Bayesian mixed model analysis uncovered 21 risk loci for chronic kidney disease in boxer dogs.

Chronic kidney disease (CKD) affects 10% of the human population, with only a small fraction genetically defined. CKD is also common in dogs and has been diagnosed in nearly all breeds, but its genetic basis remains unclear. Here, we performed a Bayesian mixed model genome-wide association analysis for canine CKD in a boxer population of 117 canine cases and 137 controls, and identified 21 genetic regions associated with the disease. At the top markers from each CKD region, the cases carried an average of 20.2 risk alleles, significantly higher than controls (15.6 risk alleles). An ANOVA test showed that the 21 CKD regions together explained 57% of CKD phenotypic variation in the population. Based on whole genome sequencing data of 20 boxers, we identified 5,206 variants in LD with the top 50 BayesR markers. Following comparative analysis with human regulatory data, 17 putative regulatory variants were identified and tested with electrophoretic mobility shift assays. In total four variants, three intronic variants from the MAGI2 and GALNT18 genes, and one variant in an intergenic region on chr28, showed alternative binding ability for the risk and protective alleles in kidney cell lines. Many genes from the 21 CKD regions, RELN, MAGI2, FGFR2 and others, have been implicated in human kidney development or disease. The results from this study provide new information that may enlighten the etiology of CKD in both dogs and humans.

The ESR1 gene is associated with risk for canine mammary tumours.

BACKGROUND: The limited within-breed genetic heterogeneity and an enrichment of disease-predisposing alleles have made the dog a very suitable model for the identification of genes associated with risk for specific diseases. Canine mammary cancer is an example of such a disease. However, the underlying inherited risk factors for canine mammary tumours (CMTs) are still largely unknown. In this study, 52 single nucleotide polymorphisms (SNPs) in ten human cancer-associated genes were genotyped in two different datasets in order to identify genes/alleles associated with the development of CMTs. The first dataset consisted of English Springer Spaniel (ESS) CMT cases and controls. ESS is a dog breed known to be at increased risk of developing CMTs. In the second dataset, dogs from breeds known to have a high frequency of CMTs were compared to dogs from breeds with a lower occurrence of these tumours. RESULTS: We found significant associations to CMT for SNPs and haplotypes in the estrogen receptor 1 (ESR1) gene in the ESS material (best PBonf = 0.021). A large number of SNPs, among them several SNPs in ESR1, showed significantly different allele frequencies between the high and low risk breed groups (best PBonf = 8.8E-32, best PBPerm = 0.076). CONCLUSIONS: The identification of CMT-associated SNPs in ESR1 in two independent datasets suggests that this gene might be involved in CMT development. These findings also support that CMT may serve as a good model for human breast cancer research.

A 1 bp deletion in HACE1 causes ataxia in Norwegian elkhound, black.

A number of inherited ataxias is known in humans, with more than 250 loci implicated, most of which are included in human ataxia screening panels. Anecdotally, cases of ataxia in the Norwegian elkhound black have been known for the last 40 years. Affected puppies from three litters were clinically and neurologically examined, and postmortem samples were collected for morphological studies, including ultrastructural analyses. The puppies displayed vestibulocerebellar neurological signs and had degenerative histopathological alterations in cerebellum and brain stem. Three affected dogs, each from different litters, as well as both parents and one healthy littermate from each litter, were whole genome sequenced. Through variant calling we discovered a disease-associated 1 bp deletion in HACE1 (CFA12), resulting in a frameshift at codon 333 and a premature stop codon at codon 366. The perfect association combined with the predicted significant molecular effect, strongly suggest that we have found the causative mutation for Norwegian elkhound black ataxia. We have identified a novel candidate gene for ataxia where dogs can serve as a spontaneous model for improved understanding of ataxia, also in human.

Serum biochemistry and haematology in wild and captive bearded seals (Erignathus barbatus) from Svalbard, Norway.

