Case Report: Subclinical Verminous Pneumonia and High Ambient Temperatures Had Severe Impact on the Anesthesia of Semi-domesticated Eurasian Tundra Reindeer (Rangifer tarandus tarandus) With Medetomidine-Ketamine.

Semidomesticated Eurasian tundra reindeer (Rangifer tarandus tarandus, n = 21) were scheduled twice for chemical immobilization with medetomidine-ketamine as part of a scientific experiment in June 2014. During the first round of immobilizations, seven animals developed severe respiratory depression (RD). Three individuals died, and 4 recovered. The ambient temperature during the 2 days of immobilization (June 3 and 4) was high (mean 13.9-17.6°C) compared to the normal mean temperature for these 2 days (7-8°C) based on statistical records. During the second round of immobilizations, using the same anesthetic protocol for the remaining animals as in the first round but conducted under cooler conditions (mean 6.6°C for the period June 9-18), no signs of RD were observed. Clinical and pathological investigations indicated that the animals suffered from circulatory changes possibly caused by high ambient temperatures and granulomatous interstitial pneumonia due to Elaphostrongylus rangiferi larvae. These conditions, together with the cardiovascular effects of medetomidine, were likely causes of RD and the fatal outcome. We conclude that chemical immobilization of reindeer with medetomidine-ketamine should be avoided in May-June due to the potential risk when animals partly in winter coats encounter rising ambient temperatures and usually have parasites developing in their airways.

Yew (Taxus) intoxication in free-ranging cervids.

Wild ruminants, including deer species (cervids) have incorrectly been regarded as refractory to yew (Taxus) intoxication. This assumption has been based upon anecdotal observations of individual deer browsing on yew over time without apparent adverse effect. A single case of yew intoxication was reported in a free-ranging Norwegian moose (Alces alces) in 2008. The current report describes five additional cases of yew toxicosis in moose, seven in roe deer (Capreolus capreolus) and two in reindeer (Rangifer tarandus tarandus), all in Norway. The animals were found dead during the winter, close to or within gardens containing yew plants showing signs of browsing. Gross findings included lung congestion and edema, thoracic and pericardial effusion, bilateral heart dilatation, epi- and endocardial hemorrhage, and enlarged (congested) spleen. Yew plant remnants were detected in the rumen of all animals with the exception of a single moose. Histology revealed multifocal acute myocardial degeneration and necrosis with hemorrhage in roe deer, but not in the two other species. A qualitative high performance liquid chromatography-ion trap mass spectrometry analysis was used to tentatively identify five major Taxus alkaloids (taxines) in crude yew extracts and in heart and liver samples from the moose cases. All five major taxines were detected with good signal/noise ratio in tissue samples from the four moose with visible ruminal yew content, whereas lower levels of taxines were detected in the moose without visible ruminal yew content. Possible differences in interspecies tolerance to taxines and role of individual protective adaptation are discussed.

Molecular characterisation of Sarcocystis lutrae n. sp. and Toxoplasma gondii from the musculature of two Eurasian otters (Lutra lutra) in Norway.

