Report of the 4th DFO Atlantic Parasitologist's Workshop 29 September, 1994 Northwest Atlantic Fisheries Centre St. John's, Newfoundland

Editor J. Richard Arthur

Fish and Marine Mammals Division Department of Fisheries and Oceans Maurice Lamontagne Institute P.0 Box 1000 Mont-Joli, Quebec G5H 3Z4

Canadian Manuscript Report of Fisheries and Aquatic Science 231 6

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1995

Report of the 4th DFO Atlantic Parasitologist's Workshop 29 September 1994 Northwest Atlantic Fisheries Centre St. John's, Newfoundland

Edited by

J. Richard Arthur

Fish and Marine Marnmals Division Department of Fisheries and Oceans Maurice Lamontagne Institute P.0 Box 1000 Mont-Joli (Quebec) G5H 324 O Minister of Supply and Services Canada 1995 Cat. No. Fs 97-410000E ISSN 0706-6473

Correct citation for this publication:

Arthur, J.R. (ed.) 1995. Report of the 4th DFO Atlantic Parasitologist's Workshop, 29 September, 1994, Northwest Antantic Fisheries Centre, St. John's Newfoundland. Cm. Manuscr. Rep. Fish. Aquat. Sci. 2316: IV + 50 p. ... Ill

TABLE OF CONTENTS

ABSTRACT ...... iv

&SU& ...... iv

EXECUTIVE SUMMARY ...... 1

REPORTS OF THE REGIONS:

Gulf Region. S.E. McGladdery and M.F. Stephenson ...... 6

Newfoundland Region. J. Brattey...... 10

Quebec Region. JJI. Arthur, D J. Marcogliese and L. Measures ...... 18

Scotia-Fundy Region. G. McClelland and D.J.Martel1 ...... 36

ANNEXES:

ANNEX 1: List of participants ...... 46

ANNEX II: Schedule ...... 47

ANNEX III: Report of the meeting to update sealworm dynamics ...... 48 iv ABSTRACT

Arthur, J.R. (ed.) 1995. Report of the 4th DFO Atlantic Parasitologist 's Workshop, 29 September, 1994, Northwest Antantic Fisheries Centre, St. John's Newfoundland. Can. Manuscr. Rep. Fish. Aquat. Sci. 2316: IV + 50 p.

The 4th DFO Atlantic Parasitologist's Workshop, held on the 29 September 1994 at the Northwest Atlantic Fisheries Centre, St. John's, Nfld., brought together researchers from four of the Department's Atlantic laboratories to review work accomplished since Septem ber 1993 and to discuss and coordinate activities planned for 1995/1996. This report summarizes by region the large amount of research and related activities accomplished by DFO's Atlantic parasitologists during the past year and outlines that planned for the near future. An executive summary provides highlights of these accomplishments and outlines the research priorities for the Atlantic Canada zone and the individual regions.

Arthur, J.R. (ed.) 1995. Report of the 4th DFO Atlantic Parasitologist's Workshop, 29 September, 1994, Northwest Antantic Fisheries Centre, St. John's Newfoundland. Can. Manuscr. Rep. Fish. Aquat. Sci. 2316: IV + 50 p.

Le 4ième atelier de travail des parasitologistes du MPO de la Zone atlantique s'est tenu au Centre des Pêches de l'Atlantique du Nord-Ouest à St. John's, Terre-Neuve , le 29 septembre 1994. Des chercheurs provenant de quatre des laboratoires du Ministère. de la Zone atlantique se sont réunis pour passer en revue le travail accompli depuis septembre 1993 et pour discuter et coordonner les activités pour 1995-1996. Ce rapport résume, par région, la grande quantité de travail de recherche et les activités connexes réalisés par les parasitologistes durant la dernière année et trace les grandes lignes de la planification pour le futur immédiat. Un sommaire met en lumière ces accomplissements et donne un aperçu global des priorités de recherche du MPO pour la Zone atlantique canadienne ainsi que pour chacune des régions. EXECUTIVE SUMMARY

The fourth meeting of DFO's parasitologists from the Atlantic Regions was organized by the Parasitology CODE of the Maurice Lamontagne Institute and hosted on 29 September, 1994 by the Northwest Atlantic Fisheries Centre, St. John's, Nfld. A total of eight scientists and technical support staff representing all of DFOYsAtlantic laboratories and one international laboratory were in attendance (see Annex 1).

The goal of the meeting was to review parasitological research accomplished by the DFO in Atlantic Canada since September 1993 and that proposed for the coming year (1995-1996).

During the workshop, scientists from the various laboratories presented information related to these goals (see Reports of the Regions), and discussed possible studies for the more distant future. A special guest at this meeting was Dr. Alistair McVicar of the Marine Laboratory, Scottish Office Agriculture and Fisheries Department, Aberdeen, who presented a review of the research in progress at his laboratory.

Below we highlight some of the accomplishments of members the Parasitology Working Group and outline the research priorities for the Atlantic Canada zone and the regions.

ACCOMPLISHMENTS

During 1993194, significant progress was made towards completing DFOYsgoals for sealworm research as outlined under the former SeaVSealworm Ecology Program (SSEP). A special effort is under way to produce a new sealworm model. To this end, a separate premodelling meeting of scientists involved in SSEP and others was held at the NAFC on 28 September to review the status of research (see Annex III).

Some of the noteworthy activities undertaken in 199311994 include:

For the Gulf Region:

- Studies on a viral disease of American oyster (with IFREMER) and transplant experiments to investigate Malpeque disease of oysters

- Work towards completion of a major survey of bivalve parasites

- Work towards completion of protocols for shellfish health diagnostics (part of the Shellfish Health Regulations) and preparation of a Synopsis of Diseases of Shellfish For the Newfoundland Region:

- Assessment of the role of various seal species and of harbour porpoise in the transmission of to commercial fishes

- Studies on the genetics of nematodes of fish and seals

- Analysis of temporal and geographical variations of in

For the Quebec Region:

- Studies on the use of parasites as biological tags for Greenland halibut, redfish, and capelin

- Studies towards improving knowledge of the sealworm (aspects of the life cycle, transmission dynamics, larval development, abundance and distribution in fish and seals)

For the Scotia-Fundy Region:

- Research related to the sealworm problem (sealworm index surveys in fish, seai prey surveys, infections in seais, life cycle studies, including flatfish diet studies, effects of fish on size and morphometry, serial transmission, larvd longevity and host response in plaice)

- Use of nematodes as natural tags for fish and seal hosts

The large amount of work accomplished each year by DFO's Atlantic parasitologists indicates that parasitology is one of the most productive areas of research undertaken by the Department. During the past year, scientists at the four laboratories published or have had accepted for publications, a total of 41 scientific papers and reports, including 30 peer reviewed primary publications. Together, they also made a total of 34 presentations at scientific meetings, workshops and invited seminars.

DFO's parasitologists provide a wide variety of services to a diverse group of clients. Major research programs are designed to provide information to aid managers in more accurate stock management, and to assist the fishing industry, fish processors and the aquaculture industry in finding solutions to parasite-caused losses. DFO's specialized expertise in fisheries parasitology is tapped by a wide variety of Canadian and international clients, ranging from other DFO programs (fisheries research, aquaculture, toxicology) to international agencies (IDRC, FAO), provincial ministries, universities, museums, native organizations, hospitals, veterinarians, individual fishermen and the general public.

To assist management, an attempt has been made to prioritize research activities for both the Atlantic zone as a whole and on a regional basis, the results of which are presented below (regions where individual priorities are of key importance are given in parentheses):

Completion of a new sealworm modelling exercise. (Que, S-F, Nfldf

Much of the research outlined under the former SSEP Program has now been completed, providing substantial new data for a sealworm model. Although a predictive model is not yet possible, a modelling effort will identify key areas where additional information is needed and may be able to indicate the probable effects (positive or negative) of fluctuations in key components (e.g., seals, fish, worm abundances).

Completion of key elements of sealworm research. (Que, S-F)

Key areas of research identified in a recent review of the status of sealworm research (Marcogliese and McClelland. 1994. Cm. Manusc. Rep. Fish. Aquat. Sci. No. 2260) or identified via priority #1 (above) which must be completed to improve the sealworm model should be addressed. These include determination of:

- worm fecundity and its relation to worm density (Que, S-F)

- longevity of sealworm and host response in seal prey species (Que, S-F)

- the distribution and abundance of sealworm in macroinvertebrates (Que)

- the importance of small benthophagous fish in the life cycle (S-F)

- effects of temperature on transmission and development (Que)

- the effects of on behaviour of invertebrates and fish (Que)

Additionally, the long-term monitoring of sealworm infections in fish via the American plaice survey conducted by Scotia-Fundy should be continued. Without data provided by this survey, DFO will not be able to state with confidence whether infections are increasing or decreasing or determine success or failure of any attempt to regulate seal populations. Research related to aquaculture develoument. (Gulf, Nfld, S-Fr

Research related to the development of marine finfish and shellfish culture is needed to provide support to this growing industry. Areas of primary importance include:

-Research to support disease diagnostics and the Canadian Fish Health Protection Regulations.

-Applied research on parasites of current or potential economic importance to the mariculme industry (e.g., identities, life cycles, pathogenicities, distributions, control and prevention measures). Prograrns for the development of new species for culture should include parasitological examination of stocks from which broodstock will be collected; experimentally cultured species should be monitored for parasite-caused diseases. Sites proposed for aquaculture development should be surveyed to identify potential parasite problems.

-Research on cnistacean parasites and diseases to fil1 the lack of knowledge on normal parasite patterns for lobsters and commercially exploited crabs. This is especially important in light of their potential inclusion in the revised Fish Health Protection Regulations.

Research on the use of uarasites as biolonical indicators. (Nfld, Que, S-F)

Parasites have been successfully used to study the migrations and stock structure of commercial fish species. Using these techniques, parasitologists can contribute valuable information to interdisciplinary studies of key management problems (e.g., recruitment patterns of juvenile cd to adult populations, movements and distincmess of adult stocks).

Parasites are also of value as indicators of environmental quality, a developing field of research which has yet to be addressed by DFO. Parasitological components should be included in programs monitoring the health of aquatic systems.

Research on the effects of parasites on natural populations of aquatic . (Nfld, Que, S-F)

The role of parasites as causes of mortality in wild populations of commercially important fish and marine mammals and their long term impacts on commercial fisheries are poorly known.

Extensive data bases, collected over many years by DFO ground fish cruises, exist on infection levels of the parasitic Sphyrion lumpi in redfishes and Lernaeocera branchialis in . Analysis of these data may provide important insight into the role copepods play as causes of mortality of Atlantic cod and the factors which determine their abundance. Analysis of -sh infection data will indicate long term infection trends and areas most severely affected, information valuable to managers and the commercial fishery.

The parasites of marine marnmals and their role in the regulation of host populations are also poorly known. Basic information on the species infecting and seals, their geographic distributions and their pathogenicities is needed to determine to what extent parasites are responsible for mortalities in natural populations. A number of marine mammal parasites have the potential to infect humans, and may occasionally cause disease or death.