BACKGROUND: Health assessment of seals in captivity include haematology and serum biochemistry measurements. Because such parameters differ between species, it is crucial to have species-specific reference values for the interpretation of clinical samples. Furthermore, differences in nutrition and environment, life cycles as well as seasonal/annual cycles and varying physiological conditions can potentially affect serum chemistry and haematology parameters. Blood samples from four captive adult bearded seals (initially caught as pups in Svalbard, Norway, now held at Polaria, an Arctic experience centre in Tromsø, Norway) collected over a 16-month period were analysed for haematology (n = 22) and serum chemistry (n = 25) parameters. Serum chemistry analyses were also conducted on blood samples from 74 wild bearded seals (1995-2007) collected from Svalbard, Norway. RESULTS: We found higher activity of creatine kinase (CK) and higher concentrations of cortisol in the wild animals when compared to the captive seals, probably reflecting the physical restraint and concomitant stress induced during sampling. For the captive bearded seals, we did not find marked differences in haematology or serum chemistry parameters throughout the different seasons of sampling. CONCLUSIONS: This study presents haematology and serum chemistry reference values for captive and wild bearded seals. Comparing physiological parameters for captive seals with wild seals indicated that having wild-caught bearded seals under the conditions offered at Polaria for several years did not markedly affect physiological parameters of the animals, and that training may have helped to alleviate stress associated with blood sampling and veterinary inspection.

Hematology and serum clinical chemistry reference intervals for free-ranging Scandinavian gray wolves (Canis lupus).

BACKGROUND: Scandinavian free-ranging wolves (Canis lupus) are endangered, such that laboratory data to assess their health status is increasingly important. Although wolves have been studied for decades, most biological information comes from captive animals. OBJECTIVES: The objective of the present study was to establish reference intervals for 30 clinical chemical and 8 hematologic analytes in Scandinavian free-ranging wolves. METHODS: All wolves were tracked and chemically immobilized from a helicopter before examination and blood sampling in the winter of 7 consecutive years (1998-2004). Seventy-nine blood samples were collected from 57 gray wolves, including 24 juveniles (24 samples), 17 adult females (25 samples), and 16 adult males (30 samples). Whole blood and serum samples were stored at refrigeration temperature for 1-3 days before hematologic analyses and for 1-5 days before serum biochemical analyses. Reference intervals were calculated as 95% confidence intervals except for juveniles where the minimum and maximum values were used. RESULTS: Significant differences were observed between adult and juvenile wolves for RBC parameters, alkaline phosphatase and amylase activities, and total protein, albumin, gamma-globulins, cholesterol, creatinine, calcium, chloride, magnesium, phosphate, and sodium concentrations. CONCLUSION: Compared with published reference values for captive wolves, reference intervals for free-ranging wolves reflected exercise activity associated with capture (higher creatine kinase activity, higher glucose concentration), and differences in nutritional status (higher urea concentration).

Effects of dietary deoxynivalenol or ochratoxin A on performance and selected health indices in Atlantic salmon (Salmo salar).

Post-smolt Atlantic salmon (Salmo salar) were fed with standard feed added one of five concentrations of either pure deoxynivalenol (DON; 0.5-6 mg/kg) or pure ochratoxin A (OTA; 0.2-2.4 mg/kg), or no added toxins for up to 8 weeks. Performance effects (feed intake, feed efficiency, gain, length and condition factor), various clinical biochemical parameters, packed cell volume and vaccination response against Aeromonas salmonicidae were all inversely correlated with DON dose, whereas relative liver weight increased with DON dose. In fish fed OTA, however, the effects at the doses tested were rather small. We observed no effects of OTA exposure on performance parameters, but some clinical biochemical parameters tended to increase with OTA dose primarily at 3 weeks, and compared with controls OTA exposure caused increased mRNA expression of two immune markers in the spleen. No liver histopathological effects were found from DON or OTA exposure. For DON, we derived a BMDL20 of 0.3 mg/kg feed for reduced total protein in plasma, a BMDL5 of 0.5 mg/kg feed for reduced condition factor, and a NOAEL of 1 mg/kg feed for DON. For OTA, a BMDL or NOAEL could not be derived (>2.4 mg/kg).

