Detection of bovine herpesvirus type 1 in blood from naturally infected cattle by using a sensitive PCR that discriminates between wild-type virus and virus lacking glycoprotein E.
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In the present study, we report for the first time on the detection of bovine herpesvirus type 1 (BHV-1) in whole-blood samples derived from naturally infected cattle. Sensitive PCR assays specific for glycoprotein B (gB), gC, and gE of BHV-1 allow the detection of one BHV-1 DNA copy in 10(5) to 10(7) peripheral blood leukocytes (PBLs). The incidence of BHV-1-positive PBLs in naturally infected cattle appears to be quite high (92.2% positive PBLs among all samples tested), although in most cases only between 10(-5) and 10(-7) positive leukocytes were present. The results demonstrate that the viral DNA is detectable not only in the peripheral blood of acutely infected animals but, more importantly, also in the peripheral blood of subclinically infected cattle. The gE-specific PCR described in the report allows discrimination between wild-type (WT) virus-infected and vaccinated animals, which is of importance for control programs that use the recently introduced vaccination strategy with a gE-negative virus. The results further show that doubtful serological results can be verified or falsified and that individual animals can be monitored for the presence or absence of WT BHV-1 or gE-negative virus in cattle herds. The PCR protocols allow the detection of BHV-1 prior to seroconversion or in BHV-1-seronegative cattle. Finally, the results indicate the simultaneous presence of WT and gE-negative vaccine virus in the PBLs of several cattle. Therefore, investigations of viremia in naturally and experimentally infected cattle and on the identification of infected cell types of bovine PBLs can be now performed. (+info)
Listeriosis in a Holstein cow.
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Bovine listeriosis is a sporadie bacterial infection most commonly manifested by encephalitis or meningoencephalitis in adult animals. Cattle fed poorly fermented haylage or corn silage are at increased risk. Mortality is high without early antibiotic and supportive therapy. Listeria monocytogenes can affect many species and is a public health concern. (+info)
Human Ehlers-Danlos syndrome type VII C and bovine dermatosparaxis are caused by mutations in the procollagen I N-proteinase gene.
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Ehlers-Danlos syndrome (EDS) type VIIC is a recessively inherited connective-tissue disorder, characterized by extreme skin fragility, characteristic facies, joint laxity, droopy skin, umbilical hernia, and blue sclera. Like the animal model dermatosparaxis, EDS type VIIC results from the absence of activity of procollagen I N-proteinase (pNPI), the enzyme that excises the N-propeptide of type I and type II procollagens. The pNPI enzyme is a metalloproteinase containing properdin repeats and a cysteine-rich domain with similarities to the disintegrin domain of reprolysins. We used bovine cDNA to isolate human pNPI. The human enzyme exists in two forms: a long version similar to the bovine enzyme and a short version that contains the Zn++-binding catalytic site but lacks the entire C-terminal domain in which the properdin repeats are located. We have identified the mutations that cause EDS type VIIC in the six known affected human individuals and also in one strain of dermatosparactic calf. Five of the individuals with EDS type VIIC were homozygous for a C-->T transition that results in a premature termination codon, Q225X. Four of these five patients were homozygous at three downstream polymorphic sites. The sixth patient was homozygous for a different transition that results in a premature termination codon, W795X. In the dermatosparactic calf, the mutation is a 17-bp deletion that changes the reading frame of the message. These data provide direct evidence that EDS type VIIC and dermatosparaxis result from mutations in the pNPI gene. (+info)
Phenotypic analysis of local cellular responses in calves infected with bovine respiratory syncytial virus.
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Changes in lymphocyte subsets in the trachea, pulmonary tissue, bronchoalveolar lavage (BAL), peripheral blood and bronchial lymph node (BLN) of gnotobiotic calves infected with bovine respiratory syncytial virus (BRSV) were analysed by flow cytometry. Following BRSV infection, virus titres in the nasopharynx reached a peak between days 5 and 7 and infection was resolving from day 10. Although calves did not develop signs of clinical respiratory disease, there was evidence of gross pneumonia and histological changes typical of BRSV bronchiolitis, which were most extensive from day 710 of infection. Following BRSV infection there was a recruitment of CD8+ T cells into the trachea and lung, which peaked on day 10 after infection. Thus, there were approximately equal numbers of CD8+ and CD4+ T cells in the lung and trachea of uninfected calves, whereas by day 10 of infection, CD8+ cells outnumbered CD4+ cells by 3:1 in the lungs and 6:1 in the trachea of the infected calves. Although the increase in CD4+ T cells into the lungs was less marked than that of CD8+ T cells, changes in expression of CD45R, CD45RO, L-selectin and interleukin-2 receptors all suggested that CD4+ T cells were activated during BRSV infection. Changes in gamma delta T cells were not observed in BRSV-infected calves. There was a marked increase in B cells in the BLN after infection and BLN CD4+ T cells changed from the majority expressing L-selectin and CD45R in uninfected calves to a predominance of L-selectin- CD45R- CD45RO+ phenotype, 10 days after infection. In conclusion, CD8+ T cells constitute the major lymphocyte subpopulation in the respiratory tract of calves recovering from BRSV infection. (+info)
Evaluation of quantitative latex agglutination for detection of Cryptosporidium parvum, E. coli K99, and rotavirus in calf feces.
