Medroxyprogesterone acetate in reindeer bulls: testes histology, cfos activity in the brain, breeding success, and semen quality.

A previous study reported that a 400-mg dose of medroxyprogesterone acetate (MPA) reduced male reindeer aggression and blocked development of secondary sexual characteristics but did not completely impair fertility. Here we have repeated that protocol in two separate trials. In 2017, tissues and blood samples, collected from MPA and control (CTL) reindeer bulls, euthanized at 30 and 60 d post-treatment were used to evaluate testes histology and morphometrics, cfos activity in the brain and androgen levels. While testes weight tended to decline from August to September in both groups, indices of spermatogenesis remained high. By September, indices of spermatogenesis were declining in both groups with sperm density lower (P = 0.05) in MPA compared to CTL bulls. Aug CTL bulls had the highest concentrations of androstenedione (A4) (P = 0.009) and testosterone (T) (P = 0.08), whereas these androgens were baseline in Aug MPA bulls. By September, A4 and T levels in CTL bulls declined to levels measured in MPA bulls. Cfos activity had a greater number (P = 0.02) of cfos positive neurons in the central amygdala in MPA compared to CTL bulls, suggesting a heightened fear response among the MPA bulls. In the second trial (2019), MPA-treated bulls, with (E, n = 4) and without (IE, n = 4) breeding experience, were blood sampled at key points from July through September when they were put in individual harems with estrous-synchronized cows. Concentrations of T were greatest (P < 0.001) among E bulls prior to MPA treatment but 1 mo after treatment, both T and A4 were baseline in all eight reindeer. Semen collected by electroejaculation at 60 d post-MPA treatment revealed only minor differences in sperm abnormalities between E and IE bulls using both fresh and frozen/thawed semen. Only three bulls (2 E and 1 IE) sired offspring. Breeding success was not related to previous breeding experience, body weight, or bull age. The failure of some MPA bulls to breed appears to be a behavioral, not a physiological, limitation. Limited application of MPA is clearly a useful tool for managing rut-aggression in non-breeding reindeer. However, the possibility that semen could be collected from MPA-treated bulls using restraint and mild sedation rather than general anesthesia should be investigated. This could improve the quality of semen collection while enhancing the safety of both handlers and animals.

A single 400 mg dose of MPA given to reindeer bulls just before the onset of rut eliminates aggressive behavior and suppresses androgen concentrations without dramatic differences in the gross or histological structure of the testes within the first 30 d of treatment. By 60 d post-treatment, there is evidence of smaller testes size and decreased sperm density in treated bulls. However, if given the opportunity, some treated bulls can still successfully breed. Breeding success in MPA bulls was not solely related to previous breeding experience, body weight, or bull age. Androgen concentrations and semen characteristics did not vary with previous breeding experience. Failure of some treated bulls to breed appears to be a behavioral limitation. Differences in brain activity between control and treated bulls were few except for increased cfos activity in the central amygdala of MPA bulls, potentially increasing the fear response in these reindeer.

Novel insights into serodiagnosis and epidemiology of Erysipelothrix rhusiopathiae, a newly recognized pathogen in muskoxen (Ovibos moschatus).

BACKGROUND: Muskoxen are a key species of Arctic ecosystems and are important for food security and socio-economic well-being of many Indigenous communities in the Arctic and Subarctic. Between 2009 and 2014, the bacterium Erysipelothrix rhusiopathiae was isolated for the first time in this species in association with multiple mortality events in Canada and Alaska, raising questions regarding the spatiotemporal occurrence of the pathogen and its potential impact on muskox populations. MATERIALS AND METHODS: We adapted a commercial porcine E. rhusiopathiae enzyme-linked immunosorbent assay to test 958 blood samples that were collected from muskoxen from seven regions in Alaska and the Canadian Arctic between 1976 and 2017. The cut-off between negative and positive results was established using mixture-distribution analysis, a data-driven approach. Based on 818 samples for which a serological status could be determined and with complete information, we calculated trends in sample seroprevalences in population time-series and compared them with population trends in the investigated regions. RESULTS: Overall, 219/818 (27.8%, 95% Confidence Interval: 24.7-31.0) samples were classified as positive for exposure to E. rhusiopathiae. There were large variations between years and regions. Seropositive animals were found among the earliest serum samples tested; 1976 in Alaska and 1991 in Canada. In Alaskan muskoxen, sample seroprevalence increased after 2000 and, in two regions, peak seroprevalences occurred simultaneously with population declines. In one of these regions, concurrent unusual mortalities were observed and E. rhusiopathiae was isolated from muskox carcasses. In Canada, there was an increase in sample seroprevalence in two muskox populations following known mortality events that had been attributed to E. rhusiopathiae. CONCLUSION: Our results indicate widespread exposure of muskoxen to E. rhusiopathiae in western Canada and Alaska. Although not new to the Arctic, we documented an increased exposure to the pathogen in several regions concurrent with population declines. Understanding causes for the apparent increased occurrence of this pathogen and its association with large scale mortality events for muskoxen is critical to evaluate the implications for wildlife and wildlife-dependent human populations in the Arctic.