Sarcocysts were detected in routinely processed histological sections of skeletal muscle, but not cardiac muscle, of two adult male otters (Lutra lutra; Mustelidae) from northern Norway following their post-mortem examination in 1999 and 2000. The sarcocysts were slender, spindle-shaped, up to 970 µm long and 35-70 µm in greatest diameter. The sarcocyst wall was thin (∼ 0.5 µm) and smooth with no visible protrusions. Portions of unfixed diaphragm of both animals were collected at the autopsies and kept frozen for about 14 years pending further examination. When the study was resumed in 2013, the thawed muscle samples were examined for sarcocysts under a stereo microscope, but none could be found. Genomic DNA was therefore extracted from a total of 36 small pieces of the diaphragm from both otters, and samples found to contain Sarcocystidae DNA were used selectively for PCR amplification and sequencing of the nuclear 18S and 28S ribosomal (r) RNA genes and internal transcribed spacer 1 (ITS1) region, as well as the mitochondrial cytochrome b (cytb) and cytochrome c oxidase subunit 1 (cox1) genes. Sequence comparisons revealed that both otters were infected by the same Sarcocystis sp. and that there was no genetic variation (100 % identity) among sequenced isolates at the 18S and 28S rRNA genes (six identical isolates at both loci) or at cox1 (13 identical isolates). PCR products comprising the ITS1 region, on the other hand, had to be cloned before sequencing due to intraspecific sequence variation. A total of 33 clones were sequenced, and the identities between them were 97.9-99.9 %. These sequences were most similar (93.7-96.0 % identity) to a sequence of Sarcocystis kalvikus from the wolverine in Canada, but the phylogenetic analyses placed all of them as a monophyletic sister group to S. kalvikus. Hence, they were considered to represent a novel species, which was named Sarcocystis lutrae. Sequence comparisons and phylogenetic analyses based on sequences of the 18S and 28S rRNA genes and cox1, for which little or no sequence data were available for S. kalvikus, revealed that S. lutrae otherwise was most closely related to various Sarcocystis spp. using birds or carnivores as intermediate hosts. The cox1 sequences of S. lutrae from the otters were identical to two sequences from an arctic fox, which in a previous study had been assigned to Sarcocystis arctica due to a high identity (99.4 %) with the latter species at this gene and a complete identity with S. arctica at three other loci when using the same DNA samples as templates for PCR reactions. Additional PCR amplifications and sequencing of cox1 (ten sequences) and the ITS1 region (four sequences) using four DNA samples from this fox as templates again generated cox1 sequences exclusively of S. lutrae, but ITS1 sequences of S. arctica, and thus confirmed that this arctic fox had acted as intermediate host for both S. arctica and S. lutrae. Based on the phylogenetic placement of S. lutrae, the geographical location of infected animals (otters, arctic fox) and the distribution of carnivores/raptors which may have interacted with them, the white-tailed eagle (Haliaeetus albicilla) seems to be a possible definitive host of S. lutrae. Some of the muscle samples from both otters were shown to harbour stages of Toxoplasma gondii through PCR amplification and sequencing of the entire ITS1 region (five isolates) and/or the partial cytb (eight isolates) and cox1 (one isolate). These sequences were identical to several previous sequences of T. gondii in GenBank. Thus, both otters had a dual infection with S. lutrae and T. gondii.

Experimental parapoxvirus infection (contagious ecthyma) in semi-domesticated reindeer (Rangifer tarandus tarandus).

Contagious ecthyma (contagious pustular dermatitis, orf) occurs world-wide in sheep and goats and is caused by orf virus (genus Parapoxvirus, family Poxviridae). Contagious ecthyma outbreaks have been described in semi-domesticated reindeer (Rangifer tarandus tarandus) in Sweden, Finland and Norway, occasionally with high mortality. Fourteen one-year-old reindeer were corralled in mid-April. One week after arrival, two animals received a commercial live orf virus vaccine for sheep (Scabivax(®)) on scarified skin of the medial thigh. Four weeks later, the two vaccinated and six additional animals were inoculated in scarified oral mucosa with parapoxvirus obtained from reindeer with clinical contagious ecthyma. The remaining six reindeer were kept as sentinels, sharing feed and water with the inoculated animals. A small whitish lesion appeared on the inoculation site and the labial skin-mucosa junction of three animals five days post inoculation (p.i.). Twelve days p.i., typical ecthyma lesions were visible on the inoculation site in six of eight animals, including both vaccinees. Four inoculated animals (including both vaccinees) and one sentinel seroconverted 12 days p.i., and five animals (including one sentinel) seroconverted 20 days p.i. No contagious ecthyma-like lesions were detected in the sentinels. All animals were euthanized at 26-29 days p.i. Histological examination of lesions showed proliferative dermatitis with epidermal hyperplasia, hyperkeratosis, intra-epithelial pustules and ulcers. Orf virus DNA was detected in mandibular lymph nodes, tonsils and mucosal lesions of four animals, including one sentinel, which showed that virus transmission took place. The commercial orf virus vaccine may be difficult to administer due to the need for close-cropping and its zoonotic nature, and did not indicate significant protection, although the latter has to be verified with a larger number of animals.

Persistent organic pollutants and mercury in dead and dying glaucous gulls (Larus hyperboreus) at Bjørnøya (Svalbard).

