Experimental validation of in silico predicted RAD locus frequencies using genomic resources and short read data from a model marine mammal.

BACKGROUND: Restriction site-associated DNA sequencing (RADseq) has revolutionized the study of wild organisms by allowing cost-effective genotyping of thousands of loci. However, for species lacking reference genomes, it can be challenging to select the restriction enzyme that offers the best balance between the number of obtained RAD loci and depth of coverage, which is crucial for a successful outcome. To address this issue, PredRAD was recently developed, which uses probabilistic models to predict restriction site frequencies from a transcriptome assembly or other sequence resource based on either GC content or mono-, di- or trinucleotide composition. This program generates predictions that are broadly consistent with estimates of the true number of restriction sites obtained through in silico digestion of available reference genome assemblies. However, in practice the actual number of loci obtained could potentially differ as incomplete enzymatic digestion or patchy sequence coverage across the genome might lead to some loci not being represented in a RAD dataset, while erroneous assembly could potentially inflate the number of loci. To investigate this, we used genome and transcriptome assemblies together with RADseq data from the Antarctic fur seal (Arctocephalus gazella) to compare PredRAD predictions with empirical estimates of the number of loci obtained via in silico digestion and from de novo assemblies. RESULTS: PredRAD yielded consistently higher predicted numbers of restriction sites for the transcriptome assembly relative to the genome assembly. The trinucleotide and dinucleotide models also predicted higher frequencies than the mononucleotide or GC content models. Overall, the dinucleotide and trinucleotide models applied to the transcriptome and the genome assemblies respectively generated predictions that were closest to the number of restriction sites estimated by in silico digestion. Furthermore, the number of de novo assembled RAD loci mapping to restriction sites was similar to the expectation based on in silico digestion. CONCLUSIONS: Our study reveals generally high concordance between PredRAD predictions and empirical estimates of the number of RAD loci. This further supports the utility of PredRAD, while also suggesting that it may be feasible to sequence and assemble the majority of RAD loci present in an organism's genome.

Organohalogen Contaminants and Vitamins in Northern Fur Seals (Callorhinus ursinus) Collected During Subsistence Hunts in Alaska.

During native subsistence hunts from 1987 to 2007, blubber and liver samples from 50 subadult male northern fur seals (Callorhinus ursinus) were collected on St. Paul Island, Alaska. Samples were analyzed for legacy persistent organic pollutants (POPs), recently phased-out/current-use POPs, and vitamins. The legacy POPs measured from blubber samples included polychlorinated biphenyl congeners, DDT (and its metabolites), chlorobenzenes, chlordanes, and mirex. Recently phased-out/current-use POPs included in the blubber analysis were the flame retardants, polybrominated diphenyl ethers, and hexabromocyclododecanes. The chemical surfactants, perfluorinated alkyl acids, and vitamins A and E were assessed in the liver samples. Overall, concentrations of legacy POPs are similar to levels seen in seal samples from other areas of the North Pacific Ocean and the Bering Sea. Statistically significant correlations were seen between compounds with similar functions (pesticides, flame retardants, vitamins). With sample collection spanning two decades, the temporal trends in the concentrations of POPs and vitamins were assessed. For these animals, the concentrations of the legacy POPs tend to decrease or stay the same with sampling year; however, the concentrations of the current-use POPs increased with sampling year. Vitamin concentrations tended to stay the same across the sampling years. With the population of northern fur seals from St. Paul Island on the decline, a detailed assessment of exposure to contaminants and the correlations with vitamins fills a critical gap for identifying potential population risk factors that might be associated with health effects.

Life-history stage influences immune investment and oxidative stress in response to environmental heterogeneity in Antarctic fur seals.

Immune defenses are crucial for survival but costly to develop and maintain. Increased immune investment is therefore hypothesized to trade-off with other life-history traits. Here, we examined innate and adaptive immune responses to environmental heterogeneity in wild Antarctic fur seals. In a fully crossed, repeated measures design, we sampled 100 pups and their mothers from colonies of contrasting density during seasons of contrasting food availability. Biometric and cortisol data as well as blood for the analysis of 13 immune and oxidative status markers were collected at two key life-history stages. We show that immune responses of pups are more responsive than adults to variation in food availability, but not population density, and are modulated by cortisol and condition. Immune investment is associated with different oxidative status markers in pups and mothers. Our results suggest that early life stages show greater sensitivity to extrinsic and intrinsic effectors, and that immunity may be a strong target for natural selection even in low-pathogen environments such as Antarctica.

