Environmental sampling to assess the bioburden of Mycobacterium avium subspecies paratuberculosis in drylot pens on California dairies

Tapakorn Chamchoy, Deneice R. Williams, John M. Adaska, Randall J. Anderson, Sharif S. Aly

Research output: Contribution to journalArticle

Abstract

Mycobacterium avium subspecies paratuberculosis (MAP) is a bacterium that can cause substantial economic losses in infected dairy herds due to reduced milk production and increased cow-replacement costs. In order to control MAP in dairies with drylot pens, a standardized environmental sampling protocol to quantify MAP in fecal slurry was developed based on an existing protocol for freestall pens. Specifically, following a 24 h hold of the flush, a grab sample of approximately 10 ml of fecal slurry was collected every 1 malong the flush lane of the drylot pens, avoiding individual cow fecal pats. To determine the reliability and repatability of the new environmental sampling protocol for estimation of MAP bioburden at the pen level, two collectors simultaneously collected fecal slurry samples every day for 3 days from six drylot cow pens on two Central California dairies. During the study period no cow movement between pens was allowed with the exception of sick cows. The study herds had MAP seroprevalence of 5.8% and 3.2%, respectively, based on whole pen serum ELISA results. Variance components models for quantitative real-time PCR (qPCR) results showed samples collected from different pens on different dairies accounted for greater variablitiy in MAP concentration (65%), while samples collected by different collectors had the least variability (0.1%). In contrast, variability in MAP concentration in environmental samples collected on different days had 25% variability. The intraclass correlation coefficient showed high reliability (93%) of environmental sampling simultaneously by different collectors. In contrast, the reliability of environmental sampling at different days was 65%, which was similar to the reliability for sampling by different collectors on different days. Investigators can expect high reliability when employing the new environmental sampling protocol along with qPCR testing of environmental samples from drylot pens.

Original languageEnglish (US)
Article numbere8081
JournalPeerJ
Volume2019
Issue number11
DOIs
StatePublished - Jan 1 2019

Fingerprint

Mycobacterium avium subsp. paratuberculosis
microbial load
Mycobacterium avium
Dairies
paratuberculosis
dairies
Sampling
collectors
sampling
cows
Seroepidemiologic Studies
Bacteria
Real-Time Polymerase Chain Reaction
Milk
Enzyme-Linked Immunosorbent Assay
Economics
Research Personnel
Costs and Cost Analysis
Testing
dairy herds

Keywords

  • Drylot pen
  • Environmental sampling
  • Intraclass correlation coefficient
  • Mycobacterium avium subspecies paratuberculosis
  • Quantitative real-time PCR

ASJC Scopus subject areas

  • Neuroscience(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Environmental sampling to assess the bioburden of Mycobacterium avium subspecies paratuberculosis in drylot pens on California dairies. / Chamchoy, Tapakorn; Williams, Deneice R.; Adaska, John M.; Anderson, Randall J.; Aly, Sharif S.

In: PeerJ, Vol. 2019, No. 11, e8081, 01.01.2019.

Research output: Contribution to journalArticle

Chamchoy, Tapakorn ; Williams, Deneice R. ; Adaska, John M. ; Anderson, Randall J. ; Aly, Sharif S. / Environmental sampling to assess the bioburden of Mycobacterium avium subspecies paratuberculosis in drylot pens on California dairies. In: PeerJ. 2019 ; Vol. 2019, No. 11.
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abstract = "Mycobacterium avium subspecies paratuberculosis (MAP) is a bacterium that can cause substantial economic losses in infected dairy herds due to reduced milk production and increased cow-replacement costs. In order to control MAP in dairies with drylot pens, a standardized environmental sampling protocol to quantify MAP in fecal slurry was developed based on an existing protocol for freestall pens. Specifically, following a 24 h hold of the flush, a grab sample of approximately 10 ml of fecal slurry was collected every 1 malong the flush lane of the drylot pens, avoiding individual cow fecal pats. To determine the reliability and repatability of the new environmental sampling protocol for estimation of MAP bioburden at the pen level, two collectors simultaneously collected fecal slurry samples every day for 3 days from six drylot cow pens on two Central California dairies. During the study period no cow movement between pens was allowed with the exception of sick cows. The study herds had MAP seroprevalence of 5.8{\%} and 3.2{\%}, respectively, based on whole pen serum ELISA results. Variance components models for quantitative real-time PCR (qPCR) results showed samples collected from different pens on different dairies accounted for greater variablitiy in MAP concentration (65{\%}), while samples collected by different collectors had the least variability (0.1{\%}). In contrast, variability in MAP concentration in environmental samples collected on different days had 25{\%} variability. The intraclass correlation coefficient showed high reliability (93{\%}) of environmental sampling simultaneously by different collectors. In contrast, the reliability of environmental sampling at different days was 65{\%}, which was similar to the reliability for sampling by different collectors on different days. Investigators can expect high reliability when employing the new environmental sampling protocol along with qPCR testing of environmental samples from drylot pens.",
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