal alphastat -] out, larger relative concentrations of other buffer species, behavior, we have compiled data on plasma [HC03 and such as bicarbonate or phosphate, with lower intrinsic PCO') from selected species and for subgroups of ecto­ dpH/ dT characteristics, will produce a more shallow thennic vertebrates as a function of temperature (Table slope than that predicted from pure imidazole behavior. 6). Adequate data on these relationships are much less Turtle plasma, for example, has a lower protein concen­ abundant than for pH, but the assembled data do reveal -] tration and a higher [HC03 than does dog plasma, and some general trends that tend to support the generaliza­ this could lead to a smaller dpH/dT slope. Second, the tions stated above. Air-breathers (amphibians and rep­ predicted behavior of blood based on imidazole chemis­ tiles), consistent with in vitro behavior, show little change '] try requires an accurate knowledge of the enthalpy of in plasma [HC03 with temperature, but do show an ionization (~Ho) for imidazole since ~Ho is related to the increase in blood PCO') with temperature. Exceptions -] change of pK with temperature according to the van't with regard to constant [HC03 include Amb),stoma Hoff equation, tig;rinwn larvae, which can be regarded as a water­ breather, and Coluber constrictOl; both of which have a pI<-r\ - pI<-r2 = (~H()/2.303R) (l/T) -1/T2) significantly negative slope for the [HC03-] vs. tempera­ ture relationship. Note also that the [HC0 -] of reptiles as where R is the gas constant and TI and T2 are different 3 a group also decreases significantly with temperature, but temperatures. No apparent general agreement exists for this is an artifact due to the preponderance at low tem­ the value of ~Ho in plasma (Heisler, 1986), and in the peratures of data from turtles and the fact that turtles have intracellular compartment with its multitude of proteins high [HC0 -] 'so For example, neither turtles nor non­ and microenvironments, the situation is even less clear 3 turtle reptiles plotted separately exhibit a significant (Cameron, 1989). A third explanation for the discrepancy change in their [HC0 -] with temperature (Fig. 12). Am­ between predicted and observed behavior, emphasized by 3 phibians and turtles, as well as non-turtle reptiles and Heisler (1986), concerns regulatory adjustments of body reptiles as a group all showed a significant increase in fluid acid-base state superimposed on the simple behavior PCO') with temperature, although the individual non­ of the buffer systems. Acid-base-related transfers of ions turtle reptiles (the crocodilians, lizards, and snakes) did between body f1uid compartments and between the body not. The mean values for these reptiles, however, did show and the environment have been observed following tem­ upward trends with temperature (Fig. 12) and it is likely perature change, especially in fishes (Heisler, 1986), but that additional data would make some or all of these also in amphibians (Stinner et aI., 1994) and reptiles slopes significant. (Stinner and Wardle, 1988). These transfers, in addition The relationship of [HC0 -] and PC0 with tempera­ to possibly adjusting body f1uid pH values away from the 3 2 ture for water-breathers was less consistent. Two of 3 condition predicted by ideal alphas tat behavior, may also species (accepting the data of Cameron and Kormanik as produce changes in compartmental CO') concentration, the best estimate of the situation in ir/alurus jJUnrtatus) and thereby violate the second condition-of the alphastat exhibited a significant fall in [HC0 -] with temperature hypothesis, that compartmental CO,) concentration is 3 and these same 2 species also showed a significant rise in temperature-independent. - PCO') with temperature (Table 6). In the well-studied rainbow trout (Oncorh)'nclttts 111),ltiss) , neither of these vari­ ables changed significantly with temperature although CO2 CONTENT-The decrease in body fluid pH values of ectothermic vertebrates as temperature increases is asso­ blood pH values did show a highly significant fall as presented earlier (Table 2). Freshwater teleosts as a ciated With some combination of increased PC02 and/or 28 BULLETIN 18 December 1,1996

-] Table 6. Regression equations for the relationships of arterial plasma [HC03 and PC02, and for arterial plasma and intracellular (a-imidazole and pH-pNw' with temperature among several well-studied species and among various groups of ectothermic vertebrates. In addition, in the accounting of number of species, the larval and adult forms of the salamander Ambystoma tigrinum are counted separately, as are the subspecies of Chrysemys pieta (bellii and dorsalis).

Temp- Number Number 95% confidence erature of of interval P Species and Groups range ("C) species data points Slope of slope Intercept r (slope)

[HCO:;-]

Cyjninus rarjJio 5-25 17 -0.291 -0.141 to -0.441 16.83 0.729 0.0009 /rtaluTUs punctatus 15-31 11 -0.152 0.113 to -0.417 9.48 0.398 0.2257 I IetaluTUs punctatus (Cameron & Kormanik, 1982a) -14-33 -51 -0.086 7.58 0.60 <0.01 Oncorhynchus mykiss 5-20 67 -0.016 0.135 to -0.167 7.35 0.027 0.8303 Freshwater teleosts (excluding air-breathers) 5-31 10 36 -0.118 0.056 to -0.292 10.51 0.230 0.1779 Marine fishes (teleosts and elasmobranchs) 9-28 14 20 -0.243 0.091 to -0.577 9.80 0.338 0.1466 AmlYystoma tigrinum (larval) 5-25 31 -0.393 -0.121 to -0.665 27.28 0.481 0.0062 Bufo marinus 10-30 27 -0.321 0.068 to -0.710 30.14 0.321 0.1024 Rana catesbeiana 5-34 27 -0.002 0.223 to -0.226 26.93 0.003 0.9883 Amphibians 5-34 212 40 -0.114 0.143 to-0.371 23.98 0.144 0.3744 Chrysemys piela bellii 3-30 22 0.039 0.197 to -0.119 39.09 0.114 0.6123 Turtles 3-36.2 14 30 -0.124 0.124 to -0.372 35.03 0.190 0.3140 Coluber constrictor 10-40 15 -0.156 -0.013 to -0.299 21.86 0.546 0.0353 Snakes 15-40 5 11 -0.213 0.339 to -0.765 24.80 0.279 0.4062 Lizards 16-42 8 25 0.025 0.302 to -0.253 21.79 0.038 0.8558 Crocodilians 15-35 2 4 0.035 1.524 to -1.454 16.46 0.071 0.9290 Reptiles (excluding turtles) 15-42 15 40 0.002 0.235 to -0.231 21.00 0.003 0.9857 Reptiles 3--42 29 70 -0.315 -0.124 to -0.506 33.64 0.371 0.0016

PCO.)

CY/Jrinus car/Jio 5-25 17 0.053 0.105 to 0.001 3.07 0.486 0.0479 /rtalurus /JUnctatus 15-31 11 0.045 0.140 to -0.050 1.69 0.336 0.3124 Ietalurus punctatus (Cameron & Kormanik, 1982a) -14-33 -51 0.095 0.455 0.98 <0.01 Onrorhynchus mylliss 5-20 67 0.045 0.098 to -0.008 1.89 0.208 0.0920 Freshwater teleosts (excluding air-breathers) 5-31 10 36 0.070 0.244 to -0.104 1.69 0.504 0.0017 Marine fishes (teleosts and elasmobranchs) 9-28 14 20 -0.056 0.055 to -0.167 3.24 0.243 0.3022 A mlYystoma tigrinum (larval) 5-25 31 0.121 0.245 to -0.003 6.46 0.347 0.0559 Bufo marinus 10-30 27 0.428 0.557 to 0.298 0.402 0.806 *** Rana catesbpiana 5-34 27 0.452 0.569 to 0.335 2.63 0.848 *** Amphibians 5-34 12 40 0.403 0.517 to 0.289 2.30 0.758 *** Chrysemys jJicta bellii 3-30 22 0.840 0.941 to 0.739 6.80 0.968 *** Turtles 3-36.2 14 30 0.888 1.089 to 0.687 5.56 0.862 *** Coluber constrictor 10-40 15 0.588 0.711 to 0.465 2.00 0944 *** Snakes 15-40 5 11 0.528 1.238 to -0.182 5.98 0.489 0.1269 Lizards 16-42 8 25 0.318 0.744 to -0.108 11.25 0.306 0.1373 Crocodilians 15-35 2 4 0.798 2.859 to -1.272 0.52 0.761 0.2389 Reptiles (excluding turtles) 15-42 15 40 0.412 0.726 to 0.098 8.90 0.396 0.0115 Reptiles 3--42 29 70 0.520 0.710 to 0.330 8.81 0.551 *** Ultsch & Jackson pH AND TEMPERATURE IN ECTOTHERMS 29

Table 6. (ron/inu{!(l)

Temp- Number Number 95% confidence erature of of interval P Species and Groups range (0C) species data points Slope of slope Intercept r (slope)

pH - pN"

C),/Jlinus ((II/Jio 5.25 17 0.012 0.019 to 0.004 0.585 0.670 0.0033 Irtalurlts /lUnc/atus 15-31 11 0.004 0.011 to -0.002 0.675 0.445 0.1707 Onrorhyndl1ls lIIy/{iss 5-20 67 0.010 0.Ql5 to 0.004 0.547 0.414 0.0005 Freshwater teleosts (excluding air-breathers) 5-31 10 36 0.007 0.012 to 0.003 0.628 0.505 0.0017 Marine fishes (teleosts and elasmobranchs) 9-28 14 20 0.012 0.018 to 0.007 0.468 0.722 0.0003 Amb)'s/ol1la /igrinU11l (larval) 5-25 31 0.006 0.010 to 0.002 0.623 0.523 0.0025 Bufo lllaril1US 10-30 27 0.000 0.003 to -0.003 0.823 0.003 0.9874 Rana calesbl'iana 5-34 27 0.003 0.006 to 0.001 0.746 0.451 0.0182 Amphibians 5-34 12 40 0.002 0.005 to 0.000 0.704 0.287 0.0721 ChFysemys /Jic/a bellii 3-30 22 0.005 0.007 to 0.003 0.560 0.776 *** Turtles 3-36.2 14 30 0.002 0.004 to 0.000 0.556 0.339 0.0672 Coluber cons/lie/or 15-40 15 0.007 0.009 to 0.004 0.382 0.856 *** Snakes 15-40 5 11 0.006 0.012 to -0.001 0.403 0.568 0.0680 Lizards 16-42 8 25 0.004 0.008 to 0.001 0.518 0.442 0.0271 Crocodi lians 15-35 2 4 0.003 0.007 to -0.001 0.430 0.922 0.0779 Reptiles (excluding turtles) 15-42 15 40 0.005 0.009 to 0.002 0.455 0.459 0.0029 Reptiles (= amniotes) 3-42 29 70 0.003 0.005 to 0.001 0.530 0.371 0.0016 Anamniotes (excluding air-breathing fishes) 5-34 36 96 0.006 0.008 to 0.003 0.631 0.451 *** All ectotherms (excluding air-breathing fishes) 3-42 65 166 0.001 0.003 to -0.001 0.656 0.074 0.3461 "\lhite or skeletal muscle (all ectotherms) 3-42 ~23 63 0.004 0.009 to 0.000 0.000 0.262 0.0382 Heart muscle (all ectotherms) 3-42 15 38 0.010 0.015 to 0.005 0.024 0.560 0.0003 Brain (all ectotherms) 9-31 7 14 -0.004 0.003 to -0.011 0.477 0.347 0.2236 Liver (all ectotherms) 3-37 7 16 0.000 0.009 to -0.010 0.322 0.014 0.9580 Erythrocytes (all ectotherms) 6-24 ~13 23 0.003 0.014 to -0.007 0.125 0.154 0.4841

a-imidaLOle

C),/Jri IlUS rm/Jio 5.25 17 0.003 0.005 to 0.001 0.812 0.686 0.0024 Ir/aluFlts /nl11c/a/us 15-31 11 0.001 0.002 to 0.000 0.843 0.639 0.0345 Onrorhyncll1ls lIIylliss 5-20 67 0.002 0.004 to 0.000 0.806 0.303 0.0127 Freshwater teleosts (excluding air-breathers) 5-31 \0 36 0.002 0.014 to -0.011 0.831 0.435 0.0081 Marine fishes (teleosts anel elasmobranchs) 9-28 14 20 0.004 0.006 to 0.002 0.770 0.698 0.0006 AlIlbys/ol1la lig.,.illum (larval) 5-25 31 0.002 0.003 to 0.001 0.819 0.521 0.0027 Bufo IIIIIIillllS 10-30 27 0.000 0.001 to -0.001 0.878 0.021 0.9156 Rana ((l/I'sbeialla 5-34 27 0.001 0.002 to 0.000 0.858 0.448 0.0190 Amphibians 5-34 '12 40 0.001 0.002 to 0.000 0.843 0.302 0.0579 Chl)'SI'III)'S /Jic/a bellii 3-30 22 0.002 0.002 to 0.001 0.799 0.756 *** Colubl'/" rollsllic/or 15-40 15 0.002 0.003 to 0.001 0.738 0.813 0.0002 Turtles 3-36.2 14 30 0.001 0.002 to 0.000 0.795 0.374 0.0419 Snakes 15-40 5 II 0.002 0.005 to 0.000 0.737 0.571 0.0664 Litards 16-42 8 25 0.002 0.003 to 0.001 0.776 0.520 0.0078 Crocodilians 15-35 2 4 0.001 0.004 to -0.001 0.744 0.828 0.1719 Reptiles (excluding turtles) 15-42 15 40 0.002 0.003 to 0.001 0.754 0.512 0.0007 Reptiles (= amniotes) 3-42 29 70 0.001 0.002 to 0.000 0.785 0.406 0.0005 Anamniotes (excluding air-breathing fishes) 5-34 36 96 0.002 0.002 to 0.00 I 0.825 0.419 *** 30 BULLETIN 18 December 1, 1996

Table 6. (continued)

Temp- Number Number 95% confidence erature of of interval P Species and Groups range (DC) species data points Slope of slope Intercept T (slope)

All ectotherms (excluding air-breathing fishes) 3-42 65 166 0.000 0.001 to 0.000 0.829 0.090 0.2510 White or skeletal muscle (all ectotherms) 3-42 ~23 63 0.002 0.005 to 0.000 0.527 0.237 0.0610 Heart muscle (all ectotherms) 3-42 15 38 0.005 0.007 to 0.003 0.542 0.553 0.0003 Brain (all ectotherms) 9-31 7 14 0.001 0.008 to -0.007 0.693 0.060 0.8467 Liver (all ectotherms) 3-37 7 16 0.000 0.005 to -0.004 0.692 0.029 0.9156 Erythrocyte (all ectotherms) 6-24 ~13 23 0.001 0.007 to -0.004 0.595 0.115 0.6005

*** p < 0.0001 I Data are for total CO" content 2 Counting seawater- a~d freshwater-adapted OlicoriL),chus /IIy1/iss separately

group, however, did show a slight, but significant, increase 40 . T in PC0 with temperature, suggesting that acid-base regu­ I- =SIGNIFICANT SLOPE, P<:O.OS 2 lation, at least with changes in temperature, may not be

30 strictly limited to changes in [HC0 -]. These limited data c 3 "'"0> reveal a diversity of response in the water breathers and :r: suggest that fishes are able to use both respiratory and E E 20 ionic mechanisms to accomplish temperature-dependent o pH regulation. This conclusion has previously been made () by Heisler (1986) in his review of acid-base regulation in Q. 10 fishes.

E+MT SIGNIFICANCE OF INTER-GROUP pH DIFFERENcEs-Our analysis 0 of temperature-pH relations in ectothermic vertebrates 0 10 20 30 40 has revealed significant inter-group differences, particu­ FT~FRESHWATER TELEOSTS T=TURTLES E+MT=ELASMOBRANCHS AND L-liZARDS larly in blood pH. The consistency of these differences MARINE TELEOSTS C-CROCODILIANS between, for example, freshwater fishes and amphibians S=SNAKES 40 - A=AMPHIBIANS on the one hand and reptiles on the other hand, begs for a functional or adaptive interpretation. At the outset, however, we acknowledge that we do not have an explana­ 30 -T -1 tion for these differences, nor indeed are we confident ...... r;j that an "explanation" even exists. One reason for our w E uncertainty is that parallel differences within the intracel­ 20 ~A lular fluid compartments either were not detected or were ,,.,~ 0 by no means as clear or convincing, although it is within () ---s :r: the cells that important functional impacts of a pH differ­ 10 ence would be expected to exist. As we emphasized ear­ -FT lier, however, the smaller sample size, the measurement E+MT uncertainty, and the sample heterogeneity of the intracel­ o fl--~-,_-~--,-~--~-~-_.------=== lular data make conclusions for this compartment less o 10 20 30 40 solid. Our postulations about pH differences will there­ TEMPERATURE tC) fore be limited to the blood pH results.

'] Figure 12. Regression curves for PC02 and [HC03 as PLASMA [NA+] OR OSMOlALITY-A major inter-group differ­ functions of temperature for several groupings of ence in pHh we identified exists between freshwater spe­ ectotherms. See Table 6 for details of the regression cies, both teleosts and amphibians, and marine or terres­ analyses. trial species, including teleosts, elasmobranchs, and rep- Ultsch & Jackson pH AND TEMPERATURE IN ECTOTHERMS 3 1 ------

tiles. An obvious expected correlated difference between ship based on our own survey of the literature as shown these two general groupings is in body fluid osmolality or for data at 15 and 25()C in Fig. 13. We assembled values of plasmasodium concentration. Of particular interest to us, both variables from species of fishes, reptiles, and am­ therefore, was a recent analysis by Burton (1994) that phibians, using our stated criteria for pHb' and criteria for showed an inverse relationship between plasma pH and [Na+] data as described in the figure legend, and did not plasma [Na+] in various reptiles and amphibians at 25°C. find a significant relationship between plasma pH and In general he found amphibians to have a low [Na+] and [Na+]. Although we used some of the same data plotted by high pHb' non-turtle reptiles to have a high [Na+] and low Burton, some his data did not meet our criteria and are pHb' and turtles to be intermediate in both variables. therefore not included in our analysis, and we included Although not presented in the paper, the same relation­ other data he did not use. At 25°C, for example, our data ship was also reported to hold at other temperatures. A include amphibians with lower pHb and reptiles with theoretical interpretation for this finding was presented higher pHb compared to those in the Burton compilation, that was based on the indirect effect of solution [Na+] on and we also include some data on freshwater fishes. Simi­ bulk fluid pH via an effect on cell surface net negative lar analyses of available data from ectothermic vertebrates charge. Burton postulated that the pH at the cell surface at 5, 10,20, and 30°C likewise failed to reveal the consis­ is the same in all species. tent relationship reported by Burton. We unfortunately were unable to verity this relation- An inverse relationship between plasma pH and [Na+]

7.9 l B.O • 12

/5 7.8 -! 3 •• .11 4 ·6 4 ,,! • 1 9 • • 1 m--0.0002 5 •• .10 6 m=-0.0007

.2 . 13 .2 ~ 7.81 • 3 ·7 ~ "7. 6 1 .7 .14 .8 7.7l 7.5 -! 115°c I ~ .15

7 .4 : 76 80 100 120 140 160 \ 8 0 80 100 120 140 160 180 [Net] (mEq/L) [Ncr] (mEq/L)

Figure 13. Plasma pH at 25 ± 2°C and at 15 ± 2°C as a function of [Na+] in a variety of ectotherms. The slopes of the regression lines are not significantly different from zero (25°C, P= 0.57, r = 0.159; 15 C, P = 0.88, r = 0.069). Only pH data from temperature-acclimated, arterially-catheterized, resting animals were used, as shown directly or by interpola­ tion from Table 1. Data on [Na+] were from temperature-acclimated animals only, although blood samples were not always drawn through catheters nor where they always from the same studies from which pH data were obtained. When multiple studies of pH or [Na+] were available, the points shown represent a mean of these studies. Animals and references for data at 25°C: Salamanders-(I) Cryptobra1lchlls allegalliellsis (Moalli et aI., 1981); (2) NecrnrllS maclilosus (Stiffler et aI., 1983); (3) larval Ambystoma tigrillum (Rohrbach & Stiffler, 1987; Stiffler, 1991; Stiffler & Bachoura; 1991; Stiffler et aI., 1983, 1987; Talbot & Stiffler, 1992); (4) Sire1llacerti1la (Heisler et aI., 1982); (5) Amphiuma mea1ls (Heisler et aI., 1982); (6) adult Ambystoma tigrilll1111 (Stiffler, 1991 )-Freshwater fishes-(7) Protopterus aethiopicus (DeLaney et aI., 1977); (11) Ictalurlls pllllctatus (Cameron, 1980 )-Anurans-(8) Bufo marilllls (Hillman, 1980; Tufts & Toews, 1985); (9) Bufo viridis (Gordon, 1962; Katz, 1980a,b, 1981); (10) Xellopus laevis (Boutilier, 1984; Boutilier & Shelton, 1986; Boutilier et aI., 1987; Emilio & Shelton 1980; Hillman, 1980)-Lizard-(12) Dipsosaurus dorsalis (Bickler, 1984)­ Snakes-( 13) Pituophis melalloleucas (Komadina & Solomon, 1970; Stinner, 1982); (14) Nerodia siped011 (Dantzler, 1967; Dean & Gratz, 1983; Gratz, 1984; Komadina & Solomon, 1970; Wasser, 1990); -Alligator (15) Alligator mississippiensis (Coulson etaI., 1950; Davies, 1978; Davies etaI., 1982). For data at 15°C, the animals and references are: Salamanders­ (1) Cryptobrallchus allegalliellsis (Moalli et aI., 1981); (2) Ambystoma tigrillltl11 larvae (DeRuyter & Stiffler, 1988 );­ Turtle-(3) Chrysemys picta bellii (Butler & Knox, 1970; Herbert & Jackson, 1985)-Freshwater fishes-(4) Cyprimls carpio (Houston & Madden, 1968; Jensen et aI., 1987; Ultsch et aI., 1981); (5) Anguilla allguilla (Bornancin et aI., 1977); (6) 01lcorhYllchus mykiss (Curtis & Wood, 1992; Holeton et aI., 1983a,b; Ishimatsu et al., 1992; Munger et al., 1991; Thomas & Poupin, 1985; Thomas et al., 1986; Turner, Wood & Clark, 1983; Wang et aI., 1994; Wilkie & Wood, 1991; Wood & LeMoigne, 1991 );-Snake-(7) PitllOphis melalloleucas (Komadina & Solomon, 1970; Stinner, 1982). 32 BULLETIN 18 December 1, 1996

is likely to exist for teleosts. As discussed earlier in this the study of comparative aspects of acid-base balance paper, pH" values are significantly higher in freshwater among ectothermic vertebrates. teleosts than in marine teleosts, and plasma [Na+] is gen­ erally lower in the freshwater species (Schmidt-Nielsen, Acknowledgements 1990, page 313); however, insufficient data exist where We thank P. J. Butler, A. Ishimatsu, S. F. Perry, D. F. pH" and [Na+] are available for the same marine species Stiffler, P. L. M . van Dijk, and J. S. Wasser for providing to permit a reliable comparison. Elasmobranchs, on the details on their published acid-base data, and J. B. Gra­ other hand, have pH" values that do not differ signifi­ ham for information on the behavior of air-breathing cantly from marine teleosts, yet their [Na+] is significantly fishes. G. R. U. appreciates the hospitality and space higher. Burton's suggested explanation for plasma pH provided him during his sabbatical at the Department of variability, therefore, is an intriguing one, but does not Physiology at Brown University, during which time this appear to provide a consistent basis for the observed inter­ review was finalized. group differences. Literature Cited WATER-BREATHERS vs. AIR BREATHERS-Another possible reason for consistent pH" differences could relate to res­ Ackerman, R.A., and F.N. ""hite. 1980. The effects of tempera­ ture on acid-base balance and ventilation of the marine piratory mode. Reptiles, the most terrestrial and commit­ iguana. Respiration Physiology 39: 133-147. ted air-breathers among the ectothermic vertei)rates, Aguinaga, E.M., and D.F. Stiffler. 1993. Interrenal function in have a consistently lower blood pH than do freshwater larval Ambys{o/lw Iigrinu1Il. III. Acid-base balance responses. teleosts or amphibians. Compared to water-breathers, Ceneral Comparative Endocrinology. 90: 100-108. reptiles have greatly reduced breathing volumes because Andreasen, P. 1985. Free and total calcium concentrations in of the higher oxygen content of air, but perhaps these the blood of rainbow trout, Saimo gairtineri, during 'stress' volumes are reduced as far as possible to maximize water conditions. journal Experimental Biology 118: 111-120. conservation. Under the assumption that a range of pH" Austin,j.H., F.W. Sunderman, andj.C. Camack. 1927. Studies in values would be "normal" for a particular temperature, serum electrolytes. II. The electrolyte composition and the then the reptiles might be regulating their blood pH at pH of serum of a poikilothermous animal at different tem­ the lower end of this range, while the freshwater teleosts peratures. journal Biological Chemistry 72: 677-685. Barber, M.L., and P..J. Walsh. 1993. Interactions of acid-base and amphibians are operating closer to the upper end of status and nitrogen excretion and metabolism in the this range. For the fishes this may be governed by their ureogenic teleost O/JSal1'llS bpta. journal Experimental Biol­ requirement to move large volumes of water over their ogy 185: 87-105. gills to supply oxygen; for the amphibians, the important Bickler, P.E. 1981 . Effects of temperature on acid-base balance aspect may be the ease with which they can lose C09 and ventilation in desert iguanas. journal Applied Physiol­ cutaneously. - ogy 51: 452-460. One could argue in opposition to this idea from several Bickler, P.E. 1982. Intracellular pH in lizard Di/JSOS(lUrlIS dOl;mlis standpoints. First, it is not clear why the animals in each of in relation to changing body temperature. journal Applied these groups could not achieve the same blood pH, de­ Physiology 53: 1466-1472. spite their special considerations, by adjusting the ionic Bickler, P.E. 1984. Effects of temperature on acid and base composition of their plasma (i.e., [HCO -]). This com­ excretion in a lizard, Di!Jso.I· auru~ dOl:lalil'. Journal Compara­ g tive Physiology B 154: 97-104. pensation clearly played a role in the transition from Bickler, P.E. (1986). Day-night variations in blood and intracel­ aquatic to terrestrial life, and a small further change lular pH in a lizard, Dj/iso.mlll'lil dorsa/il.journal Comparative should certainly be feasible. Second, marine teleosts and Physiology B 156: 833-857. elasmobranchs are also water-breathers and have de­ Booth,.J.H., C.F.jansz, and C.F.Holeton. 1982. CI-, K+, and acid­ mands for gill ventilation comparable to freshwater teleo­ base balance in rainbow trout during exposure to, and recov­ sts, yet their blood pH is significantly lower then that of ery from, sublethal environmental acidification. Canadian freshwater teleosts. Journal Zoology 60: 1123-1130. Bornancin, M., C, DeRenzis, and.J. Maei/. 1977. Branchial CI CONCLUSIONs-We have assembled what we believe is the transport, anion-stimulated ATPase and acid-base balance in majority of data available on the acid-base status of ecto­ Anguilla anguilla adapted to freshwater: effects of hyperoxia. therms in an attempt to provide an organized data base Journal Comparative Physiology 117: 313-322. Boutilier, R.C. 1984. CharacterLmtion of the intermittent for investigators in the area. Several trends not previously breathing pattern in Xeno/J//\ lrwuis. Journal Experimental recognized have emerged, such as a consistently higher Biology 1\0: 291-309. pH at a given temperature among anamniotes when com­ Boutilier, R.C., C. Dobson, U. Hoeger, and OJ Randall. 1988. pared with reptiles. At present, we are not able to deduce Acute exposure to graded levels of hypoxia in rainbow trout mechanisms that may be responsible for such differences, (Sa/1l1O gairdueri): metabolic and respiratory adaptations. but hope that this review will stimulate further interest in Respiration Physiology 71: 69-82. Ultsch & jackson pH AND TEMPERATURE IN ECTOTHERMS 33