Inter-annual variation of persistent organic pollutants (POPS) in an Antarctic top predator Arctocephalus gazella.

Persistent organic pollutants (POPs), contaminants that may bioaccumulate in upper trophic level organisms, were detected in the milk of a top predator, the Antarctic fur seal (Arctocephalus gazella). Multiparous females had significantly lower concentrations of certain POPs (trans-nonachlor, p,p'-DDE, and several PCBs) in their milk than primiparous females, likely due to the annual lactational transfer of the POP burden from mother to pup. Furthermore, there were significant interannual differences in POP concentrations in multiparous females' milk from five breeding seasons between 2000 and 2011. Decreasing trends in concentrations of certain POPs over the recent decade coincide with declining global emissions, yet atmospheric concentrations in the Antarctic are not always consistent with global trends, suggesting that additional factors may contribute to temporal trends of POPs in fur seals. Climate shifts and corresponding availability of krill over the past decade were not consistent with trends observed in POP concentrations in fur seal milk, suggesting that climate may not be a key factor. Additional mechanisms, such as variability in the geographic ranges of individual seals during overwintering migrations are discussed and should be explored further.

Hormonal responses to extreme fasting in subantarctic fur seal (Arctocephalus tropicalis) pups.

Surviving prolonged fasting implies closely regulated alterations in fuel provisioning to meet metabolic requirements, while preserving homeostasis. Little is known, however, of the endocrine regulations governing such metabolic adaptations in naturally fasting free-ranging animals. The hormonal responses to natural prolonged fasting and how they correlate to the metabolic adaptations observed, were investigated in subantarctic fur seal (Arctocephalus tropicalis) pups, which, because of the intermittent pattern of maternal attendance, repeatedly endure exceptionally long fasting episodes throughout their development (1-3 mo). Phase I fasting was characterized by a dramatic decrease in plasma insulin, glucagon, leptin, and total l-thyroxine (T(4)) associated with reductions in mass-specific resting metabolic rate (RMR), plasma triglycerides, glycerol, and urea-to-creatine ratio, while nonesterified fatty acids (NEFA) and ß-OHB increased. In contrast, the metabolic steady-state of phase II fasting reached within 6 days was associated with minimal concentrations of insulin, glucagon, and leptin; unchanged cortisol and triiodothyronine (T(3)); and moderately increased T(4). The early fall in insulin and leptin may mediate the shift to the strategy of energy conservation, protein sparing, and primary reliance on body lipids observed in response to the cessation of feeding. In contrast to the typical mammalian starvation response, nonelevated cortisol and minimal glucagon levels may contribute to body protein preservation and downregulation of catabolic pathways, in general. Furthermore, thyroid hormones may be involved in a process of energy conservation, independent of pups' nutritional state. These original hormonal settings might reflect an adaptation to the otariid repeated fasting pattern and emphasize the crucial importance of a tight physiological control over metabolism to survive extreme energetic constraints.

Persistent organic pollutant exposure and associations with gene expression in northern fur seals (Callorhinus ursinus) from St. Paul Island, Alaska.

Persistent organic pollutants (POPs) are highly lipophilic compounds that accumulate at increased concentrations in high tropic level organisms like marine mammals. Marine mammals' reliance on blubber makes them susceptible to accumulating POPs at potentially toxic concentrations. In this study, we analyzed POP concentrations, (polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs), polybrominated diphenyl ethers (PBDEs), and methoxylated-BDE (MeOBDE), in the blubber of 16 subsistence harvested sub-adult, male northern fur seals as well as assessed changes in mRNA gene expression of nine relevant biomarkers including the aryl hydrocarbon receptor, thyroid receptor-α, and adiponectin. PBDE and MeOBDE concentrations were significantly lower than PCB and OCP concentrations. A negative relationship was observed between percent lipid in the blubber and contaminant concentrations, both individual and sum. Expression changes in eight biomarkers were correlated with individual and sum contaminant concentrations. This study shows that contaminant concentrations measured are correlated to changes in expression of genes from different physiological systems, metabolism and endocrine, that are important for the regulation of blubber metabolism. Northern fur seals are reliant on blubber as an energy source during times of low food intake. Potential contaminant induced changes in blubber metabolism pathways could have significant impacts on the health of individuals during critical periods.

