ANTIBODIES AGAINST INFLUENZA VIRUS TYPES A AND B IN CANADIAN SEALS.

Influenza viruses have been reported from marine mammals worldwide, particularly in pinnipeds, and have caused mass mortalities of seals in North America and Europe. Because influenza viruses in marine mammals can be zoonotic, our objective was to examine Canadian phocids for exposure to influenza A and B viruses in order to understand health risks to wild populations as well as to humans who consume or handle these animals. Blood was collected from 394 seals in eastern Canada from 1994 to 2005. Sera were screened for exposure to influenza viruses in three resident species of seals: harbour, Phoca vitulina (n=66); grey, Halichoerus grypus (n=82); ringed, Phoca hispida (n=2); and two migrant species: harp, Pagophilus groenlandica (n=206) and hooded, Cystophora cristata (n=38). Included were samples from captive grey (n=1) and harbour seals (n=8) at two aquaria. Sera were prescreened using indirect enzyme-linked immunosorbent assay (ELISA), and antibodies against influenza A virus were confirmed using a commercial competitive ELISA (IDEXX Europe B.V.). A subset of influenza A virus positive sera was used to determine common virus subtypes recognized by sera using reference strains. All positive sera in the indirect ELISA reacted with influenza A virus subtypes H3, H4, and H10 using a hemagglutination inhibition assay. Sera from harbour, grey, harp, and hooded seals had antibodies against influenza A and influenza B viruses (some cross-reactivity occurred). Overall, 33% (128/385) of wild seals were seropositive to influenza viruses, with the highest seroprevalence in harp (42%) followed by harbour (33%), grey (23%), and hooded (11%) seals. Antibodies were detected in both sexes and most age classes of wild seals. Two of eight captive harbour seals were seropositive to influenza B virus and four had cross-reactions to influenza A and B viruses. This study reports antibodies against influenza A and B viruses in four seal species from the same geographic area in eastern Canada.

Toxoplasma gondii, Sarcocystis sp. and Neospora caninum-like parasites in seals from northern and eastern Canada: potential risk to consumers.

Zoonotic parasites of seals that are harvested for food may pose a health risk when seal meat or organ tissues of infected animals are eaten raw or undercooked. In this study, 124 tissue samples from 81 seals, comprising four species, were collected from northern and eastern Canada. Tissues from 23 ringed seals (Pusa hispida), 8 hooded seals (Cystophora cristata), 21 harp seals (Pagophilus groenlandicus), and 29 grey seals (Halichoerus grypus) were tested for parasites of the Sarcocystidae family including Toxoplasma gondii, Sarcocystis spp., and Neospora spp. using nested PCR followed by Sanger sequencing. Toxoplasma gondii DNA was present in 26% of ringed seals, 63% of hooded seals, 57% of harp seals, and 31% of grey seals. Sarcocystis sp. DNA was found in 9% of ringed seals, 13% of hooded seals, 14% of harp seals, and 4% of grey seals, while N. caninum-like DNA was present in 26% of ringed seals. While it is unclear how pinnipeds may become infected with these protozoans, horizontal transmission is most likely. However, one harp seal pup (4 days old) was PCR-positive for T. gondii, suggesting vertical transmission may also occur. Phylogenetic analysis of the 18S gene region indicates that Sarcocystis sp. in these seals belongs to a unique genotype. Furthermore, this study represents a new host report for T. gondii in harp seals, a new host and geographic report for N. caninum-like parasites in ringed seals, and four new hosts and geographic reports for Sarcocystis sp. These results demonstrate that parasites of the Sarcocystidae family are prevalent in northern and eastern Canadian seals. While the zoonotic potential of Sarcocystis sp. and the N. caninum-like parasite are unclear, consumption of raw or undercooked seal meat or organ tissues pose a risk of T. gondii infection to consumers.

Toxoplasma gondii infection in stranded St. Lawrence Estuary beluga Delphinapterus leucas in Quebec, Canada.

