Mike O. Hammill

Levels and patterns of PCBs and OC pesticides in harbour and grey seals from the St Lawrence Estuary, Canada

Blubber samples from harbour (Phoca vitulina) and grey seals (Halichoerus grypus) captured in the St Lawrence Estuary were analysed for PCBs and OC pesticides. Concentrations of sigma PCB, sigma DDT, sigma CHLOR and mirex were higher in harbour than in grey seals, while sigma HCH and HCB were similar in the two species. Age vs. concentration plots showed that sigma PCB, sigma DDT and sigma CHLOR concentrations increased with age in males, but plateaued at sexual maturity in females. Concentrations of sigma HCH decreased and mirex increased with age for post-weaning animals, regardless of gender. HCB did not show age-related trends. PCB congener and OC pesticide patterns varied within harbour seals (based on gender and maturity) and between harbour and grey seals. PCB and OC concentrations in harbour seals were lower than those seen in a sample of significantly older beluga whales. Both species remain in the Estuary year-round. PCB and OC pesticide concentrations were higher in resident harbour seals than in either grey or harp seals that visit the Estuary seasonally. PCB and OC pesticide concentrations in harbour seals from the Estuary were generally higher than in harbour seals from other regions of North America. They were comparable to, or higher than those from the NE Atlantic Ocean and the North Sea, and lower than those from the Wadden and Baltic Seas.

Effects of individual polybrominated diphenyl ether (PBDE) congeners on harbour seal immune cells in vitro.

Effects of polybrominated diphenyl ethers (PBDEs) on the immune system of marine mammals are poorly understood. One important innate immune function of granulocytes is the respiratory burst which generates reactive oxygen species (ROS) used to kill engulfed microorganisms. The present study investigates in vitro the effects of BDE-47, -99 and -153, on the formation of ROS, on intracellular level of thiols, on activity and efficiency of phagocytosis and on apoptosis in granulocytes of harbour seals. Compounds were tested at four different concentrations ranging from 1.5 to 12 microM. Results showed that ROS levels, thiol levels and phagocytosis were all affected when harbour seal cells were exposed to the highest concentration (12 microM) of PBDE congeners. Apoptosis was not affected by PBDEs. The observed effects were similar in adults, pups and in the 11B7501 cell line of harbour seals.

Energetics during nursing and early postweaning fasting in hooded seal (Cystophora cristata) pups from the Gulf of St Lawrence, Canada.

Abstract In this study we measured growth and milk intake and calculated energy intake and its allocation into metabolism and stored tissue for hooded seal (Cystophora cristata) pups. In addition, we measured mass loss, change in body composition and metabolic rate during the first days of the postweaning fast. The mean body mass of the hooded seal pups (n = 5) at the start of the experiments, when they were new-born, was 24.3 ± 1.3 kg (SD). They gained an average of 5.9 ± 1.1. kg · day−1 of which 19% was water, 76% fat and 5% protein. This corresponds to an average daily energy deposition of 179.8 ± 16.0 MJ. The pups were weaned at an average body mass of 42.5 ± 1.0 kg 3.1 days after the experiment was initiated. During the first days of the postweaning fast the pups lost an average of 1.3 ± 0.5␣kg of body mass daily, of which 56% was water, 16% fat and 28% protein. During the nursing period the average daily water influx for the pups was 124.6 ± 25.8 ml · kg−1. The average CO2 production during this period was 1.10 ± 0.20 ml · g−1 · h−1, which corresponds to a field metabolic rate of 714 ± 130 kJ ·  kg−1 · day−1, or 5.8 ± 1.1 times the predicted basal metabolic rate according to Kleiber (1975). During the postweaning fast the average daily water influx was reduced to 16.1 ± 6.6 ml · kg−1. The average CO2 production in␣this period was 0.58 ± 0.17 ml · g−1 · h−1 which corresponds to a field metabolic rate of 375 ± 108 kJ · kg−1 · day−1 or 3.2 ± 0.9 times the predicted basal metabolic rate. Average values for milk composition were 33.5% water, 58.6% fat and 6.2% protein. The pups drank an average of 10.4 ± 1.8␣kg of milk daily, which represents an energy intake of 248.9 ± 39.1 MJ · day−1. The pups were able to store 73.2 ± 7.7% of this energy as body tissue.