Boutilier, R.G., R.A. Ferguson, R.P. Henry, and B.L. Tufts. ] 993. base balance in the salamander, Ambystomn tigJinu1I!: influ­ Exhaustive exercise in the sea lamprey (Petrolll),ZOI1 IIUlrinus) : ence of temperature acclimation and metamorphosis. jour­ relationship between anaerobic metabolism and intracellu­ nal Comparative Physiology 144: 241-246. lar acid-base balance. journal Experimental Biology] 78: 7]- Burton, R.F. 1994. The dependence of normal plasma pH on 88. sodium concentration in amphibians, reptiles and man. Boutilier, R.G., M.L. Glass, and N. Heisler. ] 987. Blood gases, Comparative Biochemistry Physiology 108A: 1-5. and extracellular/intracellular acid-base status as a function Bushnell, P.G., P.L. Lutz, J.F. Steffensen, A. Oikari, and S.H. of temperature in the anuran amphibians XenojJ1o {arois and Gruber. 1982. Increases in arterial blood oxygen during Rrma mtl>sbeirl11a.journal Experimental Biology 130: 13-25. exercise in the lemon shark (Nfgaj)rion brevirostris). journal Boutilier, R.G., and N. Heisler. 1988. Acid-base regulation and Comparative Physiology ]47: 4]-47. blood gases in the anuran amphibian, Buro I/lminw, during Butler, D.G., and W.H. Knox. ]970. Adrenalectomy of the environmental hypercapnia. journal Experimental Biology painted turtle (Chysemis j)irta belli) : effect on ionoregulation ]34: 79-98. and tissue glycogen. General Comparative Endocrinology Boutilier, R.G., G.K Iwama, anel DJ. Randall. ] 986. The promo­ ]4,55]-566. tion of catecholamine release in rainbow trout, Salmo Butler, PJ., and N. Day. 1993a. The relationship between intrac­ gainlneri, by acute acidosis: interactions between red cell pH ellular pH and swimming performance of brown trout ex­ and haemoglobin oxygen-carrying capacity. journal Experi­ posed to neutral and sublethal pH. journal Experimental mental Biology 123: 145-]57. Biology 176: 271-284. Boutilier, R.G., and CJ. Lantz. ] 989. The effects of forced and Butler, PJ., and N. Day. ] 993b. The relationship between intrac­ voluntary diving on plasma catecholamines and erythrocyte ellular pH and seasonal temperature in the brown trout pH in the aquatic anuran, Xel10jJlIS {aevis. Experimental Biol­ Salmo truUa. journal Experimental Biology 177: 293-297. ogy 48: 83-88. Butler, PJ., and E.W. Taylor. ] 975. The effect of progressive Boutilier, R.G., D.G. McDonald, and D.P. Toews. ] 980. The hypoxia on respiration in the dogfish (Sryliorhinus caniwln) effects of enforced activity on ventilation, circulation and at different seasonal temperatures. journal Experimental blood acid-base balance in the aquatic gill-less urodele, Biology 63: ]] 7-] 30. CI)'j)tobrnnrll1ts aUI!ganiensis; a comparison with the semi-ter­ Cameron, J.N. 1976. Branchial ion uptake in arctic grayling: restrial anuran, Bufo lIIarinus.journal Experimental Biology resting values and effects of acid-base disturbance. journal 84: 289-302. Experimental Biology 64: 7] ]-725. Boutilier, R.G., DJ. Randall, G. Shelton, and D.P. Toews. 1979a. Cameron, J.N. 1978. Regulation of blood pH in teleost fish . Acid-base relationships in the blood of the toad, 8ufo Respiration Physiology 33: 129-144. 'IIlarinus. 1. The effects of environmental CO~. journal Ex­ Cameron, J.N. ]980. Body fluid pools, kidney function, and perimental Biology 82: 33 ]-344. acid-base regulation in the freshwater catfish, letalltrus Boutilier, R.G., DJ. Randall, G. Shelton, and D.P. Toews. ] 979b. jntllctatus. journal Experimental Biology 86: ] 71-] 85. Acid-base relationships in the blood of the toad, BuJo Cameron, J.N. ] 985. The bone compartment in a teleost fish, I//arin-Its. II. The effects of dehydration. Journal Experimen­ le/alltrlls jntl1ctntus: size, composition and acid-base response tal Biology 82: 345-355. to hypercapnia.journal Experimental Biology] 17: 307-3] 8. Boutilier, R.G., DJ. Randall, G. Shelton, and D.P. Toews. 19791:. Cameron, J.N. ] 989. Acid-base homeostasis: past and present Acid-base relationships in the blood of the toad, /311fo perspectives. Physiological Zoology 62: 845-865. I/lruinus. Ill. The effects of burrowing. Journal Experimental Cameron,J.N., and N. Heisler. 1983. Studies of ammonia in the Biology 82: 357-365. rainbow trout: Physicochemical parameters, acid-base Boutilier, R.G., and G. Shelton. 1986. The effects of forced and behaviour and respiratory clearance. journal Experimen tal voluntary diving on ventilation, blood gases and pH in the Biology 105: 107-125. aquatic amphibian, XellojJUs (rll!vis. Journal Experimen tal Bi­ Cameron, J.N., and G.K. Iwama. ] 987. Compensation of pro­ ology 122: 209-222. gressive hypercapnia in channel catfish and blue crabs.Jour­ Boutilier, R.G., and D.P. Toews. ] 981. Respiratory, circulatory nal Experimental Biology ]33: ]83-]97. and acid-base acljustments to hypercapnia in a strictly aquatic Cameron, IN., and G.A. Kormanik. ]982a. Intracellular and and predominantly skin-breathing urodele, CI)'j)lobrrl11r/t-us extracellular acid-base status as a function of temperature in a {{1!ganil'llsis. Respiration Physiolq,,")' 46: 177-192. the freshwater channel catfish, /ctalllrus jmnrtatlls. journal Branco, L.G.S., M.L. Glass" T. Wang, and A. Hoffmann. 1993. Experimental Biology 99: 127-142. Telll perature and cen tral chemoreceptor drive to ven tilation Cameron, .J.N., and G.A. Kormanik. 1982b. The acid-base re­ in toad (Buro j)((mnll>mis). Respiration Physiolq,'")' 93: 337- sponses of gills and kidneys to infused acid and base loads in 346. the channel catfish, /clnlnrlls /mnclatus. journal Experimen­ Burggren, W.W. 1978. Gill ventilation in the sturgeon, Adj)(!lIsl>r tal Biology 99: ]43-160. Imnsl//olllrlllUS: unusual adaptations to bottom dwelling. Res­ Cameron,J.N., and DJ. Randall. ]972. The effect of increased piration Physiology 34: 153-170. ambient CO., on arterial CO., tension, CO., content and pH Burggren, W.W., J.N. Cameron. 1980. Anaerobic metabolism, in rainbow lI~ul.journal Experimental Bi;logy 75: 673-680. gas exchange, and acid-base balance during hypoxic expo­ Cameron,J.N., and C.M . Wood. ]978. Renal function and acid­ sure in the channel catfish, irlalunts IJU11ctallls. Journal Ex­ base balance in two Amazonian erythrinid tishes: HOj)lias perimental. Zoology 213: 40!l-4l6. lIutlabmiclIs, a water breather, and Hnj)li!I),t/uinlls unitfUmilttus, a Burggren, W.W., and S.c. Wood. 1981. Respiration and acid- facultative air breather. Canadian Journal Zoology 56: 9] 7-930. 34 BULLETIN 18 December 1,1996

Cech, JJ., D.W. Bridges, D.M. Rowell, and PJ. Balzer. 1976. Delaney, R.G., S. Lahiri, and A.P. Fishman. 1974. Aestivation of Cardiovascular responses of winter flounder Pselldo­ the African lungfish Protopterus aethio/Jieus : cardiovascular jJlellmnertes allll'ricanlls (Walbaum), to acute temperature in­ and respiratory function. Journal Experimental Biology 61: crease. Canadian Journal Zoology 54: 1383-1388. 111-128. Cech,JJ., D.M. Rowell, andJ.S. Glasgow. 1977. Cardiovascular Delaney, R.G., S.Lahiri, R. Hamilton, and A.P. Fishman, A.P. responses of the winter flounder Pselldo/Jleuronl'rtes 1977. Acid-base balance and plasma composition in the aes­ ameriwnusto hypoxia. Comparative Biochemistry Physiology tivating lungfish (Proto/Jtems). American Journal Physiology 57A: 123-125. 232: RI0-RI7. Claiborne, J.B., and D.H. Evans. 1992. Acid-base balance and DeRuyter, M.L., and D.F. Stiffler. 1988. Acid-base and electrolyte ion transfers in the spiny dogfish (Squalus acanthias) during responses to low pH maintained by phosphate-citrate buffers hypercapnia: a role for ammonia excretion. Journal Experi­ in the bathing medium of the larval salamander Ambystom(l mental Zoology 261: 9-17. tigrinum. Canadian Journal Zoology 66: 2383-2389. Claiborne,J.B., and N. Heisler. 1984. Acid-base regulation and Dill, D.B., and H.T. Edwards. 1935. Properties of reptilian ion transfers in the carp (Cyprinus rar/Jio) during and after blood. IV. The alligator (Alligator mississi/Jiensis) .Journal Cel­ exposure to environmental hypercapnia. Journal Experi­ lular Comparative Physiology 6: 243-254. mental Biology 108, 25-43. Dill, D.B., H. T. Edwards, A.V. Bock, and J.H. Talbott. 1935. Claiborne,J.B., and N. Heisler. 1986. Acid-base regulation and Properties of reptilian blood. Ill. The chuckawalla ion transfers in the carp (Cyprinus cmlJio): pH compensation (Saummalus obesus). Journal Cellular Comparative Physiol­ during graded long- and short-term environmental hyper­ ogy 6: 37-42. capnia, and the effect of bicarbonate infusion. Journal Ex­ Eddy, F.B. 1974. Blood gases of the tench (Tinra tinra) in well perimental Biology 126: 41-61. aerated and oxygen-deficient waters. Journal Experimental Coulson, R.A., T. Hernande7, and F.G. Brazda. 1950. Biochemi­ Biology 60: 71-83. cal studies on the alligator. Proceedings Society Experimen­ Eddy, F.B., J.P. Lomholt, R.E. Weber, and K Johansen. 1977. tal Biology Medicine 73: 203-206. Blood respiratory properties of rainbow trout (Salmo

Courtice, G.P. 1981. Respiration in the eastern water dragon, gairrineri) kept in water of high CO2 tension. Journal Experi­ Physignathus lesueurii (Agamidae). Comparative Biochemis­ mental Biology 67: 37-47. try Physiology 68A: 429-436. Emilio, M.G., and G. Shelton. 1980. Carbon dioxide exchange Crocker, C.E., and D.F. Stiffler. 1991. The effects of amiloride on and its effects on the pH and bicarbonate equilibria in the electrolyte transport and acid-base balance in the larval sala­ blood of the amphibian, Xeno/ms trwvis. Journal Experimen­ mander, Ambystoma tigrinum. Journal Comparative Physiol­ tal Biology 85: 253-262. ogy B 161: 460-464. Ferguson, R.A., and R.G. Boutilier. 1988. Metabolic energy Currie, S., and B.L. Tufts. 1993. An analysis of carbon dioxide production during adrenergic pH regulation in red cells of transport in arterial and venous blood of the rain bow trout, the Atlantic salmon, Salmo sa la 1". Respiration Physiology 74, (Oncorhynchus my/liss) , following exhaustive exercise. Fish 65-76. Physiology Biochemistry 12: 183-192. Ferguson, R.A.,J.D. Kieffer, and B.L. Tufts. 1993. The effects of Curtis, BJ., and C.M. Wood. 1992. Kidney and urinary bladder body size on the acid-base and metabolite status in the white responses of freshwater rainbow trout to isosmotic NaCI and muscle of rainbow trout before and after exhaustive exercise.

NaHCD:1 infusion. Journal Experimental Biology 173: 181- Journal Experimental Biology 180: 195-207. 203. Fuchs, D.A., and C. Albers. 1988. Effect of adrenaline and blood Dantzler, W.H. 1967. Glomerular and tubular effects of arginine gas conditions on red cell volume and intra-erythrocytic vasotocin in water snakes (Natrix sipedon). AmericanJournal electrolytes in the carp (CY/Jl7n'llS Cfll/Jio) .Journal Experimen­ Physiology 212: 83-91. tal Biology 137: 457-477. Davies, D.G. 1978. Temperature-induced changes in blood acid­ Garey, W. F., and H. Rahn. 1970. Normal arterial gas tensions base status in the alligator, Alligator mississi/Jiensis. Journal and pH and the breathing frequency of the electric eel. Applied Physiology 45: 922-926. Respiration Physiology 9: 141-150. Davies, D.G.,J.L. Thomas, and E.N. Smith, E.N. 1982. Effect of Garland, R.J., and D.P. Toews. 1992. Acid-base regulation in body temperature on ventilatory control in the alligator. response to hypercapnia in the lymphatic and circulatory Journal Applied Physiology. 52: 114-118. systems of the toad Bufo IIUlrin'lls.Journal Experimental Biol­ Dean,.J.B., and R.K Gratz. 1983. The effect of body t~mperature ogy 170: 271-276.

and CO2 breathing on ventilation and acid-base status in the Gilmour, KM., DJ. Randall, and S.F. Perry. 1994. Acid-base northern water snake Nerodia sij){!({on. Physiological Zoology disequilibrium in the arterial blood of rainbow trout. Respi­ 56: 290-301. ration Physiology 96: 259-272. Dejours, P. 1973. Problems of control of breathing in fishes. In Gilmour, KM., and S.F Perry, S.F. 1994. The effects ofo hypoxia, L. Bolis, K Schmidt-Nielsen & S.H.P. Maddrell (eds.). Com­ hyperoxia or hypercapnia on the acid-base disequilibrium in parative Physiology: Locomotion, Respiration, Transport the arterial blood of rainbow trout. Journal Experimental and Blood (eel., pp. 117-133. North Holland Publishing Co., Biology 192: 269-284. Amsterdam. Glass, M.L., N.A. Andersen, M. Kruh0ffer, E.M. Williams, and N.

Dt:jours, P., andJ. Armand. 1973. L'equilibre acide-base du sang Heisler. 1990. Combined effects of environmen tal P02 and chez la Carpe en fonction de la tempeature.Journal Physiol­ temperature on ventilation and blood gases in the carp ogy (Paris) 67: 264A. (CY/Jl7nltS cmlJio).Journal Experimental Biology 148: 1-17. Ultsch & Jackson pH AND TEMPERATURE IN ECTOTHERMS 35

Glass, M.L., R.G. Boutilier, and N. Heisler. ] 983. Ventilatory intracellular pH with changes of temperature in the dogfish control of arterial PO~ in the turtle Ch,}'selll),s jJieta bellii: Scyliorhinus slelim1s. Respiration Physiology 26: 249-263. effects of temperature and hypoxia. Journal Comparative Henderson, L.J. ] 928. Blood. A Stud)' in General Physiology. Physiology 15]: ]45-]53. 397 pp. Yale Univ. Press, New Haven. Glass, M.L., R.G. Boutilier, and N. Heisler. 1985. Effects of body Herbert, C.V., and D.C. Jackson. 1985. Temperature effects on temperature on respiration, blood gases and acid-base status the responses to prolonged submergence in the turtle in the turtle Chryselll)'s jJicta bellii. Journal Experimental Biol­ Chr)'sem)'s picta bellii. I. Blood acid-base and ionic changes ogy] ]4: 37-51. during and following anoxic submergence. Physiological Glass, M.L., and N. Heisler. ] 986. The effects of hypercapnia on Zoology 58: 655-669. the arterial acid-base status in the tegu lizard, TitjJina1llbis Hicks, G.H., and D.F. Stiffler. 1980. The effects of temperature nigrojmnctatus. Journal Experimental Biology] 22: 13-24. and hypercapnia on acid-base status of larval Amb),stoma Gleeson, T.T., and A.F. Bennett. ] 982. Acid-base imbalance in tigrinulIl. Federation Proceedings 39: 1060. lizards during activity and recovery. Journal Experimental Hicks, G.H., and D.F. Stiffler. ] 984. Patterns of acid-base regula­ Biology 98: 439-453. tion in urodele amphibians in response to variations in Gordon, M.S. ]962. Osmotic regulation in the green toad (BuJo environmental temperature. Comparative Biochemistry viridis).Journal Experimental Biology 39: 26]-270. Physiology 77A: 693-697. Goss, G.G., and C.M. Wood. ] 990. Nat and Cl- uptake kinetics, Hicks, J.H., A. Ishimatsu, and N. Heisler. 1987. Oxygen and diffusive effluxes and acidic equivalent fluxes across the gills carbon dioxide transport characteristics of the blood of the of rainbow trout. Journal Experimental Biology 152: 459- Nile monitor lizard (\farrl1lus niloliells) .Journal Experimental 571. Biology] 30: 27-38. Goss, G.G., and C.M. Wood. ] 991. Two-substrate kinetic analy­ Hillman, S.S. 1980. Physiological correlates of differential dehydra­ sis: a novel approach linking ion and acid-base transport at tion tolerance in anuran amphibians. Copeia 1980: ]25-]29. the gills of freshwater trout, Oncorhynchus lIly/dss. Journal Hitzig, B.M. 1982. Temperature-induced changes in turtle CSF Comparative Physiology B ]6]: 635-646. pH and central control of ventilation. Respiration Physiology Gottlieb, G., and D.C. Jackson. ] 976. Importance of pulmonary 49: 205-222. ventilation in respiratory control in the bullfrog. American Hitzig, B.M., and E.E. Nattie. 1982. Acid-base stress and central Journal Physiology 230: 608-613. chemical control of ventilation in turtles. Journal Applied Graham, M.S., J.D. Turner, and C.M. Wood. 1990. Control of Physiology 53: ]365-]370. ventilation in the hypercapnic skate Raja oceliala: I. Blood Hitzig, B.M., W. Perng, T. Burt, P. Okunieff, and D.C.Johnson. and extradural fluid. Respiration Physiology 80: 259-277. 1994. IH-NMR measurement of fractional dissociation of Gratz, R.K. 1984. Effect of bilateral vagotomy on the ven tilatory imidazole in intact animals. American Journal Physiology responses of the water snake Nerodia sijJedon. American Jour­ 266: R]008-RlO]5. nal Physiology 246: R221-R227. Hobe, H., P.R.H Wilkes, R.L. Walker, C.M. Wood, and B.R. Heisler, N. ]982a. Transepithelial ion transfer processes as McMahon. 1983. Acid-base balance, ionic status, and renal mechanisms for fish acid-base regulation in hypercapnia and function in resting and acid-exposed white suckers (Catostomus lactacidosis. Canadian Journal Zoology 60: ] ]08-] ]22. commerso11i). Canadian Journal Zoology 61: 2660-2668. Heisler, N. ] 982b. Intracellular and extracellular acid-base regu­ Hobe, H., C. M.Wood, and M.G. v\'heatly_ ]984. The mecha­ lation in the tropical fresh-water teleost fish S),nbrrl1lrlws nisms of acid-base and ionoregulation in the freshwater marmoratus in response to the transition from water breath­ rainbow trout during environmental hyperoxia and subse­ ing to air breathing. Journal Experimental Biology 99: 9-28. quent normoxia. I. Extra- and intracellular acid-base status. Heisler, N. ] 986. Comparative aspects of acid-base regulation. In Respiration Physiology 55: 139-154. N. Heisler (ed.). Acid-base Regulation in Animals, p. 397- Holeton, G.F. 1977. Constancy of arterial blood pH during CO­ 450. Elsevier, Amsterdam. induced hypoxia in the rainbow trout. Canadian Journal Heisler, N.,G. Forcht, G.R. Ultsch, and J,F. Anderson. ]982. Zoology 55: 1010-]0]3. Acid-base regulation in response to environmental hyper­ Holeton, G.F., P. Neumann, and N. Heisler. ] 983. Branchial ion capnia in two aquatic salamanders, Simn {a(('rti11a and exchange and acid-base regulation after strenuous exercise A mjJ/tiuma means. Respiration Physiology 49: ] 4] -158. in rainbow trout (Salmo gairdneri). Respiration Physiology 5]: Heisler, N., P. Neumann, and G.F. Holeton. ]980. Mechanisms 303-318. of acid-base adjustment in dogfish (Scyliorhinus stl'ilruis) sub­ Holeton, G.F., J.H Booth, and G.F. Jansz. 1983. Acid-base bal­ jected to long-term temperature acclimation. Journal Ex­ ance and Nat regulation in rainbow trout during exposure perimental Biology 85: 89-98. to, and recovery from, low environmental pH. Journal Ex­ Heisler, N., D.P Toews, and G.F. Holeton. 1988. Regulation of perimental Zoology 228: 21-32. ventilation and acid-base status in the elasmobranch Houston, A.H., and J.A. Madden. 1968. Environmental tem­ Sl),liorhinlls stellmis during hyperoxia-induced hypercapnia. perature and plasma electrolyte regulation in the carp, Respiration Physiology 71: 227-246. Cyjninlls rm/Jio. Nature 217: 969-970. Heisler, N., H. Weitz, and A.M. Weitz. 1976a. Hypercapnia and Howell, Bj., F.W. Baumgardner, K. Bondi, and H. Rahn. ] 970. resul tan t bicarbonate transfer processed in an elasmobranch Acid-base balance in cold-blooded vertebrates as a function fish. Bulletin Europeen Physiopathologie Respiratoire 12: of body temperature. American Journal Physiology 2] 8: 600- 77-85. 606. Heisler, N., H. Weitz, and A.M . Weitz. ]976b. Extracellular and Hughes, G.M., andI.A.Johnston. ]978. Some responses of the 36 BULLETIN 18 December 1,1996

electric ray (TOIpedo 11lannomla) to low ambient oxygen ten­ Johansen, K., C. Lenfant, K. Schmidt-Nielsen, andJ.A. Petersen. sions.Journal Experimental Biology 73: 107-117. 1968. Gas exchange and control of breathing in the electric Ishimatsu. A, and Y. Itazawa. 1983. Blood oxygen levels and acid­ eel, Electro/J/WTIIS electl7Clls. Zeitschrift Vergleichende Physio­ base status following air exposure in an air-breathing fish, logie 61: 137-163. Channa argus: the role of air ventilation. Comparative Bio­ Johnson, D.C., C.T. Burt, W. Perng, and B.M. HiLLig, B.M. 1993. chemistry Physiology 74A: 787-793. Effects of temperature on muscle pHi and phosphate me­ Ishimatsu, A., G.K. Iwama, T.B. Bentley, and N. Heisler. 1992. tabolites in newts and lungless salamanders. American Jour­ Contribution of the secondary circulatory system to acid­ nal Physiology 265: Rl162-R1167. base regulation during hypercapnia in rainbow trout Katz, U. 19800. Acid-base status in the toad Bufa vil7dis in vivo. (Oncorh),l1fhus lIly/liss). Journal Experimental Biology 170: Respiration Physiology 41: 105-115. 43-56. Katz, U .. 1980b. The effect of dehydration on the in vivo acid­ Itazawa, Y., and T. Takeda. 1978. Gas exchange in the carp gills base status of the blood of the toad 8ufo viridis. Journal in normoxic and hypoxic conditions. Respiration Physiology Experimental Biology 88: 403-405. 35: 263-269. Katz, U. 1981. The effect of salt adaptation and amiloride on the Jackson, D.C. 1982. Strategies of blood acid-base control in in vivo acid-base status of the euryhaline toad 8u[0 viridis. ectothermic vertebrates. In C.R. Taylor, K. Johansen, & L. Journal Experimental Biology 93: 93-99. Bolis (eds.). A Companion to Animal Physiology, pp 73-90. Kieffer, J.D., S. Currie, and B.L. Tufts. 1994. Effects of environ­ Cambridge University Press, Cambridge. mental temperature on the metabolic and acid-base re­ Jackson, D.C., and B.A. Braun. 1979. Respiratory control in sponses of rainbow trout to exhaustive exercise. Journal bullfrogs: cutaneous versus pulmonary response to selective Experimental Biology 194: 299-317.

