A comparison of three rapid diagnostic methods for the detection of rotavirus infection in calves

Three techniques for the detection of rotavirus in faecal samples from calves with neonatal gastroenteritis were compared. A preliminary study indicated that reverse passive haemagglutination (RPHA) was at least as sensitive as the enzyme-linked immunosorbent assay (ELISA). These two immunoassays were compared with the detection of viral RNA by polyacrylamide gel electrophoresis (PAGE) on 209 field samples. Of the 77 samples in which at least one test gave a positive result, 69 were positive by both RPHA and PAGE, but only 49 were also positive by ELISA, indicating a lower sensitivity for the latter test. The overall agreement between RPHA and PAGE was 96%. The reasons for the discrepancies between the tests are discussed.     

Virulence factors in Escherichia coli isolated from piglets with neonatal and post-weaning diarrhea in Japan

A total of 567 strains of Escherichia coli were isolated from piglets with neonatal diarrhea (ND) or post-weaning diarrhea (PWD) in Japan. They were investigated for enterotoxigenicity and possession of adhesins and O antigens. There were clear differences between the strains of ND origin and those of PWD origin in the occurrence of enterotoxigenic E. coli (ETEC) strains, type of enterotoxin and frequency of adhesins: ETEC was found in 77 (25.7%) of 300 strains of ND origin and in 137 (51.3%) of 267 strains of PWD origin. ETEC strains producing heat-labile enterotoxin (LT) or heat-stable enterotoxin (STa), alone or in combination were evenly distributed among the strains of PWD origin. In contrast most of the ETEC strains of ND origin produced LT alone. Adhesins appeared in 42 (54.5%) of 77 ETEC strains of ND origin and in 36 (26.3%) of 137 ETEC strains of PWD origin. Adhesins were less common in ETEC strains of PWD origin than in those of ND origin. Some K99-positive ETEC strains of PWD origin produced both LT and STa. There was a similarity in the distribution of O antigens, particularly O149 and O157, between the strains of ND origin and those of PWD origin.     

Antigenic variation of porcine transmissible gastroenteritis virus detected by monoclonal antibodies

Mouse myeloma cells (SP2/O) were fused with spleen cells from BALB/c mice immunized with detergent-solubilized antigen of purified virus, and 21 monoclonal (MC) antibodies reactive in enzyme-linked immunosorbent assay with the TO-163 strain of porcine transmissible gastroenteritis (TGE) virus were obtained. Of these MC antibodies, 14, 6 and 1 were IgG1, IgG2a and IgM, respectively. All of the MC antibodies contained light chains of the kappa type. Of these MC antibodies, 8 were found to have neutralization (NT) activity against the TO-163 strain. Comparison of 7 strains of TGE virus by NT tests using our panel of MC antibodies confirmed their close antigenic relationships, but also revealed the occurrence of distinct antigenic differences. These results suggest that there may be at least 6 different epitopes involved in NT reaction on the virion of the TO-163 strain. This notion was confirmed by the competitive binding assay.     

Characterisation of the outer membrane protein antigens of British field isolates of Moraxella bovis

Outer membranes of a single isolate of Moraxella bovis were prepared and their purity evaluated by a comparison of the iodinated proteins from whole cells and on outer membranes. The protein patterns of the outer membranes of 10 isolates were examined by SDS-polyacrylamide gel electrophoresis, and the antigenic relationship between proteins of different isolates determined by immunoblotting, using antiserum produced against the outer membrane of the single isolate of M. bovis. The overall protein pattern of the different isolates was similar although not identical, but, significantly, only three separate immunogenic proteins were common to all the isolates under examination.     

Exploration of the cellular mediated immunity by the blastogenesis test during chronic brucellosis in human

The diagnosis of the chronic, human brucellosis is frequently difficult and usually needs experimental methods. This paper describes a lymphocyte stimulation test with a Brucella antigen and the results of this test concerning 45 brucellic or not brucellic patients. It is concluded that this test is interesting, especially for the chronic and sero-negative Brucellosis diagnosis.     

Decrease in catalase activity of cultured cells by Mycoplasma gallisepticum infection

The effect of Mycoplasma gallisepticum infection on the host cell catalase activity was histochemically examined in cultured chicken embryo fibroblasts (CEF) and kidney cells. The activity in normal cells was detected as fine, brown granules in the cytoplasm, which appeared ultrastructurally to correspond to anucleoid microbodies. By infecting cultured cells with a CEF-passaged strain of M. gallisepticum, the catalase-positive granules clearly decreased in amount, whereas the UV light-killed mycoplasma and the original strain failed to decrease the granules. The cell-passaged strain was able to induce cytopathic effects and this appeared to be due to its enhanced adherent ability as compared with the original strain. These findings suggest that attachment of viable organisms to cells is crucial to decrease the catalase activity and that the decreased activity may be an important process for the subsequent development of cytopathic effects.     

Cytotoxic capabilities of bovine lymphocytes after lectin stimulation

Bovine peripheral blood mononuclear leukocytes (PBML) were stimulated in vitro with the mitogenic lectins concanavalin A (Con A) and phytohemagglutinin. Their cytotoxic capabilities were evaluated in a 51Cr release assay. Lectin-activated bovine effector cells did not mediate antibody dependent cellular cytotoxicity (ADCC) nor direct killing against cultured tumor target cells. Nevertheless, activation of PBML with lectins consistently generated effector cells able to mediate lectin-dependent cellular cytotoxicity. Cultivation of Con A stimulated-PBML for 3 to 4 weeks in the presence of lymphokines-containing IL-2 generated cells with the ability to mediate lysis without using Con A-coated target cells. However, cytotoxic cultures capable of mediating direct lysis of target cells were not able to mediate ADCC.     

Correlation between adherence of Erysipelothrix rhusiopathiae strains of serovar 1a to tissue culture cells originated from porcine kidney and their pathogenicity in mice and swine

Adherence of four virulent and four avirulent strains of Erysipelothrix rhusiopathiae, serovar 1a, to porcine kidney cell lines, PK-15 and ESK cells, was examined in an in vitro system. The virulent strains adhered well to the cells (range of means, 9.95 +/- 0.87-36.01 +/- 1.10 per cell). In contrast, the avirulent strains showed negligible adherence to the cells (range of means, 0.11 +/- 0.04-1.41 +/- 0.13 per cell). Pretreatment of bacteria with heat, trypsin, or antiserum resulted in a marked decrease in adherence. Scanning electron microscopic examination revealed that the bacteria attached directly to the microvilli of cells.