Correction: Novel insights into serodiagnosis and epidemiology of Erysipelothrix rhusiopathiae, a newly recognized pathogen in muskoxen (Ovibos moschatus).

[This corrects the article DOI: 10.1371/journal.pone.0231724.].

Heavy metal and mineral concentrations and their relationship to histopathological findings in the bowhead whale (Balaena mysticetus).

The bowhead whale (Balaena mysticetus) is a species endangered over much of its range that is of great cultural significance and subsistence value to the Inuit of Northern Alaska. This species occupies subarctic and arctic regions presently undergoing significant ecological change and hydrocarbon development. Thus, understanding the health status of the Bering-Chukchi-Beaufort Sea (BCBS) stock of bowhead whales is of importance. In this study, we evaluated the concentrations of six essential and non-essential elements (Zn, tHg, Ag, Se, Cu and Cd) in liver and kidney of bowhead whales (n=64). These tissues were collected from the Inuit subsistence hunt in Barrow, Wainwright and Kaktovik, Alaska between 1983 and 2001. Reference ranges of these elements (including previously reported data from 1983-1997) were developed for this species as part of a health assessment effort, and interpreted using improved aging techniques (aspartic acid racemization and baleen isotopic (13)C methods) to evaluate trends over time with increased statistical power. Interactions between element concentrations and age, sex and harvest season were assessed. Age was found to be of highest significance. Sex and harvest season did not effect the concentrations of these elements, with the exception of renal Se levels, which were significantly higher in fall seasons. In addition, histological evaluation of tissues from whales collected between 1998-2001 was performed. Associations between concentrations of Cd in kidney and liver and scored histopathological changes were evaluated. Liver Cd concentration was strongly associated with the degree of lung fibromuscular hyperplasia (P=0.001) and moderately associated with the degree of renal fibrosis (P=0.03). Renal Cd concentration influenced the degree of lung fibromuscular hyperplasia and renal fibrosis (P=0.01). A significant age effect was found for both pulmonary fibromuscular hyperplasia and renal fibrosis, indicating age may be a causative factor. Improvements in aging techniques and the addition of histological indices help clarify the relationships between elements and the influence of life history parameters on concentrations of these elements and potential impacts on health. These data provide essential baseline input useful for monitoring the effects of arctic ecosystem change as it relates to global climate change and industrial development, as well as help inform epidemiological studies examining the public health implications of heavy metals in subsistence foods.

Assessment of necropsy findings in sled dogs that died during Iditarod Trail sled dog races: 23 cases (1994-2006).

OBJECTIVE: To describe the character and frequency of causes of death and associated lesions in long-distance racing sled dogs. DESIGN: Retrospective case series. ANIMALS: 23 dogs. PROCEDURES: Medical records of dogs that died during or soon after competition in the Iditarod Trail sled dog races (1994 through 2006) were examined for fi ndings of gross necropsy and histologic evaluation of tissue samples. From the data, descriptive and comparative statistics were obtained. RESULTS: Recognized causes of death included aspiration of gastric contents (n = 4), aspiration pneumonia (4), acute blood loss secondary to gastric ulceration (3), and sled dog myopathy (2). A cause of death was not established for 7 dogs. Prevalent lesions among the study population included rhabdomyolysis (n = 15), enteritis (10), gastritis (10), aspiration pneumonia (8), and gastric ulceration (8). All dogs with aspiration pneumonia had concurrent gastric mucosal lesions. Subjective biventricular cardiac hypertrophy was evident in most dogs; other lesions detected frequently included centrilobular hepatic fibrosis, gastric dilatation, and mild cardiac myodegeneration and necrosis. CONCLUSIONS AND CLINICAL RELEVANCE: Unexpected death is a rare event among conditioned sled dogs during competition in endurance races. Potentially life-threatening conditions of dogs that are associated with periods of long-distance physical exertion include aspiration pneumonia, gastric mucosal lesions, and severe rhabdomyolysis. Dogs that develop clinical signs suggestive of these conditions should be excluded from strenuous activities. Epidemiologic investigations are required to clarify the risk for death associated with these lesions in dogs competing in endurance races.