CO2 exposure. Journal Comparative Physiology 129: 339- Kinkead, R., and W.K. Milsom. 1994. Chemoreceptors and con­ 342. trol of episodic breathing in the bullfrog (Rana catl!sbl!iana). Jackson, D.C., and N. Heisler. 1983. Intracellular and extracellu­ Respiration Physiology 95: 81-98. lar acid-base and electrolyte status of submerged anoxic Kinney,J.L., D.T. Matsuura, and F.N. White. 1977. Cardiorespi­ turtles at 3()C. Respiration Physiology 53: 187-201. ratory effects of temperature in the turtle, PseurlelllYs Jackson, D.C., and R.D. Kagen. 1976. Effects of temperature jlUlidrl1la. Respiration Physiology 31: 309-325. transients on gas exchange and acid-base status of turtles. Komadina, S., and S. Solomon. 1970. Comparison of renal American Journal Physiology 230: 1389-1393. function of bull and water snakes (PiIUO/J/tis JIlelmlOlf?!tCllsand Nollix Jackson, D.C., S.E. Palmer, and W.L. Meadow. 1974. The effects sijJet/ol1). Comparative Biochemisu'y Physiology 32: 333-343. of temperature and carbon dioxide breathing on ven tilation Kraus, D.R., and D.C. Jackson. 1980. Temperature effects on and acid-base status of turtles. Respiration Physiology 20: ventilation and acid-base balance of the green turtle. Ameri­ 131-146. can Journal Physiology 239: R254-R258. Jackson, D.C., and H.D. Prange. 1979. Ventilation and gas ex­ Kruh0ffer, M., M.L. Glass, A.S. Abe, and K. Johansen, K. 1987. change during rest and exercise in adult green sea turtles. Control of breathing in an amphibian Bu!o /JllrrtClWmi.l: e!~ Journal Comparative Physiology 134: 315-319. fects of temperature and hypoxia. Respiration Physiology 69: Jackson, D.C., and H. Silverblatt. 1974. Respiration and acid­ 267-275. base status of turtles following experimental dives. American Lahiri, S.,JP. Szidon, and A.P. Fishman. 1970. Potential respira­ Journal Physiology 226: 903-909. tory and circulatory adjustments to hypoxia in the African Jackson, D.C., J.S. Wasser, and R.B. Silver. 1988. Effect of in­ lungfish. Federation Proceedings 29: 1141-1148.

duced hypercapnia on anaerobic metabolic rate of anoxic Larsen, B.K., and F.B. Jensen. 1993. Arterial P02, acid-base musk turtles. American Journal Physiology 254: R944-R948. status, and red cell nucleoside triphosphates in rainbow Janssen, R.G., and DJ. Randall. 1975. The effects of changes in trout transferred from fresh water to 20% sea water. Journal

pH and PC02 in blood and water on breathing in rainbow Fish Biology 42: 611-614. trout, Sallllo gairdneri. Respiration Physiology 25, 235-245. Lenfant, c., K.Johansen, and G. C.Grigg. 1966/67. Respiratory Jensen, F.B. 1987. Influences of exercise-stress and adrenaline properties of blood and pattern of gas exchange in the upon intra- and extracellular acid-base status, electrolyte lungfish NeoCl!mtodu5 fOl:~tm7 (Krefft). Respiration Physiology composition and respiratory properties of blood in tench 2: 1-21. , (Tinm tinm) at differen t seasons.Journal Comparative Physi­ Lillo, R.S. 1978. The effect of arterial-blood P02 PCO~, and pH ology B 157: 51-60. on diving bradycardia in the bullfrog Raila ratl'sbl'ia/la. Physi­ Jensen, F.B., N.A. Andersen, and N. Heisler. 1987. Effects of ological Zoology 51: 340-346. nitrite exposure on blood respiratory properties, acid-base Lindinger, M.I. , and D.G. McDonald. 1986. Cutaneous and and electrolyte regulation in the carp (CY/Jl711'1lS fmjJio) .Jour­ renal responses to intravascular infusions or \-ICI and NH"Cl nal Comparative Physiology B 157: 533-541. in the bullfrog (Rami catl!sbeimw). Comparative Biochemistry Jensen, F.B., and R.E. Weber. 1985. Kinetics of the acclimational Physiology 84A: 113-122. responses of tench to combined hypoxia and hypercapnia. Lutc<\vage, M.E., and P.L. Lutz. 1991. Voluntary diving metabo­ Journal Comparative Physiology B 156: 205-211. lism and ventilation in the loggerhead sea turtle. Journal Jensen, F.B ., and R.E. Weber. 1987. Internal hypoxia-hypercap­ Experimental Marine Biology Ecology 147: 287-296. nia in tench exposed to aluminium in acid water: effects on Mackenzie,J.A., and D.C.Jackson. 1978. The efrect of tempera­

blood gas transport, acid-base status and electrolyte composition ture on cutaneous CO2 loss and conductance in the bullfi-c>g. in arterial blood. Journal Experimental Biology 127: 427-442. Respiration Physiology 32:313-323. Ultsch & Jackson pH AND TEMPERATURE IN ECTOTHERMS 37 ------

Maginniss, L.A., and B.M. Hitzig. ] 987. Acid-base status and ity in the inactive benthic starry flounder Platicllth),s stellatus. electrolytes in red blood cells and plasma of turtles sub­ Physiological Zoology 60: 54-68. merged at 3"C. American Journal Physiology 253: R64-R70. Mitchell, G.S., and T.T. Gleeson. 1985. Acid-base balance during Malan, A., T.L. Wilson, and R.B. Reeves. 1976. Intracellular pH lactic acid infusion in the lizard \~ranus salvatoJ: Respiration in cold-blooded vertebrates as a function of body tempera­ Physiology 60: 253-266. ture. Respiration Physiology 28: 29-47. Mitchell, G.S., T.T. Gleeson, and A.F. Bennett. 1981 . Ventilation Maxime, v.,J. Pennec, and C. Peyraud, C. ] 991. Effects of direct and acid-base balance during graded activity in lizards. Ameri­ transfer from freshwater to seawater on respiratory and cir­ can Journal Physiology 240: R29-R37. culatory variables and acid-base status in rainbow trout.Jour­ Moalli, R, R. Meyers, G.R. Ultsch, and D.C.Jackson. 1981. Acid­ nal Comparative Physiology B 161: 557-568. base balance and temperature in a predominantly skin­ Maxime, v., M. Peyraud-Waitzenegger, G. Claireaux, and C. breathing salamander, O'yptobmnchus alleganiensis. Respira­ Peyraud. 1990. Effects of rapid transfer from sea water to tion Physiology 43: 1-11. fresh water on respiratory variables, blood acid-base status Munger, R.S., S.D. Reid, and C.M. Wood. 1991 . Extracellular and 02 affinity of haemoglobin in Atlantic salmon, (Sal11l0 fluid volume measurements in tissues of the rainbow trout salarL.).Journal Comparative Physiology B 160: 31-39. (Oncorh),nchus 1I!ykiss) in vivo and their effects on intracellu­ McDonald, D.G. 1983. The interaction of environmental cal­ lar pH and ion calculations. Fish Physiology Biochemistry 9: cium and low pH on the physiology of the rainbow trout, 313-323. Sal-Illo gairlineJi. 1. Branchial and renal net ion and H+ fluxes. Nicol, S.C., M.L. Glass, and N. Heisler. 1983. Comparison of Journal Experimental Biology 102: 123-140. directly determined and calculated plasma bicarbonate con­ McDonald, D.G., R.G. Boutilier, and D.P. Toews. 1980. The centration in the turtle Chr)lselll)IS pic/a bellii at different tem­ effects of enforced activity on ventilation, circulation and peratures. Journal Experimental Biology 107: 521-525. blood acid-base balance in the semi-terrestrial anuran, BuJo Nolan, W.F., and H. M. Frankel. 1982. Effects of temperature on lIUl J'in us. Journal Experimental Biology 84: 273-287. ventilation and acid-base status in the black racer snake, Coluber McDonald, D.G., H. Hobe, and C.M. Wood. 1980. The influence constJielOJ: Comparative Biochemistry Physiology 73A: 57-61. of calcium on the physiological responses of the rainbow Perry, S.F. 1982. The regulation of hypercapnic acidosis in the trout, Sallllo gairrl11eJi, to low environmental pH. Journal two salmon ids, the freshwater trout (Salmo gairdneri) and the Experimental Biology 88: 109-131. seawater salmon (Oncorh),nehus Idsllleh). Marine Behavior McDonald, D.G., and E.T. Prior. 1988. Branchial mechanisms of Physiology 9: 73-79. ion and acid-base regulation in the freshwater rainbow trout, Perry, S.F., M.S. Haswell, OJ. Randall, and A.P. Farrell. ] 981 Sal1llo gairrilleri. Canadian Journal Zoology 66: 2699-2708. Branchial ionic uptake and acid-base regulation in the rain­ McDonald, D.G., R.L. Walker, P.R.H. Wilkes, and C.M. Wood. bow trout, Sal1l!0 gairdneri.Journal Experimental Biology 92: 1982. H+ excretion in the marine teleost Pam/Jlzr),s vetlllus. 289-303. . Journal Experimental Biology 98: 403-414. Perry, S.F., R. Kinkead, P. Gallaugher, and D.J. Randall. 1989. McDonald, D.G., and C.M. Wood. 1981. Branchial and renal Evidence that hypoxemia promotes catecholamine release acid and ion fluxes in the rainbow trout, Sal1l!o gaiJ"{l1leJi, at during hypercapnic acidosis in rainbow trout (S(llmo low environmental pH. Journal Experimental Biology 93: gairr/neJi). Respiration Physiology 77, 351-364. 101-118. Perry, S.F., and S.Thomas. 1991 . The effects of endogenous and Milligan, C.L., and T.P. Farrell. 1986. Extracellular and intracel­ exogenous catecholamines on blood respiratory status dur­ lular acid-base status following strenuous activity in the sea ing acute hypoxia in rainbow trout (Oncorhynchus Inylliss) . raven (HelllitJijJtm"ltS m1//!Jicrmus). Journal Comparative Physi­ Journal Comparative Physiology B 161 : 489-497. ology B 156: 583-590. Perry, S.F., PJ. Walsh, T.P. Mommsen, and T.W. Moon. 1988. Milligan, C.L., D.G. McDonald, and T. Prior. 1991. Branchial Metabolic consequences of hypercapnia in the rainbow acid and ammonia tluxes in response to alkalosis and acido­ trout, SalJllo gainlneri: (p-adrenergic effects. General Com­ sis in two marine teleosts: coho salmon (Olleorh),l1chu5 Itilll.lrh) parative Endocrinology 69: 439-447. and starry flounder (Plaliehth),s stellatus). Physiological Zool­ Piiper,J.,M. Meyer, F. Drees. 1972. Hydrogen ion balance in the ogy 64: 169-192. elasmobranch Sryli01i1inus stellmis after exhausting activity. Milligan, C.L., and C. M. Wood. 1986(1. Intracellular and extra­ Respiration Physiology 16: 290-303. cellular acid-base status and H+ exchange with environment Piiper,J., and D. Schumann. 1967. Efficiency of 02 exchange in after exhaustive exercise in the rain bow trout. Journal Ex­ the gills of the dogfish, Scyliorhinlts slellmis. Respiration Physi­ perimental Biology 123: 93-121. ology 2: 135-148. Milligan, C.L., and C. M. Wood. 1986b. Tissue intracellular acid­ Pinder, A.W. 1987. Cutaneous diffusing capacity increases dur­ base status and the fate oflactate after exhaustive exercise in ing hypoxia in cold submerged bullfrogs (Rmw mtesbeimw) . the rainbow trout. Journal Experimental Biology 123: 123- Respiration Physiology 70: 85-95. 144. Portner, H.O., R.G. Boutilier, Y. Tang, and D.P. Toews. 1990. Milligan, C.L., and C. M. Wood. 1987 a. Effects of strenuous Determination of intracellular pH and PC02 after metabolic activity on intracellular and extracellular acid-base status and H+ inhibition by fluoride and nitrilotriacetic acid. Respiration exchange with the environment in the inactive benthic starry Physiology 8] : 255-274. flounder Pla/iell/lI)'s s/ellatus. Physiological Zoology 60: 37-53. Portner, H.O., L.M. MacLatchy, and D.P. Toews. 1991. Acid-base Milligan, C.L., and C. M. Wood. 1987b. Muscle and liver extra­ regulation in the toad 811fo 1lum'nus during environmental cellular acid-base and metabolite status after strenuous activ- hypoxia. Respiration Physiology 85: 217-230. 38 BULLETIN 18 December I, 1996

Primmett, D.R.N., DJ. Randall, M. Mazeaud, and RG. Boutilier. Short, S., E.W. Taylor, and PJ. Butler. 1979. The effectiveness of 1986. The role of catecholamines in erythrocyte pH regula­ oxygen transfer during normoxia and hypoxia in the dogfish tion and oxygen transport in rainbow trout (Salmo gainineri) (Scyliorhinlls canicula L.) before and after cardiac vagotomy. dming exercise. Journal Experimental Biology 122: 139-148. Journal Comparative Physiology 132: 289-295. Prosser, C.L. 1991. Temperatme. In C.L. Prosser (ed.) Envi­ Silver, R.B., and D.C. Jackson. 1985. Ventilatory and acid-base ronmental and Metabolic Animal Physiology, pp. 109-165. responses to long-term hypercapnia in the freshwater turtle, Wiley-Liss, New York. Chr)'sem)'s {Jieta bellii. Journal Experimental Biology 114: 661- Rahn, H. 1966. Gas exchange from the external environment to 672. the cell. In A.y'S. de Reuck and R Porter (eds.) Develop­ Silver, R.B., and D.C.Jackson. 1986. Ionic compensation with no ment of the Lung, pp. 3-23. Churchill, London. ren;-I response to chronic hypercapnia in Chr)'se11l)'s jJieta Rahn, H., and W.F. Garey. 1973. Arterial CO"' 0", pH, and bellii. American Journal Physiology 251: RI228-RI234. HCOs· values of ectotherms living in the Am;zon~ American Smatresk, NJ., andJ.N. Cameron. 1982a. Respiration and acid­ Journal Physiology 225: 735-738. base physiology of the spotted gar, a bimodal breather. I. Randall, DJ., andJ.N. Cameron. 1973. Respiratory control of Normal values, and the response to severe hypoxia. Journal arterial pH as temperature changes in rainbow trout Salmo Experimental Biology 96: 263-280. gairdneri. American Journal Physiology 225: 997-1002. Smatresk, NJ., andJ.N. Cameron. 1982b. Respiration and acid­ Randall, DJ., J.N. Cameron, C. Daxboeck, and N. Smatresk. base physiology of the spotted gar, a bimodal breather. II. 1981. Aspects of bimodal gas exchange in the bowfin, A mia Responses to temperature change and hypercapnia. Journal calva L. (Actinopterygii: Amiiformes). Respiration Physiol­ Experimental Biology 96: 281-293. ogy 43: 339-348. Snyder, G.K., andJ.R. Nestler. 1991. Intracellular pH in the toad Randall, DJ., A.P. Farrell, and M.S. Haswell. 1978a. Carbon Bufo marinus following hypercapnia. Journal Experimental dioxide excretion in the jeju, HojJlerytll1inus unitaeniatus, a Biology 161: 415-422. facultative air-breathing teleost. Canadian Journal Zoology Stiffler, D.F. 1991. Partitioning of acid-base regulation between 56: 970-973. renal and extrarenal sites in the adult, terrestrial stage of the Randall, DJ., A.P. Farrell, and M.S. Haswell. 1978b. Carbon salamander Amb),stoma tif51inulIl during respiratory acidosis. dioxide excretion in the pirarucu (Amj)(li11la gigas), an obli­ Journal Experimental Biology 157: 47-62. gate air-breathing fish. Canadian Journal Zoology 56: 977- Stiffler, D.F., and N. Bachoura, N. 1991. Metabolic alkalosis in 982. the larval salamander, A1Ilbyst01l!a tigrinwll: partitioning regu­ Reeves, RB. 1972. An imidazole alphastat hypothesis for verte­ latory responses between the skin and kidneys. Journal Ex­ brate acid-base regulation: tissue carbon dioxide content perimental Zoology 258: 196-203. and body temperatme in bullfrogs. Respiration Physiology Stiffler, D.F., M.E. Kopecky, M.L. Thompson, and R.G. Boutilier. 14: 219-236. 1990. Acid-base-electrolyte balance responses to catechola­ Reeves, R.B. 1976. Temperature-induced changes in blood acid­ mine antagonists in A1Ilbyst01l!a tif51inu1I!. American Journal base status: pH and PCO" in a binary buffer. Journal Applied Physiology 258: RI363-RI370. Physiology 40: 752-761. - Stiffler, D.F., S.L. Ryan, and R.A. Mushkot. 1987. Interactions Reeves, R.B. 1977. The interaction of body temperature and between acid-base balance and cutaneous ion transport in acid-base balance in ectothermic vertebrates. Annual Re­ larval Amb),st01lla tif51inum (Amphibia: Caudata) in response views Physiology 39: 559-586. to hypercapnia. Journal Experimental Biology 130: 389-404. Robin, E.D. 1962. Relationship between temperature and Stiffler, D.F., and D.P. Toews. 1992. Acid-base-electrolyte bal­ plasma pH and carbon dioxide tension in the turtle. Nature ance responses of Bufo 1llurim15 to aminoglutethimide, corti­ 195: 249-251. costerone, and aldosterone during hypercapnia. General. Rohrbach ,J.w. , and D.P. Stiffler. 1987. Blood-gas, acid-base, and Comparative Endocrinology 86: 152-16l. electrolyte responses to exercise in larval A11lbyst01lla ligrinu11l. Stiffler, D.F., B.L. Tufts, and D.P. Toews. 1983. Acid-base and Journal Experimental Zoology 244: 39-47. ionic balance in Am/Jystomu ligrinu1l! and NfrturllS l1laCUlOSllS Rosenthal, T.B. 1948. The effect of temperature on the pH of during hypercapnia. American Journal Physiology 245: the blood and plasma in vitro. Journal Biological Chemistry R689-R694. 173: 25-30. Stinner,J.N. 1982. Ventilation, gas exchange and blood gases in Schmidt-Nielsen, K. 1990. Animal Physiology: Adaptation and the snake, Pitlto/Jhis melunoleums. Respiration Physiology 47: Environment. 602 pp, 4th ed. Cambridge University Press, 279-298. Cambridge. Stinner, J.N. 1987. Thermal dependence of air convection re­ Seymour, R.S., A.F. Bennett, and D.F. Bradford. 1985. Blood gas quirement and blood gases in the snake Colu/Jfr ronslrirlOl: tensions and acid-base regulation in the salt-water crocodile, American Zoologist 27: 41-47. Crocodyl1l5jJOroSUS, at rest and after exhaustive exercise.Jour­ Stinner, .J.N., D.L. Newlon, and N. Heisler. 1994. Extracellular nal Experimental Biology 118: 143-159. and intracellular carbon dioxide concentration as a function Seymour, R.S., G.P. Dobson, and J. Baldwin. 1981. Respiratory and of temperature in the toad Buro IIUlrinus.Journal Experimen­ cardiovascular physiology of the aquatic snake, ArrorilOrrl1ls tal Biology 195: 345-360. amJitme.Journal Comparative Physiology 144: 215-227. Stinner, J.N., and Y.H. Shoemaker. 1987. Cutaneous gas ex­ Seymom, R.S., and M.E.D. Webster. 1975. Gas transport and change and low evaporative water loss in the frogs blood acid-base balance in diving sea snakes. Journal Experi­ Ph),li011lPr/usu sauvagei and Chirol1l(l11lis xemm/}(dina. Journal mental Zoology 191: 169-182 Comparative Physiology B 157: 423-427. Ultsch & Jackson pH AND TEMPERATURE IN ECTOTHERMS 39 ------

Stinner,J.N., and R.L. Wardle. 1988. Effect of temperature upon hypoxia on acid-base balance in trout: role of ion transfer carbon dioxide stores in the snake Coluber constrictor and the processes. American journal Physiology 250: R319-R327. turtle Chl)'sel/lYs scri/lla. Journal Experimental Biology 137: Thomas, S., and C.M. Hughes. 1982a. Effects of hypoxia on 529-548. blood gas and acid-base parameters of sea bass. Journal Takeda. T. 1990. Ventilation, cardiac output and blood respira­ Applied Physiology 53: 1336-1341. tory parameters in the carp, C)'prinus ca/pio, during Thomas, S., and C.M. Hughes. 1982b. A study of the effects of hyperoxia. Respiration Physiology 81: 227-240. hypoxia on acid-base status of rainbow trout blood using an Takeda, T. 1991. Regulation of blood oxygenation during short­ extracorporeal blood circulation. Respiration Physiology 49: term hypercapnia in the carp, CyjJrinus cm/Jio. Comparative 371-382. Biochemistry Physiology 98A: 517-521. Thomas, S., and H. Le Ruz. 1982. A continuous study of rapid Talbot, C.R., and D.F. Stiffler. 1990. Effects of hypoxia on acid­ changes in blood acid-base status of trout during variations . base balance, blood gases, cathecholamines, and cutaneous of water PC02 journal Comparative Physiology 148: 123- ion exchange in the larval tiger salamander (Ambystoma 130. tigrinulll). Jouranl Experimental Zoology 257: 299-305. Thomas, S., andJ. Poupin. 1985. A study of the effects of water Talbot, C.R., and D.F. Stiffler. 1991. Cutaneous ion exchange, carbonate alkalinity on some parameters of blood acid-base and renal and extrarenal partitioning of acid and ammonia status in rainbow trout (Sallllo gainlneri R.). Journal Com­ excretion in the larval tiger salamander, Ambystoma tigrinu1/l, parative Physiology B 156: 29-34. following ingestion of ammonium salts. Journal Compara­ Toews, D.P., and N. Heisler. 1982. The effects of hypercapnia on tive Physiology B 162: 416-423. intracellular and extracellular acid-base status in the toad, Tang, Y, and R.C. Boutilier. 1988a. Clearance of lactate and BuJo IIlminus.Journal Experimental Biology 97: 79-86. protons following acute lactacidosis: a comparison between Toews, D.P., C.F. Holeton, and N. Heisler. 1983. Regulation of seawater- and freshwater-adapted rainbow trout (5al1ll0 the acid-base status during environmental hypercapnia in gaircineri) . Experimental Biology 48: 41-44. the marine teleost fish Conger congel: Journal Experimental Tang, Y, and R.C. Boutilier. 1988b. Correlation between cat­ Biology 107: 9-20. echolamine release and degree of acidotic stress in trout. Toews, D.P., and S. Kirby. 1985. The ventilatory and acid-base AmericanJournal Physiology 255: R395-R399. physiology of the toad, BuJo 1/l(l1inus, during exposure to envi­ Tang, Y, and R.C. Boutilier. 1991. White muscle intracellular ronmental hyperoxia. Respiration Physiology 59: 225-229. acid-base and lactate status following exhaustive exercise: a Toews, D.P., and D.F. Stiffler, D.F. 1990. Compensation of pro­ comparison between freshwater- and seawater-adapted rain­ gressive hypercapnia in the toad (BuJo IIIwinus) and the bow trout. journal Experimental Biology 156: 153-171. bullfrog (Rana catesbeiana). Journal Experimental Biology Tang, Y, D.C. McDonald, and R.C . Boutilier. 1989a. Acid-base 148: 293-302. regulation following exhaustive exercise: a comparison be­ Truchot,J., A. Toulmond, and P. Dejours. 1980. Blood acid-base tween freshwater- and seawater-adapted rainbow trout (5al1ll0 balance as a function of water oxygenation: a study at two

gairdnl'li).Journal Experimental Biology 141: 407-418. different ambient CO2 levels in the dogfish, 5cyliorhinus Tang, Y, D.C. McDonald, and R.C. Boutilier. 1989b. Adrenergic canicula. Respiration Physiology 41: 13-28. regulation of blood acid-base status following exhaustive Tufts, B.L. 1991. Acid-base regulation and blood gas transport exercise in seawater-adapted rainbow trout, 5almo gairdneri. following exhaustive exercise in an agnathan, the sea lam­ Physiological Zoology 62: 950-963. prey Petrolll),zo/l. 1/lfllinus.Journal Experimental Biology 159: Tang, Y., S. Nolan, and R.C. Boutilier. 1988. Acid-base regula­ 371-385. tion following acute acidosis in seawater-adapted rainbow Tufts, B.L., B. Bagatto, and B. Cameron. 1992. In vivo analysis of trout, Sallllo gairdneri: a possible role for catecholamines. gas transport in arterial and venous blood of the sea lamprey Journal Experimental Biology 134: 297-312. Petrom),zon l/ul1imts.journal Experimental Biology 169: 105- Tazawa, H., M. Mochizuki, andJ. Piiper. 1979. Respiratory gas 119. transport by the incompletely separated double circulation Tufts, B.L., R.A. Ferguson, and R.C. Boutilier.. 1988. In vivo and in the bullfrog, Rana catesbeialla. Respiration Physiology 36: in vitro effects of adrenergic stimulation of chloride/bicar­ 77-95. bonate exchange in rainbow trout erythrocytes. journal Ex­ Thomas, S. 1983. Changes in blood acid-base balance in trout perimen tal Biology 140: 301-312. (Sallllo g(lir(i1!e17 Richardson) following exposure to com­ Tufts, B.L., D.C. Mense, and DJ. Randall. 1987. The effects of bined hypoxia and hypercapnia. Journal Comparative Physi­ forced activity on circulating catecholamines and pH and ology 152: 53-57. water content of erythrocytes in the toad. Journal Experi­ Thomas, S., A. Belaud., L. Barthelemy, and C. Peyraud, C. 1980. mental Biology 128: 411-418. Acid-base status in plasma of trout and eel in hypocapnic and Tufts, B.L., and D.P. Toews. 1985. Partitioning of regulatory sites normocapnic conditions. Journal Comparative Physiology in BuJo I/lfllinus during hypercapnia. Journal Experimental 140: 249-254. Biology 119: 199-209. Thomas, S., B. Fievet, L. Barthelemy, and C. Peyraud. 1983. Tufts, B.L., and D.P. Toews. 1986. Renal function and acid-base Comparison of the effects of exogenous and endogenous balance in the toad BIlJo II/.winus during short-term dehydra­ hypercapnia on ventilation and oxygen uptake in the rain­ tion. Canadian Journal Zoology 64: 1054-1057. bow trout (Sallllo gairrlneri R.). Journal Comparative Physiol­ Turner, .J.D., C.M. Wood, and D. Clark. 1983. Lactate and pro­ ogy 151 : 185-190. ton dynamics in the rainbow trout (Salmo gairtineri).journal Thomas, S., B. Fievet, and R. Motais. 1986. Effect of deep Experimental Biology 104: 247-268. 40 BULLETIN 18 December 1, 1996