The St. Lawrence Estuary (SLE) beluga Delphinapterus leucas in Quebec, Canada, is endangered due to intensive hunting in the 19th and 20th centuries and subsequent anthropogenic contamination and human activities in the region. Infectious disease is a primary cause of death in this population. The protozoan parasite Toxoplasma gondii is reported in numerous marine mammal species, including beluga. In the present study, 55 tissue samples (heart and brain) collected from 34 stranded SLE beluga were analysed by PCR followed by DNA sequencing and restriction fragment length polymorphism analysis (RFLP) to determine the PCR prevalence and genotypes of T. gondii in these beluga. Of 34 beluga tested, 44% were positive for T. gondii by PCR, with males having a higher prevalence of infection than females and with more infected neonates and juveniles than adults. Molecular analyses indicated that all T. gondii infecting stranded SLE beluga grouped into genotype II, which predominates in humans. While our results indicate that a high prevalence of stranded beluga are PCR-positive for T. gondii infection, very few deaths are attributed to toxoplasmosis based on published necropsy results. Toxoplasma gondii can cause a range of diseases, including neurological deficits, and more data are needed to investigate this parasite's effect on population recovery.

Multispecies mass mortality of marine fauna linked to a toxic dinoflagellate bloom.

Following heavy precipitation, we observed an intense algal bloom in the St. Lawrence Estuary (SLE) that coincided with an unusually high mortality of several species of marine fish, birds and mammals, including species designated at risk. The algal species was identified as Alexandrium tamarense and was determined to contain a potent mixture of paralytic shellfish toxins (PST). Significant levels of PST were found in the liver and/or gastrointestinal contents of several carcasses tested as well as in live planktivorous fish, molluscs and plankton samples collected during the bloom. This provided strong evidence for the trophic transfer of PST resulting in mortalities of multiple wildlife species. This conclusion was strengthened by the sequence of mortalities, which followed the drift of the bloom along the coast of the St. Lawrence Estuary. No other cause of mortality was identified in the majority of animals examined at necropsy. Reports of marine fauna presenting signs of neurological dysfunction were also supportive of exposure to these neurotoxins. The event reported here represents the first well-documented case of multispecies mass mortality of marine fish, birds and mammals linked to a PST-producing algal bloom.

HERPESVIRUSES INCLUDING NOVEL GAMMAHERPESVIRUSES ARE WIDESPREAD AMONG PHOCID SEAL SPECIES IN CANADA.

Little is known about herpesviruses in Canadian pinnipeds. We measured prevalence of antibodies to herpesviruses in the sera from Canadian phocid seals by an indirect enzyme-linked immunosorbent assay. Wild harbor seals (Phoca vitulina) and captive harbor seals were positive for antibodies to Phocid herpesvirus 1 (PhoHV-1) at prevalences of 91% and 100%, respectively. Sera from wild hooded seals (Cystophora cristata), harp seals (Pagophilus groenlandica), and grey seals (Halichoerus grypus) were positive for antibodies to PhoHV-1 antigenically related herpesvirus antigens at 73%, 79%, and 96%, respectively. We isolated new herpesviruses in cell culture from two hunter-harvested ringed seals (Pusa hispida) in poor body condition from Ulukhaktok, Northwest Territories, Canada; one lethargic hooded seal from the St. Lawrence Estuary, Québec, Canada; and one captive, asymptomatic harp seal from the Magdalen Islands, Québec. Partial sequencing of the herpesvirus DNA polymerase gene revealed that all four virus isolates were closely related to PhoHV-2, a member of the Gammaherpesvirinae subfamily, with nucleotide similarity ranging between 92.8% and 95.3%. The new seal herpesviruses were genetically related to other known pinniped herpesviruses, such as PhoHV-1, Otariid herpesvirus 3, Hawaiian monk (Monachus schauinslandi) seal herpesvirus, and Phocid herpesvirus 5 with 47-48%, 55%, 77%, and 70-77% nucleotide similarities, respectively. The harp seal herpesvirus and both ringed seal herpesviruses were almost identical to each other, whereas the hooded seal herpesvirus was genetically different from the three others (92.8% nucleotide similarity), indicating detection of at least two novel seal herpesviruses. These findings are the first isolation, partial genome sequencing, and identification of seal gammaherpesviruses in three species of Canadian phocid seals; two species of which were suspected of exposure to one or more antigenically related herpesviruses based on serologic analyses.

