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Brucellosis in captive Rocky Mountain bighorn sheep (Ovis canadensis) caused by Brucella abortus biovar 4.

Nine (four female, five male) captive adult Rocky Mountain bighorn sheep (Ovis canadensis) contracted brucellosis caused by Brucella abortus biovar 4 as a result of natural exposure to an aborted elk (Cervus elaphus) fetus. Clinical signs of infection were orchitis and epididymitis in males and lymphadenitis and placentitis with abortion in females. Gross pathologic findings included enlargement of the testes or epididymides, or both, and yellow caseous abscesses and pyogranulomas of the same. Brucella abortus biovar 4 was cultured in all bighorn sheep from a variety of tissues, including testes/epididymides, mammary gland, and lymph nodes. All bighorn sheep tested were positive on a variety of standard Brucella serologic tests. This is the first report of brucellosis caused by B. abortus in Rocky Mountain bighorn sheep. It also provides evidence that bighorn sheep develop many of the manifestations ascribed to this disease and that infection can occur from natural exposure to an aborted fetus from another species. Wildlife managers responsible for bighorn sheep populations sympatric with Brucella-infected elk or bison (Bison bison) should be cognizant of the possibility of this disease in bighorn sheep.

Abortion, Veterinary↗

[Polymorphism of the kappa-casein gene in populations of the subfamily Bovinae].

Polymorphism of the 5'-untranslated region and exon 4 of kappa-casein (kappa-casein) gene was studied in Yakutian and Black Pied cattle, yak, European bison, and buffalo by means of a polymerase chain reaction and subsequent restriction fragment length polymorphism analysis (PCR-RFLP). In the species studied, restriction polymorphism by the endonucleases AluI and Bg/II in the 5'-untranslated region of the gene is absent. Four restriction endonucleases testing nucleotide substitutions in 136 codon (TaqI), 148 codon (HinfI and HindIII), and 167 and 168 codons (PstI) were used to study polymorphism of exon 4. The use of several restriction endonucleases allowed three alleles of kappa-casein (kappa-CnA, kappa-CnB, kappa-CnF) to be typed and new allele variants in yak, European bison, and buffalo to be revealed. Nucleotide sequences of the fragments of exon 4 studied were determined for two new alleles of the gene: kappa-CnG in yak and European bison and kappa-CnH in buffalo. Nucleotide substitutions determining new alleles were localized. In kappa-CnG, 148 and 168 codons coincide with the corresponding codons of kappa-CnB, and 136 and 167 codons correspond to kappa-CnA. Stop codons of kappa-CnG in yak are different from stop codons of other alleles of the gene: TGA, instead of TAA. The nucleotide sequence of exon 4 of kappa-CnH differs from bovine kappa-CnA by 15 nucleotide substitutions, causing 10 amino acid changes in the protein sequence, which coincide with the corresponding known amino acid sequence of kappa-casein in buffalo. Interbreed and interspecies differences in the profile of allele frequencies of the species studied were revealed. Aspects connected with evolution of the alleles of kappa-casein are discussed.

Alleles↗

Long-term viral serology of semi-free-living and captive ungulates.

Between 1973 and 1994, blood samples were collected at Whipsnade Wild Animal Park (UK) from three ungulate species kept in enclosures, including 28 European bison (Bison bonasus), 37 scimitar-horned oryx (Oryx dammah), and 49 Père David's deer (Elaphurus davidianus), and also from 144 semi-free-living Chinese water deer (Hydropotes inermis). These samples were tested for the presence of antibodies against three bovine viral diarrhea virus (BVDV)-like strains, three alpha-herpesvirus strains, enzootic hemorrhagic disease virus (EHDV) of deer, bovine respiratory syncytial virus (BRSV), bovine adenovirus 3 (BAV-3), and enzootic bovine leucosis virus (EBLV). Thirty-three individuals (13.1%) had antibodies to one or more of the three BVDV-like viruses, with titers ranging from 1:5 to 1:16, and 17 individuals (6.6%) were positive for antibodies to one or more of the three alpha-herpesviruses, with titers between 1:4 and 1:80. The highest titers and greatest proportion of seropositivity were against SH9/11, a recently isolated cytopathogenic pestivirus from wild roe deer (Capreolus capreolus). There were no positive reactors to BRSV and EHDV, and there was only one BAV-3 positive reactor, a scimitar-horned oryx, and one EBLV reactor, a European bison. There is no serologic evidence that semi-free-ranging Chinese water deer are important reservoirs or transmitters of the viral diseases investigated.