Turner, J.D., C.M. Wood, and H. Hobe. 1983. Physiological trout white muscle: acid-base, phosphogen, carbohydrate, consequences of severe exercise in the inactive benthic flat­ lipid, ammonia, fluid volume and electrolyte metabolism. head sole (Hippoglossoides elassodon): a comparison with the Journal Experimental Biology 195: 227-258. active pelagic rainbow trout (Salmo gairdneri). Journal Ex­ Warburton, SJ.,J.S. Wasser, and D.C. Jackson. 1989. Cardiovas­ perimental Biology. 104: 269-288, cular and metabolic responses during anoxic submergence Ultsch, G.R. 1987. The potential role of hypercarbia in the in the bullfrog with and without maintained extracellular transition from water-breathing to air-breathing in verte­ pH. Journal Experimental Zoology 251: 13-19. brates. Evolution 41: 442-445. Wasser,J .S. 1990. Seasonal variations in plasma and tissue chem­ Ultsch, G.R. 1988. Blood gases, hematocrit, plasma ion concen­ istry in water snakes, Nerodia siJJe(lon. Copeia 1990: 399-408. trations, and acid-base status of musk turtles (Sternotherus Wasser,J.S., and D.e.Jackson. 1988. Acid-base balance and the odoratus) during simulated hibernation. Physiological Zool­ control of respiration during anoxic and anoxic-hypercapnic ogy 61: 78-94. gas breathing in turtles. Respiration Physiology 71: 213-226. Ultsch, G.R. 1989. Ecology and physiology of hibernation and Wasser,J.S., D.e.Jackson, S.Y. Chang, and Sj. Warburton. 1993. overwintering among freshwater fishes, turtles, and snakes. Maintenance of high extracellular pH does not influence Biological Reviews 64: 435-516. cell pH or metabolism in submerged anoxic bullfrogs. Jour­ Ultsch, G.R., and B.M. Cochran. 1994. Physiology of northern nal Experimental Zoology 265: 619-626. and southern musk turtles (Stemotherus odoratus) during Wasser,J.S., Sj. Warburton, and D.e.Jackson. 1991. Extracellu­ simulated hibernation. Physiological Zoology 61: 263-281. lar and in tracellular acid-base effects of submergence anoxia Ultsch, G.R., R.W. Hanley, and T.R. Bauman. 1985. Responses to and nitrogen breathing in turtles. Respiration Physiology 83: anoxia during simulated hibernation in northern and south­ 239-252. ern painted turtles. Ecology 66: 388-395. West, N.H., A.W. Smits, and W.W. Burggren. 1989. Factors ter­ U1tsch, G.R., C.Y. Herbert, D.e. Jackson, D.C. 1984. The com­ minating nonventilatory periods in the turtle, Chelydra parative physiology of diving in North American freshwater se/1mltina. Respiration Physiology 77: 337-350. turtles-I. Survival, gas exchange, and acid-base balance. White, F. N., and G. Somero. 1982. Acid-base regulation and Physiological Zoology 57: 620-631. phospholipid adaptations to temperature: time courses and Ultsch, G.R., and D.C. Jackson. 1982. Long-term submergence physiological significance of modifying the milieu for pro­ at 3°C of the turtle Chrysemys jJicta bellii in normoxic and tein function. Physiological Reviews 62: 40-90. severely hypoxic water-I. Survival, gas exchange, and acid­ Wilkie, M.P., and C.M. Wood. 1991. Nitrogenous waste excre­ base status. Journal Experimental Biology 96: 11-28. tion, acid-base regulation, and ionoregulation in rainbow Ultsch, G. R., and D. C. Jackson. 1995. Acid-bas status and ion trout (Oncorhynchus lIl)'lliss) exposed to extremely alkaline balance during simulated hibernation in freshwater turtles water. Physiological Zoology 64: 1069-1086. from the northern portions of their ranges. Journal Experi­ Wilson, R.W., and E.W. Taylor. 1992. Transbranchial ammonia mental Zoology 273: 482-493. gradients and acid-base responses to high external ammonia Ultsch, G.R., M. Ott, M., and N. Heisler. 1981. Acid-base and concentration in rainbow trout (OnrorhYllchus lIlylliss) accli­ electrolyte status in carp (Cyjn1nltS cm1Jio) exposed to low mated to different salinities. Journal Experimental Biology environmental pH. Journal Experimental Biology 93: 65-80. 166: 95-112. van den Thillart, G., D. Randall, and L. Hoa-Ren. 1983. CO2 and Wilson, R.W., P.M. Wright, S. Munger, and C.M. Wood. 1994. H+ excretion by swimming coho salmon, Oncorhynchus Ammonia excretion in fj·eshwater rain bow trout (OncOlilynchus llisutch.Journal Experimental Biology 107: 169-180. mylliss) and the importance of gill boundary layer acidification: van Dijk, P.L.M., G.E.E.J.M. van den Thillart, P. Balm, and S.E. lack of evidence for Na+ /NH/ exchange. Journal Experimen­ Wendelaar Bonga. 1993. The influence of gradual water tal Biologyl91: 37-58. acidification on the acid-base status and plasma hormone Wood, e.M., and E.B.Jackson. 1980. Blood acid-base regulation levels in carp. Journal Fish Biology 42: 661-671. during environmental hyperoxia in the rainbow trout (SallllO Waddell, Wj., and T.C. Butler, T.e. 1959. Calculation of intrac­ gairdneri). Respiration Physiology 42: 351-372. ellular pH from the distribution of 5,5-dimethyl-2,4- Wood, e.M., and J. LeMoigne. 1991. Intracellular acid-base oxazolidindione (DMO). Application to skeletal muscle of responses to environmental hyperoxia and normoxic recov­ the dog. Journal Clinical Investigation 38: 720-729. ery in rainbow trout. Respiration Physiology 86: 91-113. Walsh, Pj., and T.W. Moon. 1982. The influence of temperature Wood, e.M., B.R. McMahon, and D.G. McDonald. 1977. An on extracellular and intracellular pH in the American eel, analysis of changes in blood pH following exhausting activity Anguilla rostmta (Le Sueur). Respiration Physiology 50: 129- in the starry flounder, Platirhthys stellatus. Journal Experi­ 140. mental Biology 69: 173-185. Wang, T., L.G.S. Branco, and M.L. Glass. 1994. Ventilatory Wood, e.M., ~.R. McMahon, and D.G. McDonald. 1979. Respi­ responses to hypoxia in the toad BuJo j}(lmrnPlnis before and ratory gas exchange in the resting starry flounder, Platirhthys after a decrease in haemoglobin oxygen-carrying capacity. stellatll.l: a comparison with other teleosts. Journal Experi­ Journal Experimental Biology 186: 1-8. mental Biolob'Y 78: 167-179. Wang, T., W. Fernandes, and A.S. Abe. 1993. Blood pH and 02 Wood, e.M., and C.L. Milligan. 1987. Adrenergic analysis of

homeostasis upon CO2 anesthesia in the rattlesnake (Crota­ extracellular and intracellular lactate and H+ dynamics after lus durissus). The Snake 25, 21-26. strenuous exercise in the starry Hounder Platirhthys stellatlls. Wang, Y., Gj.F. Heigenhauser, and e.M. Wood. 1994. Integrated Physiological Zoology 60: 69-81. responses to exhaustive exercise and recovery in rainbow Wood, e.M., and R.S. Munger. 1994. Carbonic anhydrase injec- Ultsch & Jackson pH AND TEMPERATURE IN ECTOTHERMS 41

tion provides evidence for the role of blood acid-base status Wood, S.C., KJohansen., M.L. Glass, and R.W. Hoyt. 1981. Acid­ in stimulating ventilation after exhaustive exercise in rain­ base regulation during heating and cooling in the lizard, bow trout. Journal Experimental Biology 194: 225-253. Varanus exanthematicus. Journal Applied Physiology 50: 779- Wood, C.M., R.S. Munger, and D.P. Toews. 1989. Ammonia, 783. urea, and H+ distribution and the evolution of ureotelism in Wood, S.C., G. Lykkeboe, KJohansen, R.E. Weber, and G.M.O. amphibians. Journal Experimental Biology 144: 215-233. Maloiy. 1978. Temperature acclimation in the pancake tor­ Wood, C.M., B.P. Simons, D.R. Mount, and H.L. Bergman. 1988. toise, l'v[alacochel'sus tomieri: metabolic rate, blood pH, oxy­ Physiological evidence of acclimation to acid/aluminum gen affinity and red cell organic phosphates. Comparative stress in adult brook trout (Salvelinus Jontillalis). 2. Blood Biochemistry Physiology 59A: 155-160. parameters by cannulation. Canadian Journal Zoology 45: Wood, S.C., and W.R. Moberly. 1970. The influence of tempera­ 1597-1605. ture on the respiratory properties of iguana blood. Respira­ Wood, C.M.,J.D. Turner, R.S. Munger, and M.S. Graham. 1990. tion Physiology 10: 20-29. Control of ventilation in the hypercapnic skate Raja oceliata: Wright, P.A., DJ. Randall, and C.M. Wood. 1988. The distribu­ II. Cerebrospinal fluid and intracellular pH in the brain and tion of ammonia and H+ between tissue compartments in other tissues. Respiration Physiology 80: 279-298. lemon sole (PaTOphl')'s vetulus) at rest, during hypercapnia Wood, C.M., P.J. Walsh, S. Thomas, and S. F.Perry. 1990. Control and following exercise. Journal Experimental Biology 136: of red blood cell metabolism in rainbow trout after exhaus­ 149-175. tive exercise. Journal Experimental Biology 154: 491-507. Wright, P.A., and C.M. Wood. 1985. An analysis of branchial Wood, S.C., R.N. Gatz, and M.L. Glass. 1984. Oxygen transport ammonia excretion in the freshwater rainbow trout: effects in the green sea turtle. Journal Comparative Physiology B of environmental pH change and sodium uptake blockade. 154: 275-280. Journal Experimental Biology 114: 329-353. Wood, S.C., M.L. Glass, and K Johansen. 1977. Effects of tem­ Wright, P.A., C.M. Wood, and DJ. Randall. 1988. An in vitTO and perature on respiration and acid-base balance in a monitor in vivo study of the distribution of ammonia between plasma lizard. Journal Comparative Physiology 116: 287-296. and red cells of rainbow trout (Salmo gail'cineri). Journal Wood, S.C., and KJohansen. 1973. Blood oxygen transport and Experimental Biology 134: 423-428. acid-base balance in eels during hypoxia. American Journal Ye, X., DJ. Randall, and X. He. 1991. The effect of acid water on Physiology 225: 849-851. oxygen consumption, circulating catecholamines and blood Wood, S.C., and K Johansen. 1974. Respiratory adaptations to ionic and acid-base status in rainbow trout (Salmo gairdneri, diving in the Nile monitor lizard, Vrll,(lnllS nilolicu5. Journal Richardson). Fish Physiology Biochemistry 9: 23-30. Comparative Physiology 89: 145-158.

Life Histories of Noturus baileyi and N. flavipinnis (Pisces: Ictaluridae), Two Rare Madtom Catfishes in Citico Creek, Monroe County, Tennessee

Gerald R. Dinkins

3D/ International, Inc. 7039 Maynardville Highway Knoxville, TN 37918

and

Peggy W. Shute

Regional Natural Heritage Program Tennessee Valley Authority Norris, TN 37828

ABSTRACT: Dinkins, Gerald R. and Peggy W. Shute. 1995. The Life Histories of Nohlrus baileyi and N. flavipim,is (Pisces: Ictaluridae), Two Rare Madtom Catfishes in Citico Creek, Monroe County, Tennessee. Bulletin Alabama Museum of Natural History, Number 18:43-69, 8 tables, 18 figures. The life histories of the federally endangered smoky madtom (N. baileyi) and the federally threatened yellowfin madtom (N. flavipi'IIIis) were studied in Citico Creek, a tributary to the lower Little Tennessee River in eastern Tennessee. Most of the observations and measurements were made between May 1981 and June 1984. Live and preserved individuals were observed for information on distribution, macro- and microhabitat occurrences, age and growth, reproduction and nesting, larval development, feeding ecology, and parasit­ ism. Notllrlls baileyi lives in only a 10.8 km reach of Citico Creek, beginning at creek km 6.0. From late spring to late fall, N. baileyi occur underneath flat, palm-sized rocks (slabrocks) in riffles, especially riffle crests. During the colder months, pools are occupied and slabrocks similar in physical dimension to those used as shelter in the riffles are used. Nohlrlls baileyi lives approximately two years, and sexual maturity is reached in the second summer of life (one year of age). Nohlrlls baileyi nests under large, flat rocks where one or both parents excavate a cavity for the egg mass. Nesting takes place from May to July, and an average of 36.3 eggs were found in four nests. Larval development is described for hatchlings and subsequent stages. Nohlrlls flavipi'IIIis is found in only three widely distributed locations in the upper Tennessee River system. In Citico Creek, N. flavipilll,is occurs year-round in pools in a 3.6 km reach beginning above a small concrete darn at creek km 13.7. Movement between these pools is limited. The species lives three to four years, and sexual maturity is reached in the third summer of life (two years of age). NOhlrllsflavipilll,isnests under large, flat rocks that are slightly larger in physical dimensions than those used by N. baileyi. Nesting takes place from May to July, and an average of 55 eggs were found in ten nests. Evidence supporting polyandry is given for both species. Larval development is described for several larval stages of N. flavipilll,is. Both species are almost exclusively insectivorous.

Bull. Alabama Mus. Nat. Hist. 18:43-69

December 1, 1996 44 BULLETIN 18 December 1, 1996

Introduction Notttnts for which there is no available biological informa­ Although Federal environmental regulations have re­ tion. In many cases, however, specific information suffi­ sulted in some relatively recent, localized improvements cient to allow protection, management, and possible re­ in water quality, many rivers and streams in the southeast­ covery has not been easily obtainable, or may be available ern United States have been permanently altered because only when it is too late to protect the species. For example, of urban, industrial, and agricultural encroachment. As a N. trautmani is probably extinct (Rohde 1978). result, numerous fish species specialized for particular Catfish are known to be highly social animals, and stream habitats have been able to survive only as frag­ bullheads (A 11leiurus) have been reported to display so­ mented remains of once more widespread populations. phisticated behaviors based on chemoreception. They This is particularly true of many species of mad tom cat­ apparently recognize each other as individuals and are fish, Notttrus (Taylor 1969). Several madtom species have thus able to coexist successfully by forming dominance relatively restricted distributions and may be naturally hierarchies (Atema et al. 1969; Todd et al. 1967; Todd rare (sensu Sheldon 1987). 1973) . As the majority of madtoms are nocturnal, they Because of these localized distributions, nine madtom presumably also rely heavily on chemoreception for sur­ species were listed in 1972 as species of concern by the vival (finding food, potential mates, and raising young). American Fisheries Society (Miller 1972) and increased to The more sensitive members of the genus may be detri­ eleven in 1989 (Williams et al. 1989). Currently five are mentally affected by small changes in water quality. Etnier listed by the Department of Interior under the Endan­ and Jenkins (1980, p. 20) speculated that "... recent gered Species Act: N. baileyi-endangered; N. flavipinnis­ extinction and extirpation of several species of madtoms threatened; N. placidus-threatened; N. stanattli-endan­ may, in addition to visible habitat degradation, be related gered; and N. trautmani-endangered, probably extinct to their being unable to cope with 'olfactory noise' being (U.S. Fish and Wildlife Service 1994a). The latest listing of added to riverine ecosystems in the form of a wide variety taxa being considered for federal status includes six spe­ of complex organic chemicals that may occur in only trace cies and one subspecies (U.S. Fish and Wildlife Service amounts." 1994b). As a group, members of the genus Noturus are This paper presents ecological data obtained by moni­ imperiled disproportionately to the percentage of the toring reproductive success, demography, and food habits ichthyofauna which they comprise (Etnier and Starnes for the only known population of the federally endan­ 1991, Warren and Burr 1994). gered smoky mad tom, N. baileyi, and one of the three Before the last two decades, there were relatively few extant populations of the federally threatened yellowfin published discussions of madtom life histories. The ear­ madtom, N. flavipinnis. These data were collected and lier works on this group of fishes included: Clugston and documented in more detail by Dinkins (1984) and Shute Cooper (1960); Curd (1960); Carlson (1966); (1984) as partial fulfillment for their M.S. degrees. Our Thomerson (1966) and Mahon (1977) . With the recent work has also been supplemented with data obtained interest and concern for rapidly declining populations of 1986 to present while monitoring and performing a cap­ Nottt1"us, numerous ecological works on the group have tive rearing and reintroduction project for both species appeared in the literature. These include: Mayden et al. (see Shute et al. 1992 for a detailed description of these (1980); Burr and Dimmick (1981); Mayden and Burr projects). Populations of both species examined are (1981); Burr and Mayden (1982a, b); Lindquist et al. highly localized and it is hoped that the information (1982); Burr and Mayden (1984); Mayden and Walsh obtained and methods used to collect the data may be (1984); Miller (1984); Starnes and Starnes (1985); Walsh useful to ensure their future protection. and Burr (1985); Whiteside and Burr (1986); Burr et al. (1989); Vives (1987); Luttrell et al. (1992); Simonson and Background Neves (1992); Baker and Heins (1994); Fuselier and Edds NOTURUS BAlLEYl--Notttms baileyi (Figure la) was described (1994); and Pfingsten and Edds (1994). One of the most by Taylor (1969) from five specimens collected during a comprehensive studies, which suggests application of life 1957 stream reclamation (poisoning) project conducted history data to management of threatened and endan­ on Abrams Creek (Figure 2), a tributary to the Little gered species is that of Mayden and Walsh (1984). Tennessee River in the Great Smoky Mountains National Several unpublished theses and status survey reports Park. This project, designed to eliminate "rough" fish and have also added knowledge about ecology of madtoms. enhance conditions for trout stocking, was done in con­ These include Bowman (1932, 1936); Gilbert (1953); junction with the closure of the gates of Chilhowee Dam Andrews (1963); Case (1970); Madding (1971); Bowen (Lennon and Parker 1959). The exact locality where N. (1980); Moss (1981); Robison and Harp (1981a, b); baileyi was captured was unknown at the time of descrip­ Dinkins (1982); Bauer and Clemmer (1983); Dinkins tion, but Dinkins (1982) concluded that the type speci­ (1984); Shute (1984); Burr et al. (1993); and Shute et al. mens were collected from the Abrams Creek campground (1992) . As a resul t, today there are only a few species of area (approximate creek km 12.9) based on habitat, Dinkins & Shute LIFE HISTORIES OF NOTURUS BAILEYI AND N. FLAVIPINNIS 45

Figure la: Noturus baileyi (Photograph by 1. R. Shute).

Figure 1b: Noturus flavipinnis (Photograph by 1. R. Shute). 46 BULLETIN 18 December 1, 1996

stream gradient, elevation, and discussions with person­ survey and make management recommendations. She nel involved with the reclamation. Collections made in determined that the Citico Creek population is appar­ Abrams Creek and other Little Tennessee River tributar­ ently confined upstream of a small concrete dam at creek ies subsequent to Taylor's monograph revealed no addi­ km 13.7. Starnes and Etnier (1986) concluded that the tional specimens of N. baileyi, and the species was believed lack of morphological intraspecific variation between the to be extinct. Citico Creek and Copper Creek populations indicated a On 23July 1980, while surveying Citico Creek for the relatively recent continuous population. spotfin chub (Cyprinella (=Hybopsis) monacha) a single Noturusjlavipinniswas listed as federally threatened on specimen of N. baileyi was collected (Bauer et al. 1983). 9 September 1977 (U. S. Fish and Wildlife Service 1977) Citico Creek is a direct tributary to the Little Tennessee with the Powell River and Copper Creek designated as River; the mouth of Citico Creek is approximately 11 river Critical Habitat. Williams et al. (1989) listed the species as km downstream from the mouth of Abrams Creek. With nationally threatened for the same reasons given for N. funding from the U.S. Fish and Wildlife Service (FWS) for jlavipinnis. NotUTUS jlavipinnis is listed as endangered by a status survey of the species, Dinkins (1982) concluded the state of Tennessee; endangered in Virginia (Burkhead that N. baileyi only existed in a 10.6 km stretch of Citico and Jenkins 1991); and presumed extirpated in Georgia Creek, and recommended the species be given federal (B.J. Freeman pers. comm.) endangered status. On 26 October 1984 N. baileyi was listed as federally endangered and Citico Creek was desig­ Study Area nated as Critical Habitat (U. S. Fish and Wildlife Service Citico Creek is a moderate sized (10 to 25 m width), 1984). Williams et al. (1989) also considered the species relatively pristine, fourth order stream located in eastern nationally endangered because of present or threatened Tennessee (Figure 2). The stream originates in the steep, destruction, modification or curtailment of its habitat or mountainous Blue Ridge physiographic province along range. NotUTUS baileyi is also listed as endangered by the the Tennessee/North Carolina border and has its state of Tennessee (Tennessee Wildlife Resources Agency confluence with the Little Tennessee River at the eastern 1994) . edge of the Valley and Ridge Province (Swingle et al. 1966). The lower 2.3 km of Citico Creek are inundated NOTURUS FLAVIPINNls-Noturus jlavijJinnis (Figure 1 b) was (since 1979) by Tellico Reservoir. Most of the heavily described by Taylor (1969) from thirteen specimens col­ forested and mountainous watershed is within the Chero­ lected between 1884 and 1893 at four locations in the kee National Forest and the Tellico Wildlife Management upper Tennessee River system. Only three of the original localities are known with certainty: West Chickamauga Creek, Georgia; North Fork Holston River, Virginia; and Hines [Hinds] Creek, Tennessee (Figure 3). In his de­ scription of the species, Taylor speculated that the histori­ cal range of N. jlavipinnis was the upper Tennessee River basin, but because of impounded or polluted conditions of historical localities, and the lack of specimens in pre­ impoundment and other numerous surveys, the species had become extinct, presumably in the early 1900's. Since Taylor's original description, three geographi­ cally isolated populations of N. jlavijJinnis have been dis­ covered. One specimen was taken in 1968 during night­ time sampling in the Powell River at McDowell Shoals (river km 171.8, Hancock County, Tennessee) by Tennes­ see Valley Authority (TVA) biologists (Taylor et al. 1971). In 1969, four localities in Copper Creek (a Clinch River tributary in Scott and Russell counties, Virginia) yielded Nanlahalc National Fcre3t " 28 specimens of N. jlavijJinnis. Because the original de­ c, scription was based on 13 faded specimens, N. jlavijJinnis TENNESSEE ~18 NORTH o(;-~~f CAROUNA was redescribed by Taylor et al. (1971) based on these f' ..f,~ ...D 1\ individuals from Copper Creek and the Powell River. In 7 1 ; June 1981, five specimens of N. jlavijJinnis were collected , '-----J 4km by the senior author while skin diving at night in search of N. baileyi in Citico Creek. Subsequent to this discovery, Shute (1984) was funded by FWS to perform a status Figure 2: Map of the middle Little Tennessee River system. Dinkins & Shute LIFE HISTORIES OF NOTURUS BAILEYI AND N. FLAVIPINNIS 47

I

.: : . ~: ....'...... 1

. . '

, ': "

" '., /-/ NF. Holston River Va

. :,~.' . "',

Tn Lyons Cnff1k ..... , ~ bJ' NC

J+\!st Chickamauga CrPJtJk·

AI -?- sc " Ga "' "

Figure 3: Map of the upper Tennessee River system.

