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Survival of Mycobacterium bovis during forage ensiling

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  • 1 Department of Large Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824.
  • | 2 Department of Pathobiology and Diagnostic Investigation, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824.
  • | 3 Department of Diagnostic Center for Population and Animal Health, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824.
  • | 4 Department of Diagnostic Center for Population and Animal Health, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824.
  • | 5 Department of Diagnostic Center for Population and Animal Health, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824.
  • | 6 Department of Diagnostic Center for Population and Animal Health, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824.
  • | 7 Michigan State University Extension, Michigan State University, East Lansing, MI 48824.
  • | 8 Department of Animal Science, College of Agriculture and Natural Resources, Michigan State University, East Lansing, MI 48824.
  • | 9 Department of Animal Science, College of Agriculture and Natural Resources, Michigan State University, East Lansing, MI 48824.
  • | 10 Department of Animal Science, College of Agriculture and Natural Resources, Michigan State University, East Lansing, MI 48824.
  • | 11 Animal Industry Division, Michigan Department of Agriculture and Rural Development, 611 W Ottawa St, Lansing, MI 48909.

Abstract

OBJECTIVE To determine whether Mycobacterium bovis remains viable in ensiled forages.

SAMPLE Alfalfa, mixed mostly grass, and corn silages.

PROCEDURES For each of 10 sampling days, six 250-g replicate samples of each feedstuff were created and placed in a film pouch that could be vacuum sealed to simulate the ensiling process. Within each set of replicate samples, 4 were inoculated with 10 mL of mycobacterial liquid culture medium containing viable M bovis and 2 were inoculated with 10 mL of sterile mycobacterial liquid culture medium (controls) on day 0. Pouches were vacuum sealed and stored in the dark at room temperature. On the designated sampling day, 1 control pouch was submitted for forage analysis, and the other pouches were opened, and forage samples were obtained for M bovis culture and analysis with a PCR assay immediately and 24 hours later.

RESULTS None of the control samples had positive M bovis culture or PCR assay results. Among M bovis-inoculated samples, the organism was not cultured from alfalfa and corn silage for > 2 days but was cultured from mixed mostly grass silage for 28 days after inoculation and ensiling initiation. Mycobacterium bovis DNA was detected by PCR assay in samples of all 3 feedstuffs throughout the 112-day observation period.

CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that properly ensiled forages would be an unlikely source for M bovis transmission to cattle. Further research is necessary to determine whether ensiling kills M bovis or forces it to become dormant and, if the latter, elucidate the conditions that cause it to revert to an infectious state.

Abstract

OBJECTIVE To determine whether Mycobacterium bovis remains viable in ensiled forages.

SAMPLE Alfalfa, mixed mostly grass, and corn silages.

PROCEDURES For each of 10 sampling days, six 250-g replicate samples of each feedstuff were created and placed in a film pouch that could be vacuum sealed to simulate the ensiling process. Within each set of replicate samples, 4 were inoculated with 10 mL of mycobacterial liquid culture medium containing viable M bovis and 2 were inoculated with 10 mL of sterile mycobacterial liquid culture medium (controls) on day 0. Pouches were vacuum sealed and stored in the dark at room temperature. On the designated sampling day, 1 control pouch was submitted for forage analysis, and the other pouches were opened, and forage samples were obtained for M bovis culture and analysis with a PCR assay immediately and 24 hours later.

RESULTS None of the control samples had positive M bovis culture or PCR assay results. Among M bovis-inoculated samples, the organism was not cultured from alfalfa and corn silage for > 2 days but was cultured from mixed mostly grass silage for 28 days after inoculation and ensiling initiation. Mycobacterium bovis DNA was detected by PCR assay in samples of all 3 feedstuffs throughout the 112-day observation period.

CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that properly ensiled forages would be an unlikely source for M bovis transmission to cattle. Further research is necessary to determine whether ensiling kills M bovis or forces it to become dormant and, if the latter, elucidate the conditions that cause it to revert to an infectious state.

Contributor Notes

Dr. Grooms’ present address is Office of the Dean, College of Veterinary Medicine, Iowa State University, Ames, IA 50011.

Ms. Plastow's present address is MPI Research, 54943 N Main St, Mattawan, MI 49071.

Dr. Lim's present address is Breathitt Veterinary Center, Huston School of Agriculture, Murray State University, Hopkinsville, KY 42241.

Address correspondence to Dr. Grooms (dgrooms@iastate.edu).