Animals↗

K-casein gene phylogeny of higher ruminants (Pecora, Artiodactyla).

To assess phylogenetic relationships among the higher ruminants (infraorder Pecora, order Artiodactyla), we analyzed K-casein DNA sequences, including 434 nucleotides of the fourth exon. The higher ruminant families Bovidae, Cervidae, Giraffidae, and Antilocapridae each have monophyletic K-casein sequences. Maximum parsimony and distance analyses identify Giraffidae as a sister group to either Cervidae or a Bovidae-Cervidae clade and Antilocapridae as a sister group to a Bovidae-Cervidae-Giraffidae clade. At a higher level these four families occur as a monophyletic clade relative to Tragulidae and Suidae. Within Cervidae, the subfamily Odocoileinae is monophyletic and Cervinae and Muntiacinae occur as independent lineages within a separate clade. Within Bovidae, the subfamilies Bovinae and Caprinae are monophyletic. Genera within Cervinae (Cervus, Elaphurus) and Bovinae (Bison, Bos) are paraphyletic. There is intraspecific allelic variation in Cervus elaphus, Odocoileus hemionus, and Bison bison. The rate of K-casein fourth exon DNA sequence evolution is estimated to be about 0.004 nucleotide substitutions per million years. The K-casein phylogeny is discussed relative to other molecular and morphological data.

Animals↗

Rumen microorganisms in buffalo from southern utah.

Rumen microbial populations from buffalo (Bison bison bison Linn.) in southern Utah were identified on the basis of their morphology and staining characteristics. The rumen bacteria and ciliate protozoa were similar in number and kind to those found in domestic livestock.

Journal Article↗

Immune responses of elk to initial and booster vaccinations with Brucella abortus strain RB51 or 19.

Previous studies have suggested that currently available brucellosis vaccines induce poor or no protection in elk (Cervus elaphus nelsoni). In this study, we characterized the immunologic responses of elk after initial or booster vaccination with Brucella abortus strains RB51 (SRB51) and 19 (S19). Elk were vaccinated with saline or 10(10) CFU of SRB51 or S19 (n=seven animals/treatment) and booster vaccinated with a similar dosage of the autologous vaccine at 65 weeks. Compared to nonvaccinates, elk vaccinated with SRB51 or S19 had greater (P<0.05) antibody responses to SRB51 or S19 after initial vaccination and after booster vaccination. Compared to nonvaccinated elk, greater (P<0.05) proliferative responses to autologous antigen after initial vaccination occurred at only a few sample times in SRB51 (6, 14, and 22 weeks) and S19 (22 weeks) treatment groups. In general, proliferative responses of vaccinates to nonautologous antigens did not differ (P>0.05) from the responses of nonvaccinated elk. Gamma interferon production in response to autologous or nonautologous Brucella antigens did not differ (P>0.05) between controls and vaccinates after booster vaccination. Flow cytometric techniques suggested that proliferation occurred more frequently in immunoglobulin M-positive cells, with differences between vaccination and control treatments in CD4+ and CD8+ subset proliferation detected only at 22 weeks after initial vaccination. After booster vaccination, one technique ([3H]thymidine incorporation) suggested that proliferative responses to SRB51 antigen, but not S19 antigen, were greater (P<0.05) in vaccinates compared to the responses of nonvaccinates. However, in general, flow cytometric and other techniques failed to detect significant anamnestic responses to autologous or nonautologous Brucella antigens in S19 or SRB51 vaccinates after booster vaccination. Although some cellular immune responses were detected after initial or booster vaccination of elk with SRB51 or S19, our data suggest that responses tend to be transient and much less robust than previously reported in SRB51-vaccinated cattle (Bos taurus) or bison (Bison bison). These data may explain why the vaccination of elk with S19 and SRB51 induces poor protection against brucellosis.

Animals↗

A simpler, more sensitive competitive inhibition enzyme-linked immunosorbent assay for detection of antibody to malignant catarrhal fever viruses.