Area. Although the watershed receives an average of 152 '<\There the stream enters the higher elevations of the Blue cm of precipitation annually (Anonymous 1972), the Ridge physiographic province, pools become deeper, creek water is seldom turbid and the bottom is relatively shorter, and bedrock intrusions dominate the substrate. free of organic debris and silt. Fishes are relatively diverse in Citico Creek. Fifty-six There are two, small concrete dams located at creek km (56) species were documented during this study (Table 13.7 and 25.3 which were built in 1973 to impede the 1) . In addition to N flavipinnis and N baileyi, the fish vernal migration of redhorses (Moxostoma spp.) into up­ fauna also includes the dusky tail darter (Etheostoma stream reaches where hatchery-raised trout are released. percnumm) listed as federally endangered and considered These structures inundate two short stretches of creek endangered by the states of Tennessee (Tennessee Wild­ (approximately 91 m and 18 m respectively, during nor­ life Resources Agency 1994) and Virginia (Burkhead and mal water level). Jenkins 1991), and threatened by the American Fisheries Citico Creek proper has a gradient drop of 6.8 m per Society (Williams et al. 1989). km with a noticeable gradient change at creek km 16.7. When Citico Creek is separated into two contiguous sec­ Materials and Methods tions-the upper being creek km 26.5 to 16.7, and the COLLECTION OF FISHEs-Several collections of fishes using lower being 16.7 to the mouth-the gradient is 14.4 m per standard techniques were made in Citico Creek prior to km and 2.5 m per km, respectively. the discovery of N baileyi (Etnier 1978; University of In the lower section, the creek consists mainly of riffles Michigan, unpublished collection records) . The first with flat, palm-sized rocks (slabrocks) and cobble sub­ specimen was collected with an electroshocker and seine strates alternating with long, shallow pools with medium­ net, but this collecting technique used on many subse­ sized gravel, pebble, slabrock, and bedrock substrates. quent trips to Citico Creek between September 1980 and 48 BULLETIN 18 December I, 1996

Table 1. Species List of Fishes in Citico Creek

Species Common Name I Species Common Name

Ichthyomyzon castaneus chestnut lamprey Noturns baileyi smoky mad tom Lampetra appendix American brook lamprey N. flavipinnis yellowfin mad tom Lepisosteus oculatus spotted gar Pylodictis olivaris flathead catfish Campostoma anomalllm central stoneroller Oncorhynchus myhiss rainbow trout Clinostomus funduloides rosyside dace Salmo IrnUa brown trout Cyprinelia galactura whitetail shiner Salvelinus Jontinalis brook trout C. monacha1 spotfin chub Fundulus catenatus northern studfish C. spiloptera spotfin shiner Coitus caTolinae banded sculpin Cyprinus carpio common carp A mblopliles rnpestris rock bass Eri11lystax insignis blotched chub Lepomis auritus redbreast sunfish Luxilus coccogenis warpaint shiner L. C)lanelius green sunfish L. chrysocephalus striped shiner L. gulosus warmouth Lylhrnrns lirns mountain shiner L. maCTochirus bluegill Nocomis 11liCTopogon river chub L. megalotis longear sunfish Notropis sp. sawfin shiner MiCToplerus dolomieu smallmouth bass N. amblops bigeye chub M. punctulalus spotted bass N. leuciodus Tennessee shiner M. salmoides largemouth bass N. photogenis silver shiner Etheosloma blennioides greenside darter N. telescopus telescope shiner E. chlorobranchium greenfin darter Phenacobius uranops stargazing minnow E. jessiae blueside darter Phoxinus lennesseensis Tennessee dace E. /JerCntlrn11l dusky tail darter Rhinichthys alralulus blacknose dace E. rufilineatum redline darter R. cataractae longnose dace E. simoterum Tennessee snubnose darter Ictiobus sp. buffalo E. zonale banded darter Hypenteliu11l nigricans northern hog sucker Percina eaprodes logperch Moxosloma carinalum river redhorse P. evides gilt darter A meillrns natalis yellow bullhead Perea jlavescens yellow perch IClalurus punctalus channel catfish Apliodinotus grunniens freshwater drum

ICY/Jlillel/a lIlollarha is represented by a single 1936 record and is probably extirpated from Citico Creek.

June 1981 yielded only 16 specimens (13 catalogued at the amining the substrate while floating downstream. Care University of Tennessee Research Collection of Fishes and was taken during the breeding season to avoid distur­ three others released). Direct underwater observations bance of potential nesting habitat. using skin and SCUBA diving gear ultimately proved to be Attempts to collect N. flavipinnis during the day were the most efficient method for collecting life history data unsuccessful, except during the breeding season. After and capturing specimens in all habitats. This approach dark, however, individuals were often observed in the provided the additional benefits of minimizing the distur­ open. To collect them, snorkelers were spaced from bank bance to the benthic habitats where both of these rare to bank across a pool. Each observer swam the length of species occur, and made it possible for a single collector to the pool and searched in the open, under rocks, and capture several specimens in a short period of time. Day­ among debris. When N. flavipinniswere spotted, they were time and nighttime searches were conducted from creek easily captured with small dip nets. Very few individuals km 26.6 to the area inundated by Tellico reservoir (creek evaded capture, and ones that did were often seen again km 2.3). and captured during a second swim through the pool. Daytime riffle searches were conducted by lying head Flow-through minnow buckets were used to temporarily forward in the current, and moving slowly across the hold both mad tom species. Individuals collected were channel while all substrate within an arm's reach was measured and weighed (some were marked) and released examined. By moving approximately one arm's length in the same general area from which they were taken. upstream when the stream edge was reached and then Both species use large, flat rocks as cover for nests. returning across the channel, a riffle area from lower to Searches for nests were made during the breeding season upper end could be thoroughly examined. Daytime by carefully lifting appropriate rocks. After a few nests searches for N. baileyi in pools were accomplished by ex- were observed in this manner, however, this type ofsurvey Dinkins & Shute LIFE HISTORIES OF NOTURUS BAILEYI AND N. FLAVIPlNNIS 49 was discontinued for fear of dislodging egg masses or different population estimates (Schumacker and modi­ disturbing larvae which might be lost to current or con­ fied Schnabel) were determined for each pool in the sumed by minnows. There was also the possibility that the manner described by Ricker (1975). guardian mad tom would not return after being disturbed, Each pool within the 3.2 km ofCitico Creek inhabited although our observations indicated that it usually did not by N. flavipinnis was characterized and designated as be­ move far from the nest. ing similar to the upstream or the downstream study pool. The Citico Creek N. flavipinnis population was studied The total length of each pool type was then summed, primarily by observing individuals inhabiting two adja­ stream width was fixed at 20.7 m, and a population esti­ cent pools. These pools, located at creek km 16.4, were mate for the 3.2 km was extrapolated. representative of two different pool habitats found in the stretch of stream inhabited by N. flavipinnis. Physical char­ Reproduction and Early Life History acteristics of these pools were measured in March 1994. Annual fecundity was determined by examining ova­ The downstream pool was long (147 m) and shallow, and ries from preserved specimens with the aid of a dissecting averaged about 1 m midstream depth. Substrate consisted microscope. Ova were excised from the ovary and sorted mainly of gravel and pebbles with occasional slabrocks. into various size classes, the diameters measured with an Boulder and bedrock predominated at the head of the ocular micrometer, and counted. In the summer of 1983, pool, and bedrock shelves lined the northern shores of one nest of each species was removed from the field, taken the pool. Current velocity was slow (0.33 m/sec). The to the laboratory, and placed in a small aerated container upstream pool was long (173 m) and deeper, with a large of creek water. The container was maintained at 18 to proportion of the pool exceeding 2.5 m. The substrate 20 ac. Larvae were periodically removed, preserved in 5% was large boulders and bedrock. Current was somewhat buffered formalin and illustrated. Larval terminology fol­ swifter in this pool. These two pools were separated by a 45 lows that of Snyder (1976). m riffle. Stream discharge in this area averaged 4.9 m3/sec. Between 1986 and present, 79 smoky and 16 N. Most collections of preserved material and observa­ flavipinnis nests containing eggs or larvae were removed tions oflive individuals were made between May 1981 and from Citico Creek and reared in captivity as part of a June 1984. Preserved specimens used for this paper in­ recovery project designed to reestablish extirpated popu­ cluded 13 N. baileyi and 12 N. flavipinnis from Ci tico lations of both species (Shute et a\. 1992; Shute et al. 1993; Creek, and 30 N. flavipinnis from Copper Creek, Virginia. Shute and Rakes 1994; Rakes et al. 1990; Rakes et al. 1995). Some data from the recovery project are included Tagging and Population Estimation in this paper. Between June and November 1982, 46 adult N. baileyi were individually marked with a mixture of water and Diet acrylic paint injected under the skin. However, the diffi­ Stomachs and intestinal tracts from preserved speci­ culty in handling individuals of this diminutive species, mens were examined for gut contents. Prey items were fear of causing mortality, and the lack of recaptures led us identified to the lowest practical taxonomic unit. Diffi­ to abandon efforts to mark and recapture N. baileyi. How­ culty was encountered in identifYing some mayfly ever, adult N. flavipinnis are larger and the same tech­ nymphs due to digestion of soft body parts. For this nique was much easier to perform on this species. Data on reason, madtom capture localities were surveyed for individual growth rates, longevity, movement, and popu­ Ephemeroptera. Results of the survey were used to devise lation density were gathered from this effort. a diagrammatic key for identifYing mayfly fragments in Forty-five N. flavipinnis were individually marked with the stomachs and intestines of madtoms. In this way, acrylic paint using different combinations of colors and almost all prey items were identified to the generic level. body locations. In addition to the acrylic marks, the left Percent of occurrence was determined by dividing the pelvic fin was clipped on 21 other N. flavipinnis from the total number of madtoms in which the prey item was upper pool, and the right pelvic fin was clipped on 32 found by the total number of prey items. Percent fre­ mad toms from the lower poo\. Yearling N. flavijJinniJ were quency of occurrence was determined by dividing the neither marked nor fin-clipped for fear of potential in­ total number of madtoms in which the prey item was jury. found by the total number of stomachs. In order to meet the multiple census restrictions of negligible recruitment or mortality, mark-recapture took Results and Discussion place after the breeding season was over and before win­ Range Within Citico Creek ter began (between August and October 1983). During NOTURUS BAlLEYf--The habitat change at creek km 16.8, as each census subsequent to the initial tagging, the number the creek descends from the higher elevations of the Blue of recaptured fish and location of marks were recorded, Ridge physiographic province, marks the upstream extent and any unmarked fish were tagged. From these data two of N. baileyi (Figure 4). Upstream, the substrate consists 50 BULLETIN 18 December 1, 1996

Tellico Citico Creek at creek km 11.4. On several occasions fol­ ; •• :'~ ., ' r , . Reservoir lowing heavy rains, we have watched Caney Branch release heavy sediment loads into Citico Creek. Caney Branch's . ~ 2 '-- .. ~ Forest headwaters drain approximately 0.4 km of a private prop­ Boundary erty inholding within the National Forest. The presence of N. baileyi in this lower stretch of Citico Creek adds further complexity to this question. Because CHEROKEE NATIONAL of their preference for riffles during most of the year, N. FOREST baileyi may be more tolerant of siltation than the year­ round pool-dwelling N. jlavipinnis. Occasional periods of turbidity are common, however, at the other two localities where N. jlavipinnis occurs (Powell River and Copper Creek). Finally, because of the popularity of Citico Creek with local campers and fishermen, it is possible that N. jlavipinniswas introduced into Citico Creek and has been l J unable to expand its range downstream of the dam. This is an appealing theory because it would indicate that an­ other, most likely local, population may exist.

N. flavipinnis study poolS (kin 16 4)

upslreamD~lanl Habitat of N.fl.svipinnis (km 17.3) NOTURUS BAlLEYl--From late May to early November non­ nesting N. baileyi were found underneath slabrocks in CHEROKEE NATIONAL swift to moderate currents in all parts of riffles, especially FOREST riffle crests. At three riffle crest localities where 48 speci­ mens were captured, current ranged from 0.52 to 0.67 m/ sec (X = 0.58), and depths ranged from 30 to 41 cm (X = 34.0). In these areas, the substrate was comprised largely ~ Dam (km 25.3) SCAl..E.: ,~" - 1.6km of slabrocks lying in a pea-sized gravel matrix. Physical dimensions of riffle area slabrocks used as shelter by Figure 4: Map of lower Citico Creek. N. baileyiare presented in Table 2. Some of these slab rocks (18%) had slight depressions on the undersurface. largely of boulders and bedrock, and short cascades alter­ Between early November and late May, N. baileyi were nate with short pools. The 10.8 km stretch of creek occu­ found in shallow pools. The shift in habi tat from the riffles pied by N. baileyibetween creek km 6.0 and 16.8 is typified they inhabit in warmer months occurred in less than one by shallow riffles composed of abundant slabrocks, long week and coincided with a drop in water temperature to shallow pools with pea-sized gravel and occasional 7-8 °C. Mayden and Walsh (1984) also suggested a late slabrocks, and a few deeper pools with large boulders and fall/early winter habitat shift for the least madtom, silty/sandy bottoms. The Ranger-Citico-Fletcher soil asso­ N. hildebmndi, based on a shift in gut contents. ciation roughly delineates that section of the stream occu­ Water depth at three localities where 41 N. baileyi were pied by N. bailp)'i (Hall et al. 1981). Below creek km 6.0, collected during late fall and winter ranged from 60 to 68 the substrate is dominated by bedrock and sand. During cm (X= 62.7); current ranged from 0.15 to 0.43 m/sec (X this study, water temperatures in the reach occupied by = 0.27). Substratum was comprised primarily of large N. baileyi ranged from a low of 5°C (January) to a high of boulders, with occasional slabrocks, and a sand/gravel 23°C (July). matrix. Roughly half of the pool slabrocks used by N. baileyi were either entirely concave (side view) or had a NOTURUS FlAV1PINNI~In Citico Creek, N. jlavipinnis occurs depression on the undersurface. Analyses of variance tests in a 3.6 km reach upstream of a small concrete dam at on slabrock dimensions examined for the two habitat creek km 13.7 (Figure 4). The absence of N. jlavij)innis types produced low F-ratio values (Table 2), indicating below the dam is puzzling, as pools in the lower reaches that the slabrocks N. baile),i used for cover in the riffles are plentiful and habitat is apparently similar to that were not significantly different from those used for cover above the dam. Absence of the species from the lower in the pools. No statistical tests were conducted to deter­ reaches of Citico Creek may be related to short-term mine if the slab rocks used for cover by N. bailrryi differed degradation that occurred during dam construction or significantly from other available slabrocks in the same may be related to agricultural practices in the headwaters area. Our observations indicate a wide range of rock sizes of Caney Branch, a second order stream that flows into occur in the riffles and pools and rocks of the preferred size and shape are less abundant in the pools than in the Dinkins & Shute LIFE HISTORIES OF NOTURUS BAILEYI AND N. FLAVIPINNIS 51

Table 2. Dimensions of Slabrocks Occupied by Non-Nesting Noturus baileyi with Standard Lengths, by Habitat, from Citico Creek,June 1982 to June 1983

SL (mm) Rock Surface Area (mm) Rock Thickness (mm2) N X±SE Range F-Ratio X±SE Range F-Ratio X ± SE Range F-Ratio

Untransformed Data

Rift1es 46 39.78 ± ].35 ~ 0.00] 8300.65 ± 638.67 2796-]8265 0.]30 19.89 ± 1.42 4-50 0.010 Pools 30 39.83 ± 3.] 6 27-58 8422.47 ±] ]80.68 3]]7-33006J 20.93 ± 1.70 8-50 Both 76 39.80 ± 1.08 22-58 8348.74 ± 600.79 2796-33006 20.30 ± ].09 4-50

Log J!! Transformed Data

Riffles 46 1.59 ± 0.02 1.34-1.76J 0.003 3.86 ± 0.03 3.45-4.26 J 0.] 15 ].25 ± 0.03 0.60-1.70l 0.042 Pools 30 1.59 ± 0.03 1.43-1.76 3.85 ± 0.05 3.49-4.52 ].28 ± 0.03 0.90-1.70 Both 76 1.59 ± 0.0] 1.34-1.76 3.85 ± 0.03 3.45-4.52 1.26 ± 0.02 0.60-1.70 riffles. Thus, competition for slabrocks may have in­ fish individually tagged with acrylic paint, seven were creased as N. baileyi left the riffle areas and entered the recaptured between seven and 32 months after tagging pools. While N. baileyi selected for a certain size of protec­ (Table 3). All of these recaptures were found in the same tive cover rock in both habitats, multiple regression re­ pool where originally tagged. The 13 recaptured fin­ sults showed no relationship between fish length, rock clipped individuals also had not moved from their pool of thickness, and rock surface area. An analysis of variance original capture. test on the standard lengths (SL) of madtoms captured in That localization of small populations may be common the two habitats produced low F-ratio values, indicating an in mad toms is further supported by other evidence. In a equally collected distribution of sizes. Thus, there was no study of N. gyrinus, Case (1970) showed that movement of selection for age class in the areas sampled. this species is very limited; of 693 adult fish marked, only 5 individuals were recaptured at sites other than where NOTURUS FLAVIPINNIs--According to Jenkins (1978), N. they were first captured and released. Fuselier and Edds jlavipinnis is an inhabitant of pools and backwaters in (1994) reported that only one of twelve marked Copper Creek. In Citico Creek, N. jlavijJinnis were usually N. placidus had moved from the locality of original cap­ found in shallow pools (less than one m deep); in deeper ture. pools individuals were almost always observed at depths of Limited movement patterns observed in this study ap­ less than two m. Thorough searching beneath movable ply to adult N. jlavipinnis only, since small individuals were cover in the main stream bed rarely revealed any individu­ not marked. Proper censusing ofthe young by snorkeling als during daylight hours or at dusk. Presumably, they requires a great deal more time than searching for adults, were diurnally associated with other cover types such as and a much differen t search image is needed to detect the stream banks, bedrock ledges, or tree roots. During cryptic juveniles. It is possible that juveniles are the dis­ spring, summer, and fall, adult N. jlavipinnis were noctur­ persal agents for the species. nally associated with open benthic areas and occasionally, Jenkins (1975) reported that all life stages of N. cover. Adults were increasingly scarce in open benthic jlavipinnis in Copper Creek occupy the same habitat. The areas as the water temperature decreased in late fall, and same appears to be true for N. jlavipinnis in Citico Creek; we presume that very little activity occurred during winter none were found beneath rocks in riffles or runs while months. Young N. jlavijJinnis may be more active at cooler searching for N. baileyi. Free-swimming juveniles as small temperatures than adults. This is based on a November as 19 mm SL were observed over fine-grained substrates in observation when five young and only one adult were pools as early as mid-August. Perhaps the reason two such observed (water temperature was 6°C). Substrate in pools closely related congeners can occur sympatrically in occupied by N. jlavijJinnis consists mainly of gravel and Citico Creek is that adult N. jlavipinnis, unlike N. baileyi, pebbles, with occasional slabrocks, boulders, and bed­ have never been observed to switch habitats. Therefore, rock. there is little interaction during warmer months when both species are most active. Movement Recapture data from marked fish indicate that adult Population Estimate N. .flavipinnis do not ordinarily move between pools. Of 45 Recaptures of fin-clipped individuals were used to esti- 52 BULLETIN 18 December 1,1996

Table 3. Recapture Data from Individually Tagged Noturus flavipinnis from Citico Creek

Locality of Size When Locality of Size at Tagging Date of Tagged Recapture Date of Recapture Growth (pool) Tagging (mm SL) Sex (pool) Recapture (mmSL) (mm)

Lower 10/ 1/ 81 39 Lower 6/30/84 108 69 Upper 10/ 1/ 81 110 0 Upper 5/ 29/ 82 114 4 Upper 10/ 1/ 81 103 'i' Upper 5/ 8/ 83 108 5 Lower 10/ 1/ 81 73 Lower 7/ 15/ 83 106 33 Lower 10/ 1/ 81 80 ? Lower 6/ 27/ 83 93 13 Upper 10/ 4/ 81 83 0 Upper 5/ 29/ 82 104 21 Lower 6/ 27/ 82 100 Lower 8/ 9/ 82 106 6

Table 4. Recapture and Population Estimation Data for Noturusflavipinnis Based on Mark-Recapture (Fin-clipping) in the Upper and Lower Study Pools

Confidence Date C, R, M, C,M, M,R, C,M,2 R2/C, Estimate 95%

Upper Pool

8/ 25/ 83 6 0 0 0 0 0 0 9/ 1/ 83 6 0 6 36 0 216 0 9/ 28/ 83 11 4 12 132 48 1584 1.4545 33" 13-132 34" (Poisson) 38e 10/ 16/ 83 2 0 19 38 0 722 0 53e 20-206 21 41 " (Poisson) Lower Pool

8/ 16/83 11 0 0 0 0 0 0 8/25/83 6 2 11 66 22 726 0.6667 33" 9-330 (Poisson) 9/ 1/83 11 4 15 165 60 2475 1.4545 33" 18-105 39" 30-56 9/ 28/ 83 11 3 21 231 63 4851 0.8181 46" 27-116 56e 35-160 10/16/83 5 2 29 145 58 4205 0.800 51" 41-114 32 60" 31-112

RI = total recaptures on day l. t. el = total fish captured on day l MI = effective number of tags at large 011 start of t " day (number previously marked). "Petersen estimate. "Modified Schnabel estimate. 'Schumaker estimate. Dinkins & Shute LIFE HISTORIES OF NOTURUS BAILEYl AND N. FLAVlPlNNlS 53

I ~ Jur.e H-18 .L -1.1 I • : JLlI~ H_Zl - • 1 ... .J I .... ~ Augu,t N_4 I .. I • ~ S'Jlt,mbe!' N.. l. JI •• i I I •• I ·: Octooer N-Jl j • .1 M..I ~ .. ·I ~ N07YtImb", N.. 2 ~ I 15 I • • ~ : Jonl.lC1')' N .. !I I • • ~ I ~ i : f.br""lry N_S ••• • • I ·: ~lIrch N-17 • L • I I •• ~ APril N .. 1 , • : lIay N_1.3 • I. • 1. ~. .10 .. STA~D L.£NaTH (mm) • '20 Figure 5: Length frequency histograms for specimens of Figure 6: Length frequency histograms for specimens of Noturlls baileyi from Citico Creek. Noturus flavipinnis from Citico Creek. mate a population size of 41 and 53 adult Njlavipinnis for hildebrandi species group as defined by Grady and the upper pool (modified Schnabel and Schumaker LeGrande (1992), of which N baileyi, N hildebrandi, and methods, respectively; Table 4). Estimates obtained for N siana1lli are members, are the smallest and shortest­ the lower study pool were 51 and 60. By extrapolating lived madtoms. Etnier and Jenkins (1980) noted two age these figures to the total length of each pool type within classes in N stana1lli, indicating a lifespan of 1+ years. the range of N jlavijJinnis in Citico Creek, estimates of 451 Mayden and Walsh (1984) and Baker and Heins (1994) (modified Schnabel) and 549 (Schumaker) with confi­ reported a lifespan of 1 + years for N hildebrandi. dence limits of 312-1453 were obtained. Using these figures, the density of adult N jlavijJinnis in this stretch of NOTURUS FLAVIPINNIs--Length frequency histograms of 233 creek is 1.4 and 1.8 madtoms per 10 m 2, respectively. The N jlavipinnis in Citico Creek indicated a three-year density of the rare, riffle-dwelling NjJIaciri1lswas estimated lifespan (Figure 6). Jenkins (1978) reported that at 3.3 madtoms per 100 m2 (Fuselier and Edds 1994). N jlavipinnis lives three to four years in Copper Creek. Juvenile N jlavipinnis may be free-swimming and re­ Age and Growth leased from paternal guardianship as early as August. W. NOTURUS BAILEYI-Length-frequency distribution of 131 N C. Starnes (pers. comm.) reported a 16.5 mm SL indi­ baileyi suggests an average lifespan of two years (Figure 5). vidual taken from Copper Creek on 9 August 1975. In The proportion of small individuals in the population Citico Creek, ten juveniles captured in August 1983 aver­ indicates healthy recruitment. Data from individuals aged 24.9 mm SL (19-29 mm SL) and five individuals raised in captivity under conditions simulating those in captured in November 1983 averaged 37.8 mm SL (34-45 Citico Creek (Shute et al. 1992) support these field obser­ mm SL). Three juveniles captured from Copper Creek in vations. Thirteen 9-month old, five 14-month old, 21 21- November averaged 23.7 mm SL (31 to 35 mm SL). month old and 12 24-month old captive reared N baileyi Captive-reared young average 40 mm SL by November (P. averaged 52.9, 61.4, 63.3 and 68.9 mm SL, respectively. L. Rakes pers. comm). Juveniles may be free-swimming and released from Data on individual growth rates were also obtained by paternal care as early asJuly. In Citico Creek, six juveniles recapturing madtoms that had been tagged with acrylic captured on 30 August 1994 measured 12-29 mm SL (X = paint. These data indicate that N jlavijJinnis may reach a 18). Two captive reared individuals from the 1990 breed­ certain maximum size limit after which growth ceases or ing season, when measured on 27 November 1990, were occurs at a much slower rate. There may be a genetically 30 and 32 mm SL. predetermined maximum length of 110-115 mm SL, at A maximum lifespan of 18 months to nine years has least for the Citico Creek population (see Table 2 and been reported for various species of madtoms (Mayden Figure 6). and Walsh 1984; Mayden and Burr 1981). However, the Data from captive-reared individuals which support the average lifespan of most mad toms is two or three years field observations include the following: one five-month (Case 1970; Mahon 1977; Thomerson 1966; Clark 1978; old individual was 40 mm SL, four nine-month old indi­ Burr and Mayden 1982b; and others). Members of the viduals averaged 60.8, five 14-month old individuals avel'" 54 BULLETIN 18 December 1, 1996

growth in either weight or length, although the largest female collected during their study was 51.6 mm SL while the largest male was 48.6 mm SL; the female was 15 months old and the male was 18 months old. No such information exists for N. stanauli. Given that N. stanauli is probably an annual species (Etnier and Jenkins 1980), it would be difficult to discern sexual size dimorphism as a function of longevity. Male N. baileyi in breeding condition show more body A c color (yellow) than immature males and gravid females. No similar pattern has been reported for N. hildebrandi (Mayden and Walsh 1984) or N. stanauli (Etnier and Jenkins 1980). Secondary sexual characteristics and length frequency data from wild and captive-reared fish (Figure 5, in part), indicate that individuals of N. baileyi can become sexually mature in their second summer of life (at one year of age). Two females collected and released in July 1982

> .~~~ --...... S _--:-" Ill' . . .' .~ ..s..'~. ~JIIL:er.: .:'. :..;.';;_.'

B D .. ..-~ , ......

Figure 7: Genital papillae of Noturus baileyi. (A) Breeding female 52mmSL. (B) Non-breeding female 37mmSL. (C) Breeding male 52 mm SL. (D) Non-breeding male 34 mm SL. A

aged 74.8 mm SL and one 26-month old individual was 96 mmSL.