Introduction to ISO 15189: a blueprint for quality systems in veterinary laboratories.

BACKGROUND: A trend in human and veterinary medical laboratory management is to achieve accreditation based on international standards. The International Organization for Standardization (ISO) 15189 standard is the first developed especially for accreditation of medical laboratories, and emphasizes the laboratory-client interface. European veterinary laboratories seeking to train candidates for the certification examination of the European College of Veterinary Clinical Pathology (ECVCP) require approval by the ECVCP Laboratory Standards Committee, which bases its evaluation in part on adherence to quality systems described in the ISO 15189 standards. OBJECTIVE: The purpose of this article was to introduce the latest ISO quality standard and describe its application to veterinary laboratories in Europe, specifically as pertains to accreditation of laboratories involved in training veterinary clinical pathologists. METHODS: Between 2003 and 2006, the Laboratory Standards Committee reviewed 12 applications from laboratories (3 commercial and 9 university) involved in training veterinary clinical pathologists. Applicants were asked to provide a description of the facilities for training and testing, current methodology and technology, health and safety policy, quality assurance policy (including internal quality control and participation in an external quality assurance program), written standard operating procedures (SOPs) and policies, a description of the laboratory information system, and personnel and training. Also during this time period multiple informal and formal discussions among ECVCP diplomates took place as to current practices and perceived areas of concern with regard to laboratory accreditation requirements. RESULTS: Areas in which improvement most often was needed in veterinary laboratories applying for ECVCP accreditation were the written quality plan, defined quality requirements for the tests performed, written SOPs and policies, training records, ongoing audits and competency assessments, and processes for identifying and addressing opportunities for improvement. Recommendations were developed for a stepwise approach towards achieving ISO 15189 standards, including 3 levels of quality components. CONCLUSION: The ISO 15189 standard provides a sound framework for veterinary laboratories aspiring to meet international quality standards.

Serum chemistry of free-ranging white whales (Delphinapterus leucas) in Svalbard.

BACKGROUND: Abnormal physiological conditions and diseases can change the concentrations of enzymes, metabolites, and minerals in the body. Serum chemistry information may thus be indicative of a specific disease; interpretation of such information requires knowledge of serum chemistry reference intervals from a seemingly healthy population of the species. OBJECTIVE: The aim of this study was to obtain serum chemistry reference intervals for a population of white whales. METHODS: Blood samples were collected from 21 free-ranging white whales (beluga; Delphinapterus leucas). The whales were live-captured in nets during 1996-2001 in Storfjorden, Van Mijenfjorden, and Van Keulenfjorden (Svalbard, Norway). While the whales were briefly physically restrained, blood was collected from the caudal vein into vacuum tubes without anticoagulant. The blood was left to clot for 4-6 hours before serum was obtained by centrifugation. The serum samples were then kept at -20 degrees C until analysis. Enzymes (aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase [ALP], creatine kinase, lactate dehydrogenase [LDH], amylase, lipase), metabolites (urea, creatinine, bilirubin, cholesterol, triglycerides, nonesterified fatty acids, glucose), and minerals (calcium, phosphate, magnesium, sodium, potassium, chloride) were analyzed in an Advia 1650 System (Bayer, Tarrytown, NY, USA). Cortisol was analyzed in an Immulite One system (Diagnostic Products Corporation, Los Angeles, CA, USA). The major blood proteins (albumin and globulins) were separated by gel electrophoresis in a Beckman Paragon electrophoresis system (Beckman Coulter, Inc., Fullerton, CA, USA). RESULTS: Serum values for all analytes were reported as median and range, and reference intervals were calculated as 10-90th percentiles. Activities of ALP and LDH and cortisol concentration were higher, and protein and bilirubin concentrations were lower compared with those previously reported for white whales from Canada; remaining results were strikingly similar in these 2 white whale populations. CONCLUSIONS: These data provide valuable serum chemistry reference intervals for future health assessments of white whales in Svalbard and other white whale populations, as well as captive individuals.