An earlier competitive inhibition enzyme-linked immunosorbent assay (CI-ELISA) was developed for detection of specific antibody against malignant catarrhal fever (MCF) viruses (MCFV) in ruminants. In this study, the indirect CI-ELISA was improved by conjugating the monoclonal antibody 15-A directly with horseradish peroxidase and by developing a method of producing precoated, dried antigen plates. This new test is referred to as a direct CI-ELISA. The reformatted test yielded a significantly improved sensitivity, and the time required was reduced to about one-sixth of the previous time. Of 37 MCF cases in cattle that were confirmed by histopathology and polymerase chain reaction (PCR) assay, 37 (100%) were positive by the new test, whereas the indirect CI-ELISA detected only 23 (62%). The direct CI-ELISA detected antibody to MCFV in 100% of 48 sheep that had been defined as infected with ovine herpesvirus 2 (OvHV-2) by PCR, whereas the indirect CI-ELISA detected only 41 (85%). Comparison of antibody titers measured by the 2 assays for sera collected from OvHV-2-infected sheep and from cattle, bison, and deer with clinical sheep-associated MCF revealed that the direct CI-ELISA offered a 4-fold increase in analytical sensitivity over the indirect format. The number of seropositive animals detected by the direct CI-ELISA among apparently normal cattle and bison was 2-3 times greater than the number detected by the indirect CI-ELISA, indicating that a significant percentage of normal cattle and bison are subclincally infected with MCFV.

Animals↗

Protein G binding to enriched serum immunoglobulin from nondomestic hoofstock species.

Quick and cost-effective serologic assays, such as those based on enzyme-linked immunosorbent assay (ELISA) technology, are useful for screening animal populations for infectious diseases. Recombinant protein G is described as an almost universal ELISA conjugate for the detection of antibodies from a wide range of animal species. However, there is limited data documenting the ability of protein G to bind immunoglobulin (Ig) from many captive and free-ranging nondomestic hoofstock (Order Artiodactyla, e.g., elk, antelope, bison). Protein G binding to Ig from 11 species within this taxonomic order (addax, antelope, bison, bontebok, elk, impala, kudu/nyala, muntjac, oryx, sheep, and white-tailed deer) and 2 control species (bovine and chicken) was assessed. A serum Ig enrichment protocol, using high-performance liquid chromatography (HPLC), was optimized in bovids (Bos taurus) and then applied to the other study species. Binding assays were performed by adding protein G to microtiter wells coated with titrated dilutions of enriched artiodactyl Ig. Optical densities were measured and binding curves generated. Differences in protein G binding were observed, both within and among species, as well as within taxonomic families. Significant intraspecies binding variation was observed for 7 species tested (antelope, oryx, sheep, muntjac, impala, bontebok, and addax). No statistically significant intraspecies differences in protein G binding were found for Ig from bison, elk, kudu/nyala, white-tailed deer, plus control species (cattle and chicken). Binding of protein G to Ig from impala, muntjac, and elk was statistically different from the positive control (cattle), with muntjac binding curves statistically comparable with the negative control (chicken). For the other 7 species tested, binding curves illustrated the ability of protein G to bind Ig as well as, or better than, the positive control. These findings expand the list of animal species whose Ig is capable of being detected using recombinant protein G, with the caveat that protein G does not bind Ig uniformly in closely related species. It is concluded that recombinant protein G conjugates may serve as useful reagents for serodiagnosis by ELISA in nondomestic hoofstock, although different assay interpretation algorithms and assay protocols may need to be developed on a per species basis for maximum diagnostic effectiveness.

Animals↗

Frequency of pathology in a large natural sample from Natural Trap Cave with special remarks on erosive disease in the Pleistocene.

Population data are presented for erosive arthritis, osteoarthritis, diffuse idiopathic skeletal hyperostosis (DISH), joint eburnation and dental injury in a fauna from Natural Trap Cave, Wyoming, represented by over thirty thousand bones from twenty-four different species. Erosive arthritis is limited to the bovids, Bison, Ovis and Bootherium. Erosive arthritis is also present in bison from the late Pleistocene Twelve Mile Creek site in Kansas and from an early Holocene site in Wisconsin. The restriction of the known Pleistocene occurrences to bovids indicate the presence of a pathogen that predisposes bovids to erosive arthritis. The pathogen was identified as Mvcobacterium tuberculosis. Osteoarthritis and DISH are rare in the Natural Trap Cave, although Bison shows a relatively high occurrence of the former. Tooth breakage due to errors in bone manipulation was a problem for carnivores and one lion, Pantera atrox, was apparently reduced by joint disease to a scavenging lifestyle.