Sexual Dimorphism NOTURUS BAILETI-SeVen of eight N. baileyi 57 mm SL or ----- larger collected during this study were females (the larg­ est measured 63 mm SL). The largest male collected was 58 mm SL. Ten reproductively mature male and 14 female N. baileyi collected during breeding seasons in 1981 to B 1983 averaged 49 mm SL (ranging from 43-58, SD = 4.8) and 52 mm SL (ranging from 38-63, SD = 7.4), respec­ tively. Although these data indicate that female N. baileyi attain greater lengths than males, the difference was not statistically significant (X 2 of 0.0675 < X2 0.05 (1) = 3.841). Within the hildebrandi species group, sexual size dimor­ phism is equivocal. D. S. Wilkins (pers. comm.) indicated that there was no significant size difference between c males and females of a Mississippi population of N. hildebrandi. Mayden and Walsh (1984) reported that Figure 8: Genital papillae of Noturusflavipinnis. (A) Breeding male and female N. hildebrandi from a population in west­ female 79 mm SL. (B) Breeding male 90 SL. (C) Non­ ern Tennessee did not differ significantly with respect to breeding male 47 SL. Dinkins & Shute LIFE HISTORIES OF NOTURUS BAILEYI AND N. FLAVIPINNIS 55

were 38 mm SL each and appeared to be gravid. Two male around the eyes and small areas anterior and posterior to N. bailcyi in breeding condition collected in June 1982 the dorsal fin, and in the adipose and caudal fins (Mayden measured 43 and 44 mm SL. That these mad toms were all and Burr 1981). likely one year old is supported by an average length of 51 Male and female N. jlavipinnis in breeding condition mm SL from 17 captive reared individuals that were 16 exhibited secondary sexual characteristics similar to months old. breeding N. bailcyi. Non-breeding male and female N. During the spawning season, male N. bailcyi possess jlavipinnis could not be distinguished in the field using secondary sexual characters as described by Taylor (1969) genital papillae as a guide (Figure 8). for other species of Noturus, i.e., enlarged genital papilla, Twen ty mature male N. jlavipinnis collected during the cephalic epaxial muscles and lips. In fact, the swollen lips 1982 and 1983 reproductive seasons averaged 4 mm of the type specimens of N. bailcyi collected from Abrams longer than 21 mature females collected during the same Creek led Taylor (1969) to describe N. bailcyi as having a time period. This size differential was not statistically lower jaw being only slightly included in the upper jaw significant (X 2 of 0.083 < X2 0.05(1) = 3.841). The mean (Bauer et al. 1983). standard length ofthe males was 98.8 mm SL (85-114, SD The genital papilla of preserved male N. bailcyi in non­ = 9.9), and the mean for the females was 94.8 mm (80- breeding condition is distinguishable from that of pre­ 112, SD = 9.1). Most of these individuals were probably served non-gravid females (Figure 7) but could not be t\vo years old at the time of capture. used as a reliable field character for sex determination of The sexual size dimorphism reported for most live, non-breeding madtoms. In preserved specimens, sex madtom species (Burr and Mayden 1984; and others) may is easily discerned by inspecting the gonadal tissue. Testes explain the observation of two specimens that were about of male N. bailcyi consist of long, whitish, finger-like pro­ the same size at marking (73 and 80 mm SL), but mark­ jections as described for catfish and other mad toms edly different lengths when recaptured 21 months later (Sneed and Clemens 1963; Mayden and Burr 1981; Burr (from 1 October 1981 to 15July 1983, and 1 October 1981 and Mayden 1982a,b; Clark 1978) . to 27 June 1983, respectively) . One had increased 33 Female N. bailcyi in breeding condition were identified millimeters and the other 13 mm in approximately the by their distended abdomens and swollen genital papillae same length of time (Table 3). (Figure 7). The enlarged cephalic muscles and swollen Differential growth rates have also been reported for lips, apparent in nuptial males, are not present in gravid the checkered madtom, N. jlavater (Burr and Mayden females. 1984). Bowen (1980) analyzed annuli formation on otoliths and vertebrae of N. miurus and found that no NOTURUS FLAVIPINNIs--Female N. jlavipinnis appear to be significant difference existed between the lengths of first sexually mature by their third summer of life (at two years and second summer males and their female counterparts. of age), as indicated by the presence of ripe or nearly ripe Third summer males were significantly longer than simi­ ova in the gonads of three females taken in May 1982 that lar age females. Bowman (1932) reported that although were 75 - 79 mm SL. Three females 49 - 58 mm SL taken both males and females of the margined madtom, inJune 1981 were probably in their second summer oflife N. insignis, live for three years, males are slightly larger. He (one year of age) and contained no mature ova. However, used length frequencies to demonstrate that the same most captive-reared individuals became gravid at one year three peaks were evident in both sexes. of age. This may be related to "winter" water temperatures Although some species of Noturus apparently do not in the laboratory which were approximately 6 DC warmer exhibit sexual size dimorphism (e.g., N. furiosus, Burr et than Citico Creek. Perhaps a more regular feeding sched­ al. 1989), males of many species attain a larger size than ule also contributed to this apparent early maturity of females. This sexual size-dimorphism is possibly due to captive-reared individuals. selective pressures favoring a larger body size for the nest­ Some male N. jlavipinnis may also become reproduc­ guarding parent thereby affording better protection of tively mature at one year of age as indicated by a 74 mm SL the brood. Also, during the seasons in which more rapid male found guarding a nest on June 1982. This male was growth (length) occurs (spring and summer), the female the smallest male observed exhibiting nesting behavior. may require proportionately more energy from food re­ Breeding male N. jlavipinnisare not colored differently sources for gonadal development. than immature males or gravid females. The coloration of An alternative explanation of Notunts size dimorphism breeding N. nocturnus males also remains unchanged was presented by Burr and Mayden (1982b) who sug­ (Burr and Mayden 1982b) . Males of the closely related gested that N. 1Iliunts males attain a larger size than fe­ N. miurus were described as more drab than gravid fe­ males because the majority of females do not live as long males (Burr and Mayden 1982a). The general coloration as males. This hypothesis was also proposed for of N. exilis males in breeding condition differs from non­ N. eleuthems (Starnes and Starnes 1985) and N. exilis breeding males in that there is more yellow on the venter, (Mayden and Burr 1981). 56 BULLETIN 18 December I, 1996

Table 5. Nesting Variables for Noturus baileyi and N. flavipimlis in Citico Creek

Length, Width I, and Sex and Total Length Nest Water Temp. Thickness of Nest Water Depth Current Clutch (mm) of the Guardian No. Date (C) Habitat/Creek Km Rock (cm) (cm) (cm/sec) Size or Breeding Pairs

N. baileyi

1 7/ 2/ 82 23° Shallow pool/9.0 26:21:4 50 40 33 o:U 2 7/ 2/ 82 23° Shallow pool/ 14.8 23:17:5 40 37 PS~ 0:55,

N. jlavijJinnis

1 5/ 29/ 82 20° Head of pool/ 16.4 52:50:10 100 33 89 0:108 2 7/ 2/ 82 23° Pool/I 6.4 51:25:- 33 45 4 0:132 3 7/ 2/ 82 23° Poo1/16.4 51:25:- 100 33 60 1 0:120 4 7/ 2/ 82 23° Poo1/16.4 51 :25:5 41 33 50 1 0:125 5 7/ 2/ 82 23° Poo1/16.4 25 :25:- 100 33 25.1 0:110 6 7/ 2/ 82 23° Poo1/16.4 51 :51 :- 33 45 1 0:130 7 7/ 8/ 82 23° Pool/ 16.4 61 :46:- 56 33 78.1 0 :102 8 6/ 25/ 83 22° Pool/I 6.4 61:36:12 46 33 100 1 0:90 9 6/25/83 22° Poo1/16.4 36:30:5 61 33 30 1 U3 10 6/25/83 22° Poo1/16.4 56:23:5 61 33 9-_!1 1 0:104

I Length and width of nest rock was based on widest and narrowest measuremenls across rock f~lce. ~ Pre-spawning. ·~ Unknown (madtom escaped capture) . IEstimate, based on visual observation of nest cavity.

Nesting Behavior rity was correlated with female size, larger females spawn NOTURUS BAILEYI-Madtoms are included in the reproduc­ earlier than smaller ones. tive guild of fishes termed speleophils by Balon (1975). Water depth at seven nest sites averaged 39 cm and Species in this guild construct a nest or choose a natural current averaged 37.5 cm/ sec. Nest rocks averaged 3.6 cm cavity for a nesting chamber and, generally, the male in thickness and 25.8 cm in length and in width. Aquatic guards the eggs. Noturus baileyi choose large, flat rectangu­ insects and crayfish were noticeably absent from the un­ lar rocks for nesting cover. Nest rocks were much larger in derside of all nest rocks. Four nests collected in 1982 and overall dimensions than those used for shelter during the 1983 were each attended by a single male. The stomach of remainder of the year. Presumably, one or both parents one of these guardian madtoms was nearly empty. Expos­ enlarge the nesting cavity by fanning with body and fins ing the nest cavity resulted in differing responses by the since the area beneath each nest rock was devoid of silt. guardian madtom. In one instance (Nest no. 1, Table 5) No nests were found beneath rocks that were lying flush the madtom was reluctant to leave the nest cavity even on the substrate with no apparent cavity underneath, even after the clutch was removed. In another (Nest no. 3), the in areas where the number of large, flat rocks was limited. mad tom fled immediately after the nest was exposed. Male N. bailryi were reproductively mature from the Several hours later this nest was re-examined and the end of May throughJuly. Gravid female N. baileyiwere also guardian madtom had not returned. Other fish or cray­ found from early May to late July. NotuTlls baile)'i in breed­ fish had evidently moved in and consumed most of the ing condition were found underneath large, flat rocks in eggs. shallow pools and riffle crests from early June to mid-:July Noturus baih,).i males were observed guarding larvae (Table 5) . Baker and Heins (1994) reported ripe females estimated to be five days old (post-hatching). Individuals in a southern population of N. hildebrandi as late as mid­ hatched in an aquarium actively fed at about the age of September. All females with ripe or ripening ova were five days, depending upon water temper-ature. Based on one-year olds. They suggested that, because ovary matu- the length of time required for a freshly fertilized batch of Dinkins & Shute LIFE HISTORIES OF NOTURUS BAILEYI AND N. FLAVIPINNIS 57

70 indicated a spawning period for N. leptacanthus that coin­ .. cided with peak water temperatures for two successive o 0 60 years, although she suggested that this correlation may have been coincidental. Breder (1935) indicated that 55 o water temperature is the determining factor in initiating "" reproductive behavior in the brown bullhead, Arneiurus .. nebulosus. o The spawning and nesting season for N. baileyi and N. '" o 0 I,. jlavipinnis in Citico Creek are essentially the same. Male ;; o ~ and female N. jlavipinnis have been found in breeding 30 N. tIQ.,ipiiufi8 nmlb N_27 condition from late May to mid:July. While they normally ,. o o are not syntopic with N. baileyi during the non-breeding 20 season, juvenile N. jlavipinnis have been found under

'5 large, flat rocks in shallow pools in close proximity to N. baileyi nests. Further, nests of both species have been '0 found in proximity to each other in shallow pools. Figure 9 iIIustrates the size of nest rocks used by each

10 15 20 25 ~ J5 ~ 45 00 ~ ~ ~ ro mad tom species. Although there is overlap in the size of l.ngtl1 (em) Figure 9: Graphic presentation of nest rock utilization these rocks, in general N. jlavipinnis uses larger rocks. (overlap) by Noturus baileyi and N. flavipinnis in Citico Adult N. jlavipinnis are larger than adult N. baileyi, and Creek. would therefore be expected to require and be able to defend a larger nesting space. Several other species of Not1lrus coexist. NotuTtts elegans, N. eleutherus, N. miurus, N. nocturnus, and N. stanauli are known to occur at a single locality in the Duck River, eggs to hatch (16 days at 20-21 0c), a guardian male may Humphreys County, Tennessee (D. A. Nieland, unpubl. attend his brood for at least three weeks. data), and Not1lrus eleutherus, N. jlavus, N. gyrin1ls, N. noct1lrnus, and N. mi1lrus are known to occur in the NOTURUS FLAVIPINNIs--Male N. jlavijJinnis in breeding condi­ same reach of the Wabash River (R. Bogardus, pers. tion were found in Citico Creek from late May to mid:July. comm.). VlThere N. albater, N. jlavate,~ and N. exilis are Water depth at eight nest sites found in late May to mid­ syntopic, N. albaternests under smaller rocks in shallower July 1982 and June 1983 ranged from 41 to 100 em. Nest water in pools and riffle crests (Burr and Mayden 1984). rocks averaged 50 cm long and 34 em wide (N = 10). In Lake Waccamaw, North Carolina, the undescribed Current measured 33 em/sec. Substrate beneath the nest broadtail madtom (subgenus Schilbeodes) coexists with rocks consisted of gravel, cobbles and sand. Ten nests with N. gyrinus, and even spawns at the same time of year and eggs in varying stages of development averaged 55 eggs under the same nesting cover (Lindquist et al. 1982); (Table 5). Three of these (Nests 1, 7 and 8) contained these two species are temporally separated by diel activity newly laid eggs with an average of89 eggs per clutch. Each differences (Reynolds et al. 1982). clutch was attended by a single male. The earliest N. Nests of N. jlavipinnis were only found in the pool jlavijJinnis nest was found on 29 May 1982, and nesting habitats normally occupied by the species. Flat rocks of continued that year through June (Table 5). the size typically used by N. jlavipinnis for nesting cover In 1983, field surveys for nests began in early May. are uncommon in some pools, and although there is Although gravid females were evident in May and June, minimal competition with N. baileyi for this resource, and four pairs of fish were observed beneath potential there may be competition with other organisms. On sev­ nest rocks on 2 June 1983, no nests were found until 25 eral occasions, single egg-guarding mudpuppies (Nect1lTlts June and nesting appeared to be almost over by mid:July. mawlosus) were found beneath large, flat rocks in pools The difference in spawning seasons between these two occupied by nesting N. jlavijJinnis. Burr et al. (1989) also years may have been due to a comparatively cold spring in suggested that syntopic Nect1lTlts compete with N. jW70SUS 1983 which delayed spawning by sustaining lower water for nest rocks. temperatures. Therefore, photoperiod may be important Guardian N. jlavipinnis males apparently do not feed, in preparing the gonads for reproduction and water tem­ based on the single parent removed from a nest offreshly perature (20-23 0c) may be the environmental trigger laid eggs during the night of 29 May 1982 (1:00-2:00 which initiates spawning. A.M.). His stomach was completely empty, and the hind­ Starnes and Starnes (1985) reported a possible delayed gut contained only a few sand grains and some well di­ reproductive period for N. eleutherus, and Clark (1978) gested chironomid and trichopteran larvae. In com pari- 58 BULLETIN 18 December 1,1996

son with other individuals collected at night, the diversity Mayden and Walsh (1984) reported a mean of 29.9 ma­ and amount of food organisms found in this male's gut ture oocytes for one of N. baileyi's closest relatives, was greatly reduced, and the larval insects present could N. hildebrandi. Baker and Heins (1994) reported a range have been ingested during nest construction or mainte­ of 16-68 ova, correlated with female size, in a southern nance. population of N. hildebrandi; further these authors empha­ Whiteside and Burr (1986) reported two N. gyrinus sized that the lifetime reproduction potential of the short­ guarding a single clutch of eggs. However, most recent life lived N. hildebrandi (one year plus) is limited to one breed­ history studies have shown that males alone assume pa­ ing season. In comparison with other madtoms, N. baileyi rental responsibility of the clutch and do not feed while in is among the least fecund. that capacity (Clark 1978; Mayden et al. 1980; Burr and Mayden 1984; Mayden and Burr 1981; Burr and Dimmick NOTURUS FLAVIPINNI5--The total number of ova contained in 1981; Mayden and Walsh 1984; Starnes and Starnes 1985). five gravid N. flavipinnis ranged from 382 to 456 (X = 408). Brown bullheads (A. nebulosus) exhibit biparental care, Oocytes of these females were separable into three size but the males are the principal care-giver and fast while classes. The mature oocytes were translucent yellow and guarding eggs and larvae (Blumer 1985).

Fecundity NOTURUS BAlLEYI-Immature female N. baileyi taken from Citico Creek during the breeding season contained ova that were undifferentiated in size. The ovaries of a nearly gravid female (52 mm SL) collected in early May con­ tained 287 oocytes of three sizes. The largest oocytes were orangish in color, spherical, and ranged in diameter from 0.9 to 1.0 mm (X= 1.0, N = 55). Intermediate-sized oocytes were either clear or whitish in color, spherical, and ranged in diameter from 0.3 to 0.5 mm (X = 0.4, N = 122). The smallest oocytes were translucent, somewhat flattened, A and ranged in diameter from 0.1 to 0.3 mm (X = 0.2, N = 110). Because this female was collected one to two months before spawning, the largest oocytes may not have been fully developed. A total of 653 eggs was found in a fully gravid female (63 mm SL) that had been reared from the egg stage in an aquarium. The abdomen of this apparently healthy fe­ male was severely distended, more so than in typically gravid females found in the wild. Her largest oocytes averaged 2.8 mm in diameter (N = 87), the intermediate­ sized oocytes averaged 0.9 mm (N = 91 mm), and the B smallest oocytes averaged 0.2 mm (N = 475). The wide difference in the number of immature ova between these two madtoms may have been the result of the environ­ ment in which each had been raised. The clutch size of four nests found in 1982 and 1983 ranged from 30 to 42 eggs (X = 36.3, Table 5). Subsequent data collected during captive rearing efforts indicates a much wider range in clutch size; between 1986 and 1994 a range of 20 - 65 eggs (X = 30) was found in 79 N. baileyi nests of varying development stages (P. L. Rakes, pers. comm.). Madtoms in general exhibit low fecundity in compari­ son with other North American freshwater fishes (Breder c and Rosen 1966). A combination of relatively large egg size and the high level of parental care given to the Figure 10: Embryo and larvae of Noturus baileyi. (A) fertilized eggs and larvae reduce early mortality and Prehatchling, chorion removed. (B) Top and (C) Side view therefore the need to produce a large number of gametes. of one-day old larvae, 3.4 mm TL. Dinkins & Shute LIFE HISTORIES OF NOTURUS BAILEYI AND N. FLAVIPINNIS 59

A A

B

Figure 12: Larvae of Noturus baileyi. (A) Top and (B) Side view of six-day old larvae, 4.3 mm TL.

for N baileyi nests as part of a project to reintroduce the B species to the type locality, a single male N baileyi was found guarding a nest comprised of two distinct age Figure 11: Larvae of Noturus baileyi. (A) Top and (B) Side classes (Rakes et al. 1990). An average of approximately view of three-day old larvae, 3.5 mm TL. 89 newly laid eggs were found in three N flavipinnis nests observed in 1982 and 1983, and an average of 99 eggs or larvae were found in 13 nests observed between 1986 and ranged in diameter from 2.5 to 3.3 mm; the average 1990 (Shute et al. 1992). Because the latter group in­ number of ripe eggs per female was 177 (150-190). Inter­ cluded larval broods, the mean number of eggs originally mediate-sized oocytes averaged 114 and ranged in diam­ deposited in each nest may have actually been higher. eter from 0.8 to 1.0 mm; the smallest-sized oocytes aver­ These average clutch sizes represent only about one-half aged 116 and ranged in diameter from 0.3 to 0.8 mm. The of the average mature oocyte count determined by exam ovaries of immature female Nflavipinniscollected during the breeding season contained ova that were undifferenti­ ated in size. Since sexual maturity apparently is not reached until age two and maximum lifespan is probably three years, most individuals probably spawn only once or twice. Menzel and Raney (1973), Clark (1978), Mayden and Burr (1981), Mayden and Walsh (1984), and Walsh and Burr (1985) have presented evidence suggesting that at least some species of Noturus may be polyandrous. How­

ever, the evidence is inconclusive because of the correla­ A tion between ova produltion and female body length. Also, some data presented in the above accounts often included larval broods and not freshly fertilized eggs. No adult female N baileyi or N flavipinnis were found in nests with eggs, either with a male, or singly, but other observations indicate that both species may occasionally exhibit polyandry. On 2July 1982, a partially spent gravid female N. baileyi was collected in a shallow pool, approxi­ mately one meter from a nest containing 33 eggs guarded by a single madtom. Several eggs were protruding from B her urogenital opening. In late July 1989 while searching Figure 13: Larvae of Noturus baileyi. (A) Top and (B) Side view of 12-day old larvae, 10.5 mm TL. 60 BULLETIN 18 December 1, 1996

B A B A Figure 16: Embryo of Noturusflavipimlis. (A) Top and (B) Figure 14: Embryo of Noturusflavipinnis. (A) Top and (B) Side view of prehatchling, 12 days post-fertilization. Side view of prehatchling, six-days post-fertilization. ining the ovaries of gravid females. In the laboratory, the number of N. baileyi and N. flavipinnis hatchlings is often only half that of freshly laid eggs (P. L. Rakes pers. comm.).

Larval Development NOTURUS BAlLEYl-On 2 July 1982, a nest contammg 33 N. baileyi eggs was removed from Citico Creek and taken to the lab for observation. The eggs were well developed, sulphur-yellow in color, and had cohesive properties so that they remained together in a clump but would not stick to other objects. Embryos could be seen rotating inside their chorion. Egg diameters ranged from 2.6 to 3.0 mm (X = 2.9), yolk diameters ranged from 0.08 to 0.10 B

Figure 17: Larvae of Noturus flavipillnis. (A) Top and (B) Side view of eight-day old larvae 12.0 mm TL.

A

Figure 15: Embryo of Noturusflavipinnis. (A) Top and (B)

Side view of prehatchling, nine-days post fertilization. A mm (X = 0.09, N = 4). The eggs were placed in a shallow petri dish containing well aerated stream water and kept at room temperature (ca. 20° C) . At the time the nest was discovered, four eggs were preserved in buffered forma­ lin for measurement, and one of these was in the process of hatching. Pre-hatchlings had rudimentary barbels, pec­ toral fins, and dorsal fin. The eyes were darkly pigmented (Figure 10). The remainder of the clutch hatched within Figure 18: Larvae of Notunls flavipi1ll1is. (A) Top and (B) the next 12 hours. Hatching success of the 33 eggs in this Side view of 24-day old larvae 17.0 mm TL. nest was 94%. N. bailf:)l i hatchlings break through the chorion tail first as noted by Mayden and Burr (1981) in N. exilis, Hatchlings had heavily pigmented eyes and scattered were present and the nares were discernible. The urogeni­ melanophores on top of the head (Figure 10). All fins tal duct and anus were also evident. Hatchlings exhibitecl were formed but only the caudal and anal fins showed any tight schooling behavior and oriented themselves toward ray differentiation. At this stage, ray primordia were evi­ the airstone. dent in the dorsal fin. Four pairs of rudimentary barbels There was increased pigmen tation on the dorsal sur- Dinkins & Shute LIFE HISTORIES OF NOTURUS BAILEYI AND N. FLAVIPINNIS 61

Table 6. Stomach Contents of 13 Smoky Madtoms from Citico Creek

Percent Percent of Frequency of Prey Item 2 3 4 5 6 7 8 9 10 11 12 13 Occurrence Occurrence

Gravel 2 3 2 3 10.7 38.5

Ephemeroptera Baetidae Pseudocloen sp. 4 2 1 6.8 23.1 Unid. Baetidae 1.9 15.4 Heptageniidae Stenonema sp. 1.0 7.7 Ritll1'ogel1a sp. 1.0 7.7 Unid. Heptageniidae 2.9 23.1 Oligoneuridae 63.1 Ison)lchia sp. 2.9 23.1 Siphlonuridae Ameletus sp. 6 6.8 15.4 Ephemerellidae Drunella sp. 2 1.9 7.7 Apliemerella sp. 8 7 20 34.0 23.1 Unid. Ephemeroptera 2 3.9 23.1

Plecoptera Perlidae 1.0 7.7

Trichoptera Hydropsychidae Hydroj)syclze sp. .0 7.7 Philopotamidae 3.9 Chimarm sp. 2 ) 2.9 15.4