Morbidity and mortality in stranded Cook Inlet beluga whales Delphinapterus leucas.

The endangered Cook Inlet (Alaska, USA) stock of beluga whales Delphinapterus leucas declined 47% between 1994 and 1998, from an estimated 653 whales to 347 whales, with a continued decline to approximately 312 in 2012. Between 1998 and 2013, 164 known dead strandings were reported by the National Marine Fisheries Service. Only 38 of these animals, or 23% of the known stranded carcasses, were necropsied. Carcasses were found between April and October. The majority of animals necropsied were adults (n=25), followed by juveniles (n=6), calves (n=3), and aborted fetuses (n=4). Eight of the 11 mature females were pregnant, post-partum, or lactating. Many (82%) of these belugas were in moderate to advanced autolysis, which hampered determination of a cause of death (COD). Each animal had a single primary COD assigned within a broad set of categories. The CODs were unknown (29%), trauma (18%), perinatal mortality (13%), mass stranding (13%), single stranding (11%), malnutrition (8%), or disease (8%). Other disease processes were coded as contributory or incidental to COD. Multiple animals had mild to moderate verminous pneumonia due to Stenurus arctomarinus, renal granulomas due to Crassicauda giliakiana, and ulcerative gastritis due to Anisakis sp. Each stranding affords a unique opportunity to obtain natural history data and evidence of human interactions, and, by long-term monitoring, to characterize pathologies of importance to individual and population health.

A twenty-one year temporal trend of persistent organic pollutants in St. Lawrence Estuary beluga, Canada.

Persistent organic pollutants (POPs) were measured in blubber from 144 stranded adult belugas (Delphinapterus leucas) found on the shores of the St. Lawrence Estuary (SLE) between 1987 and 2007. Temporal trends of POP concentrations (ln transformed) in beluga were described by three models, zero slope (ZS), linear (L) and two-segment piecewise (PW). Often two but sometimes all three models were equivalent in describing temporal trends based on Akaike's Information Criterion for small sample sizes. Over this 21-year time period, concentrations of most legacy POPs, including PCBs, DDTs and HCHs, exhibited relatively weak (≤11% per year) but significant decreasing trends in beluga. For PBDEs, temporal trends were best described by a PW model, characterizing a rapid increase until 1997-1998 followed by a slower increase for males and a steady-state for females. Potential cofactors such as blubber lipid content and carcass state of preservation did not show any significant temporal trends over the time period considered. Nitrogen stable isotope ratios (δ(15)N) in beluga liver, a proxy of trophic level, could not be associated to any effect on temporal trends of POP concentrations in beluga. Several POPs exhibited significant relationships with age of beluga and data were age-adjusted. Temporal trends of POP concentrations adjusted for age of beluga were reassessed but results were essentially identical as those obtained with the original POP data. Overall, POP temporal trends observed in SLE beluga are consistent with changes expected from regulations and restrictions in the use of these compounds in developed countries.

Marine mammal strandings and environmental changes: a 15-year study in the St. Lawrence ecosystem.