A number of administrative problems were also discussed during the meeting. In order to function effectively, zonai programs should be funded separately from regional initiatives. Without secure funding, intemgional parasitological research is difficult to accomplish (this is particularly obvious in the case of the Ottawa-mandated SSEP Program, where problems with timing and levels of funding and differing regional priorities have hindered completion of this project (see Marcogliese and McClelland 1994)). Decreasing financial support to parasitological research and loss of support staff have made it increasingly difficult for DFO's parasitologists to complete research programs and have serîously affected morale in some regions. 6 REPORT OF THE GULF REGION

Sharon E. McGladdery and Mary F. Stephenson

Gulf Fisheries Centre Moncton, New Brunswick

The parasitology mandate in the Gulf Region continues to include both service and research. The service portion includes diagnostic services and advice in support of DFO and provincially regulated introductions and transfers of shellfish (both molluscs and, more recently, species). Additional services include scientific advice for the preparation of the shellfish portion of the revised Fish Health Protection Regulations. Teaching and supervision of students (science, veterinary, and health technicians) and provincial biologists with interest in health monito~gof the Atlantic shellfish populations includes shellfish pathology techniques (as demonstrated, in part at the 2nd DFO Atlantic Parasitologist's Meeting), as well as taking on our first post-graduate student from the University of New Brunswick. The research component of our workplan includes the continued investigation of techniques applicable to examination of intracellular microbial parasites (e.g., viruses) as well as continued investigation of the aetiology of Malpeque disease of American oysters (Crassostrea virginica), which was started this summer. This is a multi-regional question which is being encouraged by DFO Scotia-Fundy (R. Lavoie, J. Pringle, J. Kean-Howie, and C. Momson).

The following summary provides details of work conducted since the last DFO Atlantic Parasitologist' s Workshop.

PLANNED ACTIVITIES

DFO-IFREMER Research Exchanne (vear 2 of 3)

Attempts were made to isolate and purify a viral agent of American oyster (Crassostrea virginica) gonads, using the facilities and expertise of two laboratories in France. The work was supported financially by the Going Global Program of the Deparmient of External Affairs, Canada, for promotion of European scientific and technical exchange. It was also supported by the DFO/IFREMER agreement for scientific exchange between laboratories. Purification of the virus was complicated by its fragility, however, preliminary results are promising and the investigation will be continued in Moncton in FY95196 using ce11 dissociation techniques recently developed by IFREMER. Malmue Disease - Transplant Exwrirnent (vear 2 of 3)

Transplant studies were put in place in June 1994 to replicate and extend those undertaken in the late 1950's which demonstrated susceptibility of Cape Breton oysters to the unknown aetiologic agent of Malpeque disease. Two size groups of oysrers from two salinity sources in Cape Breton are being held at two sites (one on PEI and another in NB) and are being monitored fortnightly for mortalities. So far no abnormal mortalities have been observed.

Musse1 Ciliate Identification

Results indicating the intracellular habitat and light microscope comparisons with other bivalve ciliates were presented at ISEP-10 in Halifax, in August, 1994. Preliminary observations indicate an affinity with phyllokinesomal ciliates, however, further EM studies, as well as kinetsomal silver staining is required. This work is planned for completion and publication in 95/96.

Bivalve Parasite Survev (year 3 of 3)

Over 2000 slides have been prepared for examination, including two years of material from soft- shell clams and quahaugs. These will be examined this winter and during the next fiscal year, since further collections of material, except for diagnostic submissions, will be temporarily discontinued.

Musse1 Shelf-life: Parasites, Diseases, and Handling (year 1 of 2)

A new Master's student, James Weldon, registered at UNB under Dr. M.D.B. Burt, started work this sumrner on the problems associated with short shelf-life of blue mussels (Mytilus edulis). This has ben a recurrent question of concem to some musse1 processors on PEI and in NB, and is designed to cover processing techniques, holding regimes, parasites and pathology.

Shellfish Health Protection Regulations (year 3 of 4)

Work on refining protocols for shellfish health diagnostics is near completion and a rough draft should be available for review by a technical committee before the next FY.

Manual Reprints

An additional 50 manuals have been printed, 30 of which have gone to DFO library circulation. The remainder will be used to fil1 outstanding requests. 8 Memorial University of Newfoundland - Marine Institute - Course

A two-day shellfish health orientation program was given this spring to both aquaculture students, as well as benthic fishery industry people, enrolled in career diversification prograrns at the Marine Institute-MUN, Newfoundland.

UNPLANNED ACïIVITIES

Microbial Diseases of Shellfish

A chapter. on microbial diseases of shellfish for an upcoming series on fish diseases is in preparation (eds. R.W. Stephenson and P.T.K. Woo). This complements the research concentrated on viral diseases of shellfish (DFOIIFREMER) described above (1) and is planned to be completed by October 1994.

Bay Scallo~Parasite

Louise Goggin is completing her post-doctoral fellowship at the University of Prince Edward Island-Atlantic Veterinary College and has determined that the biflagellate stage described as part of the life-cycle of Perkinsus karlssoni in bay scallops (Argopecten irraàians) is likely a contaminant &-living flagellate, Cafeteria roenbergensis. The subunit ribosomal RNA from flagellates isolated from bay scallop matenal was sequenced and matched identically that of C. roenbergensis. These observations are being prepared for publication as a scientific note. The identity of the thioglycollate-positive tissue lesions is under further EM investigation to establish its relationship to the post-culture prezoosporangial-like bodies observed in the original investigation.

Histopatholonv of Glochidial Infections

Electron and light microscopie studies of glochidial infections of parr (Salmo salar) were presented as a poster at the American Fisheries Society meeting in Halifax in August. Work on this project is currently on-hold until the pnnciple investigator, L. Nezlin from Russia, can complete other sabbatical research commitments at Dalhousie University.

Synopsis of Diseases of Shellfish

The comprehensive list of parasites and pathogens of shellfish compiled as the reference base for the Shellfish Health Protection Regulations is currently in press as a special edition of the Annual Review of Fish Diseases (Bower, McGladdery and Price). Animal Health Technician Course

The Nova Scotia Technical College has accepted a program designed at GFC for animal health technicians interested in internships (8 weeks) related to fmfish and shellfish pathology. The program is designed to give working experience which will be applicable to government, veterinary, or private laboratory diagnostic laboratories in service to the shellfish industries.

Canadian Aauaculture Institute - Course

AVC-UPEI has recently set up a Canadian Aquaculture Institute, which will be providing two- week intensive courses on fish health, including shellfish. Gulf Region Parasitology has agreed to provide a couse outline, which will be published and updated on a regular basis, as a foundation for these courses, which should start to be offered in 1995 (Chairman, Dr. Nigel Robbins, CAI-AVC).

New Brunswick Shellfish Biolopists: Sarnple Processinn and Evaluation - "Shellfish Patholo~v Guide"

Two one-week courses were given this year to NB DFA personnel working with shellfish. In an effort to enhance the consistent quality of samples submitted for health checks and diagnostics, a basic guide was prepared. Interest in this training and guide has been expressed by PEI DFFA as well as others working with shellfish aquaculture.

Other Related Duties

-1CES Working Group on Pathology and Diseases of Marine Organisms - hosted meeting at the Gulf Fisheries Centre, March, 1994

-Co-Chair of Aquatic Parasitic and Pathogenic Protists Workshop at the International Society of EvolutionaIy Protistologists #IO, Dalhousie University, Halifax, August 1994

-Co-Chair of Special Theme Session of the ICES Mariculture Committee entitled "Parasite Problems in Mariculture" at the ICES Statutory Meeting in St. Johns, Newfoundland, September, 1994 10 REPORT OF THE NEWFOUNDLAND REGION

John Brattey

Northwest Atlantic Fisheries Centre St. John's, Newfoundland

The parasitology research program of the Newfoundland Region examines the distribution, abundance, annual variability, and host-parasite relationships of parasites infecting commercially exploited stocks of shellfish, finfish, and marine mammals. The research program aims to develop a detailed understanding of host-parasite relationships, to contribute to the development of parasite-free fish products, and to provide advice on parasitological matters to the general public, the fishing industry, and to DFO senior management. Parasites are also used as biological tags to investigate stock relationships in commercial fisheries. The role of parasites as sources of mortality or reduced quality in fisheries is explored, and the public health significance of parasites is investigated. Current regional priorities are to investigate the role of pathogenic parasites in suppressing the growth of host populations, with particular emphasis on the effects of Lernaeocera branchialis on northern cod, and to complete studies on the genetics of the sealworm species complex.

PROGRAM REVIEW GOALS 1993-4

A-Base Funded Research

- Complete and submit for publication a MS assessing the role of various seal species (grey, harbour, bearded, ringed, hded) in the transmission of nematode parasites such as sealwoxm and to fish stocks off Newfoundland and Labrador. (with Stenson-PSMM).

This project has been completed, and a MS was submitted and published in the Canadian Journal of Zoology. Grey, harbour, and to a lesser extent, hooded seals are important in transmission of sealworm. Only grey seals appear to play a role in transmission of Anisakis, but their role is probably minor relative to that of cetaceans. - Examine the data base on the parasitic Sphyrion to investigate possible tempord and geographic variation in the abundance of the parasite on redfish (Sebastes sp.) and prepare a MS if findings are suitable for publication. (with Atkinson and Power-GF).

This activity was suspended due to mid-year funding cutbacks.

- Complete and submit a manuscript on the parasites of harbour porpoise, Phocoena phocoena, describing their role in the transmission of the nematode parasite Anisakis simplex to fish stocks off Newfoundland and Labrador. (with Stenson-PSMM)

The MS was completed and is submitted to the Journal of the Helminthological Society of Washington.

- Continue collaboration with Prof. L. Paggi (University of Rome, ) and CO-workerson the genetics of parasitic nematodes from fish and seals.

Further samples were collected and taken to Rome in September 1993. 1 spent three weeks at Prof. Paggi's laboratory learning further techniques and screening the material 1 collected. A third MS dealing with the genetics of the nematode Phocascaris from fish and seals in the North Atlantic is being prepared. Results indicate a complicated pattern. Harp and hded seals appear to be hosts to "incipient" species, but these are capable of hybridizing and hybrid forms appear to greatly outnumber either parental type in ringed seals. Also, species of Phocascaris in seals are genetically much more closely related to Contracaecum from seals than Contracaecum from birds are to Contracaecum from seals.

- Continue to coliect and examine for nematodes fish species that are commonly eaten by grey seals. These species are important in the life cycle of sealworm and are used to acquire data to monitor trends in sealwonn abundance.

Another sample of cod (n=250) and sculpins (n=130) was collected off the south Coast of Newfoundland during the 1994 spnng groundfish survey of 3Pn and 3Ps. These were examined for P. decipiens and Anisakis and the data coded, checked, and key-punched. This is the sixth sample collected since 1987, giving a good time series on trends in sealworm abundance.

- Finalize and submit for publication a MS describing a method for distinguishing larvae of parasitic nematodes found in fish off Newfoundland and Labrador. A MS entitled "Identification of larval Contracaecum osculatum and Phocascaris sp. (Nematda: Ascaridoidea) from marine fishes by allozyme electrophoresis and discriminant function analysis of morphometric data" was submitted to a special commemorative issue of the Canadian Journal of Fisheries and Aquatic Sciences. These are among the most common parasitic nematodes found in marine fishes off Newfoundland and Labrador. The manuscript gives information on their distribution and fish hosts, and describes a method for distinguishing the thespecies that occur in this area.

- Provide biological advice and data on parasites in commercial fish stocks to DFO Inspection Services and indusîry as required to contribute to the development of parasite-free fish products.

A contractual project entitled "Parasite Detection Benchmark Study" was completed by Canpolar East of St. John's, NF. The study involved assessment of the reliability of parasite removal by fish processors in Atlantic Canada.

B-Base Funded Research (Northem Cod Science Pro.gram)

- The January, 1993, Northem Cod Workshop recommended that the substantial database on Lernaeocera branchialis infestation of cod in the 2J3KL area be analyzed to determine if shifts have occurred over time and space. If available, information on other types of parasites might also be useful. Contract funds are to be provided by NCSP (Bishop-GF and Methven-MUN).