Serum chemistry values for free-ranging ringed seals (Pusa hispida) in Svalbard.

BACKGROUND: Diseases and abnormal physiologic conditions can alter the concentrations of enzymes, metabolites, minerals, and hormones in the blood of animals. The ringed seal (Pusa hispida) has been selected as a key species for environmental monitoring, but information on disease and health parameters for this species is scarce. OBJECTIVES: The aim of the study reported here was to obtain serum chemistry reference intervals for free-ranging ringed seals in Svalbard, and then to evaluate serum chemistry values in relation to age, body condition, and sex. METHODS: Blood samples were collected after death from ringed seals in Wijdefjorden and Billefjorden, Svalbard (2002-2003; n = 75). Serum was analyzed for 24 selected serum chemistry parameters (enzymes, protein, metabolites, minerals, and cortisol). RESULTS: Compared with younger or older animals, seals between 7 and 16 years of age had larger variations in the activities of alanine transaminase, aspartate transaminase, lactate dehydrogenase, and creatine kinase (CK). Animals classified as having low body condition status had more variation in the serum activity of these enzymes, compared with that in animals with higher condition scores. Serum cortisol concentration was higher in young animals (1-5 years) than in older animals. Serum CK activity was higher in males than in females. CONCLUSION: The data reported here may be useful in monitoring the health of ringed seals and for tracking the impact of environmental changes in the Arctic.

Genetics and epidemiology of hypothyroidism and symmetrical onychomadesis in the Gordon setter and the English setter.

BACKGROUND: Hypothyroidism is one of the most common endocrine disorders, whereas symmetrical onychomadesis is a rare claw disease in the general dog population. The aims of this study were to estimate the prevalence of hypothyroidism and symmetrical onychomadesis in a birth cohort of 291 Gordon setters at eight years of age. Further, to describe the age at diagnosis of hypothyroidism in the 68 Gordon setters and 51 English setters included in the DLA study. Finally, to elucidate potential associations between dog leukocyte antigen (DLA) class II and hypothyroidism and/or symmetrical onychomadesis in the Gordon setter and the English setter. RESULTS: In the birth cohort of eight years old Gordon setters, 2.7 % had hypothyroidism and 8.9 % had symmetrical onychomadesis, but only one out of these 291 dogs (0.3 %) had both diseases. Mean age at diagnosis of hypothyroidism for dogs included in the DLA study was 6.4 years (95 % CI: 5.6-7.2 years) in the Gordon setters and 7.7 years (95 % CI: 7.2-8.2 years) in the English setters. The DLA alleles most associated with hypothyroidism in the Gordon setter and English setter were DLA-DQB1*00201 (OR = 3.6, 95 % CI: 2.1-6.4, p < 0.001) and DLA-DQA1*00101 (OR = 2.9, 95 % CI: 1.3-6.6, p < 0.001), respectively. In the Gordon setter, the haplotype DLA-DRB1*01801/DQA1*00101/DQB1*00802 was significantly associated with both symmetrical onychomadesis (OR = 2.9, 95 % CI: 1.7-5.2, p < 0.001) and with protection against hypothyroidism (OR = 0.3, 95 % CI: 0.2-0.5, p < 0.001). CONCLUSION: Hypothyroidism is a complex disease where DLA genes together with other genes may be involved in the pathogenesis of the disease. In the Gordon setter, one DLA haplotype that was associated with protection against hypothyroidism was also associated with symmetrical onychomadesis. These findings indicate that closely linked genes, instead of or together with the DLA genes themselves, may be associated with hypothyroidism and symmetrical onychomadesis. In a breed where several autoimmune diseases are prevalent all possible associations between DLA genes and actual diseases need to be investigated before DLA is considered used as a tool for marker-assisted selection.