Understanding the effects of climatic variability on marine mammals is challenging due to the complexity of ecological interactions. We used general linear models to analyze a 15-year database documenting marine mammal strandings (1994-2008; n = 1,193) and nine environmental parameters known to affect marine mammal survival, from regional (sea ice) to continental scales (North Atlantic Oscillation, NAO). Stranding events were more frequent during summer and fall than other seasons, and have increased since 1994. Poor ice conditions observed during the same period may have affected marine mammals either directly, by modulating the availability of habitat for feeding and breeding activities, or indirectly, through changes in water conditions and marine productivity (krill abundance). For most species (75%, n = 6 species), a low volume of ice was correlated with increasing frequency of stranding events (e.g. R(2)adj = 0.59, hooded seal, Cystophora cristata). This likely led to an increase in seal mortality during the breeding period, but also to increase habitat availability for seasonal migratory cetaceans using ice-free areas during winter. We also detected a high frequency of stranding events for mysticete species (minke whale, Balaenoptera acutorostrata) and resident species (beluga, Delphinapterus leucas), correlated with low krill abundance since 1994. Positive NAO indices were positively correlated with high frequencies of stranding events for resident and seasonal migratory cetaceans, as well as rare species (R(2)adj = 0.53, 0.81 and 0.34, respectively). This contrasts with seal mass stranding numbers, which were negatively correlated with a positive NAO index. In addition, an unusual multiple species mortality event (n = 114, 62% of total annual mortality) in 2008 was caused by a harmful algal bloom. Our findings provide an empirical baseline in understanding marine mammal survival when faced with climatic variability. This is a promising step in integrating stranding records to monitor the consequences of environmental changes in marine ecosystems over long time scales.

First description of the gravid female of Philometra rubra (Leidy, 1856) (Nematoda: Philometridae), a parasite of the abdominal cavity of temperate basses Morone Spp. (Moronidae: Perciformes) in North America.

Gravid females of the little-known nematode species Philometra rubra ( Leidy, 1856 ) (Philometridae) are described from specimens from the abdominal cavity of the striped bass, Morone saxatilis (Walbaum), in South Carolina and Canada. The specimens were studied with the use of light and scanning electron microscopy. This species is mainly characterized by the distribution and different sizes of cephalic papillae from the external circle, which is a unique feature compared to other philometrids. Other characteristic features are the shape of the posterior end of body, size and location of caudal projections, and the presence of a well-developed anterior esophageal inflation. The morphology of the male of P. rubra and the life cycle of this nematode are still unknown.

Bronchoalveolar lavage and pulmonary histopathology in harp seals (Phoca groenlandica) experimentally infected with Otostrongylus circumlitus.

The objective of this study was to characterize pathologic changes associated with experimental infection of harp seals (Phoca groenlandica) with the lungworm Otostrongylus circumlitus (Metastrongyloidea: Crenosomatidae). The leukocyte differential cell count in samples obtained by unguided bronchoalveolar lavage (BAL) and the intensity of the histologic lesions in the lungs were assessed in seven harp seals experimentally exposed to 300 infective, third-stage O. circumlitus larvae. Seven unexposed harp seals were used as controls. First-stage larvae were observed in the feces of three of the seven exposed seals at 38, 42, and 45 days postexposure (dpe). Adult nematodes were found in the right primary bronchi of two of these three seals at necropsy 53 dpe. Fifty-six BALs were performed on the 14 seals. No statistical difference was observed between the exposed and control seals and among the four sampling times in percentage of neutrophils and macrophages in the BAL fluid. A significant difference was observed between the exposed and control seal groups in the percentage of eosinophils (P<0.0001), the count of eosinophils having increased by a factor of 70.4 in exposed seals. Significant statistical differences were observed between exposed and control seals in intensity of interstitial inflammation (P=0.001), bronchitis (P=0.02), bronchiolitis (P=0.04), alveolitis (P=0.03), and interstitial granulomatous inflammation (P=0.04). Our findings showed that harp seals are susceptible to infection with O. circumlitus. However, parasitic infections were transient and of low intensity, at least under our experimental conditions.

Infectivity of Toxoplasma gondii in northern traditional (country) foods prepared with meat from experimentally infected seals.

