Highly divergent 18S rRNA gene paralogs in a Cryptosporidium genotype from eastern chipmunks (Tamias striatus).

Cryptosporidium is an apicomplexan parasite that causes the disease cryptosporidiosis in humans, livestock, and other vertebrates. Much of the knowledge on Cryptosporidium diversity is derived from 18S rRNA gene (18S rDNA) phylogenies. Eukaryote genomes generally have multiple 18S rDNA copies that evolve in concert, which is necessary for the accurate inference of phylogenetic relationships. However, 18S rDNA copies in some genomes evolve by a birth-and-death process that can result in sequence divergence among copies. Most notably, divergent 18S rDNA paralogs in the apicomplexan Plasmodium share only 89-95% sequence similarity, encode structurally distinct rRNA molecules, and are expressed at different life cycle stages. In the present study, Cryptosporidium 18S rDNA was amplified from 28/72 (38.9%) eastern chipmunks (Tamias striatus). Phylogenetic analyses showed the co-occurrence of two 18S rDNA types, Type A and Type B, in 26 chipmunks, and Type B clustered with a sequence previously identified as Cryptosporidium chipmunk genotype II. Types A and B had a sister group relationship but shared less than 93% sequence similarity. In contrast, actin and heat shock protein 70 gene sequences were homogeneous in samples with both Types A and B present. It was therefore concluded that Types A and B are divergent 18S rDNA paralogs in Cryptosporidium chipmunk genotype II. Substitution patterns in Types A and B were consistent with functionally constrained evolution; however, Type B evolved more rapidly than Type A and had a higher G+C content (46.3% versus 41.0%). Oocysts of Cryptosporidium chipmunk genotype II measured 4.17 µm (3.73-5.04 µm) × 3.94 µm (3.50-4.98 µm) with a length-to-width ratio of 1.06 ± 0.06 µm, and infection occurred naturally in the jejunum, cecum, and colon of eastern chipmunks. The findings of this study have implications for the use of 18S rDNA sequences to infer phylogenetic relationships.

Odontogenic myxoma in an 8-year-old Labrador Retriever dog.

Odontogenic myxoma (OM) was diagnosed in an 8-year-old Labrador Retriever dog with an ulcerohemorrhagic mass located on the caudal area of the right maxillary gingiva. The neoplasm was characterized by a low mitotic index and moderate numbers of spindle, stellate, and round cells that were sparsely distributed in an alcian blue reactive myxomatous matrix. Individual neoplastic cells were characterized by small amounts of faintly eosinophilic staining cytoplasm, prominent nucleoli, and stippled amphophilic staining chromatin that was immunoreactive for vimentin but negative for cytokeratin and actin. To the authors' knowledge, this is the first reported case of canine OM from North America, and it shares histomorphologic and histochemical features with 3 other cases reported in dogs elsewhere. Whereas, a literature review suggests untreated canine OM is insidious and locally aggressive, the prognosis in the present dog remains unknown. These findings support previous recommendations for inclusion of canine OM on the World Health Organization list of odontogenic tumors.

Escherichia coli, Salmonella, and Mycobacterium avium subsp. paratuberculosis in wild European starlings at a Kansas cattle feedlot.

The prevalence of Escherichia coli, Salmonella spp., and Mycobacterium avium subsp. paratuberculosis isolated from the feces of wild European starlings (Sturnus vulgaris) humanely trapped at a feedlot in central Kansas was assessed. All E. coli and Salmonella isolates recovered were tested for antimicrobial susceptibility using National Antimicrobial Resistance Monitoring System panels and the E. coli isolates were classified as to their content of genes associated with pathogenic E. coli of birds and cattle, including cvaC, iroN2, ompTp, hlyF2, eitC, iss, iutA, ireA, papC, stxI, stxII, sta, K99, F41, and eae. Escherichia coli O157:H7 and Mycobacterium avium subsp. paratuberculosis were not detected and Salmonella was isolated from only three samples, two of which displayed antimicrobial resistance. Approximately half of the E. coli isolates were resistant to antimicrobial agents with 96% showing resistance to tetracycline. Only one isolate was positive for a single gene associated with bovine pathogenic E. coli. An interesting finding of this study was that 5% of the E. coli isolates tested met the criteria established for identification as avian pathogenic E. coli (APEC). Thus these findings suggest that starlings are not a significant source of Salmonella spp., Mycobacterium avium subsp. paratuberculosis, E. coli O157, or other shiga toxin-producing E. coli in this feedlot. However, they may have the potential to spread APEC, an important pathogen of poultry and a potential pathogen to human beings.