Toxoplasma gondii seroprevalence in pet cats in Norway and risk factors for seropositivity.

OBJECTIVES: The aims of the study were to estimate Toxoplasma gondii seroprevalence in pet cats in Norway and to evaluate risk factors for seropositivity. Additionally, serum biochemistry and haematological variables for T gondii seropositive and seronegative cats were compared. METHODS: A convenience sample of surplus sera submitted to the Central Laboratory, Norwegian University of Life Sciences, was collected. The samples were from healthy cats and cats with a variety of diseases. Analyses for IgG antibodies to T gondii were performed with a commercial direct agglutination test, with 1:40 as the threshold value. For risk factor analysis a logistic regression model of the relationship between predictors and the outcome was applied. RESULTS: One hundred and ninety-six of 478 cats were seropositive for T gondii, and the estimated seroprevalence in the study sample was 41.0% (95% confidence interval 36.6-45.4). Compared with domestic cats, pedigree cats had reduced risk for Toxoplasma seropositivity (odds ratio [OR] 0.42). Males had increased risk (OR 1.63) compared with females. The effect of age was highly significant, and an increase in the cats' age across the interquartile range (IQR; 52-160 months/4-13 years of age) doubled the risk of Toxoplasma seropositivity (OR 2.11). The risk for Toxoplasma seropositivity among cats living in Oslo was significantly reduced (OR 0.51) when compared with the rest of Norway. CONCLUSIONS AND RELEVANCE: Pet cats in Norway appear to be commonly exposed to T gondii. Signalment and geographical region influenced the odds of Toxoplasma seropositivity, whereas health status did not.

Age-related changes in hematologic and serum biochemical variables in dogs aged 16-60 days.

BACKGROUND: The literature reporting hematologic and serum biochemical variables in puppies is limited. As puppies are physiologically different from adult dogs, an age effect would be expected. OBJECTIVES: We aimed to describe age-related changes in hematologic and serum biochemical variables in puppies aged 16-60 days and compare the results to reference intervals (RI) for adults. Our second aim was to determine RI for this age group. METHODS: A total of 227 blood samples were collected from 101 clinically healthy puppies, mainly mixed breeds. To assess the effect of age, the results were compared to RI for adult dogs, and variations within the age period 16-60 days were studied. Reference intervals for the groups 16-24, 28-45, and 46-60 days of age were determined. RESULTS: Lower values in puppies compared to adults were found for RBC, HGB, HCT, concentration of albumin, globulin, total protein, creatinine, and sodium:potassium ratio. Higher values in puppies compared to adults were found for activities of ALP and CK, and concentrations of inorganic phosphorus, calcium, and potassium. For MCV, MCHC, albumin:globulin ratio, and glucose concentration, different values in puppies compared to adults were found for some of the age groups. No age-specific differences were found compared to RI for adults regarding WBC, absolute counts of lymphocytes, neutrophils, monocytes, eosinophils, and platelets, RDW, activities for AST, ALT, amylase, lipase, and concentrations of bile acids, cholesterol, urea, sodium, and chloride. CONCLUSIONS: Our results support that age has a significant effect on several hematologic and serum biochemical values in puppies, warranting age-specific RI.

A Multi-Breed Genome-Wide Association Analysis for Canine Hypothyroidism Identifies a Shared Major Risk Locus on CFA12.