The reproductive behavior and energetics of male gray seals (Halichoerus grypus) breeding on a land-fast ice substrate

The reproductive behavior of male gray seals (Halichoerus grypus) breeding on land-fast ice at Amet Island, Nova Scotia, was studied. Data on energy expenditure (rate of mass loss over time) were collected. The average time budget of males at Amet Island was comparable to that of land-breeding males. The behavior of males showed seasonal changes, with a decrease in the proportion of time spent in the water and an increase in agonistic behavior during the peak mating period. The estimated amount of body mass lost over the season ranged between 25.6 and 77.1 kg, and the estimated percent of initial body mass lost ranged between 7.7 and 26.5% (n=10). The maximum number of observed copulations for an individual male was nine. Only 15 out of 42 males observed during 1992 and 1993 were seen copulating. The number of observed copulations per male was strongly correlated with success in remaining close to, or attending, females (r=0.91, P<0.001, n=42). The mean duration of attendance was 4.5 ± 5.54 days (n=42). Large size was not an important factor in determining attendance success, but reproductive effort (the estimated proportion of body mass lost over the season) and success in agonistic interactions with other males were both correlated with male success.

Milk intake, growth and energy consumption in pups of ice-breeding grey seals (Halichoerus grypus) from the Gulf of St. Lawrence, Canada

In this study we document growth, milk intake and energy consumption in nursing pups of icebreeding grey seals (Halichoerus grypus). Change in body composition of the pups, change in milk composition as lactation progresses, and mass transfer efficiency between nursing mothers and pups are also measured. Mass transfer efficiency between mother-pup pairs (n=8) was 42.5±8.4%. Pups were gaining a daily average of 2.0±0.7 kg (n=12), of which 75% was fat, 3% protein and 22% water. The total water influx was measured to be 43.23±8.07 ml·kg-1·day-1. Average CO2 production was 0.85±0.20 ml·g-1·h-1, which corresponds to a field metabolic rate of 0.55±0.13 MJ·kg-1·day-1, or 4.5±0.9 times the predicted basal metabolic rate based on body size (Kleiber 1975). Water and fat content in the milk changed dramatically as lacation progressed. At day 2 of nursing, fat and water content were 39.5±1.9% and 47.3±1.5%, respectively, while the corresponding figures for day 15 were 59.6±3.6% fat and 28.4±2.6% water. Protein content of the milk remained relatively stable during the lactation period with a value of 11.0±0.8% at day 2 and 10.4±0.3% at day 15. Pups drank an average of 3.5±0.9 kg of milk daily, corresponding to a milk intake of 1.75 kg per kg body mass gained. The average daily energy intake of pups was 82.58±19.80 MJ, while the energy built up daily in the tissue averaged 61.72±22.22 MJ. Thus, pups assimilated 74.7% of the energy they received via milk into body tissue. The lactation energetics of ice-breeding grey seals is very similar to that of their land-breeding counterparts.

ENERGY INTAKE AND UTILISATION BY NURSING BEARDED SEAL (ERIGNATHUS BARBATUS) PUPS FROM SVALBARD, NORWAY

In this study we measure energy intake via milk in nursing bearded seal (Erignathus barbatus) pups and determine how this energy is allocated into metabolism and storage of new tissues. This was accomplished using longitudinal mass gain records and the doubly labelled water technique on nursing pups in combination with cross-sectional data on changes in milk composition from bearded seal mothers. The pups (n=3) were all less than a week old at the start of the experiments. Pups gained 3.3±0.4 kg·day-1 of which 50% was fat, 14% protein and 36% water. Average daily water influx for the pups was 69.5±9.0 ml · kg-1· day-1. Average CO2 production during the study period was 0.99±0.10 ml·g-1·h-1, which corresponds to a field metabolic rate of 642±67 kJ·kg-1· day-1, or 6.0±0.5 times the predicted basal metabolic rate according to Kleiber (1975). The pups drank an average of 7.6±0.5 kg of milk daily. This corresponds to a daily energy intake of 154±8 MJ, 47±14% of which was stored as new body tissue. Despite this high energy intake bearded seal pups do not get as fat as do other nursing phocids. This is in part due to their larger body size but also due to their very active aquatic lifestyle and the lower and more consistent fat content of the milk compared to other phocid species. Bearded seal mothers forage during lactation and may also be involved in teaching their pups to feed independently. All these data suggest that the lactation strategy of bearded seals differs from the phocid norm.

Toxic effects of tributyltin and its metabolites on harbour seal (Phoca vitulina) immune cells in vitro.