Dead and dying glaucous gulls (Larus hyperboreus) were collected on Bjørnøya in the Barents Sea in 2003, 2004 and 2005. Autopsies of the seabirds only explained a clear cause of death for three (14%) of the 21 birds. A total of 71% of the birds were emaciated. Liver and brain samples were analysed for organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs), polybrominated diphenyl ether (PBDEs), hexabromocyclododecanes (HBCDs) and mercury (Hg). High levels of SigmaOCPs, SigmaPCBs, SigmaPBDEs and alpha-HBCD were found in liver and brain. Compared to the dead and dying glaucous gulls found 1989, the congeners' composition tended to change toward more persistent compounds in the 2003-2005 samples. The brain levels of OCPs and PCBs did not differ between 1989 and 2003-2005, while the liver levels were significantly lower. The brain/liver ratio for PCB and PBDE significantly decreased with halogenations of the molecule, indicating a clear discrimination of highly halogenated PCBs and PBDEs entering the brain. There was further a clear negative correlation between contaminant concentrations and body condition. The brain levels were not as high as earlier published lethal levels of p,p'-DDE or PCB. However, more recent studies reported a range of sub-lethal OCP- and PCB-related effects in randomly sampled glaucous gulls. An additional elevation of pollutants due to emaciation may increase the stress of the already affected birds. The high brain levels of OCP, PCB and PBDE of present study might therefore have contributed to the death of weakened individuals of glaucous gull.

Fatal inanition in reindeer (Rangifer tarandus tarandus): pathological findings in completely emaciated carcasses.

BACKGROUND: In a project to determine the causes of winter mortality in reindeer in Finnmark County, northern Norway, the most frequent diagnosis turned out to be complete emaciation, despite several of the reindeer having been given silage for up to 4 weeks before they died. The present paper describes autopsy results and other findings in these animals. METHODS: Autopsies were made of 32 reindeer carcasses, and 28 of these were diagnosed as completely emaciated based on lack of visible fat and serous atrophy of subepicardial and bone marrow fat. Other investigations of the carcasses included histology, bacteriology, parasitology (counting of macro parasites and faecal egg counting), analysis of vitamin E and selenium in liver, chemical and botanical analysis of rumen content, analysis of lipid content in femur bone marrow and estimation of muscle atrophy by use of a muscle index. RESULTS: Main findings were: Low carcass weight, severe muscle atrophy, hemosiderosis in liver and spleen, subcutaneous oedema (18%) and effusions to body cavities (18%). Two types of lipofuscin granula were identified in the liver: One type occurred in liver endothelial cells of all carcasses, while the other type occurred in hepatocytes, and prevailed in adult animals. Abomasal haemorrhages, consistent with previously described stress lesions, was present in 68% of the carcasses. Diarrhoea occurred in 2 cases, and loose faecal consistency was associated with silage feeding. Rumen content was low in crude protein. Grass dominated rumen content in silage-fed carcasses, while reindeer on natural pastures had mainly woody plants, mosses and litter in rumen. Stem dominated the grass fraction in rumens with high grass content, indicating ruminal indigestion as a cause of emaciation in silage fed animals. Some cases had heavy infestation of parasites such as warble fly larvae (Hypoderma tarandi), throat bot larvae (Cephenemyiae trompe) and lung nematodes. CONCLUSION: Lack of appropriate amounts and/or appropriate quality of feed has been the main cause of emaciation, though heavy infestation of parasites may have contributed to the emaciation in some cases.

Serologic survey for antibodies against Mycobacterium a vium subsp. paratuberculosis in free-ranging cervids from Norway.

Affinity between protein-G and immunoglobulins from red deer (Cervus elaphus), moose (Alces alces), roe deer (Capreolus capreolus), and reindeer (Rangifer tarandus tarandus) was tested in a competition binding assay. Sera from red deer, reindeer, and moose inhibited the assay less than sera from cattle (less affinity), whereas sera from roe deer showed a slightly higher affinity to protein-G than did sera from cattle. The conclusion was made that protein-G could be used instead of anti-species antibodies for these cervid species, where the aim of the screening was to look for exposure or lack of exposure to mycobacteria in the tested populations. Serologic screening of 1,373 free-ranging cervids for antibodies against Mycobacterium avium subsp. paratuberculosis was conducted. All sera were tested by a protein-G-based antigen-absorbed enzyme-linked immunosorbent assay (ELISA). Seropositive moose (10/537; 1.9%), red deer (14/371; 3.8%), roe deer (6/49; 12.2%), and semidomesticated reindeer (11/325; 3.4%) were found, whereas wild reindeer (n = 91) were seronegative. In addition, the red deer sera were tested with a commercial ELISA, by which two animals tested positive and nine were suspicious of having M. avium subsp. paratuberculosis antibodies. Tissue samples and feces from 10 moose originating from a population with a clustering of seropositive animals were investigated by histology and bacteriology with negative results. Paratuberculosis has never been diagnosed in free-ranging or farmed cervid species in Norway. Thus, further studies are indicated to prove that the present findings reflect an infection with M. avium subsp. paratuberculosis.