Hypothyroidism is a complex clinical condition found in both humans and dogs, thought to be caused by a combination of genetic and environmental factors. In this study we present a multi-breed analysis of predisposing genetic risk factors for hypothyroidism in dogs using three high-risk breeds--the Gordon Setter, Hovawart and the Rhodesian Ridgeback. Using a genome-wide association approach and meta-analysis, we identified a major hypothyroidism risk locus shared by these breeds on chromosome 12 (p = 2.1x10(-11)). Further characterisation of the candidate region revealed a shared ~167 kb risk haplotype (4,915,018-5,081,823 bp), tagged by two SNPs in almost complete linkage disequilibrium. This breed-shared risk haplotype includes three genes (LHFPL5, SRPK1 and SLC26A8) and does not extend to the dog leukocyte antigen (DLA) class II gene cluster located in the vicinity. These three genes have not been identified as candidate genes for hypothyroid disease previously, but have functions that could potentially contribute to the development of the disease. Our results implicate the potential involvement of novel genes and pathways for the development of canine hypothyroidism, raising new possibilities for screening, breeding programmes and treatments in dogs. This study may also contribute to our understanding of the genetic etiology of human hypothyroid disease, which is one of the most common endocrine disorders in humans.

Serum chemistry and antibodies against pathogens in antarctic fur seals, Weddell seals, crabeater seals, and Ross seals.

Information on health parameters, such as antibody prevalences and serum chemistry that can reveal exposure to pathogens, disease, and abnormal physiologic conditions, is scarce for Antarctic seal species. Serum samples from Antarctic fur seals (Arctocephalus gazella, n=88) from Bouvetøya (2000-2001 and 2001-2002), and from Weddell seals (Leptonychotes weddellii, n=20), Ross seals (Ommatophoca rossii, n=20), and crabeater seals (Lobodon carcinophagus, n=9) from the pack-ice off Queen Maud Land, Antarctica (2001) were analyzed for enzyme activity, and concentrations of protein, metabolites, minerals, and cortisol. Adult Antarctic fur seal males had elevated levels of total protein (range 64-99 g/l) compared to adult females and pups (range 52-79 g/l). Antarctic fur seals had higher enzyme activities of creatine kinase, lactate dehydrogenase, and amylase, compared to Weddell, Ross, and crabeater seals. Antibodies against Brucella spp. were detected in Weddell seals (37%), Ross seals (5%), and crabeater seals (11%), but not in Antarctic fur seals. Antibodies against phocine herpesvirus 1 were detected in all species examined (Antarctic fur seals, 58%; Weddell seals, 100%; Ross seals, 15%; and crabeater seals, 44%). No antibodies against Trichinella spp., Toxoplasma, or phocine distemper virus (PDV) were detected (Antarctic fur seals were not tested for PDV antibodies). Antarctic seals are challenged by reduced sea ice and increasing temperatures due to climate change, and increased anthropogenic activity can introduce new pathogens to these vulnerable ecosystems and represent a threat for these animals. Our data provide a baseline for future monitoring of health parameters of these Antarctic seal species, for tracking the impact of environmental, climatic, and anthropogenic changes in Antarctica over time.

Hematologic and biochemical reference intervals for Norwegian crossbreed grower pigs.

BACKGROUND: Hematologic and biochemical reference intervals depend on many factors, including environment and age. Reference intervals for Norwegian grower pigs are lacking, and previously published reference intervals for similar pigs from other countries are now outdated due to significant changes in management and breeding on the pig farms. OBJECTIVES: The aim of this study was to determine updated reference intervals for hematologic and biochemical analytes in healthy crossbred grower pigs, and to compare the results among 3 different farms. METHODS: Blood samples were collected from 104 clinically healthy pigs of the most common Norwegian crossbreed (Landrace Yorkshire sow x Landrace Duroc boar). The pigs were 12-16 weeks old, weighed 30-50 kg, of both sexes, and lived on 3 farms in eastern Norway. Automated hematologic and biochemical analysis were performed using ADVIA 2120 and ADVIA 1650 analyzers. RESULTS: Five samples were excluded because of hemolysis (1) or outliers (4). Reference intervals were calculated using parametric or nonparametric methods, depending on data distribution. Mean, median, minimum, and maximum values were tabulated. CONCLUSIONS: The reference intervals calculated in this study will be useful for the diagnosis and monitoring of disease in this widespread crossbreed pig. Compared with previously published reference values, reference intervals for total WBC count, creatine kinase and alanine aminotransferase activities, and albumin, bilirubin, and urea concentrations in this study differed notably.

