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Applied and Environmental Microbiology, October 2008, p. 6338-6347, Vol. 74, No. 20
0099-2240/08/$08.00+0     doi:10.1128/AEM.00309-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Detection of Bifidobacterium animalis subsp. lactis (Bb12) in the Intestine after Feeding of Sows and Their Piglets{triangledown}

Gloria Solano-Aguilar,1* Harry Dawson,1 Marta Restrepo,1 Kate Andrews,2 Bryan Vinyard,3 and Joseph F. Urban Jr.1

Diet, Genomics, and Immunology Laboratory, Beltsville Human Nutrition Research Center, Agricultural Research Service, U.S. Department of Agriculture, Beltsville, Maryland,1 Food and Feed Safety Research Unit, Southern Plains Agricultural Research Center, U.S. Department of Agriculture, College Station, Texas,2 Biometrical Consulting Service, Beltsville Agricultural Research Center, U.S. Department of Agriculture, Beltsville, Maryland3

Received 5 February 2008/ Accepted 4 August 2008

A real-time PCR method has been developed to distinguish Bifidobacterium animalis subspecies in the gastrointestinal tracts of pigs. Identification of a highly conserved single-copy tuf gene encoding the elongation factor Tu involved in bacterial protein biosynthesis was used as a marker to differentiate homologous Bifidobacterium animalis subsp. lactis (strain Bb12) from Bifidobacterium animalis subsp. animalis, as well as Bifidobacterium suis, Bifidobacterium breve, Bifidobacterium longum, several species of Lactobacillus, and Enterococcus faecium. Real-time PCR detection of serially diluted DNA extracted from a pure culture of Bb12 was linear for bacterial numbers ranging from 10 to 10,000 tuf gene copies per PCR (r2 = 0.99). Relative differences in Bb12 bacterial numbers in pigs fed daily with Bb12 were determined after detection of Bb12 tuf gene copies in DNA extracted from the intestinal contents. Piglets treated with Bb12 immediately after birth maintained a high level of Bb12 in their large intestines with continuous daily administration of Bb12. Piglets born to Bb12-treated sows during the last third of their gestation and also treated with Bb12 at birth (T/T group) had a higher number of Bb12 organisms per gram of intestinal contents compared to placebo-treated piglets born to placebo-treated sows (C/C group), Bb12-treated sows (T/C group), or piglets born to placebo sows but treated with Bb12 immediately after birth (C/T group). In addition, there was a significant increase in gene expression for Toll-like receptor 9 (TLR9) in piglets from the T/T group, with no change in TLR2 and TLR4. These findings suggest that the tuf gene represents a specific and functional marker for detecting Bifidobacterium animalis subsp. lactis strain Bb12 within the microbiota of the intestine.

* Corresponding author. Mailing address: Diet, Genomics, and Immunology Laboratory, Beltsville Human Nutrition Research Center, Agricultural Research Service, U.S. Department of Agriculture, 10300 Baltimore Avenue, BARC-East, Bldg. 307C, Rm. 225, Beltsville, MD 20705. Phone: (301) 504-8068. Fax: (301) 504-9062. E-mail: Gloria.SolanoAguilar{at}ars.usda.gov

{triangledown} Published ahead of print on 8 August 2008.

Applied and Environmental Microbiology, October 2008, p. 6338-6347, Vol. 74, No. 20
0099-2240/08/$08.00+0     doi:10.1128/AEM.00309-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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