Sufentanil and xylazine immobilization of Rocky Mountain elk

From October 2009 through July 2010, five captive, 3-yr-old, female Rocky Mountain elk (Cervus elaphus) and nine free-ranging elk (one male, eight female) were immobilized with 0.1 mg/kg sufentanil plus 0.5 mg/kg xylazine which was antagonized with 1 mg/kg naltrexone and 2 mg/kg tolazoline. Induction and recovery times averaged 4.9 ± 0.3 min and 3.9 ± 0.4 min, respectively. Physiologic and blood gas parameters as well as bispectral index (BIS) were measured on the captive elk every 10 min for 30 min. Immobilization induced profound hypoxemia via hypoventilation and ventilation-perfusion mismatching as demonstrated by depressed partial pressure of arterial oxygen (P(a)O(2)) and increased partial pressure of arterial carbon dioxide (P(a)CO(2)). The only values to significantly (P<0.05) change over time were base excess (BE), bicarbonate (HCO(3)), and lactate. Bispectral index is a measure of anesthetic depth. The average BIS value over the 30 min period (59.1 ± 2.4) was higher than the BIS value at the approximate point where elk lose consciousness, which indicated that this drug combination produced neuroleptanalgesia but not general anesthesia. Sufentanil and xylazine provided effective remote immobilization in elk and could be substituted for carfentanil or thiafentanil and xylazine should the need arise.     

Snakebite in captive Rocky Mountain elk (Cervus elaphus nelsoni)

Eight cases of snakebite occurred in seven of 11 captive Rocky Mountain elk (Cervus elaphus nelsoni) during June and July 1987. Severity of reactions to envenomation varied; affected elk presented with combinations of signs that included painful swelling restricted to the face and muzzle, submandibular edema, inspiratory dyspnea, epistaxis, frothy, blood-tinged nasal discharge, epiphora, anorexia and anxiousness or depression. We observed puncture wounds in only two cases. Treatment consisted of dexamethasone (about 0.1 mg/kg subcutaneously, single dose) and procaine penicillin G (about 25,000 IU/kg subcutaneously, once or twice daily, for 5 to 6 days), as well as revaccination using clostridium and tetanus toxoids. Swelling resolved and elk recovered in 3 to 5 days without complications. Using immunodiffusion, we detected serum antibodies to prairie rattlesnake (Crotalus viridis viridis) venom in six of seven affected elk, demonstrating seroconversion in three cases and anamnesis in one elk bitten twice. Venom was undetectable in any serum samples using similar techniques.     

Gene expression alterations in Rocky Mountain elk infected with chronic wasting disease

Chronic wasting disease (CWD) is an invariably fatal neurologic disease that naturally infects mule deer, white tailed deer and elk. The understanding of CWD neurodegeneration at a molecular level is very limited. In this study, microarray analysis was performed to determine changes in the gene expression profiles in six different tissues including brain, midbrain, thalamus, spleen, RPLN and tonsil of CWD-infected elk in comparison to non-infected healthy elk, using 24,000 bovine specific oligo probes. In total, 329 genes were found to be differentially expressed (> 2.0-fold) between CWD negative and positive brain tissues, with 132 genes upregulated and 197 genes downregulated. There were 249 DE genes in the spleen (168 up- and 81 downregulated), 30 DE genes in the retropharyngeal lymph node (RPLN) (18 up- and 12 downregulated), and 55 DE genes in the tonsil (21 up- and 34 downregulated). Using Gene Ontology (GO), the DE genes were assigned to functional groups associated with cellular process, biological regulation, metabolic process, and regulation of biological process. For all brain tissues, the highest ranking networks for DE genes identified by Ingenuity Pathway Analysis (IPA) were associated with neurological disease, cell morphology, cellular assembly and organization. Quantitative real-time PCR (qRT-PCR) validated the expression of DE genes primarily involved in different regulatory pathways, including neuronal signaling and synapse function, calcium signaling, apoptosis and cell death and immune cell trafficking and inflammatory response. This is the first study to evaluate altered gene expression in multiple organs including brain from orally infected elk and the results will improve our understanding of CWD neurodegeneration at the molecular level.     