Animals↗

Sarcocystis in wild ungulates in Alberta.

Muscle samples from 557 wild ungulates in Alberta, comprising seven species, were examined grossly and/or histologically for cysts of Sarcocystis. Sarcocystis was found in 100, 96, 94, 75, 75, 73, and 49% of the wapiti (Cervus canadensis), moose (Alces alces), bison (Bison bison), mule deer (Odocoileus hemionus), bighorn sheep (Ovis canadensis), mountain goat (Oreamnos americanus), and white-tailed deer (O. virginianus), respectively.

Age Factors↗

Application of a competitive ELISA for the detection of bluetongue virus antibodies in llamas and wild ruminants.

A competitive enzyme-linked immunosorbent assay (C-ELISA), using a group-specific monoclonal antibody against bluetongue virus (BTV), was applied to detect anti-BTV antibodies in serum samples from two llamas (Llama glama) experimentally infected with BTV serotype 10. Antibodies were detected in both llamas by 1 wk or 2 wk post-infection. Antibodies to BTV increased exponentially during the first 4 wk in both llamas and stabilized at an elevated level during the remaining 5-wk-period of the experiment. We evaluated the C-ELISA for 1,442 field sera from bluetongue-free areas, collected from 398 llamas in New Zealand as well as 451 elk (Cervus elaphus canadensis), 323 bison (Bison bison) and 270 reindeer (Rangifer tarandus tarandus) in Canada. Based on the frequency distribution of the C-ELISA values, we propose that the current negative cut-off value of 50% inhibition established for bovine field sera also can be applied to the sera from these wild ruminants. The C-ELISA values for other wild ruminant field sera collected in bluetongue-free areas of Canada from 98 native caribou (Rangifer tarandus caribou), 32 white-tailed deer (Odocoileus virginianus), 14 moose (Alces alces), and nine musk-oxen (Ovibos maschatus) and 15 yak (Bos grunniens) also were less than 50%, with the exception of three caribou samples. Based on our results, we propose that the C-ELISA be used as a rapid and specific test for serodiagnosis of BTV infection in llamas and possibly other wild ruminants.

Animals↗

Prevalence of antibody to malignant catarrhal fever virus in wild and domestic ruminants by competitive-inhibition ELISA.

A competitive-inhibition ELISA (CI-ELISA), based on a monoclonal antibody to an epitope conserved among malignant catarrhal fever virus (MCFV) strains of both wildebeest and sheep origin, was used to determine the prevalence of antibody to MCFV in selected domestic and wild ruminants, both free-ranging and captive, from the USA. We evaluated 2528 sera from 14 species between 1990 and 1995, including 80 pronghorn antelope (Antilocapra americana), 339 bighorn sheep (Ovis canadensis), 103 biston (Bison bison), 17 black-tailed deer (Odocoileus hemionus columbianus), 395 domestic cattle (Bos taurus), 291 domestic goats (Capra hircus), 680 domestic sheep (Ovis ammon), 323 elk (Cervus elaphus), 41 llamas (Lama glama), 21 mouflon sheep (Ovis musimon), 54 mountain goats (Oreamnos americanus), 101 mule deer (Odocoileus hemionus), 20 muskox (Ovibos moschatus), and 63 white-tailed deer (Odocoileus virginianus). A high seroprevalence (37 to 62%) was observed in domestic sheep, domestic goats, muskox, and some bighorn sheep populations. Seroprevalence in these species was generally age-related: a very low seroprevalence was present in these animals under one year of age. A low seroprevalence (2% to 13%) was found in clinically-susceptible species such as domestic cattle, deer, elk and bison, supporting the concept that significant numbers of non-lethal infections occur among clinically susceptible ruminants.

Age Factors↗

Experimental infection of nontarget species of rodents and birds with Brucella abortus strain RB51 vaccine.

