Zentralblatt fur Veterinarmedizin. Reihe B. Journal of veterinary medicine. Series B最新文献

The mechanism of BLV-induced tumorigenesis has not been clear up to now. Changes of viral protein expression in infected cells may be involved in the molecular events leading to BLV-induced leukaemogenesis. In this study Western blot investigations of cells transfected with plasmid DNA containing the complete Japanese BLV tumour clone provirus demonstrate that this provirus is unable to express gag and env proteins. Following this an attempt was made to express the genes from this provirus in eukaryotic and prokaryotic cells using the phagemid pBK-RSV (Stratagene), but not as fusion proteins. The protein patterns expressed from the 5' and the 3' region of the BLV genome were compared with those of FLK/BLV cells. The results indicate that there is a defect in this provirus located in the genome region between the gag and env gene.

The whole-cell fatty acid methyl ester (FAME) profiles of Salmonella (S.) gallinarum and Salmonella (S.) pullorum strains were compared in the computer-linked gas-liquid chromatography. The profiles of whole cellular FAMEs allow the separation and identification S. gallinarum and S. pullorum by the Microbial Identification System and so can be used for their differentiation.

The efficacy of doramectin treatment on arrested A. caninum larvae during early pregnancy of bitches was examined. Four bitches were percutaneously infected with 20,000 third-stage larvae of A. caninum on the day of conception and treated subcutaneously with 1 mg doramectin per kg body weight on day 30 of pregnancy. Four infected untreated pregnant bitches served as controls. A single application of dormectin substantially reduced the number of somatic larvae in bitches and the number of intestinal stages in bitches and puppies. However, it did not completely prevent lactogenic transmission of A. caninum larvae because five out of 23 puppies from three litters of the treated bitches harboured adult worms in their intestines, two of them shed eggs with the faeces. Although clinical disease did not occur in puppies from treated bitches the efficacy of the treatment was not satisfactory from an epidemiological point of view. Despite the treatment puppies with patent infections contaminated their environment with high numbers of eggs thus producing an intolerable infection risk for dogs and humans. No fetotoxic side-effects of the early treatment with doramectin were seen.

In situ hybridization, (ISH) using a digoxigenin-antisense RNA-probe to detect chlamydial rRNA was applied to post mortem tissue of birds. The technique was optimized and validated using tissue from experimentally-infected chicken embryos. Tissue sections were also tested by immunohistochemistry (peroxidase-antiperoxidase reaction, PAP) for the presence of chlamydial antigen using a genus specific monoclonal antibody. In the chicken embryo tissue, ISH and PAP were comparably sensitive and specific (100% and 100%, respectively). ISH and PAP in general were correlated to microscopic lesions. For further comparison, ISH with PAP was applied retrospectively to tissues of 82 birds from which Chlamydia had been isolated, or which were suggestive of chlamydiosis. Using in situ hybridization 47 of 82 birds were found to be positive, and as were 23 of 82 birds with PAP. None of the ISH-only positive cases were found to be strongly positive. On the other hand, cases which were found positive with the ISH were also positive with other methods (PAP and isolation of Chlamydiae from chicken embryos). There was no close correlation between the positive cells and histological lesions. In spite of the higher sensitivity and specificity of the ISH, this technique is not suitable for routine diagnostic investigations. ISH is expensive, laborious, and time consuming.

Experimental intramuscular infection of hens with Pseudomonas pseudomallei, strain 2796 (1 x 10(9) CFU from a 24-h culture) was reproduced. Clinical, paraclinical and pathomorphological findings were followed from 1 to 30 days after challenge. Haemagglutinin titre, bacterial dissemination in the viscera, number of leucocytes, alveolar (aMa) and peritoneal (pMa) macrophages and their phagocytic activity in vitro were studied. During the course of infection a leucocytosis as well as an increased haemagglutinin titre (1:256) were established. The number of bacteria per gram tissue in the spleen and liver was highest at 1 day post-infection (p.i.). Melioidose bacteria from egg yolk were isolated at 15 and 30 days p.i. Leucocyte and pMa phagocytic activity was maximal at 3 days p.i. unlike the activity of aMa which increased gradually until the end of the study. Inflammatory-necrotic changes were found in the viscera and brain at 3 and 15 days p.i. The investigation of experimental melioidosis infection in hens showed that they are susceptible to P. pseudomallei and this disease takes a generalized subacute course.