A contract for preliminary analysis of the database was awarded to John McClintock of SeaConsult, St. John's, NF. Preliminary analyses indicate that the prevalence of the parasite has remained quite stable over the past several decades and is similar in each NAFO region. Infection is age-dependant and appears to peak in 4-6-year-old cod. Analysis will continue and new data based on recent examination of juvenile cod, which are under-represented in this database, will be included.

Other Onrroing Research

Consultations

As in other years, there have been numerous requests from various clients to provide advice on parasitological matters. These included:

-Testimony given in Newfoundand Provincial Court on behalf of DFO conceming the identity of a parasite (Lepeophtheirus salmonis) found in a bag on board a vesse1 operated by suspected salmon poachers. The parasite was subsequently used as evidence and the individuals were convicted of several fishery violations.

-Advice given to FPI concerning the occurrence of nematodes in the fillets of Icelandic flatfish and lesions in the flesh of Barents Sea cod. Advice on the occurrence of Kudoa sp. in the flesh of Pacific flathead sole, and on the occurrence of black lesions in the flesh of snow crab from Newfoundland was also provided.

-Advice was provided to the Newfoundland Department of Fishenes staff on the occurrence of parasitic nematodes found in capelin by members of the public.

-Advice was given to Habitat Research staff on parasitological aspects of a proposed water transferlhydro-electric development.

-Advice was provided to numerous members of the public via regional enforcement and inspection service staff concerning the public health significance of various parasites found in brook trout and lake trout (e.g., Cystidicola, Triaenophorus, Eubothrium, Echinorhynchus, , Diplostomum).

-Advice was provided to the public concerning the high prevalence of hypobranchial tumours (X-ce11 lesions) observed in cod caught during the recreational fishery.

Collaborative studies

(a) Studies on the population genetics of sealworm by RFLP analysis (with W. S. Davidson, MUN Biochemistry Dept.)

Initial difficulties with extracting DNA have now been resolved. Preliminary results indicate that the DNA fragments generated by enzymes EcoR1, Pst- 1, Hpa- 1, Hae 11, and Pvu-11 and visualized using a P32-labelled nbosomal DNA probe (PDM238) yield interpretable and informative restriction site differences among specimens from various hosts and localities. The results indicate that specimens from the bearded seal are distinct, whereas both harbour and grey seals carry the sarne species, thus corroborating results from allozyme studies. However, in contrast to the allozyme studies, several polymorphisms were also detected among specimens from harbour and grey seals and these could prove useful in investigating geographic stmcturing in sealworm populations in Atlantic Canada. This work is ongoing. Other studies

(a) Survey of parasites of Atlantic salmon and rainbow trout at the hatchery and sea-cage sites at Bay D'Espoir.

Samples of Atlantic salmon and rainbow trout were collected from within and surrounding the sea cages at various sites to give preliminary information on parasite infestation. Fish at the inshore site, which is characterized by wide fluctuations in salinity, were parasite free, but those at a more distant high-salinity site harboured several parasites, including curtus, Echinorhynchus gadi, Eubothrium sp., and another unidentified cestode. Operators of the site also report seeing nematodes and cestodes during processing. Samples from the freshwater hatchery were also collected but these have yet to be examined. This work is exploratory and continuation will depend on results from remaining samples.

Presentations (1993- 1994)

Brattey, J., and D. Methven. Trends in the prevalence of Lernaeocera branchialis (Copepoda: Pennellidae) on the gdls of Atlantic cod, Gadus morhua. Presented at the Canadian Society of Zoologists 33rd Annual Meeting, 11-14 May, University of Manitoba, Winnipeg, MA.

Brattey, J., C. Bishop, E. Dalley, and D. Methven. Long-term trends in the occurrence of Lernaeocera branchialis (Copepoda: Pennellidae) on the gills of Atlantic cd, Gadus murhua, from Newfoundland and Labrador. Presented at the Atlantic Canada Association of Parasitologists Annual Meeting, 30 September, NAFC, St. John's, NF.

Mattiucci, S., B. Berland, J. Brattey, P. Orecchia, L. Paggi, P. Arduino, and L. Bullini. Introgressive hybridization between Phocascaris phocae Host, 1932 and P. cystophorae Berland, 1964 (Nematoda: Ascaridoidea). Presented at the PARAQUA Symposium, 2-6 July, Westrnann Islands, Iceland.

Brattey, J., D. Methven, C. Bishop, and E. Dalley. Long-term trends in the occurrence of Lernaeocera branchialis (Copepoda: Pennellidae) a pathogenic gill parasite of Atlantic cod, Gadus morhua. Presented at the Northem Cod Science Program/OPEN/MUN Symposium, 24-28 October, Radisson Plaza Hotel, St. John's, NF. Publications ( 1993-94)

Brattey, J. Identification of larval Contracaecum osculatum and Phocascaris sp. (Nematoda: Ascaridoidea) fiom marine fishes by allozyme electrophoresis and discriminant function analysis of morphometric data. Can. J. Fish. Aquat. Sci. (submitted).

Brattey, J., and G. B. Stenson. 1993. Host specificity and abundance of parasitic nematodes (Ascaridoidea) fiom the stomachs of five phocid species from Newfoundland and Labrador. Can. J. Zool. 7 1: 2156-2166.

Brattey, J., and G. B. Stenson. Helminth parasites of the alimentary tract of harbor porpoise, Phocoena phocoena, from Newfoundland and Labrador. J. Helminthol. Soc. Wash. (submitted).

Nascetti, G., R. Cianchi, S. Mattiucci, S. D'Amelio, P. Orecchia, L. Paggi, J. Brattey, B. Berland, J. W. Smith and L. Bullini. 1993. Three sibling species within Contracaecum osculanun (Nematoda, Ascaridida, Ascaridoidea) from the Atlantic Arctic-Boreal region: reproductive isolation and host preferences. Int. J. Parasitol. 23: 105-120.

NEXT REVIEW PERIOD (1994-95)

Parasito1og;ical Studies of Northern Cod

- Continue with analysis of the extensive database on the parasitic copepod Lernaeocera branchialis on the gills of cod from NAFO Divisions 2J-3KL to determine whether substantial temporal or geographic changes in the level of infection have occurred. Determine whether these data can provide information on parasite-induced cod mortality, or stock mixing and migrations. Prepare a MS for publication and presentation at the Northern Cod Science Symposium (October 1994)(with Bishop and Dalley, GF and Methven, OSC)

- Examine data on occurrence of L. branchialis on juvenile cod (< 35cm) collected at various inshore trap sites of NE Newfoundland to provide information on parasite-induced mortality of cod, and seasonal and geographic aspects of parasite infection among cod size groups previously examined. Present findings at the annual meeting of the Canadian Society of Zoologists, Université du Québec à Rimouski (May 1995). (with Methven, MUN) Redfish parasites

- Continue with exploratory analysis of the extensive database on the parasitic copepod Sphyrion lumpi that infects the flesh of redfish, rendering it unmarketable. Analyze the parasite data with respect to providing insights into redfïsh stock mixing and possible reasons for recent increases in S. lumpi infection arnong -sh in the Laurentian Channel. Prepare a MS for primary publication if findings are suitable. (with Atkinson and Power, GF)

- Arrange collections of Sphyrion-infected oceanic redfishes from areas not covered by the above activity. Examine specimens, and code and key-punch data to provide additional information on stocks not covered by existing research surveys. An attempt will be made to establish collaborative contact with Russian scientists at Mmansk. (with Atkinson and Power, GF)

Nematode genetics and ecolom

- Collect nematodes from various species of fish and seal at locations throughout Atlantic Canada to investigate the population genetics of sealworm (P. decipieas) in Atlantic Canada by analysis of Restriction Fragment Length Polymorphisms. (with W. S. Davidson, Biochemistry Dept., MUN)

- Continue with the multinational study on the genetics of marine ascaridoid nematodes infecting seals and fish in the North Atlantic, the main objective being to determine the number of species, hosts, and geographic distributions of nematodes within the genera Anisakis, Pseudoterranova, Phocascaris, and Contracaecum. Prepare a MS on the genus Phocascaris in fish and seals in the North Atlantic. (with L. Paggi, Rome University)

- miblish a MS entitled "Identification of larval Contracaecum osculatum and Phocascaris sp. (Nematoda: Ascaridoidea) from marine fishes by allozyme electrophoresis and discriminant function analysis of morphomemc data" in a special commemorative issue of CJFAS.

- Publish a MS entitled "Helrninth parasites of the alimentary tract of harbor porpoise, Phocoena phocoena, from Newfoundland and Labrador" (with G. Stenson, Marine Mammals) in the Journal of the Helminthological Society of Washington.

Sealworm Program

- Continue to collect and examine for nematodes fish species that are eaten by grey seals to monitor trends in sealworm (P. decipiens) abundance within the region. Collect further samples of Atlantic cod and sculpins during spnng research vesse1 surveys to 3Pn and 3Ps, examine fish, and code and key-punch the data. Begin preparation of a MS on trends in sealworm abundance in cod stocks.

Parasite Detection and Removal Technology

- Provide biological advice and data on parasites in commercial fish stocks to DFO Inspection Services and industry as required to contribute to the development of parasite-free fish products. 18 REPORT OF THE QUEBEC REGION

J.R. Arthur, D.J. Marcogliese, and L. Measures

Maurice Lamontagne Institute Mont-Joli, Quebec

Established in 1989, the Parasitology Section of the Maurice Lamontagne Institute currently has a staff of five permanent employees (one technical post was lost in 1993). These include three scientists (R. Arthur (Section head), D. Marcogliese, and L. Measures), one biologist (Elaine Albert) and one technician (J. Guerin). The Section is designated as a DFO CODE (center of disciplinaIy expertise) and as such takes a lead role in fisheries parasitological research, plays a coordinating role in research planning and priority setting, and provides assistance to other DFO labs as requested. The Section's research programs are thus not restricted to the Quebec Region. Current research strengths within the Section are in the areas of parasite taxonomy, life cycles and ecology.

RESEARCH COMPLETED SINCE SEPTEMBER 1993

A-Base Funded Research

Under its first five year plan (1990-1995) the Section is undertaking research in three core areas:

Parasitolomcal surveys of fish and marine mammals and their use in stock assessment. (J.R. Arthur, L. Measures, and E. Albert)

(a) Parasites of marine fishes.

1. Parasites of Greenland halibut (Reinhurdtius hippoglossoides) and their use as biological tags. (R. Arthur, E. Albert)

Purpose: This study will determine the parasite fauna of Greenland halibut in Canadian Atlantic waters and will provide information on discreteness of stocks occurring in the Gulf of St. Lawrence, the Saguenay Fjord and off Labrador. Results: Two papers, one dealing with the results of stock identification studies and another presenting survey data and an analysis of the zoogeography of Greenland halibut parasites, have been published. A third paper dealing with the first record of a marine leech, Notostomum lavae, from North American waters is in press. Examination of additional collections to verify our results has been completed and data analysis is in progress. Work on the taxonomy of myxosporeans will be continued.

2. Parasites of redfish (Sebastes spp.) and an evaluation of their usefulness as indicators of dish species. (J.R. Arthur, with J. Moran)

Purpose: This project was designed to determine whether species specific differences in the parasite fauna exist between two closely related Mishes, S. mentella and S. fasciatus. Because no easy method is available to separate these species, the resource is currently managed as a multi-species fishery.