Serological and clinical evidence of human toxoplasmosis in the Canadian Arctic indicates a food safety risk associated with the consumption of wild game meat. Such meat often is eaten raw or partially cooked in locally prepared traditional (country) foods, but no data have been collected to describe survival of Toxoplasma gondii forms in these foods. The muscle of grey seals (Halichoerus grypus) experimentally infected with T. gondii oocysts was used to prepare three country foods: igunaq, a fermented product; nikku, a dried product; and sausage, a salted and spiced product. Igunaq and nikku were stored at 4 degrees C and bioassayed in cats at 49, 95, and 140 days postpreparation (DPP) and 41, 84, and 132 DPP, respectively. Raw and cooked sausages were stored at -20 degrees C and bioassayed at 50, 92, and 141 DPP. The source seal meat was infective for cats, but none of the foods prepared with this meat were infective for cats. Some cooked sausages did not reach internal temperatures considered lethal for T. gondii. Data from studies in domestic animals suggested that the negative results in this experiment were due to temperature and duration of storage. Because of the possibility that T. gondii of arctic origin might be more freeze tolerant than the swine-origin isolate used in this experiment, additional studies are necessary to clarify the risks of toxoplasmosis associated with consumption of arctic country foods.

Dioctophyme renale (Goeze, 1782) in the abdominal cavity of a domestic cat from Brazil.

This study reports a case of parasitism by the giant kidney worm, Dioctophyme renale (Goeze, 1782), in the abdominal cavity of a domestic cat from Brazil. A female adult cat presenting prostration, dehydration, physical debility, pronounced jaundice and ascitis, was taken to the Department of Animal Parasitology of the Veterinary Institute of the Universidade Federal Rural do Rio de Janeiro, Brazil. Clinical signs suggested a case of peritonitis. The cat's clinical condition was grave and death occurred within a few days. During necropsy, a brownish-red nematode, 24.9cm long, was found in the abdominal cavity and was identified as a male adult D. renale. This study reports the first confirmed case of dioctophymatosis in the domestic cat. The parasite's aberrant location in the abdominal cavity suggests that the domestic cat is not a suitable host.

Use of a SLAM transfected Vero cell line to isolate and characterize marine mammal morbilliviruses using an experimental ferret model.

Two ferrets (Mustela putorius furo) were experimentally infected with phocine distemper virus (PDV), from the 1988 seal epizootic in Europe, in order to determine whether the stable transfected Vero cell line (Vero.DogSLAMtag) expressing canine "signaling lymphocyte activation molecules" (SLAM; CD150) receptors, was more suitable for isolating and characterizing PDV when compared with Vero (American Type Culture Collection # C1008) and primary seal kidney (PSK) cells. Both ferrets displayed characteristic clinical signs of distemper, including fever and rash, 10 days postinoculation (dpi) and, due to increased morbidity, they were euthanized 12 dpi. Histologic lesions, suggestive of infection with morbilliviruses, were observed in tissues from both ferrets, and the tissues stained positive using immunohistochemistry. Isolation of PDV from isolated peripheral blood lymphocytes (PBLs), taken at 5 and 10 dpi, was achieved by cocultivation with Vero and PSK cells, following several passages. Cytopathic effects (CPE) were observed in Vero cell cultures at 29 dpi and in PSK cell cultures at 22 dpi. Phocine distemper virus was isolated from frozen infected ferret lung tissue within 48 hr, when isolation was attempted using the Vero.DogSLAMtag cell line. In addition, a reverse transcriptase polymerase chain reaction (RT-PCR) test was developed to detect a 114 base pair (bp) portion of the nucleocapsid gene found only in PDV. This RT-PCR methodology was used to confirm the identity of the virus subsequently isolated from the ferrets. Viral isolates from the infected ferrets, as well as cultures of virus originally isolated from a dolphin and a porpoise and maintained in Vero cells, also replicated faster and produced higher titers of virus when propagated in Vero.DogSLAMtag cells. These results indicate that Vero.DogSLAMtag cells offer a substantial improvement (including faster viral replication resulting in primary viral isolation in a shorter period of time, and higher yield of virus finally obtained) over traditional cell culture methodologies for isolation and characterization of marine mammal morbilliviruses.