Validation of a homologous canine relaxin radioimmunoassay and application with pregnant and non-pregnant Northern fur seals (Callorhinus ursinus).

The primary objectives of this study were to validate a canine relaxin RIA for use in otariids and phocids and consider practical applications. For 6 captive Northern fur seal females, serum samples were grouped and examined according to pregnancy (n=13), post-partum (n=8) and non-pregnancy (n=6), and, for 2 captive Northern fur seal males, serum samples were grouped and examined together regardless of age (2 mo-15 yrs, n=6). Placental tissue was available for examination from one Northern fur seal, Steller sea lion and harbor seal. The validation process involved several steps using an acid-acetone extraction process to isolate a relaxin-containing fraction in pools of serum from each group of fur seals and placental tissue from each seal species. A relaxin-like substance was detected in extracts of pregnant, non-pregnant and male serum and placental tissue in a dose-responsive manner as increasing volumes of respective extracts or amounts of canine relaxin were introduced into the assay. In raw serum samples, mean immuno-reactive relaxin concentrations were higher (P<0.05) during pregnancy than post-partum and non-pregnancy, and lower (P<0.05) in male than female fur seals. During pregnancy, mean serum concentrations of relaxin progressively increased (P<0.05) over Months 4-10 and, in serial samples collected from the same fur seals before and after parturition, mean concentrations were higher (P<0.06) pre-partum than post-partum. In conclusion, validation of a homologous canine relaxin RIA for use in otariids and phocids resulted in the discovery of a relaxin-like substance in extracted and raw serum and placental tissue from Northern fur seals, a Steller sea lion and harbor seal. Distinctly higher immuno-reactive concentrations during pregnancy indicated the potential for relaxin to serve as a hormonal marker to differentiate between pregnant and non-pregnant or pseudopregnant pinnipeds.

Tissue distribution of polychlorinated biphenyls and organochlorine pesticides and potential toxicity to Alaskan northern fur seals assessed using PCBs congener specific mode of action schemes.

The fur seal (Callorhinus ursinus) population has decreased in their primary breeding grounds in the Bering Sea; contamination is among suspected causes. Our goal was to better understand the extent of contamination of seal tissues with certain organochlorine compounds by measuring the concentrations of polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) in fur seal tissues from St. Paul Island, to gain a better perspective of tissue congener distribution and to evaluate the observed PCB levels against toxicologically significant levels for modes of action. Concentrations of 145 PCB congeners (Sigma(145)PCBs) and 12 OCPs were measured with gas chromatography-ion trap mass spectrometry in 8 different tissues of 10 male northern fur seals. The mean concentrations of SigmaOCPs [in ng/g lipid weight (lw)] were 1180 in blubber, 985 in the heart, 1007 in the liver, 817 in the kidney, 941 in muscle, 660 in reproductive tissues, 204 in the brain, and 322 in the lung. The mean concentrations of Sigma(145)PCBs (in ng/g lw) were 823 in blubber, 777 in the liver, 732 in the heart, 646 in reproductive tissues, 638 in muscle, 587 in the kidney, 128 in the lung, and 74.3 in brain tissues. Concentrations of PCBs affecting the aryl hydrocarbon receptor expressed as total PCB toxic equivalents (SigmaPCB-TEQs) ranged from 0.3 to 545 pg/g lw for the various tissues. The major contributors to SigmaPCB-TEQs are CB-118 in muscle, brain, lung, kidney, and liver, CB-126 in blubber, and CB-118 and CB-126 equally in the heart and reproductive tissues. Concentrations of PCBs affecting Ca(2+) homeostatsis expressed as the neurotoxic equivalent (NEQ) showed SigmaPCB-NEQs ranged from 17.7 to 215 ng/g lw in all tissues. Although no composite measure of perturbation of thyroid function is available, sufficient amounts of congeners with high binding to the thyroxine transport system were present to warrant consideration of this mode of action in future studies. Analyses of 145 PCBs and mode of action evaluation suggest that PCB contamination could potentially exert an effect on the Alaskan northern fur seal population although the PCB concentrations have been decreasing in the fur seals over the last decade.