Results: All work, with the exception of manuscript preparation, has been completed. Fish were identified as S. mentella, S. fasciatus or unknowns based on morphological and enzyme electrophoretic studies. Complete necropsies were performed on 50 S. mentella, 50 S. fasciatus and on 25 heterozygotes. Results indicate that parasites cannot be used with confidence to separate sharp-beaked redfishes. This work formed the basis of an M.Sc. thesis written by J. Moran (University of New Brunswick). Two papers are in preparation.

3. Parasites as biological tags for capelin (Mallotus villosus) in the Gulf of St. Lawrence. (J.R. Arthur, E. Albert, F. Boily)

This project, which is partially A-base supported, is discussed below under PFDPQ- funded projects.

(b) Parasitological surveys of marine mammals in the lower estuary and Gulf of St. Lawrence. (L. Measures)

Purpose: The objective of this survey is to document the species of parasites occurring in Canadian Atlantic waters, their host and geographic distributions, their pathogenicities and possible role in host mortalities and zoonotic potential.

Results: Since 1989, eight species of marine mammals have been examined parasitologically for specific projects. Helminth surveys have been completed for harbour seal (n=17), harbour porpoise (n=19), ringed seal (n=32), Atlantic white-sided (n=2), harp seal (n=15), blue (n=l), minke whale (n=l), and beluga (n=39). Parasites have been removed and fixed and grouped by host and parasite taxa. Nematodes have been cleared. Material hmringed seals, blue whale and beluga have been identified. Intestinal helminths only have been collected from harbour seal (n=2), grey seal (n=12) and harp seal (n=60). Other identifications are planned for 1995-96. Two papers were published and one paper has been presented at a scientific conference. The results of a collaborative project with Pierre Béland, Daniel Martineau and Sylvain De Guise, INESL on helminths of stranded St. Lawrence beluga (N = 39 of 167) collected since 1982 have been submitted for publication.

Other projects dealing with marine mammal parasites involve five M.Sc. graduate students: the from Université Laval on (1) Lungworms of phocid seals in eastern Canada, (2) Biology of Otostrongylus circumliius in ringed seals in Arctic Canada (both NSERC funded), and (3) Biology of Anisakis simplex in the St. Lawrence Estuary; and two from McGill University on (4) Stenurus minor in the cranial sinuses of harbour porpoise and (5) Anisakis sirnplex in harbour porpoises from the Gulf of St. Lawrence. Two students are writing their theses and three publications are expected to be submitted this fiscal year.

Studies on the life cycle and transmission dynamics of the sealworm (Pseudoterranova deci~iens).(D. Marconliese)

Purp: The objective of this study is to determine which macroinvertebrates are important in transmitting sealworm to fish, and to gain insights into the dynamics of transmission.

Results: Since 1989, field work has concentrated on areas of high sealworm intensity in groundfish, most notably Sable Island Bank, although some collections have been made on the Eastern Shore, and from Bras D'Or Lakes, Mingan Islands, Anticosti Island, Magdalen Islands and Georges Bay. For benthic collections, priority has been given to Sable Island Bank samples. Al1 molluscs, polychaetes and from 1989/90 cruises (>200 samples) have been identified. Only a few samples containing representatives hm,other phyla (e.g. echinoderms) remain, and will be completed in 1994. Al1 crustaceans (>23,000) and about half the polychaetes (>4,800) have been examined for parasites. No sealworm were found in any of the samples examined thus far.

One M.Sc. student (Cathy Jackson) at the University of New Brunswick, is surveying parasites of mysids and other epi- and hyperbenthic organisms from various areas in eastem Canada. Samples have been collected near Anticosti Island, the Gaspé, Sable Island, Passarnaquoddy Bay, Grand Manan, the Bras d'Or Lakes, Prince Edward Island, St. Georges Bay, and north-east Cape Breton (St. Annes Bay). Sealworm were found in mysids from the Bras d'Or Lakes and St. Annes Bay. Studies have continued in Sable Island ponds, which contain infected sticklebacks. in 1994, infection levels of sealworm in invertebrates from Wallace Lake were determined. Stickiebacks were collected to quantiq infections of sealworm and diet, in order to determine the rate of transmission from invertebrates to fish. Samples of fish and invertebrates were coilected to estimate parasite abundances. Fecal samples from harbour seals basking around the lake were also collected. Infection levels in sticklebacks were much higher than in past years, with 62% of fourspines and 44% of threespines infected with 2.6 and 1.3 worms per fish, respectively. Densities were 4,232Ag and 927/kg, respectively. One sealworm .was found in 4,000 amphipods, and 12 in 3,000 mysids. These infection levels are much higher than was previously observed.

Dr. B. Pohajdak, Dalhousie University, is examining a variety of invertebrates for sealworm using a species-specific probe for hemoglobin. Samples of invertebrates are homogenized and the DNA extracted and amplified using a DNA polymerase technique. Probes are then used to search for matching genes in the sample. This work is ongoing.

Since September 1993, one paper was published based on the results of this project, two papers were presented at scientific meetings, and two seminars were given at international institutions. In 19941'95, special funding was provided by IML to stimulate inter-regional efforts towards production of a predictive model for sealworm infections in fish and seals. Using the existing data base, prelirninary modelling efforts are being initiated this year to identify potential impacts of control measures and to prioritize sealworm research activities. An initial step towards these goals was the production of a manuscript report written with G. McClelland on the status of sealworm research in eastern Canada (See Appendk).

Studv of Pseudoterranova decipiens, Phocascaris phocae, Contracaecum osculatum, and Anisakis simplex - larval develoriment. (L. Measures)

Pur-: The overall objective is to determine how fish acquire these ascaridoid nematodes from seals and whales. The goal is to establish a laboratory model describing and comparing free-living development, parasitic development, pathogenicity, longevity and transmission dynamics in various species of invertebrates and fish. Once a suitable model is established in the laboratory various manipulations will be performed which can lead to a better understanding of transmission in the marine environment.

Free-living development - One publication is in press and three publications are in preparation. Live material of Anisakis simplex was not obtained for study of free-living development.

Development in invertebrate hosts - Cultures of amphipods (Gammarus setosus) and mysids (Mysis guspensis) have been re-established in the laboratory this summer. No experiments were carried out this past year. Analysis of previous experimental work is planned before the end of this fiscal year. Over 63,000 euphausids (99.4% Thysanoessa raschii, 0.6% Meganyctiphanes norvegica) collected in the St. Lawrence River estuary in June were examined by a M.Sc. student for larval Anisakis using pepsin digestion. Prevalence was 0.009% with four of six specimens bearing two cuticles. Calanus finmarchicas (n=135,000) were also examined for Anisakis - none was found. In August from the same region over 2,900 euphausids (99.8% T. raschii, 0.2% M. norvegica were collected from surface (1,261) and deeper waters (1,719). One larval Anisakis with one cuticle was found in the deep water sample. These data indicate that the upper St. Lawrence River estuary is an enzootic area for Anisakis.

Longevity of larval ascaridoids in naturally infected fish - American plaice collected live near Matane in 1991, 1992 and 1993 (lower estuary of the Gulf of S t. Lawrence) are king maintained in captivity to determine the longevity of naturally acquired ascaridoid larvae and endohelminths (see below). To date, over 1,000 plaice collected from Matane and maintained in captivity for varying periods of time have been examined since 1990 and ascaridoids removed and futed. Ascaridoid infections are still present in captive fish, demonstrating a longevity of at least three years. These ascaridoids consist of Pseudoterranova, Anisakis, ContracaecumlPhocascaris, and Hysterothylaciurn. Specific identifications and data entry have been completed for plaice collected in 1990 and identifications for plaice collected in 1991-1993 are on-going. This data will prove useful in modelling the epizootiology of sealworm and whaleworm in an estuarine situation where mean annual water temperatures at depth are lower than those in the southern Gulf of St. Lawrence and in the Scotia-Fundy region. American plaice muscle tissues containing larval Pseudoterranova in situ one, two and three years pst-captivity (as well as control fish) have been fixed for histopathological analysis. This data should indicate changes in host tissue in respnse to the presence of Pseudoterranova and relative health of larvae over respective time periods.

Longevity of endohelminths in naturally infected fish - Supplementary data collected since 1991 from the above study indicate that a variety of nematodes, digeneans and acanthocephalans can live in American plaice for at least two years. Data from American plaice killed and dissected immediately pst-capture (n=150) over a three-year period and after one year in captivity (n=100) and two years in captivity (n=25) have been collected. Al1 helminths have been removed and fixed. No fish killed after 1993 will be examined for al1 helminths. Specific identifications will commence when work involving nematodes from plaice is completed. Data will prove useful for aquaculture and fisheries management. PFDPQ Proiects

Survev of anisakine nematodes in moundfish and mey seals in the Gulf of St. Lawrence. p. Marconliese, with M. Hammill)

Purpose: A program to survey sealworm and other anisakine parasites in groundfish in the Gulf of St. Lawrence, and to enumerate anisakine infections in grey seals from the same area was carried out under Programme Fédéral de Développement des Pèches du Quebec (PFDPQ) funding, allowing the Quebec Region to complete important sealworm research as designated by the SeaVSealworm Ecology Program.

Results: Data describing infection levels of anisakids in cod and American plaice collected from various areas in the Gulf of St. Lawrence in 1990 and 1992 were analyzed. A manuscript describing the results is currently under revision. Another manuscript describing infection levels in 30 other species of fish is in preparation. In general, levels of sealworm are lower in the northem Gulf compared to the southem Gulf, and appear to be dropping. Levels of Contracaecinea are higher in the north, but appear to be rising throughou t the Gulf.

Identification of nematodes from seals collected in the Gulf of St. Lawrence in 1992 will be completed by the end of 1994. Over 100 samples were completed in 1993-94. A preliminary analysis suggests sealworm levels are dropping in grey seals, whereas Contracaecum osculatum is increasing.

Since September 1993, one paper was presented at an international meeting (see Appendix) .

Parasites as biological tans for capelin (Mallotus villosus) in the Gulf of St. Lawrence. (J.R. Arthur, E. Albert, F. Boily)

Purpose: This project is designed to evaluate the usefulness of parasites as biological tags for capelin in the Gulf of St. Lawrence. Because of its strategic importance in the food web, and because of the growing importance of the capelin fishery, research on this species has ben given high priority at IML. This study is funded jointly by the PFDPQ and A-base.

Results: The planned sampling program (collections from 8 spawning sites x 3 sequential collections/site) was only partially met, due to cancellation of the fishery in 1994 in the northern Gulf. As a result, samples (30 males130 females) from four sites (Ile Verte, Sept Iies, Havre-S t.-Pierre, Blanc Sablon) were obtained on three occasions (total of 720 fish). Complete dissections of 15 fish/site have been completed and we are in the process of identifying parasites and analyzing this preliminary data. In general, prevalences and intensities of infection are low, with the exception of gill monogeneans (Gyrodactyloides spp., Laminiscus gussevi) and tetraphyllidean plerocercoids, which are common and abundant in al1 collections. From examination of the raw data, we believe that parasites will not provide evidence of the presence of separate populations of Gulf capelin. Preliminary results of this study were presented at the September 1994 meeting of the Atlantic Canada Association of Parasitologists.

Other ongoing Research and Service Activities

Client consultations (al1 section staff)

Identification of parasites has been conducted during 1993-94 for various agencies, individuals, universities, etc. These include Ministère du Loisir, de la Chasse et de la Pêche, Trois-Rivières; Makivik Corporation, Kuujjuaaq; Université de Montréal, Collège Vétérinaire, Ste-Hyacinthe; INESL, Montreal; Université du Québec à Trois-Rivières; Université du Québec à Rimouski; the McGill University Tropical Disease Center, Montreal; Perch Survey, Parasitology Section Module, Canadian Society of Zoologists; Ministère de l'Agriculture, des Pêcheries et de l'Alimentation du Québec; New Brunswick Museum; and other DFO researchers. Specimens were also deposited in the Canadian Museum of Nature.