Risk factors associated with anthrax outbreak in animals in North Dakota, 2005: a retrospective case-control study.

OBJECTIVE: We identified the risk factors associated with the anthrax outbreak Of 2005 in animals in North Dakota. METHODS: Medical records of the 2005 anthrax outbreak were obtained from the Veterinary Diagnostic Laboratory at North Dakota State University. Additional data were obtained from the North Dakota state veterinarian's office, and supplemental questionnaires were administered to producers. The data obtained included ecological and environmental factors, animal health factors, and management factors. RESULTS: Anthrax occurred from July 1 to October 12, 2005. The cases were located in eastern North Dakota around the Red River Basin. Ransom, LaMoure, and Barnes counties reported most cases (71%). Species affected included cattle, bison, horses, sheep, elk, deer, pigs, and llamas. The predominant symptom was sudden death (38%) followed by bleeding from orifices (17%). Chi-square analysis indicated significant differences between case and control premises on the following variables: death reported on neighboring pasture, vaccination period, dry conditions, wet conditions, antibiotic use, multiple vaccination, and type of predator (coyote). Factors that significantly (p<0.05) predicted anthrax occurrences on the final logistic regression model were vaccination, use of antibiotics during an outbreak, and period of vaccine administration (before or during the outbreak). CONCLUSIONS: The characteristics of the anthrax outbreak regarding time and place of occurrence, animals affected, clinical signs reported, and mortality rate were consistent with previous reports of natural anthrax outbreaks in animals. A number of factors that significantly predicted anthrax occurrence in animals in the 2005 outbreak in North Dakota were identified. This information is important in planning appropriate control and prevention measures for anthrax, including recommending the right vaccination and treatment regimens in managing future anthrax outbreaks.

Molecular and immunohistochemical detection of assemblage E, Giardia duodenalis in scouring North Dakota calves.

Despite many reports on the shedding of Giardia parasites by scouring calves, the role of Giardia as a cause of calf diarrhea is still controversial. To elucidate the role of Giardia duodenalis in calf scours, diagnostic samples from 189 scouring calves were tested by different assays during a 1-year-study period. Giardia antigens were detected in 22/189 scouring calves by a fecal-based enzyme-linked immunosorbent assay and 10 of these were positive for assemblage E, G. duodenalis by polymerase chain reaction. Giardia trophozoites were demonstrated by immunohistochemistry in intestinal sections from five calves in which the parasites were spatially distributed in areas of microscopically detectable enteritis. Our data suggest that under certain circumstances, Giardia may cause intestinal lesions leading to calf scours. Gnotobiotic calf-based infectivity studies are needed if the pathogenicity of Giardia in calves is to be definitively determined.

Ethylene glycol toxicosis in adult beef cattle fed contaminated feeds.

Acute deaths of cows held in a drylot and fed several crop processing plant by-products were investigated. Clinical signs in affected cows included diarrhea, ataxia, recumbency, hypersalivation, and sunken eyes. A histological diagnosis of ethylene glycol toxicosis, based on numerous birefringent crystals in renal tubules, was supported by toxicologic findings.

Diagnostic evidence of Staphylococcus warneri as a possible cause of bovine abortion.

Following a routine necropsy of a bovine fetus aborted at 5 months of gestation, placenta, fetal tissue samples, and stomach contents were subjected to a number of laboratory tests. Staphylococcus warneri was isolated in pure culture from the lung, liver, and stomach contents, whereas the placenta yielded S. warneri and a number of contaminants. Gross evaluation of agar plates showed predominant colonies to be morphologically consistent with those of S. warneri and the identity of the agent was further confirmed on a Trek Diagnostic Systems Sensititre, gram-positive identification (GPID) plate. Microscopic evaluation of fetal tissue sections showed extensive necrotizing lesions of the tongue, lung, and placenta in which there were numerous coccoid shaped gram-positive bacteria with morphology consistent with Staphylococcus spp. These results provide strong diagnostic evidence of S. warneri as a possible cause of bovine abortion and suggest there should be further investigations into the abortivirulence of this agent.