[Comparative-enzymologic study of phosphatase activity of hydrobionts from Pacific Ocean].

Activities of acid phosphatase are studied with use as substrates of phenyl phosphate, alpha- and beta-glycerophosphates in various organs and tissues of a large group of industrial hydrobionts of the Pacific basin (12 fish species, 7 invertebrate species. and one mammalian species) and of alkaline phosphatase in various organs of the Commander (Berryteuthis magister) and the New Zealand (Nototodarus sloani sloani) squids. Intertissue and interspecies differences have been revealed in the substrate and inhibitory specificity of the studied enzyme preparations. The method of isolation and a partial purification of preparations of acid phosphatase from tissue of gonads and of alkaline phosphatase from tissues of kidney and liver of individuals of industrial squid species is described.

Preliminary study of long-range transport of halogenated flame retardants using Antarctic marine mammals.

Eight PBDE congeners, three emerging brominated flame retardants, five dechloranes and eight MeO-PBDEs were monitored in tissues (muscular, adipose, brain) and fur of southern elephant seal and Antarctic fur seal of the South Shetland Islands, Antarctic Peninsula. Total PBDEs and total dechloranes concentrations ranged between n.d.-6 ng/g lw. While PBDEs were not detected in brain tissue, Dec 602 was found in brain tissue of both seal species indicating that dechloranes -with potential neurological toxicity- could cross the blood-brain barrier. Emerging brominated flame retardants were not detected in any sample and only two MeO-PBDEs, which are of natural origin, were found. The presence of the detected compounds in biota from the Antarctic evidences their long-range transportation, being of special interest the detection of emerging compounds such as dechloranes. This is the first time that these contaminants have been detected in marine mammals from the Antarctic. BDE-47 concentrations were lower than previously reported for the same species, suggesting a successful effect of the existing regulation and bans on PBDEs. CAPSULE ABSTRACT: Halogenated flame retardants were in tissues of Antarctic seals proving long-range transport. Dechloranes showed similar behaviour to PBDEs, additionally they crossed the BBB.

First detection of plastic microfibers in a wild population of South American fur seals (Arctocephalus australis) in the Chilean Northern Patagonia.

The dramatic increase of microplastics (plastic fragments <5 mm) in marine environments is a problem that has attracted public attention globally. Within the different types of microplastics, microfibres are the least studied (size <1 mm). We examined 51 female scats from a population in Northern Patagonia. Our results showed no presence of microplastic particles, however 67% of them showed a remarkable abundance of microfibers, which until now had only been reported in animals fed in captivity. As a result of this work we propose that the examination of scats from South American Fur Seal and also other pinnipeds could be an efficient tool to monitor environmental levels of microfibres and maybe microplastics in the environment due to the easy recognition of the animals and their scats.

Accumulation patterns of organochlorines in juveniles of Arctocephalus australis found stranded along the coast of Southern Brazil.

The present study was conducted to elucidate the specific distributions of organochlorine (OC) compounds in various tissues and organs of juveniles of the South American fur seal (Arctocephalus australis) found stranded on beaches in southern Brazil. The OC residue concentrations occurred in the following order: PCBs>DDTs>CHLs>HCHs>HCB. OC concentrations in blubber were higher than other tissues (e.g. PCBs, 2480 ng g(-1) wet weight; DDTs, 660 ng g(-1) wet weight) indicating a positive association with lipid content (except for HCB). However, the poor nutritional status of these animals, possibly following a starvation period, is likely to have allowed the remobilization of organochlorines stored in lipids to other tissues throughout the body, increasing their vulnerability to toxic effects and possibly affecting their survival capability.