A trilingual poster indicating recent research results of interest to Inuit communities was prepared and displayed in Salluit, Nunavik, in September, 1994. Questions from the community were also addressed.

Section staff have reviewed a total of 32 manuscripts for the following journals: Canadian Journal of Zoology (3,Canadian Field-Naturalist (l), Fishery Bulletin (l), Journal of the Helminthological Society of Washington (l), Journal of Parasitology (IO), Journal of Wildlife Diseases (3), Marine Biology (l), Systematic Parasitology (l), Canadian Journal of Fisheries and Aquatic Science (4), Asian Fisheries Science (2),Fish Health Section, Asian Fisheries Society, Symposium Proceedings (3). Manuscripts were also reviewed for DFO internal publications, and for internal and external colleagues.

They also reviewed a total of seven grant applications for the following granting agencies: DFO subvention grants (3, NSERC Collaborative Grants Program (l), and the International Science Foundation (1); coordinated review of French-language grants for the International Science Foundation (Sweden); acted as external evaluator for the Foundation for Research Development (Government of South Africa), and as external reviewer for 1 Ph.D. thesis (University of Kerala, India). Training activities (al1 section staff)

The Parasitology Section provided short-term training of five weeks to one student (Nicole Paradis) at the request of the CEGEP de Sherbrooke.

Nine graduate students (al1 M.Sc.) are currently supervised by Section members. These are Cathy Jackson (University of New Brunswick); Nicole Dubois and Marc Bergeron (Université du Québec à Trois Rivières); Jean-François Gosselin, Emmanuelle Bergeron, Rejean Hays and Francoise Tetreault (Université Laval); and Jacqueline Faulkner and Manon Simard (McGill University). One student, Jon Moran WB), successfully completed his M.Sc. in 1994.

Blood protozoans of American plaice from the lower estuarv of the St. Lawrence. (L. Measures, with M. Siddall and S. Desser. University of Toronto)

This project, initiated in 1991, has concluded. Three publications have resulted; a fourth is in preparation.

Guide to parasites of commercially important fish in eastem Canadian waters. (L. Measures)

Photographs of some parasites in situ accompanied by a bilingual text have been completed. These parasites include Anisakis simplex in hemng, Lernaeocera branchialis on cod, Hysterothylacium aduncum in cd,Pseudoterranova decipiens in American plaice, Diplostomum sp. in tomcod and Sphyrion lumpi on redfish. Collections will continue in the coming year and additional material on hand will be prepared. Some of this matexial was presented in a poster format to a fishery industry workshop held at IML this past spring.

Studies on Ascarophis of Pacific decapods. (L. Measures, with Dr. J. Holmes, University of Alberta)

Larval material from 10 species of decapods (n=6û) was previously collected. No analysis was conducted since October. This small project is part of a larger study on parasites of Pacific rockfish by Dr. Holmes. One publication is expected to result from the decapod material. Collaboration with the International Development Research Center's Asian Fish Health Network and the Fish Health Section, Asian Fisheries Society. (J. R. Arthur)

No IDRC consultancies were undertaken in 1993194. A low level of involvement with staff f?om former IDRC-funded projects in Asia and with the Fish Health Section, Asian Fisheries Society is continuing. A key-note address on the status of quarantine measures for aquatic animals was presented at the "2nd Symposium on Diseases in Asian Aquaculture", Phuket, Thailand in October 1993 and a companion paper is in press. Co- editing of the symposium proceedings "Diseases in Asian Aquaculture II" is in progress (with Mhd. Shariff). Two taxonomic papers dealing with parasites of Philippine fishes are in press, one redescribing a rare acanthocephalan from (with S. Regidor and E. Albert) and another describing two new species of Gyrodactylus from tilapia (with M. Bondad-Reantaso and D.K. Cone) (see Appendix). Results of this work will be presented at the 1994 ACAP meeting.

Studies on the taxonomy of trichodinid ciliates. (J. R. Arthur and E. Albert, with D. Cone, D. Marcogliese and S. McGladderv)

One paper was published redescribing a species from Canadian brook trout (with D. Cone). Work on a new species from giant scallop (with S. McGladdery) has been temporarily suspended pending recollection of material. Additional fishes from Sable Island were collected and examined for mchodinids and other protozoan parasites (with D. Marcogliese).

Studies on Metorchis coniuncnts, a fish-transmitted human pathogen.(J.R. Arthur, with J. D. McLean, B. Ward, M.A. Curtis, T.W. Gvorkos, and E. Kokoskin, McGill University Center for Tropical Diseases, Montreal General Hospital, and McGill University)

A manuscript detailing the first acute case of metorchiasis in humans is cunently being prepared. Results were presented by J. D. McLean at the joint meeting of the American Society of Tropical Medicine and the American Society of Parasitologists in Atlanta, Georgia. The use of egg surface structure as a possible diagnostic tool was explored via SEM studies of the egg of Metorchis from humans and hamsters. An epidemiological study of human infections in a native population at Sault Lookout, ON is currently being conducted by medical researchers at the Montreal General Hospital. IML staff are assisting by examining white suckers from this area for metacercarial infections. Pre~arationof a srnial uarasitolom issue of the Canadian Journal of Fisheries and Aeuatic Sciences. (J. R. Arthur1

Editing of this special volume continues. A total of 40 manuscnpts were received, of which approximately 30 will be published. Of these, 14 have been finalized and the remainder are in various stages of revision. Publication is expected in the spring of 1995.

Transmission of visceral helminths between fishes via uredation. (J. R. Arthur and P. Dallaire)

Experimental transmission of digestive tract parasites between predator (Atlantic salmon) and prey (American plaice) was completed, salmon dissected and the parasites recovered. Identification of parasites and manuscript preparation remain to be accomplished.

Aauatic Parasitolow Information Network (APIN) (J. R. Arthur)

An additional 1400 citations have been entered into this literature data base, giving a total of about 4900 entered to date. Progress has been slower than anticipated due to lack of manpower.

Parasites of invertebrates. (D. Marcogliese, with M. Adamson, University of British Columbia; C. Jackson and L. Fleming, University of New Brunswick; A. Kuris, University of California)

Work is continuing on the identification of parasites of invertebrates. Parasites found in various crabs, amphipods, mysids, polychaetes and chaetognaths include Ascarophis sp., Tetrameres sp., Paracuaris adunca, Hysterothylacium aduncurn, Hemiurus levinseni, Echinorhynchus gadi, Mysidobdella borealis, and Argulus alosae. Three papers, including one review, are accepted for publication (See Appendix). Some animals await identification.

Investigations on sedimentation rates and transport of sealworm eggs. @. Marcogliese, with M. Stacey, Royal Roads Militarv College; and C. McConnell, St. Mary's University)

A manuscript is in preparation describing the transport of sealworm eggs by oceanic currents in the vicinity of Sable Island. It was found that eggs could be carried approximately 12 km given a sedimentation rate of 4 X IO4. Biogeograuhy of ara sites of fishes. @. Marcogliese, with R. Arthur; D. Cone, St. Marv's University)

A manuscript comparing the parasite fauna of ninespine sticklebacks and mummichogs inhabiting brackish and freshwater ponds on Sable Island has been submitted for publication. This work attempts to explain the distributions of the various parasites among ponds on Sable Island in terms of local versus regional effects. Sable Island fishes were also examined for protozoans (with R. Arthur). A paper was published using parasites to explain the evolutionary divergence of American and European eels (with D. Cme; see Appendix). A second paper on the biogeography of Pseudodactylogyrus anguillae on eels has been submitted for publication. Parasites from eelpout collected in the Saguenay River await identification.

Factors controlling communitv structure and com~ositionof parasites. @. Marcogliese, with D. K. Cone, St. Marv's University; and P. Magnan, N. Dubois, and M. Bergeron, Université de Ouébec à Trois Rivières)

It was demonstrated that parasite communities of eels in acid-stressed systems are impoverished, but they can recover (with D. Cone). It is suggested that parasite communities mirror food chain dynamics, and that they may be useful as indicators of pollution stress. Work is continuing examining the distribution of parasites in eels in Nova Scotia from a regional perspective. Results on a larger scale mirror those from our experimental system. A talk was prescnted to the ICES Working Group on Pathology and Diseases of Marine Organisms (see Appendix). Ms Dubois is continuing her studies of parasites of allopamc brook char, and char sympatric with white suckers. Her project will be completed in the autumn of 1994, and a manuscript is in preparation. A paper was presented at an international meeting (see Appendix). A second student, Marc Bergeron, is following up this study by examining interactions between parasites of chub and char.

Regulation of winter flounder povulations by Cn>utocotyle Zingua and Steuhanostomum baccatum. @. Marconliese, with M. D. B. Burt, University of New Brunswick)

Extremely heavy infections in flounder have been noted in New Brunswick bays. It is likely that high infection levels are detrimental to newly-metamorphosed flatfish, and may cause mortality. This mortality may influence recruitment to the fish population. A graduate student at UNB is studying this parasite system.

Parasites and biodiversity. @. Marcogliese, R. Arthur)

D. Marcogliese is the chairman of the Parasitology Module Stee~gCommittee, which is currently working towards establishing a functional and funded Parasitology Module within the Biological Survey of Canada. The module is involved in a number of projects, including a survey of perch parasites across North America. An important goal of the module is to use parasites as environmental biomonitors, and indicators of biodiversity. D. Marcogliese met twice with the Biological Survey of Canada and chaired a session of the Biodiversity Workshop held during the annual meeting of the American Society of Parasitologists in Fort Collins, CO. In addition, he has reviewed the draft of the National Biodiversity Strategy for DFO and for the Canadian Society of Zoologists. R. Arthur has received a preliminary invitation to present an invited paper on parasites and biodiversity at a special biodiversity symposium being organized as part of the Fourth Asian Fisheries Forum, king held in Bejing in October 1995.

Identification of Di~lostomumspp. infecting fishes of the lower St. Lawrence. (R.Bailey, D. McLaughlin, Concordia University)

R. Bailey, head, ,Marine Marnmals and Specialized Biology Division at IML, has coordinated a contract with D. McLaughlin to determine the species of Diplostornurn which are causing blindness in St. Lawrence fishes (particularly tomcod), a problem of concem to the province and the sport fishery. Natural infections have been studied and experimental infections established in gull chicks to determine the identity of the parasite(s) involved and their geographical distribution(s). The study involves cooperation between Concordia, IML, DFO's Habitat Branch, and Environment Canada.

Parasites of Arctic char in northem Ouebec. (R. Arthur and E. Albert, with J. Pellerin and Y. Desdevises, Université du Quebec à Rimouski)

As part of a major study looking at health risks of "country foods", section staff have been asked to provide parasitological expertise and to train a French exchangee (Y. Desdevises) in basic parasitological techniques. A total of 6û Arctic char were collected in August of 1994 and await examination.