Vitamin A and E tissue distribution with comparisons to organochlorine concentrations in the serum, blubber and liver of the bowhead whale (Balaena mysticetus).

Vitamin A and E concentrations were determined in liver (n=51), blubber (n=23) and serum (n=53) of subsistence-hunted bowhead whales (Balaena mysticetus), between 1998 and 2001. Retinol and alpha-tocopherol were the major forms of vitamins A and E detected, respectively. Liver contained the highest mean concentrations of vitamin A, followed by epidermis, blubber, and serum. Liver also contained the highest mean concentration of vitamin E, followed by serum, epidermis, and blubber. Stratification of retinol and tocopherol was examined throughout the blubber cores collected. Retinol concentrations were significantly higher in the epidermis than in the deeper blubber layers. Tocopherol concentrations were similar for epidermis and the intermediate layer of blubber. Both the epidermis and the intermediate layer of blubber had significantly higher tocopherol concentrations than the innermost and outermost blubber layers. Vitamin A and E concentrations were investigated with respect to gender and reproductive status of females (males, non-pregnant females, pregnant females), age groups and season of harvest. Certain persistent organic contaminants are known to have a negative effect on retinol concentration in serum of pinnipeds and cetaceans. Bowhead whales have relatively low concentrations of organochlorines (OCs) in comparison to other mysticete species. The relationships between serum, liver and blubber retinol and serum and blubber OC concentrations were examined with no significant correlations noted.

Seasonal hyperphagia does not reduce digestive efficiency in an Arctic grazer.

Muskoxen (Ovibos moschatus) consume fibrous plants that grow rapidly over the short Arctic summer. We studied responses of eight castrated male muskoxen to a diet of grass hay and mineral supplements during spring, autumn, and winter. Animals gained body mass in spring (239+/-39 kg) as body fat content increased from 26% to 38% of ingesta-free mass in winter without changes in lean mass and protein. Intakes of dry matter (DM) increased by 74% between spring and autumn as digestible energy increased from 554 to 923 kJ kg(-0.75) d(-1) during mass gain. Digestibility of cellulose (72%-76%) was not affected by increasing food intake between spring and autumn but was reduced to 65% in winter. Digestibility of nitrogen compounds was 61%-66%, even though intake increased by 134% between spring and autumn. Excess dietary nitrogen from hay and supplements increased urea concentrations in plasma and urine. High loads of solutes such as potassium did not affect plasma or urinary osmolality but were associated with increased rates of glomerular filtration and urinary excretion. Low intakes of sodium from grasses may limit intake and digestion during summer, but high food intakes can support deposition of nitrogen, calcium, magnesium, copper, and zinc in body tissue even when dietary concentrations are low. Seasonal increases in digestive and metabolic functions allow muskoxen to rapidly accumulate energy and nutrients in body tissue during the short season of plant growth.

Organochlorine contaminant exposure and associations with hematological and humoral immune functional assays with dam age as a factor in free-ranging northern fur seal pups (Callorhinus ursinus).

Potential effects of organochlorine contaminant (OC) exposure on humoral immune function and health of free-ranging northern fur seals were investigated. Forty-two "neonates" were captured for blood sample collection and re-sampled as "pups" 29-51 days later. OCs were extracted from whole blood to identify polychlorinated biphenyl congeners and chlorinated pesticides by high performance liquid chromatography. Humoral immune function was assessed by antibody responses to vaccination and immunoglobulin concentrations. Additional health status indicators included leukocyte counts and haptoglobin concentrations. Mean OC concentrations were higher in neonates than at recapture. Neonates of young dams had higher mean OC concentrations than neonates of older dams. A higher proportion of old dam's pups developed a twofold or greater increase in antigen-specific antibodies compared to young dam's pups. Higher OC exposure and poor immune responses in first-born pups may indicate a higher risk of secondary morbidity and mortality than pups of multiparous dams.

Copper status of muskoxen: a comparison of wild and captive populations.

We compared wild muskoxen (Ovibos moschatus) on Banks Island (Northwest Territories, Canada) with captive animals maintained on grass (Bromus sp.) hay and supplemental minerals. We measured copper (Cu) in liver, whole serum, and deproteinated serum (unbound Cu) as well as serum activity of the Cu-enzyme ceruloplasmin. Unbound serum Cu concentrations did not change with season in captive animals (n = 53). Ceruloplasmin activity was similar between seasons in females but elevated in males during breeding in autumn. Increasing concentrations of Cu in whole sera were mainly associated with protein whereas unbound Cu predominated at low concentrations of whole serum Cu. Ceruloplasmin activity and serum Cu concentration were linearly related to liver Cu in female muskoxen. Measures of copper status in females were lower in the wild (n = 19) than in captivity (n = 16): 8 vs. 160 ug Cu.g-1 of whole liver; 0.67 vs. 1.15 microgram unbound Cu.ml-1 whole serum and; 22 vs. 33 IU.l-1 ceruloplasmin activity. Bioavailability of Cu may limit the population on Banks Island especially when density of animals is high. The wide range of hepatic Cu concentrations in muskoxen indicated accumulation of Cu without apparent ill effect in captive animals. Hepatic storage of Cu may allow wild muskoxen to contend with low and fluctuating availability of Cu in small foraging areas at high latitudes.