Gene expression alterations in Rocky Mountain elk infected with chronic wasting disease

Chronic wasting disease (CWD) is an invariably fatal neurologic disease that naturally infects mule deer, white tailed deer and elk. The understanding of CWD neurodegeneration at a molecular level is very limited. In this study, microarray analysis was performed to determine changes in the gene expression profiles in six different tissues including brain, midbrain, thalamus, spleen, RPLN and tonsil of CWD-infected elk in comparison to non-infected healthy elk, using 24,000 bovine specific oligo probes. In total, 329 genes were found to be differentially expressed (> 2.0-fold) between CWD negative and positive brain tissues, with 132 genes upregulated and 197 genes downregulated. There were 249 DE genes in the spleen (168 up- and 81 downregulated), 30 DE genes in the retropharyngeal lymph node (RPLN) (18 up- and 12 downregulated), and 55 DE genes in the tonsil (21 up- and 34 downregulated). Using Gene Ontology (GO), the DE genes were assigned to functional groups associated with cellular process, biological regulation, metabolic process, and regulation of biological process. For all brain tissues, the highest ranking networks for DE genes identified by Ingenuity Pathway Analysis (IPA) were associated with neurological disease, cell morphology, cellular assembly and organization. Quantitative real-time PCR (qRT-PCR) validated the expression of DE genes primarily involved in different regulatory pathways, including neuronal signaling and synapse function, calcium signaling, apoptosis and cell death and immune cell trafficking and inflammatory response. This is the first study to evaluate altered gene expression in multiple organs including brain from orally infected elk and the results will improve our understanding of CWD neurodegeneration at the molecular level.     

Nematode-Trapping Fungi in Biological Control of Dictyocaulus viviparus

Larvae of the cattle lungworm Dictyocaulus viviparus were cultured in experimental units of 200 g cattle faeces placed in semi-transparent trays in the laboratory. In each of 4 experimental series using this experimental unit, chlamydospores (chl) of the nematode-trapping fungus Duddingtonia flagrans were admixed to half of the faecal cultures in a concentration of 50.000 chl/g. In all 4 series there was a significant reduction in the development and subsequent release of infective lungworm larvae from faecal cultures containing chlamydospores. The average reduction in larval release, caused by fungal spores, was 86%.     

Morphology of inhibited larvae of the bovine lungworm Dictyocaulus viviparus.

Some morphological features of inhibited fourth stage (L4) and fourth moult larvae (4M) of the bovine lungworm Dictyocaulus viviparus are described. Inhibition was induced by maintaining the third stage larvae (L3) for 6 weeks at 4 degrees C. Inhibited fourth stage (L4) and fourth moult larvae (4M) were collected by perfusion of the lungs of experimentally infected calves after necropsy at 15 and 68 days post infection (d.p.i.), respectively. Inhibited 4M, isolated at 68 d.p.i., were about ten times larger than inhibited L4 isolated at 15 d.p.i.     

The effects of elk on aspen in the winter range in Rocky Mountain National Park

National parks face problems in managing wild ungulates and their forage resources, including small park sizes and incomplete protection of winter ranges, absence of major predators, and influences from exterior management activities Our study focuses on the effects of elk Cervus elaphus browsing on aspen Populus tremuloides in the elk winter range of Rocky Mountain National Park Elk can prevent successful regeneration of aspen by suckers, and increase the mortality of established trees Here we quantify the effect of elk on aspen in the elk winter range of the park particularly since the adoption of a policy of “natural regulation” in 1968 We collected a stratified-random sample of aspen cohorts in the elk winter range, and also inside two long-term exclosures In each cohort we tallied live and dead trees by diameter, recorded the height and percentage of stem surface covered by bark-stripping, and estimated the date of regeneration of the cohort using increment cores We also tallied live and dead aspen suckers (young trees < 2 5 in tall) by height and the number of live and dead branches on each sucker Aspen outside the exclosures are declining as evidenced by 1) high mortality among established trees, including entire stands that are dead, 2) low density of live suckers, insufficient to regenerate the stand, 3) suckers that are excessively branched and seldom surviving to reach tree height, and 4) almost no cohort regeneration since adoption of natural regulation Exclosures and early photos of the elk winter range suggest these are anomalous conditions Over the last century, aspen cohorts regenerated only when there were fewer than ca 600 elk on the park's elk winter range, far fewer than the present estimated 1600 head Fire suppression, natural succession, fluctuations in beaver populations, climatic fluctuations, and other factors are of no or lesser importance than elk in the regeneration and survival of aspen cohorts on the elk winter range The decline of aspen is a concern when management is focused on the ecosystem scale rather than simply the scale of wildlife and their primary forage resources     

Psoroptic scabies in Rocky Mountain bighorn sheep (Ovis canadensis canadensis) from Wyoming

Thirteen Rocky Mountain bighorn sheep (Ovis canadensis canadensis) with clinical signs of psoroptic scabies were captured in Wyoming. Signs included droopy ears, depilation on the head and neck, and exudate in the ears. Mites were identified as either Psoroptes cervinus or P. equi. Two ewes with scabies at the time of original capture had no clinical signs of mite infection 1 and 2 yr later.     

