Survival of Corynebacterium renale, Corynebacterium pilosum and Corynebacterium cystitidis in soil

Survival of the causative agents of bovine pyelonephritis, Corynebacterium renale, C. pilosum and C. cystitidis, was examined at 30 degrees C in autoclaved soil. In the soil from a paddock, C. renale and C. cystitidis survived for 56 and 63 days, respectively, and C. pilosum for a longer period of at least 210 days. In soil from a pasture, sand from an athletic field and sea sand, the survival of these bacteria was of shorter duration.     

Adhesion of Corynebacterium renale and Corynebacterium pilosum to epithelial cells of bovine vulva

Corynebacterium renale and C pilosum adhered effectively to the epithelial cells of the bovine vulva; the numbers of these organisms that adhered to the vulval epithelial cells were 50 and 30/cell, respectively, which were several times as many as those that adhered to the uroepithelial cells. Of the epithelial cells of the vulva, cornified cells lacking nuclei bound more bacteria than did those with indistinct nuclei, indicating that adhesion of bacteria was most effective to the most aged cells. The marked adhesion of C renale and C pilosum to the epithelial cells of the vulva may indicate that the vulva is an important portal of entry of these bacteria.     

Isolation and characterization of monoclonal antibodies against pili of Corynebacterium renale and Corynebacterium pilosum

Five monoclonal antibodies against pili of Corynebacterium renale 115 P+ (piliated clone) and two monoclonal antibodies against pili of C. pilosum 92 P+ (piliated clone) were produced. These antibodies bound to pili of the homologous strain in in enzyme-linked immunosorbent assay (ELISA) and agglutinated P+ but not P- (non-piliated clone) of each homologous strain. The five monoclonal antibodies against C. renale 115 P+ pili were divided into 2 groups, comprising 16/5, 160/1 and 32/6 and 13/4 and B20/3, based on the results of a competitive binding assay. The results may indicate the presence of at least 2 distinct antigenic areas on the pilus of C. renale 115 P+. The monoclonal antibodies of the first group inhibited adhesion of C. renale 115 P+ bacteria to the epithelial cells of bovine vulva, while the second group did not. Two monoclonal antibodies against C. pilosum 92 P+ pili recognized the same area on the pilus of C. pilosum 92 P+, and inhibited the adhesion of C. pilosum 92 P+ bacteria to the epithelial cells of bovine vulva. The adhesion of these bacteria was inhibited by the monoclonal antibodies in the form of IgG as well as by the Fab fragment. The strains of C. renale and C. pilosum which reacted with each of the anti-C. renale 115 P+ pili and anti-C. pilosum 92 P+ pili monoclonal antibodies were small in number and of restricted distribution.     

Comparison of surface hydrophobicity of piliated and non-piliated clones of Corynebacterium renale and Corynebacterium pilosum

Piliated (P+) and non-piliated (P-) clones of Corynebacterium renale and C. pilosum were similar in hydrophobicity as measured by hydrophobic interaction chromatography, bacterial adherence to hydrocarbons and the salt aggregation test. Therefore, the previously reported adherence of P+ clone to various cells, which is more effective than that of P- clone, may be uncorrelated with the degree of hydrophobicity of both clones of these bacteria. Hydrophobicity of P+ and P- clones was found to be high when measured by hydrophobic interaction chromatography and bacterial adherence to hydrocarbons, but low when measured by the salt aggregation test.     

Cloning and expression of a pili gene of Corynebacterium renale in Escherichia coli

A plasmid gene library of Corynebacterium renale piliated strain No. 109P+ was prepared in Escherichia coli in order to study the chemical structure of the pili of C. renale. Of 3,000 recombinant clones tested, 5 reacted with anti-pili anti-serum. The gene products of these clones reacted with anti-pili monoclonal antibodies 8/4, 5/2 and B20/3 but lacked the reactivity with 13/4. SDS-PAGE analysis revealed that the expressed protein had a molecular mass of 48 kilodalton and deletion analysis showed that the encoding region for this protein was localized within a 1.4 kilobase gene including a promoter sequence. Immunoelectron microscopy showed that mouse antibodies raised to the expressed protein bound to the entire surface of the pili of C. renale. These results indicate that the cloned gene encodes a major structural protein of C. renale pili.     

Antibody coated bacteria in urine sediment from cattle infected with Corynebacterium renale.

Using the fluorescent antibody test, the presence of antibody-coated bacteria in 10 out of 17 urine sediment samples from cattle infected with Corynebacterium renale is described. These antibodies were mainly of the immunoglobulin class IgG, and to a lesser extent IgA. This finding is characteristic for infections of the upper urinary tract (pyelonephritis). In seven samples no antibody coating of the bacterial surface was detected. In these cases an infection of the lower urinary tract (cystitis) is suggested.     

Attachment of Corynebacterium renale to tissue culture cells by the pili.

One or more cells of Corynebacterium renale strains (serologic types, I, II and III), which possessed numerous pili, frequently were attached to BHK-21 cells, primary dog kidney cells, and primary rabbit kidney cells. The percentage of the cultured cells to which C renal cells were attached was about 70%. The percentage was less with cells of C renale possessing fewer pili, around 30%. After C renale was treated with the homologous anti-pili serum, the percentage of BHK-21 cells to which bacterial cells were attached was even less (22%). In electron micrographs, the pili of C renale were observed to attach themselves to the membranes of BHK-21 cells. The adhesive property of the pili of C renale to tissue culture cells was thus demonstrated.