The widespread environmental contamination, bioaccumulation and endocrine disruptor effects of butyltins (BTs) to wildlife are well documented. Although suspected, potential effects of BTs exposure on the immune system of marine mammals have been little investigated. In this study, we assessed the effects of tributyltin (TBT) and its dealkylated metabolites dibutyltin (DBT) and monobutyltin (MBT) on the immune responses of harbour seals. Peripheral blood mononuclear cells isolated from pup and adult harbour seals were exposed in vitro to varying concentrations of BTs. DBT resulted in a significant decrease at 100 and 200 nM of phagocytotic activity and reduced significantly phagocytic efficiency at 200 nM in adult seals. There was no effect in phagocytosis with TBT and MBT. In pups, the highest concentration (200 nM) of DBT inhibited phagocytic efficiency. A reduction of tumor-killing capacity of adult natural killer (NK) cells occurred when leukocytes were incubated in vitro with 50 nM DBT and 200 nM TBT for 24h. In adult seals, T-lymphocyte proliferation was significantly suppressed when the cells were exposed to 200 nM TBT and 100 nM DBT. In pups, the proliferative response increased after an exposure to 100 nM TBT and 50 nM DBT, but decreased with 200 nM TBT and 100 nM DBT. The immune functions were more affected by BTs exposure in adults than in pups, suggesting that other unsuspected mechanisms could trigger immune parameters in pups. The toxic potential of BTs followed the order of DBT>TBT>MBT. BT concentrations of harbour seal pups from the St. Lawrence Estuary (Bic National Park) ranged between 0.1-0.4 ng Sn/g wet weight (ww) and 1.2-13.4 ng Sn/g ww in blood and blubber, respectively. For these animals, DBT concentrations were consistently below the quantification limit of 0.04 ng Sn/g ww in blood and 0.2 ng Sn/g ww in blubber. Results suggest that concentrations measured in pups are considered too low to induce toxic effects to their immune system during first days of life. However, based on our in vitro results, we hypothesize that BTs, and DBT in particular, could pose a serious threat to the immune functions in free-ranging harbour seal adults.

Transfer of fatty acids from female seal blubber via milk to pup blubber

Abstract Fatty acids were determined by gas chromatography followed by principal component analysis (PCA) and soft independent modelling of class analogy (SIMCA) in the blubber of 18 female grey seals, Halichoerus grypus, in their milk and in the blubber of their 1-week-old nursing pups. Large individual differences were observed in both blubber and milk content of fatty acids. The fatty acid composition of the milk was systematically different from the composition in maternal blubber, with higher relative amounts of the saturated acids, the monounsaturated with 20 carbon atoms and the n3 polyunsaturated, except 18:3n3. The composition of the fatty acids in the blubber of the pups was different from that of the milk. The same fatty acids that were enriched in the milk were depleted in the blubber of the pups. Therefore the fatty acid composition in the blubber of the pups was similar to that in the adults, although not identical. The results from this investigation imply that the composition of the fatty acids in the blubber of female seals and in the blubber of their pups cannot be determined directly by analysis of fatty acid composition of milk.

Groups of related belugas (Delphinapterus leucas) travel together during their seasonal migrations in and around Hudson Bay

Social structure involving long-term associations with relatives should facilitate the learning of complex behaviours such as long-distance migration. In and around Hudson Bay (Canada), three stocks of beluga whales form a panmictic unit, but have different migratory behaviours associated with different summering areas. We analysed genetic variation at 13 microsatellite loci among 1524 belugas, to test hypotheses about social structure in belugas. We found significant proportions of mother–offspring pairs throughout the migratory cycle, but average relatedness extended beyond close kinship only during migration. Average relatedness was significantly above random expectations for pairs caught at the same site but on different days or months of a year, suggesting that belugas maintain associations with a network of relatives during migration. Pairs involving a female (female–female or male–female) were on average more related than pairs of males, and males seemed to disperse from their matrilineal group to associate with other mature males. Altogether, our results indicate that relatives other than strictly parents, and especially females, play a role in maintaining a social structure that could facilitate the learning of migration routes. Cultural conservatism may limit contributions from nearby summer stocks to endangered stocks such as the Eastern Hudson Bay beluga.

Postnatal development of muscle biochemistry in nursing harbor seal (Phoca vitulina) pups: limitations to diving behavior?

Adult marine mammal muscles rely upon a suite of adaptations for sustained aerobic metabolism in the absence of freely available oxygen (O2). Although the importance of these adaptations for supporting aerobic diving patterns of adults is well understood, little is known about postnatal muscle development in young marine mammals. However, the typical pattern of vertebrate muscle development, and reduced tissue O2 stores and diving ability of young marine mammals suggest that the physiological properties of harbor seal (Phoca vitulina) pup muscle will differ from those of adults. We examined myoglobin (Mb) concentration, and the activities of citrate synthase (CS), β-hydroxyacyl coA dehydrogenase (HOAD), and lactate dehydrogenase (LDH) in muscle biopsies from harbor seal pups throughout the nursing period, and compared these biochemical parameters to those of adults. Pups had reduced O2 carrying capacity ([Mb] 28–41% lower than adults) and reduced metabolically scaled catabolic enzyme activities (LDH/RMR 20–58% and CS/RMR 29–89% lower than adults), indicating that harbor seal pup muscles are biochemically immature at birth and weaning. This suggests that pup muscles do not have the ability to support either the aerobic or anaerobic performance of adult seals. This immaturity may contribute to the lower diving capacity and behavior in younger pups. In addition, the trends in myoglobin concentration and enzyme activity seen in this study appear to be developmental and/or exercise-driven responses that together work to produce the hypoxic endurance phenotype seen in adults, rather than allometric effects due to body size.