Molecular characterization of the Rocky Mountain elk (Cervus elaphus nelsoni) PRNP putative promoter

Chronic wasting disease (CWD) is a transmissible spongiform encephalopathy (TSE) affecting deer (Odocoileus spp.), moose (Alces alces), and Rocky Mountain elk (Cervus elaphus nelsoni). Leucine homozygosity at elk PRNP codon 132 has been associated with reduced CWD susceptibility. However, naturally acquired CWD has been detected in elk possessing the 132 Leu/Leu genotype. Recent human and bovine studies indicate that PRNP regulatory polymorphisms may also influence TSE occurrence. Therefore, we generated sequences for the elk PRNP putative promoter (2.2 kb), exon 1 (predicted; 54 bp), intron 1 (predicted; 193 bp), and exon 3 (771 bp). Promoter prediction analysis using CpGProD yielded a single elk PRNP promoter that was homologous to regions of known promoter activity in cow and sheep. Molecular interrogation of the elk PRNP putative promoter revealed 32 diallelic single-nucleotide polymorphisms (SNPs). No variation was detected within the predicted exon 1 or intron 1 sequences. Evaluation of elk PRNP exon 3 revealed 3 SNPs (63Y, 312R, 394W-->Met/Leu). Bayesian haplotype reconstruction resulted in 3 elk PRNP haplotypes, with complete linkage disequilibrium observed between all PRNP putative promoter SNPs and codon 132. The results of this study provide the initial genomic foundation for future comparative and haplotype-based elk PRNP studies.     

Dictyocaulus viviparus: isolation and characterization of a recombinant antigen with potential for immunodiagnosis.

Crude adult worm antigen of Dictyocaulus viviparus was examined for specific antigens by SDS-PAGE and immunoblotting using sera from cattle experimentally infected with D. viviparus, vaccinated with a normal or a reduced dosage of the commercial lungworm vaccine, and helminth-free cattle. A D. viviparus-specific region M(r) 18,000 was identified and isolated. A lambda ZAP II cDNA expression library consisting of 4.4 x 10(5) recombinant clones (88% of the total number of clones) was constructed from D. viviparus adult worm mRNA. Rabbit antiserum to the M(r) 18,000 antigen was used to screen the cDNA library and eight positive clones were picked and allocated to the same antigenic family by sibling analysis. All clones were subcloned into the plasmid pGEX-2T, and the clone with highest expression yields was expressed as a glutathione S-transferase fusion protein (DvGST3-14) or, after cleavage with thrombin, as pure recombinant parasite protein (Dv3-14). The native parasite antigen encoded by the clone was identified. The immunodiagnostic potential of the recombinant proteins was assessed by immunoblotting.     

Factors associated with pathogen seroprevalence and infection in Rocky Mountain cougars

Serological and genetic material collected over 15 years (1990-2004) from 207 cougars (Puma concolor) in four populations in the Rocky Mountains were examined for evidence of current or prior exposure to feline immunodeficiency virus (FIV), feline parvovirus (FPV), feline coronavirus (FCoV), feline calicivirus (FCV), canine distemper virus (CDV), feline herpesvirus (FHV), and Yersinia pestis. Serologic data were analyzed for annual variation in seroconversions to assess whether these pathogens are epidemic or endemic in cougars, and to determine whether family membership, age, sex, or location influence risk of exposure. FIV and FPV were clearly endemic in the studied populations, whereas exposure to FCoV, FCV, CDV, and Y. pestis was more sporadic. No evidence was found for FHV. Age was the most consistent predictor of increased exposure risk, often with no other important factors emerging. Evidence for transmission within family groups was limited to FIV and FCoV, whereas some indication for host sex affecting exposure probability was found for FIV and Y. pestis. Overall, cougar populations exhibited few differences in terms of pathogen presence and prevalence, suggesting the presence of similar risk factors throughout the study region.     

In vitro migration of Dictyocaulus viviparus larvae: migration of the infective stage inagar gel.

The infective stage of the cattle lungworm, Dictyocaulus viviparus, was able to migrate in agar gel when activated by bile. The number of larvae which penetrated the upper surface of a 2 mm thick agar layer was counted and found to be independent from the agar concentration. Larvae which had migrated out of the agar remained on the surface and did not re-enter. The agar migration process was temperature dependent. Influence of time as well as dependency of agar thickness was investigated. The significance of these findings is discussed.     

Susceptibility of two Rocky Mountain bighorn sheep to experimental infection with Anaplasma ovis.

In North America, the role of wild ruminants in the epidemiology of anaplasmosis has not been clearly defined. Such information is particularly meager in regard to bighorn sheep. We report the susceptibility of two Rocky Mountain bighorn sheep (Ovis canadensis canadensis) to experimental infection with a well characterized field isolate of Anaplasma ovis obtained from domestic sheep in Idaho. Both bighorn sheep developed infection resulting in severe clinical disease, with relatively high parasitemias, icterus and anemia. One animal required tetracycline therapy and responded well to treatment, while the other recovered uneventfully without treatment. Both bighorn sheep were spleen-intact, a condition that in A. ovis-exposed domestic sheep typically is associated with mild infection. The results indicate that bighorn sheep may be adversely affected if exposed to the organism in nature.