Comparison of the effects of different nutrient media on production of heat-stable enterotoxin-b by Escherichia coli

J. Erume, E. M. Berberov, R. A. Moxley

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Trypticase soy broth (TSB) has been commonly used to culture Escherichia coli strains for detection or harvest of heat-stable enterotoxin-b (STb); however, in our experience, the yields have been low. In this study, we compared STb yields resulting from growth in brain heart infusion broth (BHI) supplemented with 2% casamino acids (BHI-CA) with that from TSB. Since strains may concurrently express heat-labile enterotoxin (LT) and lincomycin has been reported to cause increased expression of LT, we also compared its effects on STb production. STb+ clones had significantly higher production of STb when grown in BHI-CA compared to TSB, based on enzyme-linked immunosorbent assay (ELISA) conducted on cell-free supernatants. The superiority of BHI-CA to TSB was further tested on 17 porcine enterotoxigenic E. coli (ETEC) isolates, all positive for the STb gene (estB) by PCR. Cell-free supernatants of 100% of the ETEC isolates were detectably positive for STb by ELISA when grown in BHI-CA, in contrast to only 10 strains positive (59%) when TSB supernatants were analyzed. For all the samples, the amounts of STb expressed in BHI-CA based on ELISA were significantly higher than those from TSB (p≤0.03). Incorporation of lincomycin at a concentration optimal for LT production into these media caused a significant decrease in STb levels, and use of both lincomycin and polymyxin B at these concentrations abolished detectable STb production altogether. We conclude that BHI-CA is a better medium for supporting STb production than is TSB. Culture in BHI-CA followed by testing by STb-ELISA is a useful combination for detecting STb production.

Original languageEnglish (US)
Pages (from-to)160-165
Number of pages6
JournalVeterinary Microbiology
Volume144
Issue number1-2
DOIs
StatePublished - Jul 1 2010

Fingerprint

Thermogenesis
Enterotoxins
enterotoxins
heat production
Escherichia coli
Food
nutrients
heart
brain
Brain
lincomycin
Enzyme-Linked Immunosorbent Assay
enzyme-linked immunosorbent assay
Lincomycin
Enterotoxigenic Escherichia coli
enterotoxigenic Escherichia coli
Hot Temperature
polymyxin B
heat
Polymyxin B

Keywords

  • Culture media
  • Escherichia coli
  • Heat-stable enterotoxin-b

ASJC Scopus subject areas

  • Microbiology
  • veterinary(all)

Cite this

Comparison of the effects of different nutrient media on production of heat-stable enterotoxin-b by Escherichia coli. / Erume, J.; Berberov, E. M.; Moxley, R. A.

In: Veterinary Microbiology, Vol. 144, No. 1-2, 01.07.2010, p. 160-165.

Research output: Contribution to journalArticle

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abstract = "Trypticase soy broth (TSB) has been commonly used to culture Escherichia coli strains for detection or harvest of heat-stable enterotoxin-b (STb); however, in our experience, the yields have been low. In this study, we compared STb yields resulting from growth in brain heart infusion broth (BHI) supplemented with 2{\%} casamino acids (BHI-CA) with that from TSB. Since strains may concurrently express heat-labile enterotoxin (LT) and lincomycin has been reported to cause increased expression of LT, we also compared its effects on STb production. STb+ clones had significantly higher production of STb when grown in BHI-CA compared to TSB, based on enzyme-linked immunosorbent assay (ELISA) conducted on cell-free supernatants. The superiority of BHI-CA to TSB was further tested on 17 porcine enterotoxigenic E. coli (ETEC) isolates, all positive for the STb gene (estB) by PCR. Cell-free supernatants of 100{\%} of the ETEC isolates were detectably positive for STb by ELISA when grown in BHI-CA, in contrast to only 10 strains positive (59{\%}) when TSB supernatants were analyzed. For all the samples, the amounts of STb expressed in BHI-CA based on ELISA were significantly higher than those from TSB (p≤0.03). Incorporation of lincomycin at a concentration optimal for LT production into these media caused a significant decrease in STb levels, and use of both lincomycin and polymyxin B at these concentrations abolished detectable STb production altogether. We conclude that BHI-CA is a better medium for supporting STb production than is TSB. Culture in BHI-CA followed by testing by STb-ELISA is a useful combination for detecting STb production.",
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