Nutritional development of feeding strategies in arctic ruminants: digestive morphometry of reindeer, Rangifer tarandus, and muskoxen, Ovibos moschatus.

Reindeer have been classified as intermediate feeders and muskoxen as grazers based on differences in digestive morphology and consumption of fibrous plants. We hypothesized that the digestive morphology of young (< 2 months) reindeer and muskoxen anticipates transitions in diet and determines the feeding strategy of each species at adulthood. We compared structural morphology and rates of cell division in the rumen, abomasum, duodenum and liver of reindeer and muskoxen as neonates (1 day old), during the transition from milk to forage (30-60 days old) and in adults (> 7 yr). Development in utero provided the neonate with a functioning mucosa of the gastric abomasum and duodenal mucosa with high surface enlargement for digestion and absorption of concentrated milks. Transition to forage was preceded by changes in ruminal papillae structure that increased surface area and likely contributed to active fermentation by 60 days of age. The abomasum also increased in acid-secreting parietal cells during the transition to forage, which may enhance digestion of plant and microbial proteins. Rates of cell division also indicated a sustained differentiation of tissue structure during the transitional period. Young arctic ruminants expressed digestive structures that preceded full function, which indicated the ultimate feeding strategy of each species. For example, the rumen of young muskoxen had thick cornified epithelia and muscle layers that would provide ruminal mucosa with better protection from fibrous abrasion and enhance motility of bulky diets. Conversely, young reindeer had more complex papillary shapes in the rumen and more foliate villi in the duodenum, indicating a greater absorptive capacity of these structures than in muskoxen. Ontogenetic programs, therefore, play the primary role for digestive development of reindeer and muskoxen and determine the nutritional strategies of adults.

Microanatomy of passerine hard-cornified tissues: beak and claw structure of the black-capped chickadee (Poecile atricapillus).

The microanatomy of healthy beaks and claws in passerine birds has not been well described in the literature, despite the importance of these structures in avian life. Histological processing of hard-cornified tissues is notoriously challenging and only a few reports on effective techniques have been published. An emerging epizootic of beak deformities among wild birds in Alaska and the Pacific Northwest region of North America recently highlighted the need for additional baseline information about avian hard-cornified structures. In this study, we examine the beak and claw of the Black-capped Chickadee (Poecile atricapillus), a common North American passerine that is affected by what has been described as "avian keratin disorder." We use light and scanning electron microscopy and high-magnification radiography to document the healthy microanatomy of these tissues and identify features of functional importance. We also describe detailed methods for histological processing of avian hard-cornified structures and discuss the utility of special stains. Results from this study will assist in future research on the functional anatomy and pathology of hard-cornified structures and will provide a necessary reference for ongoing investigations of avian keratin disorder in Black-capped Chickadees and other wild passerine species.

Costs of gestation in an Arctic ruminant: copper reserves in muskoxen.

The transfer of trace minerals between mother and fetus may be critical for survival of young ruminants especially among species at high latitudes, which gestate during a long winter and grow through a brief summer. We examined the distribution of copper and metalloproteins (ceruloplasmin and metallothionein) in muskoxen and their fetuses, three times during gestation. Hepatic levels of copper were high in mothers (179 microg g(-1) whole tissue) and did not change through gestation, whereas fetuses accumulated large reserves of Cu (>300 microg g(-1)), likely stored in proteins such as metallothionein, during the last third of gestation. The effect of fetal Cu demands on the pregnant female was tested by supplementation of Cu by subcutaneous injections of Cu gluconate (30 mg Cu/week) during pregnancy. Maternal copper supplementation did not significantly increase hepatic Cu in newborns (412 microg g(-1) for supplemented vs. 303 microg g(-1) for unsupplemented neonates), probably because the diet was already adequate in copper (14 microg g(-1) dry matter). Ceruloplasmin activity declined in pregnant muskoxen that had not received injections of Cu and suggested increased systemic demands for copper during late gestation. Supplies of Cu to the fetus could be limited either by low levels of Cu in the maternal liver, or in the maternal diet during late winter when fetal gains in mass and liver Cu are greatest.