Redescription of Synthesium pontoporiae n. comb. with notes on S. tursionis and S. seymouri n. comb. (Digenea: Brachycladiidae Odhner, 1905).

Synthesium pontoporiae n. comb. is redescribed, together with Synthesium tursionis and Synthesium seymouri n. comb.; the parasites were obtained from stranded and accidentally caught cetaceans. The sucker ratio (ratio between widths of the oral and ventral suckers) in S. pontoporiae was 1:1.8-3.0 (mean 1:2.2); in S. tursionis was 1:0.8-1.2; and in S. seymouri was 1:0.5-0.7. Synthesium pontoporiae differed from its congeners by additional diagnostic characters, including: oval to lobed testes; small cirrus with pyriform proximal region and flexible, tubular distal region formed by evagination of ejaculatory duct; and vitellarium in small follicles extending from the level of the seminal vesicle to the posterior extremity of the body and not forming dendritic radial bunches. Data on the morphology of adult S. pontoporiae and S. tursionis were inferred from confocal laser microscopical observations.

Temporal trends (1987-2002) of persistent, bioaccumulative and toxic (PBT) chemicals in beluga whales (Delphinapterus leucas) from the St. Lawrence Estuary, Canada.

Temporal trends of persistent, bioaccumulative and toxic (PBT) chemicals were examined in beluga whales (Delphinapterus leucas) from the St. Lawrence Estuary (SLE), Canada. Blubber samples of 86 adult belugas were collected from animals stranded on the shore of the SLE between 1987 and 2002 and analyzed for several regulated PBTs, including polychlorinated biphenyls (PCBs), p,p'dichlorodiphenyltrichloroethane (DDT) and its metabolites, chlordane (CHL) and related compounds, hexachlorocyclohexane (HCH) isomers, hexachlorobenzene (HCB) and Mirex. In addition, time trends of tris(4-chlorophenyl)methane (TCPMe) and tris(4-chlorophenyl)methanol (TCPMOH), two compounds that may origin from DDT formulations, were also examined. Concentrations of most of the PBTs examined had exponentially decreased by at least a factor of two (half-life time (t(1/2))<15 years) in beluga between 1987 and 2002 while no increasing trends were observed for any of the PBTs measured. The decreasing trends of PBT concentrations in SLE beluga may be due to a decline in contamination of its diet following North American and international regulations on the use and production of these compounds or by a change in its diet itself or by a combination of both. Some PBTs did not exhibit any significant trends in beluga possibly because the most intense elimination phase subsequent to legislative regulations occurred prior to the 1987-2002 time period. Other chemicals, such gamma-HCH, did not significantly decrease likely because they are still currently used in some restricted applications. Conversely, alpha-HCH showed a significant decreasing trend indicating that SigmaHCHs is not representative of all HCHs. Both TCPMe and TCPMOH exhibited no trends in beluga during the time period examined. The metabolic capacity of SLE beluga has apparently accelerated the depletion of at least one PBT, namely CB-28/31. A significant relationship between the half-life of PBTs in beluga and log Kow was observed for most of the chemicals examined. Several factors are expected to have influenced the temporal changes of PBT concentrations in beluga which limit the usefulness of this species as a bioindicator of changes in PBT contamination in the SLE ecosystem.

Use of end-tidal partial pressure of carbon dioxide to predict arterial partial pressure of carbon dioxide in harp seals during isoflurane-induced anesthesia.