Diptera Tipulidae 4 3.9 J 7.7 Chironomidae 2 7 3 2 3 1 _ 17.5 21.4 46.2

Total Number of Prey Items' 2 3 26 o 6 18 22 6 9 o

'Excluding gravel. face of the three-day-old larvae and melanophores ex­ full complement of caudal fin rays. Twelve-day-old larvae tended down to the anal region and dorsolaterally behind were uniformly pigmented on the dorsal region (Figure the dorsal fin (Figure 11) . Melanophores extended onto 13). Young-of-the-year less than 30 mm SL collected in the yolk sac adjacent to the embryo. At this stage, the Citico Creek during the months of June and July also dorsal fin had its adult complement of rays while the lacked this saddled pigmentation. Caudal fin-ray counts pelvic fins did not yet show any ray differentiation. did not appreciably change in larval N. baileyi in the stages The six-day-old larvae had developing rays in all fins, from day 12 to 26 (33 - 38, N = 12). Adults have 42 to 49 including the pelvics (Figure 12). By this stage, spines total caudal rays (Bauer et al. 1983); presumably adult were developing on the pectoral and dorsal fins. Pigmen­ pigmentation and full caudal development do not appear tation was more dense on the dorsum with melanophores until well after the young leave the nest. extending posteriad almost to the origin of the caudal fin. Twelve-day-old N. baileyi larvae began feeding on The yolk sac was nearly absorbed. thawed brine shrimp. Over the next 13 days periodic The twelve-day-old larvae essentially resembled adults mortalities occurred in the clutch of developing young. except they lacked the dark blotches or saddles on the Aeromonas hyd1'Ophila was cultured from the broth but not dorsum, typical of adult N. baileyi, and all other members directly from the tissue of a moribund larva frozen and of the subgenus Rabida (Taylor 1969), and they lacked the sent to Dr. John New of the College of Veterinary Medi- 62 BULLETIN 18 December 1, 1996

cine at the University of Tennessee. Pseudomonas putida, Diet Corynebacterium sp. and Lactobacillus sp. were cultured from NOTURUS BAILEYI-Like other madtoms, N. baile;,i is prima­ the tissue. These four bacteria were considered either rily insectivorous. A total of 15 taxa of invertebrates were insignificant or contaminants at the level present, and Dr. found in the stomachs ofl3 specimens (Table 6); aquatic New was unable to identify the exact cause of death. insect larvae accounted for the majority of food items (89.3%). Ephemeropteran nymphs were the most com­ NOTURUS FLAVIPINNIs-On 29 May 1982, a newly fertilized mon aquatic insect (70.7%), followed by dipterans, tri­ clutch containing 89 N. jlavipinnis was collected for ex­ chopterans, and plecopterans, which accounted for amination. The eggs averaged 3.4 mm in diameter and 23.9%,4.4%, and 1.0%, respectively. Gravel was found in had opaque or orange yolks and cohesive properties simi­ five of the stomachs and was probably ingested acciden­ lar to N. baileyi. One of the more immature of the embryos tally while feeding on benthic insect larvae. was preserved six days later. This embryo had rudimen­ A significant amount of daytime feeding appears to tary maxillary barbels and pectoral fins, dorsal and anal occur in N. baile;'i as the stomachs of six of the seven fin folds were present, and the eyes were lightly pig­ individuals collected during the daytime contained mented. At least 27 myomeres were apparent (Figure 14). freshly eaten organisms. The stomach of the seventh indi­ At nine days post-fertilization, the embryos showed vidual, a nest-guarding male, contained only the remains more pronounced eye pigmentation and maxillary and of a single mayfly nymph (Isonychia). Slabrocks used as mandibullary barbel development; on most individuals protective cover by N. baileyi may also serve as a daytime 29 myomeres were observable (Figure 15). At 12 days, feeding site; aquatic insects were devoid from the mouth development and rudimentary pectoral fins were undersurface of 98% (N = 123) of the slabrocks under eviden t (Figure 16). The maxillary barbel had increased which N. baile;'i were found. in length and the urogenital duct and anus were also Mayden and Walsh (1984) found that the closely re­ evident. There was no pigmentation and all fins lacked ray lated N. hildebrandi is also insectivorous, with chironomid, primordia. trichopteran, plecopteran and ephemeropteran larvae as On 2July 1982 four nests were found containing 20-60 the predominant food organisms. They noted a propor­ N. jlavipinnis larvae of various ages. A single larva measur­ tional deCl"ease in plecopteran nymphs and an increase in ing 8.1 mm SL (12.0 mm TL) was captured and preserved ephemeropteran nymphs in the stomach contents during from one of these nests (Figure 17). Five rudimentary autumn and early winter, suggesting a possible shift in rays were evident in the dorsal fin, ten in the anal fin, and habitat during the colder months. D. S. Wilkins (pers. 20 in the caudal fin; and 32 myomeres were discernible. comm.) reported that N. hildebrandi fed nocturnally. The eyes were darkly pigmented and melanophores were scattered over the top of the head and dorsolaterally on to NOTURUS FLAVIPINNIS-A total of 32 taxa, representing six­ the dorsal surface of the yolk sac. There was no evidence teen families of organisms were identified from the guts of of rudimentary pelvic or pectoral rays. The age of this 12 N. jlavipinnis (Table 7). The diet of N. jlavijJinnis, like individual was estimated to be at least eight days post­ N. baile;,i, is almost exclusively insect larvae. One incli­ hatching. vidual contained the remains of a crayfish. Dipterans One individual metalarva (Figure 18) measuring 12.3 were found in all 12 stomachs, and in five of these, the mm SL (17.0 mm TL) was collected from one of the other order was represented by the family Chironomidae. nests. The age of this individual was estimated to be at Ephemeroptera (mainly Ephemerellidae), Coleoptera, least 24 days. All fin rays and spines were completely and Trichoptera represented 33.8, 6.0 and 5.5% of the developed; eight dorsal, eight pelvic, 15 anal and 34 cau­ prey items, respectively. dal rays were evident. Although not yet possessing the The biomass of dipteran larvae was small in compari­ distinctive color pattern of adult N. jlavijJinnis, this indi­ son with most of the other prey items, but they may be a vidual was more completely pigmented than the younger preferred item given their numerical dominance and individual collected on the same day; melanophores were presence in all of the guts examined. Dipterans were concentrated predorsally and lightly scattered over the numerically the most dominant prey item in N. jlavijJil111is remainder of the dorsum and sides. from Copper Creek, although they wel-e found in only Burr and Mayden (l982b) described 24-day-old 48% of the guts examined. Mayflies were found in 57% N. miunts larvae with melanophores distributed in a pat­ of the Copper Creek specimens (R. E. Jenkins, pel's. tern over the body in a pattern similar to that of an adult. comm.). Burr and Dimmick (1981) described N. elegans larvae In N. jlavijJinnis from Copper Creek, water pennies (14.7 mm TL, age unknown) as having body form, fin ray, (PsejJ/zel1'lls) comprised 21 % of the total number of organ­ and spine shape similar to adults. Although pigmentation isms consumed, and were found in 23% of the guts exam­ was well advanced, the barred pigment pattern typical of ined. Two individuals had consumed a total of 33 water the adult apparently develops later. pennies, possibly indicating a preference for this prey Dinkins & Shute LIFE HISTORIES OF NOTURUS BAILEYI AND N. FLAVIPINNIS 63

Table 7. Stomach Contents of 12 YeUowfin Madtoms from Citico Creek

Percent of Percent of Frequency Prey Item 1 2 3 4 5 6 7 8 9 10 11 12 13 Occurrence Occurrence

Sand P P P P 0.8 33.3 Coleoptera Pse/I/zen'lts 5jl. 3 1.3 6.0 33.3 Unid. Coleoptera 12 7 3 4.7 25.0 Hemiptera 0.2 8.3 Ephemeroptera Baetidae CentTOptiliw/! sjJ. 2 0.8 25.0 B(/pti~ sll. 7 II II 8 7.9 33.3 Unid. Baetidae 7 2 2.3 33.3 Caenidae Caenis sll. 3 1.1 25.0 Ephemerellidae Drunella 5/1. 2 3 1.1 16.7 Seratdla 511. 3 0.6 8.3 Eurylo/lhella 511. 9 1.9 33.8 8.3 Attendla 5/1 . 4 0.8 8.3 DnnllPlla sp. 3 0.6 8.3 Unid. Ephemerellidae 16 2 4.0 25.0 Heptageniidae S/pnOllemfl 511. 2 16 4.2 33.3 Unid. Heptageniidae 0.6 25.0 Tricorythidae 2 0.4 8.3 Unid. Ephemeroptera 3 2 6 6 13 3 7.5 66.7 Plecoptera Perlidae Pl'rlpsia 5jl. 2 5 1.7 25.0 Chloroperlidae Alto/1I!rin sjl. 0.2 4.0 8.3 Unid. Plecoptenl 8 2.1 25.0 Odonata Corduliidae NWl"Ororriulia 5/1. 3 0.6 1.2 8.3 Unid. Odonata 2 0.6 16.7 Trichoptera Hydropsychidae Hydm/Is),che 5j1. 2 2 I.I 25.0 LepLOceridae 0.4 16.7 Polycentropodidae Pol)'rt'ntm/ms S/I. 0.4 5.5 16.7 Rhyacophilidae R)'ncujlhiln .Ijl. 0.2 8.3 Unid. Trichoptera 7 3 3.4 66.7 Diptera Ceratopogonidae 13 2 I 2 3.8 33.3 Chironomidae 54 47 9 2 15 36 6 92 38.2 75.0 Dytiscidae I 0.2 51.1 8.3 Unid. Diptera 6 33 2 8.9 33.3 Crayfish 0.2 8.3

Total Number of Prey Items' 19 II 8 99 102 48 3 17 62 33 58 11

'Excluding sand. 64 BULLETIN 18 December 1, 1996

Table 8. Comparison of the Ecology of Noturus baileyi and N. flavipinnis in Citico Creek

Characteristic N. baileyi N. flavipinnis

Principal non breeding habitat Riffle crests/pools Pools Age at sexual maturity (years) 1 2 Size at sexual maturity (mm SL) 38 (

*Individual reared in laboratory. type. In Citico Creek, our observations indicate that water their feeding activities. Based on the body ofliterature for pennies are more common in riffles than pools. The the genus, activity patterns vary between species. Noturus smooth, flattened exoskeleton of these coleopteran lar­ exilis, N gyrinus, and the undescribed broadtailed vae allows them to slide over rough rocks, even in swift madtom are known to be crepuscularly active species currents, without breaking contact between the rock and (Reynolds et al. 1982; Lindquist et al. 1982; Mayden and the edge of the carapace. This would appear to be an Burr 1981). Noturus eleutherus apparen tly consumes most effective predator avoidance mechanism and Murvosh of its food within four hours of sunset (Starnes and (1971) reported little evidence offish predation. Starnes 1985). Andrews (1963) reported that although Bowman (1932) also found N insignis feeding on water some N miurus collected during the daytime contained pennies and suggested that they were preferred over food in their guts, the amount was small and usually well other abundant and easier to capture prey types. These digested. Peak feeding for this species was estimated to data suggest selective feeding, although several authors have taken place between 11 :00 p.m. to 3:00 a.m. A similar (Starnes and Starnes 1985; Miller 1984; Walsh and Burr feeding pattern was observed in Noturus insignis (Bowman 1985; Mayden and Walsh 1984; Gutowski and Stauffer 1932). Curd (1960) found that although 75% of stomachs 1990) have suggested that mad toms take prey items de­ from N exilis collected during the daytime (midday) con­ pending upon their availability. tained food, most of the contents were unidentifiable, In this study, 10 of the 12 specimens examined for gut and the majority of the feeding had probably occurred at contents were collected at night. One of these was a male night. Clark (1978) reported diurnal feeding activity for guardian removed from his nest. With the exception of N leptacanthus. In contrast to our findings in Citico Creek, this male guardian, the stomachs of all individuals taken Jenkins (1975) reported Nflavipinnis in Copper Creek at night contained easily identifiable and presumably, commonly feed during the daytime as food organisms freshly eaten organisms. Two individuals captured during drift beneath their cover. the daytime had completely empty guts. There was mate­ Nocturnal drift of many insect larvae is a phenomenon rial in the hindguts of both, although it was well digested commonly observed in lotic ecosystems (Hynes 1970), and mostly unidentifiable. Starnes and Starnes (1985) and it has been suggested that these vulnerable prey are reported that complete digestion and voidance time for important food sources for madtoms (Starnes and ingested prey of N eleutherus is approximately 24 hours. Starnes 1985). However, Hynes (1970) also indicated that Assuming a similar digestion cycle, the two individuals pools do not produce as much drift as riffle areas. There­ described above had probably been feeding during the fore, pool-dwelling species such as N flavipinnis may be middle of the night. less dependent on this phenomenon for prey items. It is Taylor (1969) suggested that madtoms are nocturnal in apparent that N flavipinnis takes some of its prey from the Dinkins & Shute LIFE HISTORIES OF NOTURUS BAIlEYI AND N. FLAVIPINNIS 65

substrate as indicated by the presence of crayfish, dragon­ have been overlooked by collectors relying solely on stan­ fly nymphs and sand grains in some of the Citico Creek darel methodologies? It is en tirely likely that N. jlavipinnis specimens. In four of the 21 N. jlavijJinnis specimens from would never have been discovered in Citico Creek had Copper Creek, sand grains were found among the gut nocturnal underwater observations not been employed. contents. Obviously, direct observations will not be successful in streams that are not normally clear. Moreover, the diver Parasitism must be competent in identifying live specimens, which There are numerous reports of parasitism in ictalurids often look very different from preserved specimens, and and specifically for madtoms (Hoffman 1967; Burr and be able to visually compensate for magnification caused Mayden 1982a,b; Mayden and Burr 1981; Mayden et al. by refraction. The latter is especially important if size is a 1980; Bowman 1932; Bowen 1980). During this study, no characteristic used in species identification. Accurate internal parasites of N. baileyi or N. jlavipinnis were ob­ estimation of population size for schooling species, or for served. On 1 September 1982, three adult N. jlavijJinnis benthic species which bury beneath the substrate, may were captured, each with eroded dorsal fin rays. This fin also be difficult. erosion was later determined to have been caused by Table 8 summarizes the known life history information t:pistylis, a stalked ciliate. Hoffman (1967) reported that in for both madtom species in Citico Creek. Some of these a personal communication, Tom Wellborn had noted an characteristics, such as low fecundity, short lifespan, spe­ infestation of catfish eggs by this genus of protozoan. cialized reproductive behavior, specific breeding and However, Hoffman did not list E.pistylis as a parasi te for any non-breeding habitat requirements, have almost certainly particular ictalurids. E.pistylis has a free swimming stage added to the vulnerability of both species, and possibly to and a colonial stage which uses host fish as attachment extirpation of historical populations. sites. As such, Rogers (1971) did not consider it as an In comparison to most other cool to warmwater obligate parasite on fishes, and he suggested that an streams in the southeastern United States, the Citico outbreak of fish parasitism by this protozoan may be due Creek watershed is relatively pristine. Fortunately, the to organic enrichment. In Citico Creek, reduced flow watershed is closely monitored by the U.S. Forest Service; during the extremely dry summer of 1983 may have pro­ unfortunately, the Citico Creek populations of both duced such conditions in some of the more stagnant madtom species are small and localized. The fact that pools. N. baile),i was extirpated from Abrams Creek, while many other fish species recolonized following the 1957 reclama­ Summary and Conclusions tion project documents this species' susceptibility to a Because of the legal and ethical limitations of working single catastrophic event. Likewise, the relatively rapid with federally protected species, much of the important decline of the once more wide-ranging and relatively life history information presented herein for both species common N. jlavijJinnis is indicative of its vulnerability. was obtained by direct observations. Understandably, ev­ Not'lll'llsjlavijJinnisonce occupied 78 stream km in Copper ery effort was made to limit the number of individuals Creek and in the late 1960's and early 1970's individuals sacrificed in the interest of scientific knowledge. We be­ could be easily seined. However, in recent years, standard lieve that one of the best ichthyological applications of fish surveys (seining and electroshocking) in Copper this approach lies with the census or collection of rare Creek have produced no specimens. In the summer/fall benthic, non-schooling species; for observations of micro­ of 1993, an intensive snorkeling effort involving 52.3 habitat use and certain behavioral aspects, few other tech­ hours of underwater observations at several locations niques can compare. throughout the 78 km reach where N. jlavijJinnis was his­ Northcote and Wilkie (1963) and Goldstein (1978) torically known revealed just two specimens in a single reported comparable results between direct underwater pool. Thus, while the species is still present in Copper observation and seining, and they found numerous ad­ Creek, the population is apparently much smaller and vantages associated with the former. To name a few: more localized than it was. The only other extant popula­ amount of time and level of effort is often reduced, tion of N. jlavijJinnis occurs in the Powell River. Since numbers of species observed is often greater, estimates of 1968, only three specimens have been taken, at two locali­ relative abundances are probably more representative of ties, despite frequent and intensive sampling efforts. the actual community, and a greater variety of habitats can No museum specimens of N. jlavijJinnis exist from the be more easily surveyed. Pflieger (1978) reported that 1957 poisoning of Abrams Creek although the brindled direct observations via skin-diving was more efficient than madtom, Schilbeodes (=Notunls) 17liunts, the name given by seining for the federally threatened Nianguae darter (fish earlier collectors to N. jlavijJinnis, was reported as being observed/ effort), and collectors using only a seine net collected during the reclamation of lower Abrams were twice as likely to miss this species at a locality. How Creek (Lennon and Parker 1959). We speculate that many populations of N. jlavijJinnis or even N. baileyi may N. jlavijJinnis probably occurred in the middle and lower 66 BULLETIN 18 December 1,1996 reaches of Abrams Creek given the similarities between J. R. Shute and Patrick Rakes of Conservation Fisheries, Abrams Creek and Citico Creek in a host of habitat vari­ Inc. provided data from field surveys in Citico Creek since ables (e.g., stream size, topographical position, gradient, the time of the original theses work; they also provided and substrate composition). data from captive propagation of both madtom species. Since 1986, Conservation Fisheries, Inc, a non-profit We thank the Tennessee Wildlife Resources Agency organization located in Knoxville, with assistance and and the U. S. Fish and Wildlife Service for providing funding from several federal and state agencies, has permits which allowed us to collect the data for our theses. propagated, reared, and released into Abrams Creek a We also thank the U. S. Forest Service, in particular Jim total of 821 N baileyi and 404 N jlavipinnis (Shute et al. Herrig, for supporting this research in the Cherokee 1992; Shute et al. 1993; Shute et al. 1994; and Rakes et al. National Forest. This research was partially supported by 1995). In June 1990 and June 1991, two N baileyi were funding provided by the U. S. Fish and Wildlife Service observed guarding potential nest rocks at the transplant and the U. S. Forest Service. site in Abrams Creek; at this same location in 1994, four N baileyi and one N jlavipinnis were found. Fortunately for both species, the attitudes of southeast­ Literature ern resource managers have changed greatly since 1957, Andrews, R. D., III. 1963. Distribution, habitat, and life history when Lennon and Parker (1959, p. 1) reported on the of madtoms, Noturus, in Illinois. Unpublished M.S. Thesis, results of the Abrams Creek reclamation project, and University of Illinois. 41 pp. wrote ... "the many successes which have been achieved in Anonymous. 1972. North Fork Citico Creek watershed study. Project summary report 1960 to 1970 and report for water warm water and cold water lakes have prompted fishery years 1970 and 1971. Unpublished report TVA, U.S. Forest biologists to consider the reclamation of streams with Service, Tennessee Game and Fish Comm. 34 pp. toxicants. " Atema, j., j. H. Todd and j. E. Bardach. 1969. Olfaction and behavioral sophistication in fish. pp. 241-251. In: Olfaction Acknowledgements and taste: proceedings ofthe third international symposium. We are indebted to David Etnier for his valuable guid­ C. Pfaffman (Ed.). Rockefeller Univ. Press, New York. ance and support during the course of research and for Baker,j. A. and D. C. Heins. 1994. Reproductive life history of his advice and comments on the manuscript. Ralph the North American catfish Noturus hildebrandi (Bailey and Dimmick, Mary Ann Handel, Richard Strange, and Larry Taylor 1950), with a review of data for that genus. Ecology of Wilson also advised us during the research and provided Freshwater Fish. 3:167-175. constructive comments on the portions of this manuscript Balon, E. K. 1975. Reproductive guilds of fishes: a proposal and definition. journal of the Fisheries Research Board of that were submitted as master's theses. Barbara Dinkins, Canada 32:821-864. Steve Layman and Charles Nicholson reviewed an earlier Bauer, B. H. and G. C. Clemmer. 1983. A status report on the version of this manuscript and provided numerous sug­ frecklebelly madtom, Noturus munitus Suttkus and Taylor, a gestions and comments on ways to improve it. Steve Walsh catfish of the eastern gulf shore. Federal Aid Project-En­ and an anonymous reviewer provided helpful comments dangered Species. Unpublished Report. 20 pp. on the final version. Thomas Fields drew the original map Bauer, B. H., G. R. Dinkins and D. A. Etnier. 1983. Discovery of used for Figure 3. Thomas Hill identified the protozoan Noturus baileyi and N. jlavijJinnis in Citico Creek, Little Ten­ parasite. nessee River system. Copeia 1983:558-560. Numerous individuals assisted with field work: Bruce Blumer, L. S. 1985. Reproductive natural history of the brown Bauer, John Baxter, Hal Boles, Richard Biggins, William bullhead !ctalurus nebulosus in Michigan. The American Mid­ Chipley, Charles Dinkins, David Etnier, Michael Etnier, land Naturalist 114(2) :318-330. Bowen, C. A.,jr. 1980. The life history of the brindled madtom Richard Eager, Steve Fraley, Byron Freeman, Larry Noturus miurus Oordan) in Salt Creek, Hocking and Vinton Greenberg, Andrew Haines, Kelly Harpster, Michael counties, Ohio. Unpublished M.S. Thesis, Ohio Sate Univer­ Humphries, Greg Kauffman, Danny Lee, Steve Layman, sity. 195 pp. Richard LeDuc, Elizabeth McCullough, Jess McFarland, Bowman, H. B. 1932. A description and ecologic study of the James Negus, Wendell Pennington, Patrick Rakes, margined madtom, Rabida insignis (Richardson). Unpub­ Michael Ryon, J. R. Shute, Christopher Skelton, and lished M.S. Thesis, Cornell Univ. 40 pp. Bernie Swiney. Bowman, H. B. 1936. Further notes on the margined madtom, RobertJenkins, Noel Burkhead, Lynn Starnes, Charles Rabida insignis (Richardson), and notes on a kindred species, Saylor, Andrew Haines, and Joe Feeman provided infor­ Noturus jlavus (Rafinesque). Unpublished Ph.D. Disserta­ mation on collections of N jlavijJinnis and conditions in tion, Cornell Univ. localities other than Citico Creek. Robert Jenkins also Breder, C. M.,jr. 1935. The reproductive habits of the common catfish, Ameiurus nebulosus (LeSueur), with a discussion of provided specimens from Copper Creek. James Grady their significance in ontogeny and phylogeny. Zoologic a provided guts and gonads of two mad toms used for his 9:143-185. electrophoretic survey of the genus. Breder, C. M.,jr. and D. E. Rosen. 1966. Modes of Reproduction Dinkins & Shute LIFE HISTORIES OF NOTURUS BAILEYI AND N. FLAVIPINNIS 67

in Fishes. Natural History Press, Garden City, New Jersey. 941 Etnier, D. A. and R. E. Jenkins. 1980. Notunts stanauli, a new pp. madtom catfish (Ictaluridae) from the Clinch and Duck Burkhead, N. M. and R. E.Jenkins. 1991. Fishes. pp. 321-409. River, Tennessee. Bulletin Alabama Museum Natural History In: Terwilliger, K. (Coordinator). Virginia's Endangered 5:17-22. Species: Proceedings ofa symposium. McDonald and Wood­ Etnier, D. A. and W. C. Starnes. 1991. An analysis of Tennessee's ward Publishers Co., Blacksburg. 673 pp. jeopardized fish taxa. Journal of the Tennessee Academy of Burr, B. M. and W. W. Dimmick. 1981. Nests, eggs and larvae of Science 66:129-133. the elegant madtom Notunts elegans from Barren River drain­ Fuselier, L. and D. Edds. 1994. Seasonal variation in habitat use age Kentucky. Transacations of the Kentucky Academy of by the Neosho madtom (Teleostei: Ictaluridae: Notunts Science 42:116-118. placidus). The Southwestern Naturalist 39:217-223. Burr, B. M. and R. L. Mayden. 1982a. Life history of the freckled Gilbert, C. R. 1953. Age and growth of the yellow stone catfish, madtom, Noturus nocturntts, in Mill Creek, Illinois (Pisces: Noturus jlavlts (Rafinesque). Unpublished M.S. Thesis, Ohio Ictaluridae). Occasional Papers Museum Natural History State University. University Kansas 98:1-15. Grady, J. M. and W. H. LeGrande. 1992. Philogenetic relation­ Burr, B. M. and R. L. Mayden. 1982b. Life history of the ships, modes of speciation, and historical biogeography of brindled madtom Noturus miunts in Mill Creek, Illinois (Pi­ the madtom catfishes, genus Notunts Rafinesque sces: Ictaluridae). The American Midland Naturalist 107:25- (Siluriformes:lctaluridae). pp. 747-777. In: R. L. Mayden 41. (Ed.). Systematics, Historical Ecology, and North American Burr, B. M. and R. L. Mayden. 1984. Reproductive biology of the Freshwater Fishes. Stanford University Press, Stanford, Cali­ checkered madtom (Noturus jlavater) with observations on fornia. 969 pp. nesting in the Ozark (N. albater) and slender (N. exilis) Goldstein, R. M. 1978. Quantitative comparison of seining and madtoms (Siluriformes: Ictaluridae). The American Mid­ underwater observation for stream fishery surveys. Progres­ land Naturalist 112:408-414. sive Fish Culturist 40(3): 108-111. Burr, B. M., B. R. Kuhajda, W. W. Dimmick and J. M. Grady. Gutowski, M. J. and J. R. Stauffer, Jr. 1990. Feeding ecology of 1989. Distribution, biology and conservation status of the the margined madtom Notunts insignis (Richardson) Carolina madtom Noturus Juriosus, an endemic North Caro­ (Teleostei: Ictaluridae). Abstract 70th annual meeting ASIH lina catfish. Brimleyana 112(2):408-414. (p.95). Burr, B. M., C. A. Taylor and K. M. Cook. 1993. Status survey of Hall, W. G., B. W.Jackson, and T. R. Love. 1981. Soil survey of the coppercheek darter (Etheostoma aquali) , striated darter Monroe County, Tennessee. U.S. Department of Agriculture, (E. striatulum) and saddled madtom (Notunts sp. cf elegans) in Soil Conservation Service and U.S. Forest Service. 107 pp. the Duck River drainage, Tennessee. Un pub!, Final Report Hoffman, G. L. 1967. Parasites of North American Freshwater to the Tennessee Wildlife Resources Agency. March 1993. 77 Fishes. University of California Press. 486 pp. pp. Hynes, H. B. N. 1970. The Ecology of Running Waters. Univer­ Carlson, D. R. 1966. Age and growth of the stonecat, Notunts sity of Toronto Press. 555 pp. jlavus Rafinesque, in the Vermillion River. Proceedings of Jenkins, R. E. 1975. Status of the yellowfin madtom, Notunts the South Dakota Academy of Science 45:131-137. jlavipinnis. Unpublished report to U.S. Office of Endan­ Case, B. E. 1970. An ecological study of the tadpole madtom, gered Species. International Activities, Washington, D.C. 11 Notunts gyrinus (Mitchell), with special reference to move­ pp. ments and population fluctuations. Unpublished M.S. The­ Jenkins, R. E. 1978. Notunts jlavipinnis Taylor, yellowfin sis, University of Manitoba. 90 pp. madtom. p. 454. In: Atlas of North American Freshwater Clark, K. E. 1978. Ecology and life history of the speckled Fishes. D. S. Lee, C. R. Gilbert, C. H. Hocutt, R. E.Jenkins, D. madtom, NotUTltS lejJtacanthus (Ictaluridae). Unpublished E. McAllister, and J. R. Stauffer, Jr. (Eds.) North Carolina M.S. Thesis, University of Southern Mississippi. 133 pp. State Museum of Natural History, Raleigh, North Carolina. Clugston, J. P. and E. L. Cooper. 1960. Growth of the common Lennon, R. S., and P. S. Parker. 1959. The reclamation ofIndian eastern madtom, NotUTUS insignis in central Pennsylvania. and Abrams Creeks, Great Smoky Mountains National Park. Copeia 1960:9-16. U. S. Fish and Wildlife Service Special Scientific Report 306. Curd, M. R. 1960. On the food and feeding habits of the catfish 22 pp. Srhilbeodrs exilis (Nelson) in Oklahoma. Proceedings of the Lindquist, D. G., P. W. Shute and J. R. Shute. 1982. Spawning Oklahoma Academy of Science 40:26-29. and nest site selection by the broadtail and tadpole madtom Dinkins, G. R. 1982. Status survey of the smoky madtom catfishes: utilization of experimental spawning cover in Lake (NottlTltS bailey I) . Unpublished final report to the U.S. Fish Waccamaw, North Carolina. Abstract of paper presented at and Wildlife Service. 33 pp. American Society of Ichthyologists and Herpetologists An­ Dinkins, G. R. 1984. Aspects of the life history of the smoky nual Meeting, Northern Illinois University, Dekalb, Illinois. madtom, NotuTlts baileyi, in Citico Creek. Unpublished M.S. Luttrell, G. R., R. D. Larson, w.J. Stark, N. A. Ashbaugh, A. A. Thesis, University of Tennessee, Knoxville, Tennessee. June Eschelle and A. V. Zale. 1992. Status and distribution of the 1984.50 pp. Neosho mad tom (Noturus tJlaricius) in Oklahoma. Proceed­ Etnier, D. A. 1978. Report on the search for spotfin chub ings of the Oklahoma Academy of Science 72:5-6. (HybojJsis lIlonacha) and smoky madtom (Notunts bail!')'i) in Madding, R. S. 1971. Nutrition studies of the brindled madtom, the Little Tennessee River system, North Carolina. Unpub­ NotUTUS miunts Jordan, based on stomach content analysis. lished final report to the U.S. Forest Service. 10 pp. Unpublished M.S. Thesis, Eastern Illinois University. 22 pp. 68 BULLETIN 18 December 1, 1996

