Purification and expression of the Tf190 adhesin in Tritrichomonas foetus.

Bovine trichomoniasis is a sexually transmitted disease caused by Tritrichomonas foetus and characterized by early embryo loss. The mechanism of this loss is not known, although the parasite is known to cause inflammation and to have the ability to kill host cells by a contact-dependent cytotoxic mechanism. Antibody specific for a 190,000-Da surface complex (Tf190) was previously shown to inhibit this adhesion. In this study we used immunoaffinity chromatography to purify Tf190 from T. foetus in order to analyze its composition and examine its expression. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of purified Tf190 followed by silver staining revealed three components of Tf190. Western blotting and antibody-binding experiments showed that the 140- and 60-kDa bands were immunogenic. By using a battery of monoclonal antibodies (MAbs) periodate-sensitive epitopes were identified on Tf190, suggesting that these epitopes contained carbohydrate structures. Analyses of affinity-purified Tf190 by high-performance liquid chromatography and gas-liquid chromatography demonstrated the presence of the monosaccharides and lipids known to be prominent constituents of the lipophosphoglycan (LPG) of T. foetus. Flow cytometry experiments on several isolates of T. foetus with Tf190-specific antibodies revealed that Tf190 was present on subpopulations of all isolates but that not all epitopes were present on every isolate. This pattern of reactivities on the different parasite isolates was confirmed by Western blots of whole-parasite extracts probed with MAbs and antiserum. These results suggest that although variation in the expression of epitopes of Tf190 occurs in different strains of T. foetus, the Tf190 adhesion complex is widespread in different populations of the parasite. The data further suggest that immunogenic structures, important in the adhesion of T. foetus to mammalian cells, are located in the LPG-like component of Tf190.

Chlorine injury and the comparative performance of Colisure (TM), ColiLert (TM) and ColiQuik (TM) for the enumeration of coliform bacteria and E.coli in drinking water.

Several factors have stimulated interest in recently developed substrate specific media for the detection of coliform bacteria in water. This study compared the performance of Colisure (TM) (Millipore), ColiLert (TM) (Environetics) and ColiQuick (TM) (Hach) with accepted membrane filtration and MPN methodologies for the enumeration of total coliforms and E. coli in chlorinated water. The performance of all three media was compared, in MPN configuration, with LTB/MPN (confirmed) using a variety of drinking and source water samples, both with and without chlorination. The Cochran-Mantel-Haenszel test yielded statistical correlations between results obtained with each of the three new enzyme detection media and accepted reference methods for the detection of low numbers of total coliforms. Another series of tests compared the performance of Colisure with accepted methods (LTB/MPN confirmed with BGLB and EC-MUG) in the detection of total coliforms and E. coli in sewage-spiked samples simulating contaminated drinking water, using an USEPA/AWWA test protocol. The results demonstrated that Colisure detected these indicator bacteria with greater sensitivity than the accepted methods and that this difference increased between 24 and 28 hours of incubation. The results of this study collectively support the validity of the new enzyme detection method for the detection of low levels of coliform bacteria and E. coli in source water and contaminated drinking water.