Various factors influencing plaque formation of Chuzan virus in BHK-21 cell monolayers were studied and a practical method for plaque assay was developed. On addition of trypsin (5 micrograms/ml) and/or diethylaminoethyl (DEAE)-dextran (50 micrograms/ml) to the virus diluent as the virus adsorption medium and agar overlay medium, the number of plaques increased. When 100 micrograms/ ml DEAE-dextran was added to the diluent and overlay medium, plaques were produced in about 10-fold higher numbers than without trypsin and DEAE-dextran. Based on these results, a practical plaque assay method for Chuzan virus was established. Using this method, one-step growth of Chuzan virus was performed at an input multiplicity of 25 plaque-forming units (PFU) per cell. Cytopathic effects were first observed at 7.5 h post-inoculation (p.i.), and were complete at 12 h p.i. The titre of cell-associated virus, after gradual decline during the first 3 h of incubation, showed a rise within 4.5 h p.i. and a rise to a plateau of 10(6.3)PFU/0.2 ml at 12 h p.i. By indirect immunofluorescence, virus-specific antigen was detected in the cytoplasm of the cells at 4.5 h p.i., and all the cells fluoresced at 6 h p.i. Haemagglutination activity was first detected in infected whole cultures at 7.5 h p.i. reaching a plateau of 1:64 at 15 h p.i. Plaque formation and haemagglutination by the virus were specifically inhibited by antisera against the original and the plaque-cloned virus.

A survey of selected causative organisms of bacterial mastitis in Zebu-Holstein dairy cows was carried out on four herds in Wondogenet. Pathogens were found in 39% of udder quarters and mastitis in 16% of quarters. The main mastitis causing organisms isolated were Staphylococcus aureus (47%), Streptococcus agalactiae (15%) and Streptococcus uberis (31%). It was indicated also that udder quarters with micrococci would be less susceptible to infections by pathogenic organisms.

Experiments were designed to evaluate the effect of BLV on mitogen-stimulated peripheral blood mononuclear cells (PBMC) from naturally infected cattle. BIV was also taken into consideration due to a recent report showing that in Costa Rica, most of the BLV-infected animals are also seropositive for BIV. The methodology was based on a non-radioactive technique to determine lymphoproliferation. A colorimetric assay using XTT (formazan salt) to measure cell multiplication was adapted for bovine PBMC. ELISA and Western blotting were used to determine the serologic status of the cattle. PCR was only available for BIV detection. Our results show clearly that, dually-infected cattle (BIV-BLV) have reduced lymphoproliferative responses to the mitogen Con A. Haematological abnormalities associated with viral infections were also observed, specially leukocytosis and lymphocytosis. Cows with lymphosarcomas are severely affected. The specific antibody response to different viral proteins could not be associated with the suppressive status of the animals. Due to the high rate of dual infections observed in Costa Rica, these results are not sufficient to clarify which virus is responsible for the suppressive activity, if one or both viruses are necessary, or if they act synergistically.

Interleukin-1 (IL-1) is a key mediator in inflammatory processes. There are several immunoassays for human, but not porcine, IL-1 commercially available. Four of these were tested on porcine specimens. IL-1 was not detected, by any of the immunoassays, in cell culture supernatants with confirmed IL-1 activity in a bioassay. The results indicate that the tested antibodies to human IL-1 do not crossreact with porcine IL-1.

In this study the eyes of 15 cats in the terminal stage of FIV infection were examined. The findings were compared to those in cats, which were euthanized because of other infectious diseases for for non-infectious reasons. Thirteen FIV-infected cats showed an anterior uveitis by means of light microscopy. No accumulation of retinal lesions were found in FIV-infected cats compared to the other cats examined. Additionally, there were no indications of lesions caused by opportunistic infections. In the posterior segments of the eyes, immunohistochemical examinations proved the plasma proteins C3 and IgG to be predominantly intravascular. The eyes of 11 serologically FIV-positive cats were available for immunohistochemical examination. In all 11 cats at least one of the plasma proteins C3 or IgG could be detected in the extravascular tissue of the anterior uvea. The extravascular presence of plasma proteins within the tissue seemed to be caused by an increased permeability of the vessels due to inflammation. Furthermore, the similar extravascular distribution pattern of IgG and complement component C3 in four cases indicated that immune complexes may play a role in the anterior uveitis of FIV-infected cats.