Reproductive success is energetically linked to foraging efficiency in Antarctic fur seals.

The efficiency with which individuals extract energy from their environment defines their survival and reproductive success, and thus their selective contribution to the population. Individuals that forage more efficiently (i.e., when energy gained exceeds energy expended) are likely to be more successful at raising viable offspring than individuals that forage less efficiently. Our goal was to test this prediction in large long-lived mammals under free-ranging conditions. To do so, we equipped 20 lactating Antarctic fur seals (Arctocephalus gazella) breeding on Kerguelen Island in the Southern Ocean with tags that recorded GPS locations, depth and tri-axial acceleration to determine at-sea behaviours and detailed time-activity budgets during their foraging trips. We also simultaneously measured energy spent at sea using the doubly-labeled water (DLW) method, and estimated the energy acquired while foraging from 1) type and energy content of prey species present in scat remains, and 2) numbers of prey capture attempts determined from head acceleration. Finally, we followed the growth of 36 pups from birth until weaning (of which 20 were the offspring of our 20 tracked mothers), and used the relative differences in body mass of pups at weaning as an index of first year survival and thus the reproductive success of their mothers. Our results show that females with greater foraging efficiencies produced relatively bigger pups at weaning. These mothers achieved greater foraging efficiency by extracting more energy per minute of diving rather than by reducing energy expenditure. This strategy also resulted in the females spending less time diving and less time overall at sea, which allowed them to deliver higher quality milk to their pups, or allowed their pups to suckle more frequently, or both. The linkage we demonstrate between reproductive success and the quality of individuals as foragers provides an individual-based quantitative framework to investigate how changes in the availability and accessibility of prey can affect fitness of animals.

Regional differences in plastic ingestion among Southern Ocean fur seals and albatrosses.

We provide data on regional differences in plastic ingestion for two Southern Ocean top predators: Arctocephalus fur seals and albatrosses (Diomedeidae). Fur seals breeding on Macquarie Island in the 1990s excreted small (mainly 2-5 mm) plastic fragments, probably derived secondarily from myctophid fish. No plastic was found in the scats of these seals breeding on three islands in the southwest Indian and central South Atlantic Oceans, despite myctophids dominating their diets at these locations. Compared to recent reports of plastic ingestion by albatrosses off the east coast of South America, we confirm that plastic is seldom found in the stomachs of Thalassarche albatrosses off South Africa, but found no Diomedea albatrosses to contain plastic, compared to 26% off South America. The reasons for such regional differences are unclear, but emphasize the importance of reporting negative as well as positive records of plastic ingestion by marine biota.

Fukushima derived radiocesium in subsistence-consumed northern fur seal and wild celery.

In July 2014, our investigative team traveled to St. Paul Island, Alaska to measure concentrations of radiocesium in wild-caught food products, primarily northern fur seal (Callorhinus ursinus). The 2011 Fukushima Daiichi Nuclear Power Plant accident released radiocesium into the atmosphere and into the western Pacific Ocean; other investigators have detected Fukushima-derived radionuclides in a variety of marine products harvested off the western coast of North America. We tested two subsistence-consumed food products from St. Paul Island, Alaska for Fukushima-derived radionuclides: 54 northern fur seal, and nine putchki (wild celery, Angelica lucida) plants. Individual northern fur seal samples were below minimum detectable activity concentrations of (137)Cs and (134)Cs, but when composited, northern fur seal tissues tested positive for trace quantities of both isotopes. Radiocesium was detected at an activity concentration of 37.2 mBq (134)Cs kg(-1) f.w. (95% CI: 35.9-38.5) and 141.2 mBq (137)Cs kg(-1) f.w. (95% CI: 135.5-146.8). The measured isotopic ratio, decay-corrected to the date of harvest, was 0.26 (95% CI: 0.25-0.28). The Fukushima nuclear accident released (134)Cs and (137)Cs in roughly equal quantities, but by the date of harvest in July 2014, this ratio was 0.2774, indicating that this population of seals has been exposed to small quantities of Fukushima-derived radiocesium. Activity concentrations of both (134)Cs and (137)Cs in putchki were below detection limits, even for composited samples. Northern fur seal is known to migrate between coastal Alaska and Japan and the trace (134)Cs in northern fur seal tissue suggests that the population under study had been minimally exposed Fukushima-derived radionuclides. Despite this inference, the radionuclide quantities detected are small and no impact is expected as a result of the measured radiation exposure, either in northern fur seal or human populations consuming this species.