Behaviour of parasitically castrated Buccinium undatum in the St. Lawrence Estuarv. (L. Measures, with J. Hirnmelman, Université Laval)

An M. Sc. graduate student from Université Laval is being CO-supervisedon behaviour of parasitically castrated Buccinum undatum in the St. Lawrence Estuary. RESEARCH PLANNED FOR 1994-1995

A-base Funded Research

Parasitololrical surveys of fish and marine mammals and their use in stock assessment. (J. R. Arthur, L. Measures and E. Albert]

(a) Parasites of marine fishes (J. R. Arthur, E. Albert and F. Boily)

Work on capelin parasites (partially PFDPQ-funded) will be terminated if results show, as anticipated, homogeneity of collections within the Gulf of St. Lawrence. If so, a small amount of effort will be devoted to taxonomic studies on capelin parasites. Work will be initiated on another, yet to be determined, species of fish or invertebrate.

(b) Parasitological survey of marine mammals in the lower estuary and Gulf of St. Lawrence (L. Measures)

Identification and analysis of marine mammal material already collected and fixed will continue. Publications will be prepared for submission. Projects being accomplished by graduate students will continue (NSERC funded for the most part). Two projects investigating the value of lungworms in beluga for stock identification and investigations on the biology of Trichinella nativa in Atlantic walrus in Nunavik will be initiated late 1994-95 and continue in 1995-96, depending on availability of samples.

Studies on the life cycle and transmission dynamics of the sealworm (Pseudoterranova deciviens). (D. Marcorrliese)

Data from field work on Sable Island in 1994 to estimate transmission rates of sealworm from invertebrates to fish wili be analyzed. Results of the survey of mysids and other hyperbenthic fauna will be prepared for publication with a graduate student (C. Jackson). Identification of al1 invertebrates collected from the four research cruises in 1989-90 will be completed. Analysis of data from benthic surveys will be initiated.

An improved methodology for detecting sealworm in invertebrates, through a molecular probe technique (with Dr. B. Pohajdak, Dalhousie University) will be pursued. The DNA probe remains to be tested for specificity to P. decipiens as opposed to other anisakids. This probe is functional on samples of animals fixed and stored in ethanol. Thus, it would be possible to process large numbers of preserved animals in a short time. Density of sealwonn, as well as presence of other anisakids, can influence fecundity of individual parasites. Using sorted samples of worms from grey seals, it is planned to examine variations in fecundity of individual sealwoms under different conditions. These data will be useful for models of sealworm-grey seal-cod interactions.

Studv of Pseudoterrarwva deci~iens,Phocascaris ~hocaeand Anisakis simplex - larval develovment. (L. Measures)

Data presently on hand will be analyzed and future experiments with mysids and amphipods will continue depending on availability of infective parasitic larvae and invertebrate cultures. Emphasis will be placed on locating where the second moult of Pseudoterranova decipiens occurs (invertebrate or fish host and which host is paratenic). Studies on the longevity of larval ascaridoids in American plaice will continue into a fifth year.

APPENDIX: PUBLICATiONS AND PRESENTATIONS OFTHE PARASITOLOGY SECTION OCTOBER 1993-SEPTEMBER 1994

Articles vublished or in press:

Adamson, M. L., and D. J. Marcogliese. (In press). A possible instance of in a chromadorid nematode, Cyatholaimus cirolanae n. sp., from the marine isopod Cirolana polita. Trans. Am. Microsc. Soc.

Arthur, J.R. (In press). A history of fisheries parasitology in Southeast Asia. Asian Fish, Soc. Spec. hbl.

Arthur, J.R. (In press). Efforts to prevent the international spread of diseases of aquatic organisms, with emphasis on the Southeast Asian region. Diseases in Asian Aquaculture II. Proceedings of the 2nd Symposium on Diseases in Asian Aquaculture, 24-29 October, 1993, Phuket, Thailand. Asian Fish. Soc., Fish Health Sect., Kuala Lumpur.

Arthur, R. 1994. Meeting Reports. The Parasitology Section of the Canadian Society of Zoologists. University of Guelph, Ontario, Canada, May 1993. Int. J. Parasitol. 24: 313- 314. Arthur, J.R., and E. Albert. 1993. Use of parasites for separating stocks of Greenland halibut (Reinhardtius hippoglossoides) in the Canadian northwest Atlantic. Can. J. Fish. Aquat. Sci. 50: 2175-2181.

Arthur, J.R., and E. Albert. 1994. A survey of the parasites of Greenland halibut (Reinhardtius hippoglossoides) caught off Atlantic Canada, with notes on their zoogeography in this fish. Cm. J. Zool. 72: 765-778.

Arthur, J.R., and D. K. Cone. 1994. Redescription of Trichodina kamchatika Konovalov, Shevlyakov and Krasin, 1970 (Ciliophora: Perimchida) from the brook trout (Salvelinus fontinalis) of eastern Canada. Folia Parasitol. 41: 39-43.

Arthur, J. R., S. E. Regidor and E. Albert. (In press). Redescription of Cavisoma magnum (Southwell, 1927) (: ) from the milkfish, Chanos chanos (Forsskal), in the Philippines. J. Helminthol. Soc. Wash.

Burreson, E. M., and J.R. Arthur. (In press). First North American record of a marine leech Notostomum laeve (Hinidinea: Piscicolidae). J. Parasitol.

Cone, D.K., J.R. Arthur and M.G. Bondad-Reantaso. (In press). Description of two new species of Gyrodactylus von Nordrnann, 1832 () from cultured Nile tilapia, Tilapia nilotica (Cichlidae), in the Philippines. J. Helminthol. Soc. Wash.

Hoberg, E.P. and L.N. Measures. (In press). Anophryocephalus inuitorum sp. nov. and A. arcticensis sp. nov. (Eucestoda: Tetrabothxiidae) in ringed seals (Phoca hispida) and harp seals (Phoca groenlandica) from high latitude seas of eastem Canada and the Arctic Basin. Can. J. Zool.

Marcogliese, D. J. 1993. Larval nematodes infecting Amphiporeia virginianu (Amphipoda: Pontoporeioidea) on Sable Island, Nova Scotia. J. Parasitol. 79: 959-962.

Marcogliese, D. J. (In press). Aeginina longicornis (Amphipoda: Caprellidea), new intermediate host for Echinorhynchus gadi (Acanthocephala: Echinorhynchidae). J. Parasitol.

Marcogliese, D. J. (In press). The role of zooplankton in the transmission of helrninth parasites to fish. Rev. Fish Biol. Fish.

Marcogliese, D. J., and D. K. Cone. 1993. What metazoan parasites tell us about the evolution of American and European eels. Evolution 47: 1632-1635.

Marcogliese, D. J., and G. McClelland. (In press). The status of biological research on sealworm (Pseudoterranova decipiens) in eastem Canada. Can. Manuscr. Rep. Fish. Aquat. Sci. Measures, L.N. 1994. Synonymy of Longibucca eptesica with Longibucca lasiura (Nematoda: Rhabditoidea), and new host geographic records. J. Parasitol. 80: 486-489.

Measures, L.N. 1994. Seasonal dynamics of the bat worm, Longibucca lasiura (Nematoda: Rhabditoidea) in bats of Alberta. Cm. J. Zool. 72.

Measures, L.N. and J-F. Gosselin. 1994. Helminth parasites of ringed seal, Pbca hispida, from northem Quebec, Canada. J. Helrninthol. Soc. Wash. 61: 240 - 244.

Measures, L.N. and H. Hong. (In press). The number of moults in the egg of sealworm, Pseudoterranova decipiens (Nematoda: Ascaridoidea): an ultrastructural study. Cm. J. Fish. Aquat. Sci.

Siddall, M.E., S.S. Desser and L.N. Measures. (In press). Light and electron microscopic examination of so-called piroplasms of fishes from Atlantic Canada and systematic revision of the Haemohormiidae (incertae sedis). J. Parasitol.

Siddall, M.E., L.N. Measures and S.S. Desser. (In press). Seasonal changes in erythrocyte osmotic fragility and haematocxit in Amencan plaice (Hippoglossoides platessoides) infected with the piroplasm Haemohomidium terranovae). J. Fish Biol.

Siddall, M.E., L.N. Measures and S.S. Desser. 1994. Infection with the piroplasm Haemuhonnidium terranovae in relation to haematocrit and mortality of American plaice (Hippoglossoides platessoides). Can. J. Fish. Aquat. Sci. 51: 959 - 964.

Theses supervised bv Section Staff:

Moran, J. 1994. The parasites of sharp-beaked redfïshes (Sebastes fasciatus and S. mentella) in the Gulf of St. Lawrence. M.Sc. Thesis, Univ. New Brunswick, 142 p.

Presented Pauers:

Arthur, J.R.. Two taxonomie studies on the parasites of Philippine fishes. 4th Meeting, Atlantic Canada Association of Parasitologists, 30 September 1994, St. John's, Nfld.

Arthur, J.R. Aquatic animal quarantine: towards internatinal disease control. 2nd Symposium on Diseases in Asian Aquaculture. 25-29 October, 1993, Phuket, Thailand, Symposium abstracts, p. 65. (Invited keynote presentation)

Arthur, J.R. A brief history of fisheries parasitology in Southeast Asia. Univ. of New Brunswick, Parasitology Group, 2 December 1993, Fredricton, NB. Arthur, J.R., and E. Albert. Preliminary results of capelin stock discrimination in the Gulf of St. Lawrence using parasites. 4th Meeting, Atlantic Canada Association of Parasitologists, 30 September 1994, St. John's, Nfld.

Bergeron, E., and L.N. Measures. Biology of Otostrongylus circumlitus, a lungworm of ringed seals. 4th Annual Meeting of Atlantic Association of Parasitologists, 30 September 1994, St. John's, Nfld.

Dubois, N., D. J. Marcogliese, and P. Magnan. Effect of the introduction of white sucker, Cmstomus commersoni, on the parasite fauna of brook charr, Salvelinus fontinalis. International Chan Symposium, 13-18 June, 1994, Trondheim, Norway.

Hays, R. and L.N. Measures. The biology of Anisakis simplex, a parasite of the St. Lawrence beluga (Delphinapterus leucas). 4th Annual Meeting of Atlantic Association of Parasitologists, 30 September 1994, St. John's, Nfld.

Jackson, C. J., D. J. Marcogliese, and M. D. B. Burt. Varying roles of hyperbenthic crustaceans in the transmission of marine nematode parasites. 33rd Annual Meeting of the Canadian Society of Zmlogists, 11- 14 May 1994, Winnipeg, MA.

Marcogliese, D. J. Recent advances in sealwonn life cycle studies. 16th Symposium of the Scandinavian Society for Parasitology, 30 September-2 October 1993, Oslo, Norway.

Marcogliese, D. J. Various aspects of the biology of sealworm. Invited seminar. Zoological Museum, University of Oslo. 5 October 1993.

Marcogliese, D. J. Various aspects of the biology of sealworm. Invited seminar. Dept. of Biology, Wake Forest University, Winston-Salem, N.C. 27 October 1993.

Marcogliese, D. J. Parasite communities as indicators of environmental conditions. Invited presentation. ICES Working Group on Pathology and Diseases of Marine Organisms. 25 March 1994, Moncton, NB.

Marcogliese, D. J., and F. Boily. Changes in abundances of anisakid nematodes in Atlantic cd and grey seals in the Gulf of St. Lawrence. 69th Annual Meeting of the American Society of Parasitologists, 9-13 August 1994, Fort Collins, CO.

Marcogliese, DJ., and F. Boily. Sealworm and other anisakid nematodes in cod and seals in the Gulf of St. Lawrence. 4th Annual Meeting of the Atlantic Canada Association of Parasitologists, 30 September 1994, St. John's, Nfld.

