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  • Author or Editor: Steven Bolin x
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SUMMARY

Colostrum-deprived calves (n = 24) were fed various amounts of colostrum, colostrum substitute, or milk replacer to establish a range in titer of passively acquired viral neutralizing antibody in serum. The calves were then challenge exposed intranasally with a virulent, noncytopathic bovine viral diarrhea virus (bvdv-890). After viral challenge exposure, calves were monitored for fever, leukopenia, thrombocytopenia, and diarrhea. In addition, viral isolation and viral titration were performed on specimens of nasal secretions, buffy coat cells, and serum obtained from the calves. Fever and systemic spread of virus were detected in calves that had viral neutralizing titer of 256 or lower. Calves that had viral neutralizing titer lower than 16 developed severe clinical disease manifested by fever, leukopenia, thrombocytopenia, and diarrhea. Severity and duration of signs of disease decreased as titers of passively acquired viral neutralizing antibody increased. These results indicate that low to intermediate titers of passively acquired viral neutralizing antibody were not sufficient to fully protect calves from virulent bovine viral diarrhea virus.

Free access
in American Journal of Veterinary Research

Abstract

Objective—To determine whether an interferon (IFN)-γ response sufficient to categorize cattle as positive for tuberculosis can be detected in blood collected at commencement of exsanguination at slaughter.

Animals—15 Holstein cows.

Procedures—12 cows were experimentally sensitized by SC injection with inactivated Mycobacterium bovis in mineral oil, which induced an immune response that mimicked natural infection with M bovis. Three nonsensitized control cows were injected SC with mineral oil alone. By 5 weeks after injection, only the 12 sensitized cows had positive results for tuberculosis with whole blood IFN-γ assay. At that time, all 15 cows were sent to slaughter and samples of blood were collected from each cow immediately before stunning and at commencement of exsanguination (within 90 seconds after stunning). A whole blood IFN-γ assay was performed on the samples. Conditional probability and paired t tests were used to analyze changes in the categorical test interpretation and qualitative IFN-γ production, respectively.

Results—All 12 sensitized cows had positive results for tuberculosis in samples obtained immediately before stunning, and 9 retained positive results for samples obtained at commencement of exsanguination. There was a significant decrease in the mean background-corrected IFN-γ ELISA optical density values for samples obtained at commencement of exsanguination.

Conclusions and Clinical Relevance—IFN-γ response sufficient to classify cattle as positive for tuberculosis could be detected in blood collected at commencement of exsanguination. These findings support further development and use of the IFN-γ assay on blood samples collected at exsanguination as part of a bovine tuberculosis surveillance program.

Full access
in American Journal of Veterinary Research

Summary

A noncytopathic bovine viral diarrhea virus (bvdv), bvdv-890, isolated from a yearling heifer that died with extensive internal hemorrhages, was compared for virulence in calves with noncytopathic bvdv-TGAN, isolated from an apparently healthy persistently infected calf. After challenge exposure with bvdv-890, nonimmune calves (n = 7) developed fever > 40 C, diarrhea, leukopenia, lymphopenia, neutropenia, and thrombocytopenia. Most calves (n = 6) died or were euthanatized by 19 days after challenge exposure. Challenge exposure with bvdv-890 did not induce disease in 2 calves that had congenital persistent infection with bvdv or in 3 calves that had neutralizing antibody titer > 4 against bvdv-890. After challenge exposure with bvdv-TGAN, nonimmune calves (n = 7) developed fever > 40 C and, rarely, diarrhea or lymphopenia. All of those calves survived challenge exposure. The average maximal titer of bvdv-890 isolated from serum was 1,000 times that of bvdv-TGAN. In calves infected with bvdv-890, the average maximal percentages of lymphocytes and platelets associated with virus were greater than those found in calves infected with bvdv-TGAN. Additional findings of epidemiologic significance were prolonged shedding of virus and delayed production of viral-neutralizing antibody in 1 calf challenge-exposed with bvdv-890. Also, after production of neutralizing antibody, mutant virus that was refractory to neutralization was isolated from calves challenge-exposed with bvdv-TGAN.

