Objective—To collect and partially characterize strains of bovine viral diarrhea viruses(BVDVs) isolated from persistently infected (PI) calves born to vaccinated dams, determine genetic diversity of the isolated viruses, and identify regional distribution of genetically similar virus subpopulations.
Sample Population—17 noncytopathic (NCP) BVDVs from PI calves from 11 herds of beef or dairy cattle.
Procedures—Viral RNA was extracted from infected cell cultures, and BVDV-specific PCR primers were used to amplify > 1,000 bases of the viral genome. Derived sequences were used for molecular phylogenetic analyses to determine the viral genotype and viral genogroup and to assess genetic similarity among BVDVs.
Results—Analysis of the 17 NCP strains of BVDV failed to detect a viral genotype or viral genogroup not already reported to exist in the United States. One virus was classified as genotype 1, genogroup 1b, and 16 viruses were classified as genotype 2, genogroup 2a. Genotype 2 strains were genetically diverse, and genetic similarities were not obvious among viruses from geographic regions larger than a small locale.
Conclusions and Clinical Relevance—Viruses isolated from herds where a genotype 1, genogroup 1a BVDV vaccine was administered prior to breeding were primarily genetically diverse genotype 2, genogroup 2a BVDVs. Vaccination with multiple BVDV genotypes may be needed to improve protection. Methods used in this study to obtain and analyze field strains are applicable to assessing efficacy of current BVDV vaccines. Candidates for future vaccines are viruses that appear able to elude the immune response of cattle vaccinated against BVDV with existing vaccines.
Case Description—3 unrelated, densely populated, dynamic ferret populations with severe outbreaks of enteric coccidiosis were evaluated.
Clinical Findings—In each outbreak, morbidity rate was high, there were an appreciable number of deaths, and ferrets of all ages were affected. Affected individuals had acute onset of diarrhea, and feces often contained frank or digested blood. Other clinical signs included dehydration, weakness, lethargy, and weight loss. Fecal examinations of affected ferrets revealed sporadic and inconsistent shedding of coccidial oocysts. Necropsy findings included moderate to marked atrophic enteritis associated with numerous intraepithelial and fewer extracellular coccidial life stages. Sporulated oocysts isolated from feces were consistent with Eimeria furonis. A PCR assay was performed on formalin-fixed, paraffin-embedded sections of intestine for the gene encoding the small subunit of rRNA yielded products with sequences identical to those described for E furonis.
Treatment and Outcome—Supportive care and treatment with sulfadimethoxine over the course of these outbreaks was palliative, but long-term treatment was required and failed to completely eradicate infection as identified by the subsequent finding of oocysts in fecal samples.
Clinical Relevance—Enteric coccidiosis due to infection with E furonis has typically been reported to be subclinical rather than to cause severe gastrointestinal disease in ferrets. This report indicated that infection with E furonis may have contributed to severe enteric disease with high morbidity and mortality rates in 3 densely populated, dynamic groups of ferrets. Furthermore, long-term treatment with anti-coccidials may be required in outbreak situations, but may be ineffectual in completely eradicating infection.
CASE DESCRIPTION Within a 2-week period, 4 southern cassowaries (Casuarius casuarius) at an exhibit at a Virginia zoo died acutely subsequent to eastern equine encephalitis virus (EEEV) infection. This prompted a search for other EEEV outbreaks in cassowaries, which resulted in the identification of 2 additional cassowaries that died of EEEV infection at a conservation center in Florida.
CLINICAL FINDINGS Both juvenile and adult birds were affected. Three of the 6 birds died acutely with no premonitory signs. Clinical disease in the other 3 birds was characterized by lethargy and ataxia. Clinicopathologic findings typically included leukocytosis, hyperuricemia, abnormally high liver enzyme activities, and hyper–β globulinemia, which was indicative of acute inflammation.
TREATMENT AND OUTCOME The 3 birds with clinical disease died despite supportive treatment. Gross abnormalities commonly observed during necropsy included coelomitis and evidence of diarrhea. Frequently observed histologic abnormalities were encephalitis, vasculitis, hepatitis, nephritis, and splenitis. The diagnosis of EEEV infection was confirmed by detection of serum anti-EEEV antibodies or detection of viral RNA in brain tissue by use of a reverse-transcriptase PCR assay.
CLINICAL RELEVANCE Findings suggested that EEEV can cause high morbidity and mortality rates in southern cassowaries. Clinical disease might be reduced or prevented by vaccination, isolation of ill birds, and mosquito control strategies.
OBJECTIVE To determine whether Mycobacterium bovis remains viable in ensiled forages.
SAMPLE Alfalfa, mixed mostly grass, and corn silages.
PROCEDURES For each of 10 sampling days, six 250-g replicate samples of each feedstuff were created and placed in a film pouch that could be vacuum sealed to simulate the ensiling process. Within each set of replicate samples, 4 were inoculated with 10 mL of mycobacterial liquid culture medium containing viable M bovis and 2 were inoculated with 10 mL of sterile mycobacterial liquid culture medium (controls) on day 0. Pouches were vacuum sealed and stored in the dark at room temperature. On the designated sampling day, 1 control pouch was submitted for forage analysis, and the other pouches were opened, and forage samples were obtained for M bovis culture and analysis with a PCR assay immediately and 24 hours later.
RESULTS None of the control samples had positive M bovis culture or PCR assay results. Among M bovis-inoculated samples, the organism was not cultured from alfalfa and corn silage for > 2 days but was cultured from mixed mostly grass silage for 28 days after inoculation and ensiling initiation. Mycobacterium bovis DNA was detected by PCR assay in samples of all 3 feedstuffs throughout the 112-day observation period.
CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that properly ensiled forages would be an unlikely source for M bovis transmission to cattle. Further research is necessary to determine whether ensiling kills M bovis or forces it to become dormant and, if the latter, elucidate the conditions that cause it to revert to an infectious state.