OBJECTIVE: To evaluate the relationship between end-tidal partial pressure of CO(2) (ETCO(2)) and PaCO(2) in isoflurane-anesthetized harp seals. ANIMALS: Three 5-month-old 25- to 47-kg harp seals (Phoca groenlandica). PROCEDURES: PaCO(2) was determined in serial arterial samples from isoflurane-anesthetized seals and compared with concomitant ETCO(2) measured with a side-stream microstream capnograph. Twenty-four paired samples were subjected to linear regression analysis and the Bland-Altman method for assessment of clinical suitability of the 2 methods (ie, PaCO(2) and ETCO(2) determinations). The influence of ventilation rate per minute (VR) on the ETCO(2) to PaCO(2) difference (P[ET-a] CO(2)) was examined graphically. RESULTS: The correlation coefficient between the 2 measurements was 0.94. The level of agreement between ETCO(2) and PaCO(2) varied considerably. Values of ETCO(2) obtained with a VR of < 5 underestimated PaCO(2) to a greater degree (mean bias, -4.01 mm Hg) and had wider limits of agreement of -13.10 to 5.07 mm Hg (-4.01 mm Hg +/- 1.96 SD), compared with a VR of > or = 5 (mean bias, -2.24 mm Hg; limits of agreement, -7.79 to 3.30 mm Hg). CONCLUSIONS AND CLINICAL RELEVANCE: These results indicate that a microstream sidestream capnograph provides a noninvasive, sufficiently accurate estimation of PaCO(2) with intermittent positive ventilation at a VR > or = 5 in anesthetized harp seals.

Hematology and serum chemistry of harp (Phoca groenlandica) and hooded seals (Cystophora cristata) during the breeding season, in the Gulf of St. Lawrence, Canada.

Standard hematologic and serum chemistry parameters were determined from 28 harp seals (Phoca groenlandica) and 20 hooded seals (Cystophora cristata) sampled from 6 March 2001 to 13 March 2001 during the breeding season. Whole blood was collected immediately postmortem from harp seal mother-pup pairs and from six hooded seal pups, and from live-captured adult hooded seals and three hooded seal pups; blood was analyzed within 24 hr at a local human hospital. A certified veterinary laboratory validated subsamples of whole blood and analyzed all serum chemistry parameters. Significant interlaboratory differences in mean values of packed cell volume (PCV) and mean cell volume (MCV) were found. Significant differences were found between samples from the five seal groups (adult male hooded seals, lactating female hooded seals, unweaned hooded seal pups; lactating female harp seals, and unweaned harp seal pups) for hematology and most serum chemistry parameters. In general, age-class influenced mean values of PCV, hemoglobin (HB), red blood cell (RBC) counts, MCV, mean cell hemoglobin (MCH), mean cell hemoglobin concentration (MCHC), and nucleated red blood cell (NRBC) counts per 100 leucocytes, but most age-related variations were species specific. Harp seal pups had significantly lower mean values of HB, PCV, MCH, and MCHC than did other seal groups, and significantly lower mean RBC counts than did hooded seal pups. Mean NRBC counts per 100 leukocytes were more than three times higher in harp seal pups than in hooded seal pups, but this difference was not statistically significant. Mean MCV were significantly lower in harp and hooded seal pups compared to those of adult harp and hooded seals. Differences in hemograms between pup species were likely because of the precocious development of hooded seal pups, which are weaned within 4 days, compared to 12 days for harp seal pups. Among adult seal groups, male hooded seals had significantly higher mean values of PCV and HB than did female harp and hooded seals, and significantly higher mean RBC counts than did adult female hooded seals. Among adult females, mean values of MCH and MCHC were statistically higher in hooded seals than in harp seals. Adult female harp and hooded seals did not differ significantly in other RBC parameters and mean leukocyte counts. Mean values of glucose, blood urea nitrogen, total bilirubin, alanine aminotransferase (ALT), alkaline phosphatase (ALP), total protein, and albumin showed species-specific variations between adults and pups. Except for ALP, few significant differences in mean enzyme activities of aspartate aminotransferase (AST), ALT, creatine kinase and gamma-glutamyltransferase were found between seal groups. Mean concentrations of electrolytes (calcium, phosphorus, sodium, potassium, chloride, magnesium, and total carbon dioxide) varied with age class, but variations in potassium and magnesium were species specific. Harp seal pups had significantly higher mean phosphorus and potassium levels compared to other seal groups.