Hepatic lipidosis and other test findings in two captive adult porcupines (Erethizon dorsatum) dying from a "sudden death syndrome".

Routine postmortem examination and histologic evaluation of tissue sections demonstrated hepatic lipidosis (HL) in 2 adult captive porcupines with a history of sudden death. The male porcupine had a markedly enlarged pale liver that microscopically showed large unilocular vacuoles within hepatocellular cytoplasm. The periparturient female had similar but less marked hepatic lesions and an incidental pulmonary mycosis. These findings suggest HL as an important differential of spontaneous death in captive porcupines. It is hypothesized that in addition to the widely documented causes, HL in captive porcupines may be specifically associated with nutritional imbalances caused by the feeding of unsuitable commercial diets. The possible association of the condition with dietary and other factors in captive porcupines needs to be thoroughly investigated.

Evaluation of an outbreak of West Nile virus infection in horses: 569 cases (2002).

OBJECTIVE: To characterize an outbreak of West Nile virus (WNV) infection in horses in North Dakota in 2002, evaluate vaccine effectiveness, and determine horse characteristics and clinical signs associated with infection. DESIGN: Retrospective study. ANIMALS: 569 horses. PROCEDURE: Data were obtained from veterinary laboratory records, and a questionnaire was mailed to veterinarians of affected horses. RESULTS: Affected horses were defined as horses with typical clinical signs and seroconversion or positive results of virus isolation; affected horses were detected in 52 of the 53 counties and concentrated in the eastern and northeastern regions of the state. Among affected horses, 27% (n = 152) were vaccinated against WNV, 54% (309) were not, and 19% (108) had unknown vaccination status; 61 % (345) recovered, 22% (126) died, and 17% (98) had unknown outcome. The odds of death among nonvaccinated horses were 3 and 16 times the odds among horses that received only 1 or 2 doses of vaccine and horses that were vaccinated according to manufacturer's recommendations, respectively. Horses with recumbency, caudal paresis, and age > 5 years had higher odds of death, whereas horses with incoordination had lower odds of death, compared with affected horses without these characteristics. CONCLUSIONS AND CLINICAL RELEVANCE: Vaccination appears to have beneficial effects regarding infection and death caused by WNV.

Polymerase chain reaction and DNA sequencing for detection of ovine herpesvirus 2 in American bison (Bison bison).

Serum samples were collected at slaughter from 226 24-30-month-old ranch-raised, clinically normal American bison (Bison bison) bulls from North Dakota, Minnesota, Kansas, and Manitoba to assess the presence of antibodies to ovine herpesvirus 2 (OHV-2). Antibodies to OHV-2 were detected by competitive inhibition enzyme-linked immunosorbent assay in 10 of 226 (4.40%) samples. Polymerase chain reaction (PCR) analysis of sera positive for OHV-2 DNA demonstrated a 238 kilobase fragment. The nucleotide sequence of the PCR-positive samples in comparison to the reported OHV-2 nucleotide sequence resulted in a homology range of 82.8-95.4%.

Seroprevalence of OHV-2, BVDV, BHV-1, and BRSV in ranch-raised bison (Bison bison).

Serum samples were collected at slaughter from 226 24-30-month-old American bison (Bison bison) bulls from Kansas, Minnesota, North Dakota, and Manitoba and assayed for antibodies to ovine herpesvirus type-2 (OHV-2), bovine viral diarrhea virus (BVDV), bovine herpesvirus type-1 (BHV-1), and bovine respiratory syncytial virus (BRSV). Antibodies were detected by serum neutralization for BVDV, BHV-1, and BRSV, while antibodies to OHV-2 were detected by competitive inhibition-ELISA (CI-ELISA). Detectable antibodies were found against all viruses: 10 of 226 (4.40%) against OHV-2, 125 of 226 (55.3%) against BVDV, 99 of 226 (43.8%) against BHV-1, and 208 of 226 (92.0%) against BRSV. Titers from 93.6% of the BVDV-positive animals, 79.8% of the BHV-1-positive animals, and 98.1% of the BRSV-positive animals were > or = 1.25. These data indicate that a low percentage of clinically normal bison are seropositive for OHV-2 while a high percentage of bison sampled are seropositive for BVDV, BHV-1, and BRSV.