Mahon, R. 1977. Age and fecundity of the tadpole madtom, south central Arkansas. Unpublished final report to the U. S. NOtUTUS g)'rinus, on Long Point, Lake Erie. Canada Field­ Fish and Wildlife Service. 29 pp. Naturalist 91 :292-294. Rogers, W. A. 1971. Disease in fish due to the protoloan 1:.llil·lylil Mayden, R. L. and B. M. Burr. 1981. Life history of the slender (Ciliata: Peritrichia) in the southea~tern U.S. Proceedings of mad tom, NoluTllS exilis, in southern Illinois (Pisces: the Annual Conference of the Southea~tern Association of Ictaluridae). Occasional Papers of the Museum of Natural Game and Fish Commissions 25:493-496. History University of Kansas. 93:1-64. Rohde, F. C. 1978. Nolll./"liS Imull/llmi Taylor, Scioto madtom, Mayden, R. L., B. M. Burr and S. L. Dewey. 1980. Aspects of the p.471. In: Atlas of North American Freshwater Fishes. D. S. life history of the Ozark madtom, NOt1lTU:; albaler, in south­ Lee, C. R. Gilbert, C. H. Hocutt, R. E. Jenkins, D. E. eastern Missouri (Pisces: Ictaluridae). The American Mid­ McAlIister, andj. R. Stauffer,Jr. (Eds.) Nonh Carolina State land Naturalist 104:335-340. Museum of Natural History, Raleigh, North Carolina. Mayden, R. L. and S. j. Walsh. 1984. Life history of the least Sheldon, A. L. 1987. Rarity: patterns and consequences for madtom, Notunts hildebrandi, (Siluriformes: Ictaluridae), stream fishes. pp. 203-209 111: W. .J. Mathews and D. C. Heins with comparisons to related species. The American Midland (Eds.) Community and evolutionary ecology of North Naturalist 112 (2) :349-368. American stream !ishes. Univ. of Oklahoma Press, Norman, Menzel, B. W. and E. C. Raney. 1973. Hybrid madtom catfish, Oklahoma. Notunts g)',inus X Notu'I"Its miurlts from Cayuga Lake, New Shute, j. R. and P. L. Rakes. 1994. Captive propagation and York. The American Midland Naturalist 90: 165-176. population monitoring of rare southeastern fishes by Con­ Miller, G. L. 1984. Trophic ecology of the frecklebelly mad tom servation Fisheries, Inc. Third and founh quarterly report to NOtUTUS 1II11ni/1ls in the Tombigbee River, Mississippi. The the Tennessee Wildlife Resources Agency for fiscal year 1994. American Midland Nalllralist 111:8-15. 12 pp. Miller, R. R. 1972. Threatened freshwater fishes of the United Shute,j. R., P. W. Shute, and P. L. Rakes. 1993. Captive propaga­ States. Transactions of the American Fisheries Society tion and population monitoring of rare southeastern fishes 101 :239-252. by Conservation Fisheries, Inc. Unpublished /inal report to Moss, R. 1981. Life history information for the Neosho madtom, the Tennessee Wildlife Resources Agency for fi~cal year NoluTltS jlla(id1ls. Publications of the Kansas Fish and Game 1992-1993 and first quarterly report rOl- fiscal year 1993- Commission and Kansas Nongame Wildlife Improvement 1994.27 pp. Program. 33 pp. Shute, P. W. 1984. Ecology of the rare yellow/in madtom, Murvosh, C. M. 1971. Ecology of the water penny Pse/llu!IlIIS Notltr1ls jl(wi/lil1nis Taylor, in Citico Creek, Tennessee. Un­ henicki. Ecological Monographs 41 :79-96. published M.S. Thesis, University of Tennessee, Knoxville. Northcote, T. G. and D. W. Wilkie. 1963. Underwater census of 100 pp. stream fish populations. Transactions of the American Fish­ Shute, P. W., P. L. Rakes, and.J. R. Shute. 1992. Status report and eries Society 92(2):146-151. historical review of reintroduction efforts for the endan­ Pfingsten, D. G. and D. R. Edds. 1994. Reproductive traits of the gered smoky madtom (Nolllrlll b(/ilt~)'i) and the threatened Neosho madtom (Notunts jllacidus) (Pisces: Ictaluridae). yellow/in mad tom (Nolllrltsjl(/vi/linllil). Unpublished report Transactions of the Kansas Academy of Science 97:83-87. to the U. S. Forest Service, Cherokee National Forest Unit; Pflieger, W. L. 1978. Distribution, status, and life history of the U. S. Fish and Wildlife Service, Asheville Field Office; Ten­ Niangua darter, Elhl'os/o/l/a nial1f:,'"lulI!. Missouri Deparlmen t of nessee Wildlife Resources Agency; National Park Service, Conservation, Aquatic Series No. 16. 24 pp. Great Smoky Mountain National Park. Rakes, P. L., P. W. Shute, j. R. Shute, and D. A. Etnier. 1990. Simonson, T. D., and R. j. Neves. 1992. Habitat suitability and Reintroduction of smoky mad tom (NO/lints baileyi) and yel­ reproductive traits of orange/in madlOm, Nolums gilberti lowfin mad tom (N. jlavijlinnis) into Abrams Creek, Blount (Pisces:lctaluridae). The American Midland Naturalist County, Tennessee. Unpublished Report to the Tennessee 127:115-124. Wildlife Resources Agency, 1990 Progress Report (for 1989 Sneed, K. E. and H. P. Clemens. 1963. The morphology of the Work). 12 pp. testes and accessory reproductive glands of the catfishes Rakes, P. L., P. W. Shute, and.J. R. Shute. 1995. Captive propaga­ (Ictaluridae). Copeia 1963:606-611. tion and population monitoring of rare southeastern fishes Snyder, D. E. 1976. Terminologies for inte.-vals of larval fish by Conservation Fisheries, Inc. 1994 season. Unpublished development. pp. 41-165. Ill: John Boreman (Eel.). Great Final Report to the U. S. Forest Service. 26 pp. Lakes fish and larvae identification: proceedings of a work­ Reynolds, W. W., M. E. Casterlin and D. G. Lindquist. 1982. shop FWS/OBS-76/23. Thermal preference and diel activity patterns of fishes from Starnes, W. C. and D. A. Etnier. 1986. Drainage evolution and Lake Waccamaw. Brimleyana 7:55-60. fish biogeography of the Tennessee and Cumberland river Ricker, W. E. 1975. Computation and interpretation of biologi­ drainages, p. 325-~61. In Zoogeography of North American cal statistics of fish populations. Bulletin of the Fisheries freshwater fishes. C. H. Hocutt and E. O. Wiley (eds.) . .John Research Board of Canada, No. 191. 382 pp. Wiley and Sons, Nell' York, New York. Robison, H. W. and G. L. Harp. 1981 a. A stallls report on the Starnes, L. B. and W. C. Starnes. 1985. Ecolo!,'Y and life history of Ouachita madtom, Nolunu {a r/1Il l'I"i Taylor, an endemic the moulllain madtom, Nolu./"lls i'll,ltlhems (Pisces: ictalurid from Arkansas. Unpublished !inal report to the U. Ictaluridae). The American Midland Naturalist 114(2) :331- S. Fish and Wildlife Service. 32 pp. 341. Robison, H. W. and G. L. Harp. 1981 b. A status repon on the Swingle, G. D., R. A. Mi1\er, E. T. Luther, W. D. Hardeman, D. S. Caddo mad tom, NoillTliS Iflyloli, an endemic ictalurid from Fu1\erton, C. R. Sykes, R. K. Garman. 1966. Geologic lVlap of Dinkins & Shute LIFE HISTORIES OF NOTURUS BAILEYI AND N. FLAVIPlNNIS 69

Tennessee. State of Tennessee, Department of Conservation, ened wildlife and plants 50 CFR 17.11 and 17.12 . .July 15, Department of Geology. 1991. 37 pp. Taylor, W. R. 1969. A revision of the catfish gen us No/urns U. S. Fish and Wildlife Service. 1994b. Endangered and threat­ Ratinesque with an analysis of higher groups in the ened wildlife and plants; animal candidate review for listing Ictaluridae. Bulletin of the U.S. National Museum No. 282. as endangered or threatened species; proposed rule. Federal 315 pp. Register 50 (17): 5892-59028. Taylor, W. R., R. E.Jenkins and E. A. Lachner. 1971. Rediscovery Vives, S. P. 1987. Aspects of the life history of the slender and description of the ictalurid catfish, Noturns jlavij)innis. mad tom (Nolmus exilis) in northeastern Oklahoma (Pisces: Proceedings of the Biological Society of Washington 83:469- Ictaluridae). The American Midland Naturalist 117: 167- 476. Tennessee Wildlife Resources Agency. 1994. Endan­ 176. gered 01- threatened species and wildlife in need of man­ Walsh, S. J. and B. M. Burr. 1985. The biology of the stonecat, agement. Tennessee Wildlife Resources Agency Proclama­ Notu1'ltsjlavlts (Siluriformes: Ictaluridae), in central Illinois tion Number 94-16 and 94-17, Nashville, Tennessee. and Missouri streams, with comparisons to Great Lakes Thomerson,j. E. 1966. A collection of madtom catfish, No/unts populations and congeners. Ohio Journal of Science 85 (3): jttllP/Jris, from western Florida. Transactions of the Illinois 85-96. State Academy of Science 59:397-398. Warren, M. L. and B. M. Burr. 1994. Status offreshwater fishes of Todd, j. H. 1973. The chemical language of fishes. Scientific the United States: overview of an imperiled fauna. Fisheries American 224:99-108. 19:6-18. Todd,]. H.]. Atema and]. E. Bardach. 1967. Chemical commu­ \Vhiteside, L. A. and B. M. Burr. 1986. Aspects of the life history nication in social behavior of a fish, the yellow bullhead of the tadpole madtom, Nolurus g)'linus (Siluriformes: (/r(alu1'lts na/alis). Science 158:672-673. Ictaluridae), in southern Illinois. Ohio Journal of Science U. S. Fish and Wildlife Service. 1977. Threatened and endan­ 86(4):153-160. gered plants and animals. Federal Register 42:45527-45529. Williams,]. E.,]. E. Johnson, D. A. Hendrickson, S. Contreras U. S. Fish and Wildlife Service. 1984. Endangered and threat­ Balderas, ]. D. Williams, M. Navarro-Mendoza, D. E. ened wi ldlife and plants determination of endangered status McAllister, and]. E. Deacon. 1989. Fishes of North America and designation of critical habitat for the smoky madtom endangered, threatened or of special concern: 1989. Fisher­ (Nolul'1ts /Jailpyi). Federal Register 49:43065. ies 14:2-20. U. S. Fish and Wildlife Service. 1994a. Endangered and threat-

MUSEUM BULLETIN SERIES (1975-)

1. Systematics of the Percid Fishes of the Subgenus A1IIIIlocl)'pln, Genus AmmoClypln, with Descriptions of Two New Species. james D. Williams. 56 pp., illus.,june, 1975. $5.00

2. Endangered and Threatened Plants and Animals of Alabama. Herbert Boschung, Editor. 93 pp., illus., October, 1976. $7.50

3. Containing: A New Species of Semolilus (Pisces: Cyprinidae) from the Carolinas. Franklin F. Snelson,jr. and Royal D. Suttkus. Etheos/oma neoptemll!, a New Percid Fish from the Tennessee River System in Alabama and Tennessee. W. Mike Howell and Guido Dingerkus. Taxonomy, Ecology and Phylogeny of the Subgenus Depressicalllba'l1ts, with the Description of a New Species from Florida and Redescriptions of Cambarus graysoni, Cnlllbnrlls latilllallUS, and Call1barlls striatus (Decapoda: Cambaridae). Raymond William Bouchard. 60 pp., illus., February, 1978. $5.00

4. Systematics of the Percid Fishes of the Subgenus Microperca, Genus Etheostoma. Brooks M. Burr. 53 pp., illus.,july 1978. $5.00

5. Con taining: Notropis candidus, a New Cyprinid Fish from the Mobile Bay Basin, and a Review of the Nomenclatural History of Notro/Jis shlt11lnrdi (Girard). Royal D. Suttkus. Notropis stanauli, a New Madtom Catfish (Ictaluridae) from the Clinch and Duck Rivers, Tenessee. David A. Etnier and Robert E. jenkins. 23 pp., illus., May, 1980. $5.00

6. Containing: A New Species of Cyprinodontid Fish, Genus Fundulus (Z)lgonectes), from Lake Pontchartrain Tributaries in Louisiana and Mississippi. Royal D. Suttkus and Robert C. Cashner. Karyotypes in Populations of the Cyprinodon tid Fishes of the Fundulus nota/us species-complex: A Geographic Analysis. W. Mike Howell and Ann Black. An Isozymic Analysis of Several Southeastern Populations of the Cyprinodontid Fishes of the Fundulus /lota/us Species-Complex. Fred Tatum, Ronald Lindahl and Herbert Boschung. 35 pp., illus., April, 1981. $5.00

7. Plant Resources, Archaeological Plant Remains, and Prehistoric Plant-Use Patterns in the Central Tombigbee River Valley. Gloria May Caddell. 39 pp., February, 1982. $5.00

8. Containing: Description, Biology and Distribution ofthe Spotfin Chub, H),bopsis monacha, a Threatened Cyprinid Fish ofthe Tennessee River Drainage. Robert E.jenkins and Noel M. Burkhead. Life History of the Banded Pygmy Sunfish, Elassollla zonatwlljordan (Pisces: Centrarchidae) in Western Kentucky. Stephen]. Walsh and Brooks M. Burr. 52 pp., illus., August, 1984. $6.00

9. Systematics of Notro/Jis calwbae, a New Cyprinid Fish Endemic to the Cahaba River of the Mobile Basin. Richard L. Mayden and Bernard R. Kuhajda. 16 pp., illus., November, 1989. $3.50

10. Con taining: Nolro/Jis /'ajillesquei, a New Cyprinid Fish from the Yazoo River System in Mississippi. Royal D. Suttkus. Reproductive Behavior of ExoglosS1l11! species. Eugene G. Maurakis, William S. Woolcot, and Mark H. Sabaj. Sm/Jhirhyl1chus suUkusi, a New Sturgeon from the Mobile Basin of Alabama and Mississippi.james D. Williams and Glenn H . Clemmer. 31 pp., illus.,june 1991. $5.00

II. Containing: A New Species of H),drojJ!>J1che (Trichoptera: Hydropsychidae) from Alabama, with Additional State Records for the Curvipalpia. Paul K. Lago and Steven C. Harris. New Caddisflies (Trichoptera) from the Little River Drainage in Northeastern Alabama. Kenneth Frazer and Steven C. Harris. New Caddisflies, (Trichoptera) from Alabama and Florida. Steven C. Harris. Survey of the Trichoptera in the Little River Drainage of Northeastern Alabama. Kenneth S. Frazer, Steven C. Harris and G. Milton Ward. 22 pp., illus., September, 1991. $4.00

12 . Variation of the Spotted Sunfish, Le/Jomis /Jllllctatus Complex (Centrarchidae): Meristics, Morphometrics, Pigmentation and Species Limits. Melvin T. Warrenjr. 47 pp., illus. May 1992. $6.00 13. Containing: Effects of Impoundments on Freshwater Mussels (Mollusca: Bivalvia: Unionidae) in the Main Channel ofthe Black Warrior and Tombigbee Rivers in Western Alabama.james D. Williams, Samuel L. H . Fuller and Randall Grace. Elheosloma cltermacki, a New Species of Darter (Teleostei: Percielae) from the Black Warrior River Drainage of Alabama. Herbert T. Boschung, Richard L. Mayden, and Joseph R. Tomelleri. 21 pp., illus. September 1992. $5.00

14. Catalog of Freshwater anel Marine Fishes of Alabama. Herbert T. Boschung. 268 pp., December, 1992. $12.00

15. Containing: Archaeological Survey and Excavations in the Coosa River Valley, Alabama. Vernon james Knight, Editor. Including: Archaeological Research in the Middle Coosa Valley. Vernonjames Knight. Archaeological Research in the Logan Martin Basin. L. Ross Morrell. Lamar in the Middle Coosa River Drainage: The Ogletree Island Site. Richard Walling. The Milner Site: A Mid-Seventeenth Century Site Near Gadsden, Alabama. Marvin T . Smith, Vernonj. Knight,julie B. Smith, and Kenneth R. Turner. Seventeenth Century Aboriginal Settlement on the Coosa River. Marvin T . Smith. 87 pp., illus. , january, 1993. $10.00

16. Containing: Elassollla alabmnae, a New Species of Pygmy Sunfish Endemic to the Tennessee River Drainage of Alabama (Teleostei: Elassomatidae). Richard L. Mayden. A New Species of Percill(l (OdonlojJ/wlis) from Ken tucky and Tennessee with Comparisons to Percina r)'lIlatol(wnia (Teleostei: Percidae). Brooks M. Burr and Lawrence M. Page. Systematics of the Etlteoslomfl jon/ani Species Group (Teleostei: Percidae), with Descriptions of Three New Species. Robert M. Wood and Richard L. Mayden. 44 pp., illus.,june, 1993. $10.00

17. Containing: Ellteosloma (Ulocell 1m) scottie (Osteichtheyes: Percidae), a New Darter from the Etowah River System in Georgia. Bruce H. Bauer, David A Etnier and Noel M. Burkhead. Present and Recent Historic Habitat of the Alabama Sturgeon, ScajJ/tir/tYllr/tus sull/msi Williams and Clemmer, in the Mobile Basin. john Selden Burke and john S. Ramsey. Roland Harper, Alabama Botanist and Social Critic: A Biographical Sketch and Bibliography. L.J. Davenport and G. Ward Hubbs. 45 pp., illus., May, 1995. $10.00

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2. Ten Thousand Years of Alabama History, A Pictorial Resume. W. Phillip Krebs. 130 pp., illus., january, 1986. $10.00

3. The Mounds Awaken: Mound State Monument anel the Civilian Conservation Co rps. joy Baklanoff and Arthur Howington. 36 pp., illus. October, 1989. $3.00 MUSEUM PAPERS (1910-1960, TERMINATED)

1. Smith Hall, The New Museum and Home of the Geological Survey. E.A. Smith. 7 pp., 1 plate. Out of Print 2. The Museum as an Educator. Herbert H. Smith. 25 pp., 8 plates, 1912. Out of Print 3. Directions for Collecting Land Shells. Herbert H. Smith. 12 pp., 1912. Out of Print 4. Annotated List of the Avery Bird Collection. Ernest G. Holt. 142 pp., 1 plate, 1921. $3.00 5. Preliminary Catalogue of Alabama Amphibians and Reptiles. H.P. Loding. 59 pp., 1922. Out of Print 6. The Anculosae of the Alabama River Drainage. Calvin Goodrich. 57 pp., 3 plates, 1922 Out of Print 7. The Genus C),rotoma. Calvin Goodrich. 32 pp., 2 plates, 1924. Out of Print 8. The Terrestrial Shell-Bearing Mollusca of Alabama. Bryant Walker. 32 pp., illus., 1928. Out of Print 9. Footprints from the Coal Measures of Alabama. T.H. Aldrich, Sr. and Walter B. Jones. 64 pp., illus., 1930. $3.00 10. Goniobases of the Vicinity of Muscle Shoals. Calvin Goodrich. 25 pp., 1930. Out of Print 11. Alabama Reptiles. William L. Haltom. 145 pp., 39 plates, 57 figs., 1931. Out of Print 12. Description of a Few Alabama Eocene Species and Remarks on Varieties. T.H. Aldrich, Sr. 21 pp., 6 plates, 1931. $3.00 13. Moundville Culture and Burial Museum. Walter B. Jones and D.L. Dejarnette. 8 pp., 22 illus., 1936. Out of Print 14. The Argiopidae or Orb-Weaving Spiders of Alabama. Allan F. Archer. 77 pp., 5 plates, 1940. $3.00 15. Anthropological Studies at Moundville. Part I. Indian Skeletons from the Museum Burials at Moundville. Part II. Possible Evidence of Scalping at Moundville. C.E. Snow. 57 pp., illus. 1940. $3.00 16. Condylo-Diaphysial Angles of Indian Humeri from North Alabama. C.E. Snow. 38 pp., illus., 1940. $3.00 17. The Bessemer Site (Excavation of Three Mounds and Surrounding Village Areas near Bessemer, Alabama). D.L. Dejarnette and S.B. Wimberly. 122 pp., illus., 1941. $3.00 18. Supplement of the Argiopidae of Alabama. Allan F. Archer. 47 pp., 4 plates, 1941. $3.00 19. McQuorquodale Mound. A Manifestation of the Hopewellian Phase in South Alabama. S.B. Wimberly and H.A. Tourtelot. 42 pp., illus., (1941) 1943. $3.00 20. Mound State Monumen t. 19 pp., illus., 1941. Out of Print 21. Two Prehistoric Indian Dwarf Skeletons from Moundville. C.E. Snow. 90 pp., 2 plates, 1946. $3.00 22. The Theridiidae or Comb-Footed Spiders from Moundville. Allan F. Archer. 67 pp., 2 plates, 1946. $3.00 23. The Flint River Site, Mao48. William S. Webb and D.L. Dejarnette. 44 pp., illus., 1948. Out of Print 24. The Whitesburg Bridge Site, Ma'10. William S. Webb and D.L. Dejarnette. 44 pp., illus., 1948. Out of Print 25. The Perry Site, Luo25. William S. Webb and D.L. Dejarnette. 69 pp., illus., 1948. $3.00 26. Little Bear Creek Site, CTo8. William S. Webb and D.L. Dejarnette. 64 pp., illus., 1948. Out of Print 27. New Anophthalmid Beetles (Fam. Carabidae) from the Appalachian Region. J. Manson Valentine. 19 pp., 2 plates, 1948. $3.00 28. Land Snails of the Genus Stlmo/rema in the Alabama Region. Allan F. Archer. 85 pp., 10 plates, 1948. $3.00 29. Moundville: An Historic Documen t. Carl E. Guthe. 14 pp., 1950. Out of Print 30. A Study of the Theridiid and Mimetid Spiders with Descriptions of New Genera and Species. Allan F. Archer. 44 pp., 4 plates, 1950. $3.00 31. Carvernicolous Pselaphid Beetles of Alabama and Tennessee, with Observations on the Taxonomy of the Family. Orlando Park. 107 pp., illus., 1951. $3.00 32. Guntersville Basin Pottery. Marion D. Hemilich. 69 pp., ill us. 1952. $3.00 33. A Key to the Amphibians and Reptiles of Alabama. Ralph L. Chermock. 88 pp., illus., 1952. Out of Print 34. New Genera of Anophthalmid Beetles from Cumberland Caves (Carabidae, Trechini). J. Manson Valen tine. 41 pp., 5 plates, 1952. $3.00 35. New Genera and Species of Cavernicolous Diplopods from Alabama. Richard L. Hoffman. 13 pp., illus., 1956. $3.00 36. Archaeological Investigations in Mobile County and Clarke County, Southern Alabama. Steve B. Wimberly. 262 pp., 7 plates, 1960. $5.00

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