The effects of two dosages of midazolam on short-duration anesthesia in the harp seal (Phoca groenlandica).

The purpose of this study was to provide safe anesthesia for bronchoalveolar lavage and assess the utility of premedication with i.m. midazolam for short-duration anesthesia with isoflurane in harp seals (Phoca groenlandica). Fourteen yearling harp seal pups were anesthetized three times each as part of a prospective, cross-over, blinded study. Each animal received i.m. premedication with saline, low-dose, or high-dose midazolam (respectively 0.1 and 0.2 mg/ kg). Following premedication, anesthesia was induced with 4% isoflurane in oxygen delivered through a mask and connected to a Bain non-rebreathing system. A significantly longer time was taken from the end of general anesthesia to head movement in the high-dose group compared with the saline group (P = 0.002). A significantly longer time was taken from the end of general anesthesia to ambulation in the high-dose group compared with the saline group (P = 0.006). There were no significant differences between groups in the subjective assessment of anesthetic quality or ease of intubation. Premedication with i.m. midazolam at the dosages used did prolong recovery from anesthesia, although to a degree unlikely to be significant clinically.

Seroprevalence of Toxoplasma gondii in Canadian pinnipeds.

Sera (n = 328) collected from phocids (1995-97) from the east coast of Canada, including harp seals (Phoca groenlandica), hooded seals (Cystophora cristata), grey seals (Halichoerus grypus), and harbor seals (Phoca vitulina), were diluted 1:25, 1:50, and 1:500 and tested by a modified agglutination test for antibodies to Toxoplasma gondii. Titers equal to or greater than 1:25 were considered evidence of exposure. Grey seal (11/122, 9%), harbor seal (3/34, 9%), and hooded seal (1/60, 2%) had titers of 1:25 and 1:50. Harp seals (n = 112) were seronegative. Probable maternal antibody transfer was observed in one harbor and one grey seal pup at 10 and 14 day of age, respectively. Transmission of T gondii in the marine environment is not understood. The discovery of T. gondii in marine mammals might indicate natural infections unknown because of lack of study or might indicate recent contamination of the marine environment from the terrestrial environment by natural or anthropogenic activities.

Levels and temporal trends (1988-1999) of polybrominated diphenyl ethers in beluga whales (Delphinapterus leucas) from the St. Lawrence Estuary, Canada.

Polybrominated diphenyl ethers (PBDEs) were determined in blubber samples of 54 stranded adult beluga whales (Delphinapterus leucas) collected between 1988 and 1999 in the St. Lawrence Estuary (SLE), Quebec, Canada. Summed concentrations of 10 PBDE congeners (sigmaPBDEs) measured in beluga samples varied between 20 and almost 1000 ng/g wet weight. According to the PBDE concentrations in marine mammals reported in the scientific literature, SLE belugas appear to be relatively lightly contaminated. Only a few predominant congeners (namely, PBDE-47, -99, and -100) represent on average more than 75% of sigmaPBDEs in SLE belugas. The accumulation of sigmaPBDEs in both male and female belugas showed significant exponential increase throughout the 1988-1999 time period. The time necessary for beluga to double their blubber concentration of the most prevalent PBDE congeners was no longer than 3 years. The PBDE temporal changes reported in this study are generally faster but in agreement with the trend observed in other organisms collected in Canada, such as lake trout (Salvelinus namaycush) from the Great Lakes, ringed seal (Phoca hispida), and beluga whale from the Canadian Arctic. Some changes in the pattern of PBDEs in belugas were also observed during the time period investigated. The recent and important increase of PBDE levels in SLE belugas could explain the unexpected lack of statistical difference in PBDE contamination between males and females. This suggests that to date PBDEs tend to be accumulated by both male and female belugas, masking the elimination of PBDEs by females through post-natal transfer to their offspring. This study confirms that the growing use of PBDEs as flame retardants has resulted in rising contamination of Canadian aquatic environments. Additional studies are needed to assess the toxicological implications of the PBDE tissue levels found in SLE belugas.