The Brucella abortus vaccine strain RB51 (SRB51) is being considered for use in the management of bnucellosis in wild bison (Bison bison) and elk (Cervus elaphus) populations in the Greater Yellowstone Area (USA). Evaluation of the vaccines safety in non-target species was considered necessary prior to field use. Between June 1998 and December 1999, ground squirrels (Spermophilus richardsonii, n = 21), deer mice (Peromyscus maniculatus, n = 14), prairie voles (Microtus ochrogaster, n = 21), and ravens (Corvus corax, n = 13) were orally inoculated with SRB51 or physiologic saline. Oral and rectal swabs and blood samples were collected for bacteriologic evaluation. Rodents were necropsied at 8 to 10 wk and 12 to 21 wk post inoculation (PI), and ravens at 7 and 11 wk PI. Spleen, liver and reproductive tissues were collected for bacteriologic and histopathologic evaluation. No differences in clinical signs, appetite, weight loss or gain, or activity were observed between saline- and SRB51-inoculated animals in all four species. Oral and rectal swabs from all species were negative throughout the study. In tissues obtained from SRB51-inoculated animals, the organism was isolated from six of seven (86%) ground squirrels, one of six (17%) deer mice, none of seven voles, and one of five (20%) ravens necropsied at 8, 8, 10, and 7 wk PI, respectively. Tissues from four of seven (57%) SRB51-inoculated ground squirrels were culture positive for the organism 12 wk PI; SRB51 was not recovered from deer mice, voles. or ravens necropsied 12, 21, or 11 wk, respectively, PI. SRB51 was not recovered from saline-inoculated ground squirrels, deer mice, or voles at any time but was recovered from one saline-inoculated raven at necropsy, 7 wk PI, likely attributable to contact with SRB51-inoculated ravens in an adjacent aviary room. Spleen was time primary tissue site of colonization in ground squirrels, followed by the liver and reproductive organs. The results indicate oral exposure to SRB51 does not produce morbidity or mortality in ravens, ground squirrels, deer mice, or prairie voles.

Animals↗

Safety of Brucella abortus strain RB51 in deer mice.

Brucella abortus strain RB51 is an approved brucellosis vaccine for use in cattle that may have potential as an oral vaccine for use in elk (Cervus elaphus) and/or bison (Bison bison). This study was designed to determine effects of strain RB51 on deer mice (Peromyscus maniculatus), a nontarget species that could have access to treated baits in a field situation. In February 1994, 90 mice were orally dosed or intraperitoneally injected with 1 x 10(8) colony forming units strain RB51 and 77 controls were similarly dosed with sterile saline. At weekly intervals through early April 1994, 4 to 6 mice from each group were euthanized, gross necropsies performed, spleens and uteruses cultured, and tissues examined histologically. All orally inoculated mice cleared the infection by 6 wk post-inoculation (PI). While most of the injected mice cleared the infection by 7 wk PI, a few required 9 wk. There were minimal adverse effects attributable to strain RB51. Apparently, strain RB51 would not negatively impact P. maniculatus populations if it were used in a field situation. Also, deer mice appear to be able to clear the vaccine in 6 to 9 wk, thus the probability of these mice transmitting the vaccine to other animals is low.

Administration, Oral↗

Immune responses of elk to vaccination with Brucella abortus strain RB51.

In a study conducted from January to August 2000, elk (Cervus elaphus) were vaccinated with Brucella abortus strain RB51 (SRB51, n = 6) or injected with 0.15 M NaCl solution (n = 3) at approximately 6 mo of age. Beginning at 2 wk and continuing to 25 wk after vaccination, SRB51-vaccinated elk had greater antibody responses (P < 0.05) to SRB51 when compared to nonvaccinated elk. Peripheral blood mononuclear cells (PBMC) from SRB51-vaccinated elk had greater (P < 0.05) proliferative responses to SRB51 at 18 wk after vaccination when compared to responses of nonvaccinated elk. Strain RB51 was recovered from blood samples of all vaccinates at 2 wk, and three of six vaccinates at 4 wk after vaccination. The SRB51 vaccine strain was recovered from the superficial cervical lymph node of all vaccinates sampled at 6 wk after vaccination. but not from lymph node samples obtained from vaccinates at 12 or 18 wk after vaccination. At 34 wk after vaccination, SRB51 was recovered from the bronchial lymph node of one of five vaccinates but not from other tissues. Strain RB51 was not recovered at any time from samples obtained from nonvaccinated elk. This study suggests that following vaccination with SRB51, elk remain bacteremic for a prolonged period of time, rapidly develop high antibody titers, and are slower to develop detectable proliferative responses in PBMC when compared to responses of cattle or bison (Bison bison).