McClelland, G., D. J. Marcogliese and J. R. Arthur. Parasites as indicators of Gulf of St. Lawrence fish stocks. Fish. Oceanogr. Subcomm. Meeting, 22-24 March 1994, Dartmouth, N.S. (mimeo abstract) MacLean, J.D., J.R. Arthur, B. Ward, M.A. Curtis, T. Gyorkos and E. Kokoskin. Common source outbreak of acute metorchiasis due to the Canadian liver fluke, Metorchis conjuncncs. Joint Annual Meeting, American Society of Tropical Medicine and Hygiene and the American Society of Parasitologists, 31 October - 4 November, 1993, Atlanta, Georgia. (Presented at the session on "Late Breakers in Biology and Molecular Biology")

Measures, L.N. The number of moults in the egg of sealworm, Pseudoterrarwva decipiens, with moulting variations in the Nematoda. 4th Annual Meeting of Atlantic Canada Association of Parasitologists, 30 September 1994, St. John's, Nfld.

Measures, L.N. Helminths of a relict and endangered population of beluga in the St. Lawrence River, Canada. Canadian Society of Zoologists Annual Meeting, Winnipeg May 1994.

Moran, J.D.W., J.R. Arthur, and M.D.B. Burt. Parasites of sharp-beaked redfishes (Sebastes fasciatus and S. mentella) from the Gulf of St. Lawrence. 33 Ann. Meeting, Canadian Society of Zoologists, 11-14 May 1994, Winnipeg, Manitoba. Program and Abstracts, p. 78-79. 36 REPORT OF THE SCOTIA-FUNDY REGION

G. McClelland and D.J. Martel1

Halifax Fisheries Research Laboratory Halifax, Nova Scotia

CURRENT AND ON-GOING RESEARCH

Fish Surveys

Sealwom Index Surveys

A study descnbing temporal trends in levels of larval sealworm (Pseudoterranova decipiens) infection in an indicator host, American plaice (Hippoglossoidesplatessoides), is being prepared for primary publication. Sealwonn levels were analysed in >27,000 plaice collected in surveys conducted biennially between 1980 and 1990, throughout eastern Canada. Analyses revealed that sealwom levels in plaice from the Gulf of St. Lawrence, Breton Shelf, Scotian Shelf and Gulf of Maine have increased significantly over the course of the time series.

Examinations of plaice collected in a 1993 survey of Scotian-Fundy fisheries are also near completion. In this most recent survey, >2,000 plaice were collected from 17 locations on the Breton and Scotian shelves and in the Gulf of Maine during 1993 March and July Groundfish cruises and a dedicated cruise in August 1993. Preliminary analyses indicate that sealwom levels have stabilized or declined since the previous (1989-90) survey.

Seal Urey surveys

More than 10,000 fish collected from eastern Nova Scotia, Sable Island Bank, and southwestern Nova Scotia between 1989 and 1993 have been examined for larval anisakines. Sealwonn larvae occurred in 30 of 38 species surveyed and were most abundant in mature monkfish (Lophius americanus), cusk (Brosme brosme), cod (Gadus morhk) and sea raven (Hemitripterus americanus). Highest densities of infection @ = no. of nematodes kg-' host wt.) however, were found in small benthic consumers such as fourbeard rockling (Enchelyopus cimbrius) (D=117), hookear sculpin (Artediellus atlanticus) (D=375), juvenile H. americanus (D = 157) and longhorn sculpin (Myoxocephalus octodecemspinosus) @=187), mustache sculpin (Triglops murrayi) @=405), alligatorfish (Aspidophoroides rnonopterygius) @=285) and juvenile H. platessoides @= 186).

Additional samples of rockling, hookear and mailed sculpin, alligatorfish, spiny lumpsucker (Eumicronemus spinosus) and juvenile American plaice were collected from Sable Island Bank on a dedicated cruise in May '94. They were examine, for sealworm by pepsin-HC1 digestion.

Parasites as biological indicators of fish stocks

In collaboration with D. Marcogliese and R. Arthur of IML, a working paper on use of parasites as biological tags for groundfish stocks from the Gulf of St. Lawrence was presented at a Fisheries Oceanography Committee symposium on stock migration and mixing. A paper on the use of larval anisakine nematodes in discrimination of migrant cod fiom the southeastem and southwestem Gulf of St. Lawrence and resident Breton Shelf cod (with D. Marcogliese) was subsequently submitted for primary publication.

We also documented the prevalence and abundance of nodules of the microsporidean parasite Pleistophora hippoglossoides in the flesh of American plaice collected during the 1993 sealwom index survey. The distribution of this parasite, yet another cosmetic nuisance for fish processors, is clearly influenced by environmental temperature. It is most prevalent in plaice from the warmer waters of the southwestern Scotian Shelf and Gulf of Maine but seldom found in Gulf of St. Lawrence or Newfoundland plaice.

Consultations

Through the years we have provided parasite identifications and background information on parasitic infections in fish and shellfish for various agencies and individuals including DFO's Fish Inspection Branch (headquarters, regional and local), Benthic Fisheries and Aquaculture Division, and Freshwater and Anadromous Division, the provincial Fisheries Departrnents of Nova Scotia and Newfoundland, Health and Welfare Canada, and the US Department of Health, Education and Welfare, Food and Drug Administration, fish processors, fishermen, university researchers, students and seafood consumers.

These consultations have involved a wide variety of parasite problems. Over the past year we have identified and provided background information on:

- larval whaleworm (Anisakis simplex) in Barents Sea cod (National Sea Products) - acanthocephalans (Echinorhynchus lateralis) in brook trout from Kings Co., N.S. (recreational fisherman and N.S. Dept. of Natural Resources)

- "gullwonn", plerocercoid larvae of the economically important tapeworm Diphyllobothrium dendriticum (N.S. Dept. Natural Resources Fact Sheet)

- copepods (Ergasilus labracis) parasitic on the gills of striped bass (Morone saxatilis)

- copepods (Salmincola carpionis) parasitic on the gills of Cape Breton brook trout (recreational fishermen)

- copepods (Pennella filosa) parasitic in swordfish flesh

- cystacanths of Corynosoma wegeneri and larval sealworm and whaleworm in the body cavity and flesh of halibut (Hippoglossus hippoglossus) brood stock (Aquaculture Section, St. Andrew's Biological Station)

- harpacticoid copepods commensal on lobster (Benthic Fisheries)

During our May 1994 cruise we also collected various species of live invertebrates and fish for public display in the "Touch Tank" on the Halifax waterfront and live haddock and winter flounder as brood stock for local mariculturists.

Surveys of Parasites in Seals

Anisakine infections 'in Sable Island and mainland mey and harbour seals

A contractor, Dr. Gwyneth Jones, has identified and counted nematodes from the gastrointestinal tracts of 640 seals including the majority (350 of 416) of grey seals sarnpled on Sable Island in 1989 and >200 harbour seals collected in the Lower Bay of Fundy between 1989 and 1992. Dr. Jones is currently completing identifications of anisakines from 160 grey seals taken fi-om eastern Nova Scotia between 1989 and 1992.

Anisakine infections and diets of recently weaned mey seals

A manuscript on anisakine infections and food in the stomachs of 5-8 wk-old Sable Island grey seals is being prepared for primary publication (with B. Beck). The study shows that grey seals begin to accumulate P. decipiens, A. sirnplex and Contracaecum osculatum within 3-5 weeks of weaning, when their diet consists almost entirely of invertebrates (polychaetes, crustaceans, and echinoderms).

Life Cvcle Studies

A paper comparing the diets of sympatric flatfish species which differ radicaily in regard to larval sealworm infection levels has been published and a related paper on the natural transmission of sealworm to Sable Island Bank flatfish is now in press.

Size and momhometry of sealworm larvae from fish hosts at different trophic levels

Pepsin-HC1 digestion was employed in the recovery of larval sealworm from fourbeard rockling, hookear sculpin, mailed sculpin, alligatorfish, spiny lumpsucker and juvenile plaice collected during our August 1993 and May 1994 surveys. This approach yielded P. decipiens larvae as small as 2 mm in length. Sealworm <10 mm in length were common in 2-3-yr-old plaice (<20 cm TL), but rarely found in older (larger) fish.

To date, we have documented body lengths and other characteristic dimensions of >2,000 larval sealworm from small benthophagous fish and larger demersal fish, such as mature plaice and cd. Our results show that the parasite grows to the infective stage in small benthic fish rather than in invertebrate hosts and that larger, commercially important fish become infected primariiy through consumption of smaller fish hosts.

Serial transmission of larval sealworm to fish hosts

A manuscript describing serial transmissions of laboratory hatched sealworm larvae through benthic copepod and amphipod intermediaries to various fish species and subsequent growth and development of the nematodes in fish hosts has been accepted for publication in the Margolis commemorative volume of the Canadian Journal of Fisheries and Aquatic Sciences.

Larval sealworm lonrrevity and host response in American plaice

After more than two years in captivity, 27 plaice survive. There has been no evidence of a host response and comparisons of monthly photographs taken on a light table employing crossed-polarizing filters indicate that the nematodes continue to migrate through the musculature of the host. Twenty-one heavily infected juvenile plaice collected on our May '94 cruise are now established in a second holding tank.

Coccidial infection in seals

A recently published paper documents clinical and subclinical coccidial infections in harbour seals (Phoca vitulina) captured as recently weaned pups on Sable Island and housed in the Life Sciences Centre, Dalhousie University.

Variation in body lengths of mature anisakine nematodes from seals

A summer student and contractor measured the body lengths of mature P. decipiens and C. osculatum from the stomachs of Nova Scotian grey and harbour seals to determine if size, and by inference, fecundity of the nematodes is influenced by various factors such as species, age, and sex of host, seasonality, inter- and intraspecific parasite interactions, and parasite densities. Lengths of >7,ûûû mature nematodes from 40 harbour and 30 grey seals have been documented thus far.

Funnal lesions on cavtive Amencan plaice

As a consequence of more frequent cleaning of holding tanks and changes to their drainage systems, there have been no new incidents of infections with the opportunistic pathogen Cladosporiurn resinae in our captive plaice.

PARAOUA '94 Svmvosium

G. McClelland presented a guest lecture on recent research on life cycles of anisakine nematodes and CO-chaired the nematode session at a Scandinavian Parasitology Society symposium on economically important fish parasites, held in the Westman Islands, Iceland, July 2-6, 1994.

Sealworm Intervention Proaarn (SWIP & SWP II)

In the past DFO has employed what have become unprofitable and socially unacceptable commercial seal hunts, culls, and bounty prograrns in an effort to control levels of larval sealworm infections in groundfish and other seal-related problems, such as damage to fishing gear. SWIP was formed to investigate alternative approaches to these problems. The sealworm working group consists of scientists from universities, industry and DFO with expertise in pharmacology, irnmunology, reproductive physiology, parasitology, seal biology, and veterinary medicine.

Anthelmintics

The objective of this project was the development of an injectable anthelmintic with a sustaind release delivery vehicle which would not only purge P. decipiens from the of seals but would also prevent re-infection for several months.

EEcacious serum titres of ivermectin, an anthelmintic with broad veterinary application, have been achieved for periods of up to six months in rabbits on single injection of a freeze-dried liposome-ivermectin formulation suitable for administration by the 'Ballistivet Biobullet' system., The efficacy and biological half-life of this treatment are currently being tested on captive grey seals with experimental P. decipiens infections.

Immunolom

The main objective of immunological studies is the development of an anti-sealworm vaccine for fiee-living seals.

Significant progress has been made. When injected with "whole worm" extracts of P. decipiens larvae, captive grey seal pups produced to 45 and 105 kd sealworm proteins. Lower titres of these same antibodies were also detected in serum from free-living seals. The 45 kd protein, which has been fully sequenced, proved to be haemoglobin fiom the nematode's pseudocoelomic fluid. A second 10 kd protein with an allergenic activity has also been isolated from the pseudocoelomic fluid and sequenced. Both the allergen and the haemoglobin are ready to be cloned and produced synthetically.

