Characterization of detergent-soluble proteins of Corynebacterium pseudotuberculosis.

Whole cells and culture supernatant of isolates of Corynebacterium pseudotuberculosis were studied by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting (immunoblotting). SDS-PAGE analysis of detergent-solubilized whole cells revealed more than 20 bands in silver-stained gels. However, the SDS-soluble proteins that are present in all the isolates of the bacterium can be separated into four groups, as follows, (i) high molecular mass proteins that are greater than 119 kDa, (ii) a set of three proteins with molecular mass of 84, 64 and 58 kDa, (iii) a doublet consisting of proteins of molecular mass 33 to 30 kDa, and (iv) low molecular mass proteins of 25 to 20 kDa. SDS-PAGE analysis of ammonium sulphate concentrated culture supernatant demonstrated more than seven bands in silver-stained gels ranging in molecular mass from 64-14 kDa. Sera from goats with C. pseudotuberculosis-induced disease were used to probe immunoblots of electrophoresed SDS-soluble proteins. Ten or more SDS-soluble proteins from whole cells, ranging in molecular mass from 119-20 kDa were recognized by antibodies in sera of naturally infected goats. These sera also recognized up to five molecules ranging from 64-30 kDa, on immunoblots of ammonium sulfate concentrated culture supernatant.

Toxicological evaluation of aflatoxin and cyclopiazonic acid in broiler chickens.

The individual and combined effects of aflatoxin (AF) and cyclopiazonic acid (CPA) were evaluated in day-old Petersen x Hubbard broiler chickens to 3 wk of age. Treatments were arranged in a 2 x 2 factorial with levels of 0 and 3.5 mg AF/kg of feed, and 0 and 50 mg CPA/kg of feed. Production performance, serum biochemistry, and gross pathological observations were evaluated. Body weight gain was significantly (P less than .05) reduced by AF, CPA, and the AF-CPA combination at the end of 3 wk. Aflatoxin significantly increased the relative weight of the kidney and serum concentration of blood urea nitrogen and decreased serum concentrations of protein, albumin, cholesterol, phosphorus, and the activity of lactate dehydrogenase. The toxicity of CPA was expressed through increased relative weights of the liver, kidney, and proventriculus, increased levels of uric acid and cholesterol, and decreased serum phosphorus. The activity of AF-CPA combination was characterized by increased relative weight of the liver, kidney, pancreas, and proventriculus, decreased concentrations of serum albumin and phosphorus, increased concentrations of serum glutamic oxalacetic transaminase and blood urea nitrogen, and decreases in the relative weight of the bursa of Fabricius. Post-mortem examination revealed that the chickens fed CPA and the AF-CPA combination had thickened mucosa and dilated proventricular lumens, hard fibrotic spleen, and atrophy of the gizzard. The data from the present study demonstrate that both AF and CPA alone and the AF-CPA combination can limit broiler performance and adversely affect broiler health. In most cases the effects of AF and CPA were additive.