Serum chemistry reference values in free-ranging north Atlantic male walruses (Odobenus rosmarus rosmarus) from the Svalbard archipelago.

BACKGROUND: Information regarding health and disease is limited for walruses, a keystone species in arctic marine ecosystems. Serum chemistry analysis is a useful clinical tool for the health assessment of walruses, but only a few captive Pacific walruses have been evaluated. OBJECTIVES: The aim of this study was to determine serum chemistry reference values for free-ranging male Atlantic walruses (Odobenus rosmarus rosmarus) on Svalbard and to assess potential differences in animals with low and high tissue levels of organic pollutants. METHODS: Blood samples were collected from 17 wild, adult, male Atlantic walruses chemically immobilized with etorphine at eastern Svalbard (Norway). Serum was obtained for routine biochemical analysis as well as nonesterified free fatty acids (NEFA) and cortisol tests. Serum protein concentration was also measured by agarose gel electrophoresis. RESULTS: Reference values (ranges) included alanine aminotransferase (12-51 U/L), aspartate aminotransferase (54-137 U/L), alkaline phosphatase (42-243 U/L), creatine kinase (32-506 U/L), lactate dehydrogenase (480-1322 U/L), amylase (0-23 U/L), lipase (68-298 U/L), total protein (68-91 g/L), albumin (25.3-34.8 g/L), creatinine (84-137 mumol/L), urea (8.2-19.9 mmol/L), bilirubin (0-4 mumol/L), cholesterol (4.4-7.3 mmol/L), NEFA (0.1-0.4 mmol/L), triglycerides (0.6-2.2 mmol/L), calcium (2.0-2.7 mmol/L), phosphorus (1.7-2.8 mmol/L), sodium (147-162 mmol/L), potassium (4.7-7.4 mmol/L), chloride (102-115 mmol/L), and cortisol (<28-214 nmol/L). Walruses exposed to high levels of organic pollutants (n=6) had significantly lower (P=.022) phosphorus concentration than those with low levels of pollutants (n=6). CONCLUSIONS: The clinical chemistry reference values determined in this study can serve as baseline data for future health-related studies of walruses in a changing Arctic and may also be helpful for health evaluations of walruses in captivity. Impacts of the exposure of marine mammals to organic pollutants should be further investigated.

Effects of storage time on chemistry results from canine whole blood, heparinized whole blood, serum and heparinized plasma.

The stability and storage characteristics of 24 blood constituents from dogs including nine enzymes (ALP, ALT, amylase, AST, CK, GGT, GLDH, LDH, lipase), 15 metabolites and minerals (albumin, bile acids, bilirubin, calcium, cholesterol, creatinine, fructosamine, glucose, magnesium, phosphate, potassium, protein, sodium, triglycerides, urea) were studied. Conditions studied included storing of nonanticoagulated and heparinized whole blood for 3 days (Part A), and storing of serum and heparinized plasma for 3 days (Part B). The storage temperature for both studies was +4 degrees C from day 0 to day 1, and +20 degrees C, from day 1 to day 2 and 3. Eight of 24 analytes showed no significant differences (p > 0.05) for three days in whole blood. However, the stability of all 24 analytes greatly improved by storing serum or heparinized plasma compared to nonanticoagulated or heparinized whole blood. In stored serum or heparinized plasma, 20 of 24 analytes showed no significant differences (p < 0.05) for 3 days. Nine of 24 analytes showed significant differences (p < 0.05) between serum and heparinized plasma, where CK, LDH, GGT, and potassium showed differences of possible clinical importance. This study strongly supports the practice of separating serum/plasma from clot/cells as promptly as possible to achieve improved stability for most analytes under test.