Offspring Hormones Reflect the Maternal Prenatal Social Environment: Potential for Foetal Programming?

Females of many species adaptively program their offspring to predictable environmental conditions, a process that is often mediated by hormones. Laboratory studies have shown, for instance, that social density affects levels of maternal cortisol and testosterone, leading to fitness-relevant changes in offspring physiology and behaviour. However, the effects of social density remain poorly understood in natural populations due to the difficulty of disentangling confounding influences such as climatic variation and food availability. Colonially breeding marine mammals offer a unique opportunity to study maternal effects in response to variable colony densities under similar ecological conditions. We therefore quantified maternal and offspring hormone levels in 84 Antarctic fur seals (Arctocephalus gazella) from two closely neighbouring colonies of contrasting density. Hair samples were used as they integrate hormone levels over several weeks or months and therefore represent in utero conditions during foetal development. We found significantly higher levels of cortisol and testosterone (both P < 0.001) in mothers from the high density colony, reflecting a more stressful and competitive environment. In addition, offspring testosterone showed a significant positive correlation with maternal cortisol (P < 0.05). Although further work is needed to elucidate the potential consequences for offspring fitness, these findings raise the intriguing possibility that adaptive foetal programming might occur in fur seals in response to the maternal social environment. They also lend support to the idea that hormonally mediated maternal effects may depend more strongly on the maternal regulation of androgen rather than cortisol levels.

A colostrum trypsin inhibitor gene expressed in the Cape fur seal mammary gland during lactation.

The colostrum trypsin inhibitor (CTI) gene and transcript were cloned from the Cape fur seal mammary gland and CTI identified by in silico analysis of the Pacific walrus and polar bear genomes (Order Carnivora), and in marine and terrestrial mammals of the Orders Cetartiodactyla (yak, whales, camel) and Perissodactyla (white rhinoceros). Unexpectedly, Weddell seal CTI was predicted to be a pseudogene. Cape fur seal CTI was expressed in the mammary gland of a pregnant multiparous seal, but not in a seal in its first pregnancy. While bovine CTI is expressed for 24-48 h postpartum (pp) and secreted in colostrum only, Cape fur seal CTI was detected for at least 2-3 months pp while the mother was suckling its young on-shore. Furthermore, CTI was expressed in the mammary gland of only one of the lactating seals that was foraging at-sea. The expression of ß-casein (CSN2) and ß-lactoglobulin II (LGB2), but not CTI in the second lactating seal foraging at-sea suggested that CTI may be intermittently expressed during lactation. Cape fur seal and walrus CTI encode putative small, secreted, N-glycosylated proteins with a single Kunitz/bovine pancreatic trypsin inhibitor (BPTI) domain indicative of serine protease inhibition. Mature Cape fur seal CTI shares 92% sequence identity with Pacific walrus CTI, but only 35% identity with BPTI. Structural homology modelling of Cape fur seal CTI and Pacific walrus trypsin based on the model of the second Kunitz domain of human tissue factor pathway inhibitor (TFPI) and porcine trypsin (Protein Data Bank: 1TFX) confirmed that CTI inhibits trypsin in a canonical fashion. Therefore, pinniped CTI may be critical for preventing the proteolytic degradation of immunoglobulins that are passively transferred from mother to young via colostrum and milk.

Uridine Diphosphate-Glucuronosyltransferase (UGT) Xenobiotic Metabolizing Activity and Genetic Evolution in Pinniped Species.

