Identity and interactions of rumen microbes associated with dietary sulfate-induced polioencephalomalacia in cattle

B. A. Cummings From the Department of Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523 (Cummings, Gould, Hamar), and the Department of Molecular Biology, College of Agriculture, University of Wyoming, Laramie, WY 82701 (Caldwell).

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D. R. Caldwell From the Department of Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523 (Cummings, Gould, Hamar), and the Department of Molecular Biology, College of Agriculture, University of Wyoming, Laramie, WY 82701 (Caldwell).

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D. H. Gould From the Department of Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523 (Cummings, Gould, Hamar), and the Department of Molecular Biology, College of Agriculture, University of Wyoming, Laramie, WY 82701 (Caldwell).

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D. W. Hamar From the Department of Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523 (Cummings, Gould, Hamar), and the Department of Molecular Biology, College of Agriculture, University of Wyoming, Laramie, WY 82701 (Caldwell).

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SUMMARY

To study their role in sulfate reduction, anaerobic bacteria were cultured from rumen fluid samples of cattle fed high-carbohydrate, short-fiber diets with and without added sulfate. The steers fed the diet with added sulfate developed polioencephalomalacia. Microbiological methods included colony-type profiles, molybdate sensitivity, presence of desulfoviridin, sulfate reduction rates of pure and mixed cultures, and incubation time effects on sulfate reduction. Colony-type profiles indicated decreased diversity, but no relative change in numbers of sulfate-reducing bacteria in rumen fluid from cattle fed diets with and without added sulfate. Thirteen bacterial isolates were selected for further study on the basis of colony type, sulfate-reducing activity, and growth in lactate, sulfate, and yeast extract media. Seven of the isolates had Desulfovibrio-like characteristics (ie, they were gram-negative, motile rods that reduced sulfate, were inhibited by molybdate, and contained the pigment desulfoviridin). The remaining 6 isolates were gram-negative, nonmotile rods. Four of these released sulfide from cysteine, and 2 generated only limited amounts of sulfide from sulfate or cysteine. The 7 sulfate-reducing isolates generated sulfide in rumen fluid broth medium at greater rates than those observed in fresh rumen fluid. Sulfate reduction could be sustained in cultures for prolonged incubation times if the gas phase containing hydrogen sulfide was replaced at frequent intervals. Variations in the amount of sulfate reduced by the pure cultures were most pronounced at short incubation times. Sulfate reduction was not inhibited in mixed cultures of sulfate-reducing and nonsulfate-reducing bacteria.

SUMMARY

To study their role in sulfate reduction, anaerobic bacteria were cultured from rumen fluid samples of cattle fed high-carbohydrate, short-fiber diets with and without added sulfate. The steers fed the diet with added sulfate developed polioencephalomalacia. Microbiological methods included colony-type profiles, molybdate sensitivity, presence of desulfoviridin, sulfate reduction rates of pure and mixed cultures, and incubation time effects on sulfate reduction. Colony-type profiles indicated decreased diversity, but no relative change in numbers of sulfate-reducing bacteria in rumen fluid from cattle fed diets with and without added sulfate. Thirteen bacterial isolates were selected for further study on the basis of colony type, sulfate-reducing activity, and growth in lactate, sulfate, and yeast extract media. Seven of the isolates had Desulfovibrio-like characteristics (ie, they were gram-negative, motile rods that reduced sulfate, were inhibited by molybdate, and contained the pigment desulfoviridin). The remaining 6 isolates were gram-negative, nonmotile rods. Four of these released sulfide from cysteine, and 2 generated only limited amounts of sulfide from sulfate or cysteine. The 7 sulfate-reducing isolates generated sulfide in rumen fluid broth medium at greater rates than those observed in fresh rumen fluid. Sulfate reduction could be sustained in cultures for prolonged incubation times if the gas phase containing hydrogen sulfide was replaced at frequent intervals. Variations in the amount of sulfate reduced by the pure cultures were most pronounced at short incubation times. Sulfate reduction was not inhibited in mixed cultures of sulfate-reducing and nonsulfate-reducing bacteria.

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