Animals↗

Brucellosis in elk of eastern Idaho.

Brucellosis occurs in free-ranging elk (Cervus elaphus) and bison (Bison bison) in the Greater Yellowstone Area, which includes portions of Idaho, Wyoming, and Montana. Brucella abortus was first detected in elk in Idaho in 1998, and from 1998 to 2002, serologic surveillance of hunter-killed elk was conducted in northeastern and southeastern Idaho. Prevalence of antibodies in these elk varied annually, but averaged between 2% and 3%. Elk were also trapped in northeastern Idaho from 1998-2002 and tested for brucellosis using serology and tissue culture. In areas where artificial feeding of elk was done, antibody prevalence ranged from 12% to 80% depending on site, age, and sex. At one feeding site (Rainey Creek), a decline in the prevalence of antibodies (from 56.8% in 1999 to 13.5% in 2002) was detected after the removal of seropositive elk over 4 yr. Seropositive elk removed from two artificial winter feeding sites (Rainey Creek and Conant Creek) were euthanized and sampled or held in captivity and allowed to calve prior to euthanasia and necropsy. At necropsy, B. abortus biovar 1 and B. abortus biovar 4 were isolated from both cows and calves; however, biovar 4 was predominant. A dual infection with both biovars was found in one calf born to a seropositive cow from which biovar 4 was isolated. Abortions (16%), stillbirths (8%), and weak calves (4%) were observed in these elk. These findings confirm the presence of brucellosis in elk in eastern Idaho and provide information on disease management options.

Age Factors↗

The morphology of the hypoglossal canal and its size in relation to skull capacity in man and other mammal species.

The hypoglossal canal is a permanent element of the human skull. As well as the hypoglossal nerve, the canal also contains the venous plexus and an arterial branch leading to the dura mater. It emerged from our earlier studies that the venous plexus is a dominant component in this canal. In the present work the morphology and dimensions of the canal were studied on macerated skulls of humans and animals (rhesus monkey, European bison, fox, dog, cat, hare and rat). The hypoglossal canal was found in all the human and animal skulls examined. In both humans and animals the hypoglossal canal was frequently duplicated. The double canal was found in 43% specimens of human skulls. However, no triple division of the hypoglossal canal was found in the material under investigation. It was found that the hypoglossal canal in man, rhesus monkey and European bison had significant dimensions and in fact correlated with the size of skull capacity. This suggests that the hypoglossal canal is an essential venous emissary in man, rhesus monkey and European bison, but that in the remaining species it is of secondary importance in this respect.

Animals↗

The biology of bovine herpesvirus-4 infection of cattle.

The biology of bovine herpesvirus-4 (BHV-4) infection of cattle is reviewed. The infection is distributed worldwide. Most of isolated viruses are non-pathogenic in cattle; some of them are able to produce a genital disease. Twenty-nine structural polypeptides were described; ten of them are glycosylated. Two major glycoproteins were characterized by monoclonal antibodies. Restriction maps of BHV-4 DNA are available for the enzymes EcoRI, BamHi and HindIII. The strain variations studied by restriction analysis are very weak. The virus is able to persist in a latent state after primary infection. The identified sites of latency are nervous ganglia and mononuclear blood cells. The immune response of cattle after BHV-4 infection is characterized by low or undetectable levels of neutralizing antibodies. Four envelope proteins are recognized by convalescent sera and are the main antigenic components. Skin test remains negative in immunized cattle. Bovine herpesvirus-4 is not strictly species-specific: infection was proved in American bison (Bison bison), African buffalo (Syncerus caffer), sheep and probably cat, because feline herpesvirus-2 is in fact a BHV-4 strain. Finally BHV-4 shares antigenic and genomic relationships with alcelaphine herpesvirus-1, the causal agent of the African form of malignant catarrhal fever.

Animals↗