There are various interspecies differences in xenobiotic-metabolizing enzymes. It is known that cats show slow glucuronidation of drugs such as acetaminophen and strong side effects due to the UGT1A6 pseudogene. Recently, the UGT1A6 pseudogene was found in the Northern elephant seal and Otariidae was suggested to be UGT1A6-deficient. From the results of measurements of uridine diphosphate-glucuronosyltransferase (UGT) activity using liver microsomes, the Steller sea lion, Northern fur seal, and Caspian seal showed UGT activity toward 1-hydroxypyrene and acetaminophen as low as in cats, which was significantly lower than in rat and dog. Furthermore, UGT1A6 pseudogenes were found in Steller sea lion and Northern fur seal, and all Otariidae species were suggested to have the UGT1A6 pseudogene. The UGT1 family genes appear to have undergone birth-and-death evolution based on a phylogenetic and synteny analysis of the UGT1 family in mammals including Carnivora. UGT1A2-1A5 and UGT1A7-1A10 are paralogous genes to UGT1A1 and UGTA6, respectively, and their numbers were lower in cat, ferret and Pacific walrus than in human, rat, and dog. Felidae and Pinnipedia, which are less exposed to natural xenobiotics such as plant-derived toxins due to their carnivorous diet, have experienced fewer gene duplications of xenobiotic-metabolizing UGT genes, and even possess UGT1A6 pseudogenes. Artificial environmental pollutants and drugs conjugated by UGT are increasing dramatically, and their elimination to the environment can be of great consequence to cat and Pinnipedia species, whose low xenobiotic glucuronidation capacity makes them highly sensitive to these compounds.

Nature of adenosine triphosphatase accelerating peptide from hydrolysate of fur seal muscle.

Ultrafiltered fur seal muscle hydrolysate was divided into eleven fractions by gel filtration on Sephadex G-15. One of the fractions (Fraction G9) accelerated the ATPase activity of carp myosin B to a rate about two-fold faster than that of the control. Fraction G9 showed a single ninhydrin spot in its silica gel thin layer chromatograph, and gave a positive test for tryptophan by the p-dimethylaminobenzaldehyde method, while tests for tyrosine, and for arginine were negative. The ion exchange amino acid analysis of its acid hydrolysate showed a predominant content of lysine, nearly equivalent to the amount of tryptophan determined from its UV absorbancy and the p-dimethylaminobenzaldehyde method. The N-terminal amino acid analysis gave di-DNP-Lys as the sole DNP-amino acid. The structure of the ATPase accelerating peptide fraction, Fraction G9, was deduced to be Lys-Trp.

Factors affecting organochlorine contaminant concentrations in milk and blood of northern fur seal (Callorhinus ursinus) dams and pups from St. George Island, Alaska.

Levels of organochlorine contaminants in blood of northern fur seal (Callorhinus ursinus) pups and the blood and milk of their dams early in the lactation period are reported here. The contaminants included 15 selected individual polychlorinated biphenyl (PCB) congeners and DDT metabolites identified through high-performance liquid chromatography. Congeners CB-77, -81, -126, -169 and -189 were below the limits of detection in milk and blood samples analyzed. Congener-specific concentrations of PCBs in the blood of pups were compared based on the age of their dam (< or = 5 years or > 7 years). Pups of young (presumably primiparous) dams had significantly elevated levels of CBs-101, -118, -128, -138, -153/87, -170/194, and -180 than pups of older (multiparous) dams. Congeners CB-128 and -170/194 were detected in the blood of pups of young dams but not in the blood of pups of older dams nor in any of the dams blood. Additionally, pups had higher blood levels in seven of 10 detected PCB congeners as compared to the levels measured in milk when adjusted for lipid content. Levels of DDT metabolites and toxic equivalency quotients of dioxin-like congeners followed similar trends. Lipid-normalized concentrations of CB-101 and total PCBs were significantly higher in the blood of dams than in their milk. CB-128, -156, -157, -170/194, p,p'-DDT, o,p'-DDT, p,p'-DDD and o,p'-DDD were not detected in dam blood samples, but were detected in milk samples. Calculation of 'biomagnification factors' from milk to pup blood indicated a biomagnification of CB-101, -105, -118, -138, -153/87, and -180. Significant mean accumulation factors ranged from 1.5 to 7.5. Inter-annual differences in exposure levels and specific congener concentrations in both milk and blood were apparent. Northern fur seal pups, especially first-born, have a substantial exposure to organochlorine contaminants at a critical developmental stage.