In recent tests, again involving monitoring of faecal sealworm egg counts in control and treated seals, P. decipiens haemoglobin injected with Freund's complete adjuvant (FCA) seems to have elicited a moderate immune response resulting in partial rejection of experimental sealworm infections. Injection with a 10 kd allergen-FCA formulation, however, resulted in the complete rejection of sealworm infection in one experimental seal hos t.

Work continues on cloning the genes for the antigenic and allergenic proteins for large-scale production of recombinant protein for vaccines. Immunocontraception

In the immunocontraceptive approach to seal population control, antibodies are elicited in response to an antigen prepared from glycoproteins in the zona pellucida (ZP) layer of ova. In a ZP-immunized female, the zona pellucida is altered in such a way that it is nonreceptive to the penetration of sperm.

In recent experiments, ZP serum titres many times higher than those considered efficacious have been found in captive seals previously injected with ZP-FCA in liposome delivery vehicles. With the use of the liposome sustained release vehicle only a single injection is required.

Mature, free-living female grey seals (n=160) in the Sable Island breeding colony were injected with the ZP-FCA-liposome formulation in January '92. Immunocontraception was achieved in 15-20% of these seals and, from serum anti-ZP titres recorded in the treatment group in January 1993, 85% contraception efficiency is anticipated this year.

Publications

Marcogliese, D.J., and G. McClelland. (In press). The status of biological research on sealworm (Pseudoterranova decipiens) in eastem Canada. Can. Manusc. Rep. Fish. Aquat. Sci.

Martell, DJ., and G. McClelland. 1994. Diets of sympatric flatfishes, Hippoglossoides platessoides (Fabricius), Pleuronectes ferrugineus (Storer), Pleuronectes americanus (Walbaum), from Sable Island Bank, Canada. J. Fish Biol. 44: 821-848.

Martell, DJ., and G. McClelland. (In press). Transmission of Pseudoterranova decipiens (Nematoda: Ascaridoidea) via benthic macrofauna to sympatric flatfishes (Hippoglossoides platessoides, Pleuronectes ferrugineus, Pleuronectes americanus) on Sable Island Bank, Canada. Mar. Biol.

McClelland, G. 1993. Eimeria phocae (Apicomplexa: Eimeriidae) in harbour seals Phoca vitulina from Sable Island, Canada. Dis. Aquat. Org. 17: 1-8.

McClelland, G. (In press). Current laboratory and field research on the life cycles and population dynamics of economically important anisakine nematodes. Bull. Scand. Soc. Parasitol. (Abstrac t) McClelland, G. (In press). Experimental infection of fish with larval sealworm, Pseudoterranova decipie ns (Nematoda, Anisakinae), transmitted by arnphipods. J. Fis h. Aquat. Sci.

McClelland, G., and D.J. Marcogliese. (In press). Larval parasitic nematodes (Ascaridoidea, ) as biological indicators of cod (Gadus morhua) populations in the southern Gulf of St. Lawrence and on the Breton Shelf, eastern Canada. Bull. Scand. Soc. Parasitol.

McClelland, G., D.J. Marcogliese, and J.R. Arùiur. 1994. Parasites as indicators of Gulf of St. Lawrence fish stocks. (Fisheries Oceanography Committee (FOC) Symposium, Mar. 22-24, Dartmouth, N.S. WP94139. summarized in D'Amours, D., K.T. Frank and G. Bugden (eds.) 1994. Report of the Working Group on Oceanographic Effects on Stock Migration and Mixing - reviewed by the Fisheries Oceanography Committee (FOC). DFO Atlantic Fisheries Research Document 94/54.

FUTURE RESEARCH (1994 - )

Fish Survevs

Prepare, for primary publication, manuscripts on:

- "Long term variations in levels of sealworm (Pseudoterranova decipiens) infections in eastern Canadian cd(Gadus murhua), 1958 - 1990."

- "Geographical distribution of the microsporidean Pleistophora hippoglossoideos in American plaice, Hippoglossoides platessoides, in eastern Canada."

Prepare manuscripts for primary publication or technical reports on:

- Larval anisakine infections in gadids of eastern Canada

- Larval anisakine infections in eastern Canadian cottids

- Larval anisakine infections in pleuronectiform fish from eastern Canada

- Larval anisakine infections in miscellaneous underutilized fish species in eastern Canada 44 Seal Surveys

Complete identification and enumeration of anisakine nematodes from grey and harbour seal stomachs collected from Sable Island, eastern Nova Scotia and lower Bay of Fundy and analyze variations in nematode abundance with species, age, length and geographical origin of host and season of capture.

Complete manuscripts on:

"Variations in abundances of anisakine nematodes in the stomachs of grey (Halichoerus grypus) and harbour seals (Phoca vitulina) from the Gulf of St. Lawrence, eastern Nova Scotia and the lower Bay of Fundy."

"Discriminant analysis of anisakine nematode infections in Gulf of St. Lawrence and Atlantic grey seal (Halichoerus grypus) populations."

Life Cycle Studies

- Complete study of the morphometry of P. decipiens larvae and identification of prey of small benthophagous fish.

- Continue laboratory studies on the longevity of sealworm larvae and the fish host response and on the transmission of the microsporidean Pleistophora hippoglossoideos in live juvenile plaice.

- Continue to study variation in size and fecundity of mature anisakine nematodes from seal stomachs.

- Prepare manuscripts on:

Host diet and the transmission of the anisakine nematode Hysterothylacium aduncum to flatfish hosts.

Fungal infections (Cladosporium resinae) in captive American plaice. 45

Anthelmintics

test delivery of liposome ivermectin formulation with 'Ballistivet Biobullet' system.

begin field tests

Immunology

Complete gene cloning and synthetic production of 45 and 105 kd antigens and the 10 kd allergen for vaccines.

Test the anthelmintic activity of vaccines in captive and free-living seals.

Immunocontraception

Continue search for alternative adjuvants

Alter the liposome component of the vaccine to maximize and prolong anti-ZP antibody production

Develop a ZP-adjuvant-liposome formulation which can be contained in an 0.2 ml gelatin capsule for delivery to free-living seals with the 'Ballistivet' system

- Test seal ZP as an alternative to the porcine ZP component of the current vaccine

Clone and synthetically produce epitopes of seal ZP proteins useful in immunocontraception

Continue field testing of the ZP-FCA-liposome treatments on branded Sable Island grey seals

Test the ZP approach on captive harp seals 46 ANNEX I

LIST OF PARTICIPANTS

Gulf Region:

Sharon McGladdery

Quebec Region:

Richard Arthur Elaine Albert Dave Marcogliese Lena Measures

Newfoundland Region:

John Brattey

Scotia-Fundy Region:

Gary McClelland

Marine Laboratory, Scottish Office Agriculture and Fisheries Department:

Alistair McVicar 47 ANNEX II

4th DFO ATLANTIC PARASITOLOGIST'S WORKSHOP

NORTHWEST ATLANTIC FISHERDES CENTRE

29 SEPTEMBER, 1994

9:ûû Review of 1993/94 Activities and Future Research

Gulf Region (S. McGladdery)

Newfoundland Region (J. Brattey)

10:20 Coffee

Quebec Region (R. Arthur, D. Marcogliese, L. Measures)

Scotia-Fundy Region (G. McClelland, J. Martell)

12:ûû Lunch break

13:O Fisheries Parasitology at the SOAFD Marine Laboratory, Aberdeen (A. McVicar)

14:O Discussion

15:OO Coffee

15:20 Continuation 48 ANNEX III

REPORT OF THE MEETING TO UPDATE MODELLING OF SEALWORM DYNAMICS

ST. JOHN'S, NEWFOUNDLAND, 28 SEFTEMBER 1994

Chairman: Don Bowen (Scotia-Fundy) Rapporteur: David Marcogliese (Quebec) Participants: Elaine Albert (Quebec) Richard Arthur (Quebec) John Brattey (Nfld.) Mike Hammill (Quebec) Gary McClelland (Scotia-Fundy) Sharon McGladdery (Gulf) Lena Measures (Quebec) Becky Sjare (Nfld.) Barry Smith (Quebec) Gary Stenson (Nfld.)

Participants in the former Seal/Sealworm Ecology Program and other contributors met 28 September in St. John's, Nfld., to summarize the state of our knowledge of the biology of seals and sealworm in eastem Canada and to determine progress made in research since 1988. Gaps in our knowledge required to mode1 sealworm dynamics were outlined in des Clers and Mohn (1990). Herein, the state of current knowledge is evaluated, and preliminary groundwork is laid toward the updating of modelling efforts.

Gaps identified by des Clers and Mohn (1990), evaluation of research since 1988, and previously unidentified gaps are outlined in Table 1.

It was concluded that sufficient advances in data collection have been made to warrant another modelling effort. However, such efforts could not be fully initiated until spring 1995 when certain pertinent data sets would be available, and due to the schedules of required modelling personnel. In the meantirne, principal participants would tabulate available data for the modellers, to determine which types of models might be constructed and to develop a time table for the modelling exercise. A tentative meeting of essential personnel is scheduled for early December. It is intended to develop a suite of speculative models to point out important gaps in research, and to explore potential effects of parameters such as seal diet on parasite levels in seals, and ultimately, fish. With the data at hand, it is unlikely that predictive models could be constructed, but it should be feasible to determine the direction of change with some degree of certainty, and to use the mals to provide advice to managers.

Coordination of the modelling exercise will be assumed by the DFO CODE in Parasitology, Maurice Lamontagne Institute, Quebec, in conjunction with G. McClelland, Halifax Research Laboratory, Scotia-Fundy.

References des Clers, S., and R. Mohn (Rapporteurs). 1990. Group 4 Report: Models. p. 255-260 h:W.D. Bowen (ed.), Population biology of sealworrn (Pseudoterranova decipiens) in relation to its intermediate and seal hosts. Cm. Bull. Fish. Aquat. Sci. 222. TABLE 1. Mode1 parameters identified by des Clers and Mohn (1990). and a summary of research accompiishments since 1988.

Stage of Parameter Data Data life cycle statusa statlls' 1988 1994 Egg & crustacean Delay from egg to fish hosts SuMval egg to crustacean Crustacean mortaiity Larval mortaiity in crustacean Crustacean density Altered behaviour of crustacean Components of fish diet

Fish hosts Susceptibility to infection (habitat) ND, QN-i F Fish biomass/numbers ND, QN-iF" Time trends in fish abundance ND, QN+F Fish mortality QN+ E Fish mortaiity by parasite QL+ F Parasite martality ND, QN Proportion of infective larvae QL+ F Fish to fish transmission ND, QL+ F Time series, parasite in fish QNC Parasites in seal prey QN Parasites in benthophagous fish QNd Seal hosts Transmission rate from fish to seais Seal numbers Seal seasonal behaviour Seal mortality Larval mortality in seal Adult mdity in seal Maturation time Egg production Density dependency in egg production Prob. distribution indiv. balworm burdens Prob. dismbution, indiv. adult worm burdens Diet composition by wt/no, per fish sp., season

Abiotic Bottom temperature, seasonal and regional data

QN = quantitative estimate QL = qualitative estimate O = obtainable from existing data F = further research requird U = unlikely to be obtainable in the foreseeable future NC = not considered ND = new data available Available for flatfish on the Scotian Shelf " QN for commercially important species; U for many non-commercial ones Availability of data varies by geographic area and season