The sensitivity of an indirect enzyme-linked immunosorbent assay (ELISA) for bovine IgG serum antibody to Pasteurella haemolytica was compared with that of an indirect hemagglutination (IHA) test. Pasteurella haemolytica serotypes were grown in a chemically defined cell culture medium, and soluble antigens released into the growth medium were used in the ELISA and IHA test. An ELISA with serotype-1 antigen consistently detected antibody in sera that were positive by IHA test (correlation, 99%). Sera reacting with serotype-1 ELISA antigens also reacted with ELISA antigens prepared from other serotypes. Although ELISA titers determined by the 2 methods were approximately linear. Titer increases detected in paired serum samples by either test were similar. The ELISA was more sensitive than was the IHA in detecting colostral IgG antibody in serum of newborn calves. The ELISA uses a simple, stable antigen preparation and detects antibody to P haemolytica serotypes that commonly infect cattle.

Bovine pulmonary artery cells in cell culture were exposed to lipopolysaccharide (LPS) purified from Pasteurella haemolytica serotype A1. This resulted in severe membrane damage, which caused a time- and dose-dependent release of lactate dehydrogenase that was first detected 4 hours after exposure and reached a maximal mean release of 67% after 24 hours of exposure to 1 microgram of LPS/ml. Mean release of 51chromium followed by a similar pattern and reached a maximum of 61% following 24 hours of exposure to 10 micrograms of LPS/ml. Morphologically, endothelial cells responded to LPS by marked cell membrane retraction, the formation of numerous cytoplasmic blebs, and ruffling of the cell membrane. Subsequently, the cells became round and detached. Cell detachment reached a mean of 95% following 8 hours of exposure to 1 microgram of LPS/ml. These studies demonstrated that P haemolytica LPS is capable of causing direct damage to bovine pulmonary arterial endothelial cells, which may be important in the pathogenesis of bovine pneumonic pasteurellosis.

Pilus-mediated adherence is a virulence attribute of Moraxella bovis. Several pilus types have been shown to exist among strains of this bacterium, but correlation between pilus type and specific field cases of the disease has not been done. During the summer of 1987, an epizootic of infectious bovine keratoconjunctivitis was reported in 7 Iowa counties. Eight isolates of M bovis were secured from 12 episodes studied. All 8 of the isolates were nearly homogeneous in biochemical properties and had the same plasmid biotype. Pilus typing performed by immunofluorescence and immunogold electron microscopy identified a single new pilus type among 5 of the 8 isolates. This pilus type was identified in field cases that developed within a narrow time frame and over large distances. The implication of these findings is that infectious bovine keratoconjunctivitis epizootics may be associated with emergence of a novel pilus type, and that rapid dissemination over wide distances can occur, presumably by transportation of carrier cattle.

Antibodies to bovine serum albumin were detected in swine sera by use of an immunoblotting technique. Such sera had false-positive reactions, as determined by results of African swine fever virus serodiagnostic techniques when bovine serum albumin was a contaminant in the soluble cytoplasmic antigen obtained from infected cells cultured in the presence of bovine serum. The soluble cytoplasmic antigen obtained from cell cultures infected with African swine fever virus in the presence of porcine serum did not react with the false-positive sera and, therefore, was used for African swine fever virus serodiagnostic methods, with 0% false-positive results.

The amphotropic murine leukemia virus (MLV) has been shown to infect mammalian species other than mice. If this virus infects and expresses genes in cells of livestock species (cattle, sheep, and pigs) it has potential for use as a vector to produce transgenic livestock. Because the gene-injection technique for producing transgenic animals is inherently inefficient, our laboratory was interested in identifying or constructing retroviral vectors capable of infecting livestock embryos. The infectivity of an amphotropic MLV-based vector for ovine, bovine, and porcine cells was tested. Experiments were also conducted to test the ability of the amphotropic MLV promoter, compared with known strong promoters, to express genes in cells from these species. Results indicated that amphotropic MLV infects and expresses genes efficiently in porcine cells and is, therefore, a potential vector for producing transgenic pigs. Infection was not detected in cells from adult bovine and ovine species; however, low levels of infection, with subsequent gene expression, were detected in cells derived from bovine embryos.

In present study the transtracheal aspiration technique using a commercial polypropylene 5 fr. urinary catheter with 13-gauge 2.5 inch hypodermic needle was applied to bovine medicine in order to determine the availability of this procedure for diagnosis of lower respiratory tract disease. The subjects were forty adult cattle which consisted of thirty-one cows with respiratory signs such as cough and nasal discharge and nine cows without these signs. This procedure appeared suitable for collecting specimens of the lower respiratory tract secretions without uncontamination by the flora of the mouth, respiratory distress and any complications. The most of microorganisms isolated was Pasteurella multocida (48.7 %) and a mucopurulent inflammatory response was the most appeared accounting for 40 % of cytological basis. Most cows with severe clinical signs showed a mucopurulent inflammation (60 %) and Pasteurella multocida were isolated dominantly in severe cases (63.2 %). Cows with mild clinical signs were distributed in each of the five cytological classification and Pasteurella multocida was the most isolated accounting for 40 % of mild cases. In the cytologic examination, a few neutrophils and epithelial cells were seen in the fluid of mucoid inflammation, and many neutrophils with epithelial cells and thick mucoid exudate were seen in the fluid of mucopurulent inflammation. Various number of neutrophils, lymphocytes, macrophages and epithlial cells were examined in the fluid of mixed cell inflammation. The columnar ciliated epithelial cells were examined mainly in normal aspirates. From these results the transtracheal aspiration technique as a diagnostic aid for lower respiratory tract disease of cow is considered as a simple, safe and useful method and can be performed in the field situation

The frequency of appearance of HI antibody against Japanese encephalitis virus was investigated over the three-year period 1982 through 1984. The sample population consisted of 1306 cattle in Saitama prefecture (located in central Japan), and 536 cattle in Kagoshima prefecture (located in southern Japan). During the study period, there was no significant difference in the average antibody-positive rate between cattle in Kagoshima (68.8 %) and those in Saitama (65.5 %). Variations in the monthly antibody-positive rate and mean monthly antibody titer in the Saitama cattle followed a mountain-like pattern with the peak observed during summer. In the Kagoshima cattle, both parameters also increased in summer and these increases persisted even in winter. The distribution of antibody-positive rate as a function of age ranged from 64.0 % to 82.8 % in Saitama, though no correlation was observed between these two factors. In the Kagoshima cattle, however, the antibody-positive rate increased with age as follows; the values averaged 29.4 % in the one-year-old cattle group, 50.0 % in the two-year-old group, 47.4 % in the three-year-old group and 74.5 % in the four-year-old group. There was no significant correlation between the mean antibody titer and age in either Saitama cattle (15.3-22.5) or Kagoshima cattle (20.0-32.3)