Free access
in American Journal of Veterinary Research

Abstract

Objective—To examine effects of co-infection with porcine reproductive and respiratory syndrome virus (PRRSV) and Bordetella bronchiseptica in pigs.

Animals—Forty 3-week-old pigs.

Procedure—30 pigs (10 pigs/group) were inoculated with PRRSV, B bronchiseptica, or both. Ten noninoculated pigs were control animals.

Results—Clinical signs, febrile response, and decreased weight gain were most severe in the group inoculated with both organisms. The PRRSV was isolated from all pigs in both groups inoculated with virus. All pigs in both groups that received PRRSV had gross and microscopic lesions consistent with interstitial pneumonia. Bordetella bronchiseptica was cultured from all pigs in both groups inoculated with that bacterium. Colonization of anatomic sites by B bronchiseptica was comparable between both groups. Pigs in the group that received only B bronchiseptica lacked gross or microscopic lung lesions, and B bronchiseptica was not isolated from lung tissue. In the group inoculated with B bronchiseptica and PRRSV, 3 of 5 pigs 10 days after inoculation and 5 of 5 pigs 21 days after inoculation had gross and microscopic lesions consistent with bacterial bronchopneumonia, and B bronchiseptica was isolated from the lungs of 7 of those 10 pigs.

Conclusions and Clinical Relevance—Clinical disease was exacerbated in co-infected pigs, including an increased febrile response, decreased weight gain, and B bronchiseptica-induced pneumonia. Bordetella bronchiseptica and PRRSV may circulate in a herd and cause subclinical infections. Therefore, co-infection with these organisms may cause clinical respiratory tract disease and leave pigs more susceptible to subsequent infection with opportunistic bacteria. (Am J Vet Res 2000;61:892–899)

Full access
in American Journal of Veterinary Research

Abstract

Objective—To determine effects of intranasal inoculation with porcine reproductive and respiratory syndrome virus (PRRSV) or Bordetella bronchiseptica on challenge with nontoxigenic Pasteurella multocida in pigs.

Animals—Seventy 3-week-old pigs.

Procedure—In experiment 1, pigs were not inoculated (n= 10) or were inoculated with PRRSV (10), P multocida (10), or PRRSV followed by challenge with P multocida (10). In experiment 2, pigs were not inoculated (n = 10) or were inoculated with B bronchiseptica (10) or PRRSV and B bronchiseptica (10); all pigs were challenged with P multocida. Five pigs from each group were necropsied 14 and 21 days after initial inoculations.

ResultsPasteurella multocida was not isolated from tissue specimens of pigs challenged with P multocida alone or after inoculation with PRRSV. However, in pigs challenged after inoculation with B bronchiseptica, P multocida was isolated from specimens of the nasal cavity and tonsil of the soft palate. Number of bacteria isolated increased in pigs challenged after coinoculation with PRRSV and B bronchiseptica, and all 3 agents were isolated from pneumonic lesions in these pigs.

Conclusion and Clinical Relevance—Infection of pigs with B bronchiseptica but not PRRSV prior to challenge with P multocida resulted in colonization of the upper respiratory tract and tonsil of the soft palate with P multocida. Coinfection with PRRSV and B bronchiseptica predisposed pigs to infection of the upper respiratory tract and lung with P multocida. Porcine reproductive and respiratory syndrome virus and B bronchiseptica may interact to adversely affect respiratory tract defense mechanisms, leaving pigs especially vulnerable to infection with secondary agents such as P multocida. (Am J Vet Res 2001; 62:521–525)

Full access
in American Journal of Veterinary Research

Abstract

Objective—To determine whether vaccine virus can be detected by use of reverse transcriptase (RT)-PCR assays for pooled and individual skin samples obtained from cattle after vaccination with a commercially available modified-live bovine viral diarrhea virus (BVDV) vaccine.