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A new methodology for detection of rotavirus, Escherichia coli K99, and Cryptosporidium parvum in bovine fecal samples was developed based on a quantitative latex agglutination technique (QLAT). Calibrated microspheres coated with specific antibodies to 1 of the enteric pathogens are quantitatively agglutinated by the antigens present in diluted fecal sample. The test is performed in a 96-well flat-bottom plate. The samples were tested with a commercial enzyme-linked immunosorbent assay kit prior to being analyzed by QLAT. The calculated sensitivity and specificity are adequate for field conditions, because the amount of the pathogenic agents is generally high. The overall time to perform the test was about 20 minutes. (+info)
Evaluation of an immunofluorescent antibody test to detect bovine herpesvirus 1-specific IgM.
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An indirect immunofluorescent antibody test (IIFAT) was developed to detect bovine herpesvirus 1 (BHV-1)-specific IgM. All sera were treated with protein-G agarose prior to testing to eliminate the possibility of false-positive results due to IgM-isotype rheumatoid factor (IgM-RF). Specific IgM was first detected 8 days after experimental infection of 3 calves free of maternally derived antibody, with peak responses occurring 2-7 days later. Seroconversion was detected in all 3 calves using a single-dilution enzyme-linked immunosorbent assay. Following reinfection at 30 days postinfection, a low-level IgM response was detected in only 1 calf. Seroconversion was detected in 2 calves. There was no evidence of activation of IgM-RF by infection or reinfection with BHV-1. When 87 acute and convalescent serum pairs collected from 21 outbreaks of respiratory disease were tested, specific IgM was detected in 58 animals (66.6%) from 19 (90.5%) outbreaks. Seroconversion was detected in 44 of these animals (50.6%) from 17 outbreaks (81.0%). The correlations between these 2 assays on a calf and outbreak basis were 79.3% and 90.5%, respectively. Specific IgM was detected in 17/20 sera (85.0%) collected from an additional outbreak. No virus was detected by virus isolation or immunofluorescent staining in nasal mucus samples collected at the same time. Detection of specific IgM by IIFAT is a useful technique for the serodiagnosis of BHV-1 infection. (+info)
Factors influencing the isolation of Mycobacterium avium subsp. paratuberculosis from bovine fecal samples.
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A modified procedure was used for culture of Mycobacterium paratuberculosis (Mptb) from bovine feces. Bovine fecal samples were decontaminated with NaOH, exposed to a mixture of oxalic acid and malachite green, incubated in a mixture of neomycin and amphotericin B. Decontaminated specimens were inoculated onto modified Lowenstein-Jensen medium. Specimens processed by high-speed centrifugation showed growth earlier than specimens prepared by low-speed centrifugation. However, the overall number of positive cultures at 16 weeks was not different for the 2 methods. When infected dairy herds were sampled 4 times at 6-month intervals and culture-positive cows were culled, the prevalence of infected cattle declined over time. After selective culling, the cattle left in the herds shed low numbers of Mptb, which explains why it took longer for cultures to become positive. No heifers younger than 11 months were culture positive, but heifers 13-14 months of age were more frequently culture positive than were heifers of any other age. The 16-week culture period is needed with this method to detect cattle shedding low numbers of Mptb. High-speed centrifugation of samples does not increase the efficiency of identification of animals shedding Mptb. (+info)
DNA relatedness of Leptospira strains isolated from beef cattle in Zimbabwe.
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The DNA relatedness of 17 Leptospira strains isolated from beef cattle in Zimbabwe was determined using the hydroxyapatite method. Similarly to previously speciated African strains, all Zimbabwe isolates belonged to either Leptospira borgpetersenii or Leptospira kirschneri. All serovars within serogroups Pyrogenes (kwale, mombe and a strain closely related to serovar nigeria), Hebdomadis (marondera and mhou), Tarassovi (ngavi) and Sejroe (balcanica and hardjo) were L. borgpetersenii. L. kirschneri contained all strains in serovars of serogroups Icterohaemorrhagiae (zimbabwe), Australis (fugis), Bataviae (paidjan) and Pomona (a strain closely related to mozdok). The species designations of the Zimbabwe fugis and paidjan strains were different from those of the reference strains of these two serovars, both of which belong to Leptospira interrogans. (+info)