Animals—12 BVDV-seropositive steer calves and 7 BVDV-seronegative (antibody titer < 1:4) heifers; all cattle were free of persistent infection with BVDV.

Procedures—2 experiments were conducted. Cattle were vaccinated on day 0 with a commercially available modified-live BVDV vaccine. Skin samples were collected on days 0, 3 to 14, 16, and 18 for virus detection by use of RT-PCR assay on individual and pooled samples. In addition, blood samples and nasal swab specimens were collected for virus isolation.

Results—All cattle, regardless of serologic status, had negative results for BVDV as determined by use of RT-PCR assay of individual and pooled skin samples. Virus was detected via virus isolation in serum or the buffy coat in 5 of 7 heifers that were seronegative when vaccinated.

Conclusions and Clinical Relevance—These findings indicated that it would be unlikely to detect BVDV vaccine virus in skin by use of RT-PCR assay of individual or pooled skin samples obtained from cattle after vaccination with a commercially available modified-live BVDV vaccine. Veterinarians and producers should be confident that positive test results for BVDV on skin samples would not likely be caused by the vaccination virus after administration of a modified-live virus vaccine.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To evaluate the efficacy of a commercially available killed bovine viral diarrhea virus (BVDV) vaccine to protect against fetal infection in pregnant cattle continually exposed to cattle persistently infected with the BVDV.

Animals—60 crossbred beef heifers and 4 cows persistently infected with BVDV.

Procedures—Beef heifers were allocated to 2 groups. One group was vaccinated twice (21-day interval between the initial and booster vaccinations) with a commercially available vaccine against BVDV, and the other group served as nonvaccinated control cattle. Estrus was induced, and the heifers were bred. Pregnancy was confirmed by transrectal palpation. Four cows persistently infected with BVDV were housed with 30 pregnant heifers (15 each from the vaccinated and nonvaccinated groups) from day 52 to 150 of gestation. Fetuses were then harvested by cesarean section and tested for evidence of BVDV infection.

Results—1 control heifer aborted after introduction of the persistently infected cows. Bovine viral diarrhea virus was isolated from 14 of 14 fetuses obtained via cesarean section from control heifers but from only 4 of 15 fetuses obtained via cesarean section from vaccinated heifers; these proportions differed significantly.

Conclusions and Clinical Relevance—A commercially available multivalent vaccine containing an inactivated BVDV fraction significantly reduced the risk of fetal infection with BVDV in heifers continually exposed to cattle persistently infected with BVDV. However, not all vaccinated cattle were protected, which emphasizes the need for biosecurity measures and elimination of cattle persistently infected with BVDV in addition to vaccination within a herd.

Full access
in American Journal of Veterinary Research

SUMMARY

Enriched populations of neutrophils and mononuclear leukocytes from 9 cattle persistently infected with non-cytopathic bovine viral diarrhea virus were analyzed for frequency of association with virus, using flow cytometric procedures. Trypsinization of neutrophils decreased the frequency of viral association from 0.82% to 0.49%. Similar treatment of mononuclear leukocytes decreased the frequency of viral association from 5.53% to 4.81%. Results of immunocytochemical procedures to locate viral antigen were inconclusive for neutrophils, but viral antigen was found in the cytoplasm of mononuclear leukocytes. A distinct and highly pure population of eosinophils was identified during flow cytometric analysis of neutrophil populations from 2 of 9 cattle.

Free access
in American Journal of Veterinary Research

Abstract

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.

Full access
in American Journal of Veterinary Research

SUMMARY

Tissues from cattle that died of experimentally induced mucosal disease (n = 3), naturally acquired mucosal disease (n = 6), or naturally acquired chronic bovine viral diarrhea (n = 4) were examined. Consistent findings were lymphocytic depletion of lymphoid tissues, degeneration of myenteric ganglion cells, and mild adrenalitis. Intracytoplasmic viral antigen was detected in myenteric ganglia and in endocrine glandular cells. Noncytopathic virus was isolated from all cattle, and cytopathic virus was isolated from 12 of 13 cattle.

Free access
in American Journal of Veterinary Research