Bacterial Colonization of the Equine Gut; Comparison of Mare and Foal Pairs by PCR-DGGE

Jennifer E. Earing, Andrew C. Durig, Gloria L. Gellin, Laurie M. Lawrence, Michael D. Flythe

Agricultural Research Service, Forage-Animal Production Research Unit, USDA, Lexington, USA.
Department of Animal and Food Sciences, University of Kentucky, Lexington, USA.
Department of Animal and Food Sciences, University of Kentucky, Lexington, USA; Agricultural Research Service, Forage-Animal Production Research Unit, USDA, Lexington, USA.

DOI: 10.4236/aim.2012.22010                

Abstract

Horses, like all animals, are born without the symbiotic microbes that occupy the gastrointestinal tracts of mature animals. As grazing animals, horses rely on these microbes to fully utilize the grasses and other cellulosic feeds that they consume. Thus, colonization of the foal's gastrointestinal tract must occur between birth and weaning. The feces of nine mare and foal pairs were sampled from the day of parturition until 12 weeks of age, and the samples were analyzed by polymerase chain reaction amplification of the bacterial 16S rRNA gene and denaturing gradient gel electrophoresis (PCR-DGGE). The gels from feces of day (d) 0 foals had no or very few ( x = 3, n = 6) bands, which indicates that species richness was low. The number of bands increased during the first 4 days of life, and by d 14 the foals and mares had similar numbers of bands ( x = 28, n = 23). Some bands were present in young foals, but not in mares or in foals on d 42 or d 84, which indicated succession of bacterial species. When the PCR-DGGE profiles were compared with Dice's algorithm, all mare-foal pairwise similarities on d 14 and later were as great as the pairwise similarities between mares. These results are consistent with the idea that foals are born with a sterile gut, colonization proceeds rapidly, and a mature microbial community is present in the first few weeks of life.

Keywords

Colonization; Development; Microbial Ecology; Neonates

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	H. Tissier, “Recherches sur la Flore Intestinale des Nourrissons (e′tat Normal et Pathologique),” G Carre and C Naud, Paris, 1900.
[2]	C. F. Favier, E. E. Vaughan, W. M. De Vos and A. D. L. Akkermans, “Molecular Monitoring of Succession of Bacterial Communities in Human Neonates,” Applied and Environmental Microbiology, Vol. 68, No. 1, 2002, pp. 219-226. doi:10.1128/AEM.68.1.219-226.2002
[3]	G. Biasucci, M. Rubini, S. Riboni, L. Morelli, E. Bessi and C. Retetangos, “Mode of Delivery Affects the Bacterial Community in the Newborn Gut,” Early Human Development, Vol. 86, Suppl. 1, 2010, pp. 13-15. doi:10.1016/j.earlhumdev.2010.01.004
[4]	M. G. Dominguez-Belloa, E. K. Costellob, M. Contrerasc, M. Magrisd, G. Hidalgod, N. Fierere and R. Knight, “Delivery Mode Shapes the Acquisition and Structure of the Initial Microbiota across Multiple Body Habitats in Newborns,” Proceedings of the National Academy of Sciences, Vol. 107, No. 26, 2010, pp. 11971-11975. doi:10.1073/pnas.1002601107
[5]	C. F. Favier, W. M. De Vos and A. D. L. Akkermans, “Development of Bacterial and Bifidobacterial Communities in Feces of Newborn Babies,” Anaerobe, Vol. 9, No. 5, 2003, pp. 219-229. doi:10.1016/j.anaerobe.2003.07.001
[6]	R. Satokari, T. Groenroos, K. Laitinen, S. Salminen and E. Isolauri, “Bifidobacterium and Lactobacillus DNA in the Human Placenta,” Letters in Applied Microbiology, Vol. 48, No. 1, 2009, pp. 8-12. doi:10.1111/j.1472-765X.2008.02475.x
[7]	A. B. Onderdonk, M. L. Delaney, A. M. DuBois, E. N. Allred and A. Leviton, “Detection of Bacteria in Placental Tissues Obtained from Extremely Low Gestational Age Neonates,” American Journal of Obstetrics and Gynecology, Vol. 198, No. 1, 2008, pp. 110.e111-110.e117.
[8]	H. E. Jones, K. A. Harris, M. Azizia, L. Bank, B. Carpenter, J. C. Hartley, N. Klein and D. Peebles, “Differing Prevalence and Diversity of Bacterial Species in Fetal Membranes from Very Preterm and Term Labor,” PLoS ONE, Vol. 4, No. 12, 2009, p. e8205. doi:10.1371/journal.pone.0008205
[9]	E. Jimenez, M. L. Marin, R. Martin, J. M. Odriozola, M. Olivares, J. Xaus, L. Fernandez and J. M. Rodriguez, “Is Meconium from Healthy Newborns Actually Sterile?” Research in Microbiology, Vol. 159, No. 3, 2008, pp. 187-193. doi:10.1016/j.resmic.2007.12.007
[10]	J. S. Agerholm, M. Boye and B. Aalb?k, “Ovine Fetal Necrobacillosis,” Journal of Comparative Pathology, Vol. 136, No. 4, 2007, pp. 213-221. doi:10.1016/j.jcpa.2007.01.012
[11]	R. M. Jones, D. F. Twomey, S. Hannon, J. Errington, G. C. Pritchard and J. Sawyer, “Detection of Coxiella burnetii in Placenta and Abortion Samples from British Ruminants Using Real-Time PCR,” Veterinary Record, Vol. 167, No. 25, 2010, pp. 965-967. doi:10.1136/vr.c4040
[12]	R. E. Hungate, “Studies on Cellulose Fermentation: III. The Culture and Isolation for Cellulose-Decomposing Bacteria from the Rumen of Cattle,” Journal of Bacteriology, Vol. 53, No. 5, 1947, pp. 631-645.
[13]	M. J. Glinsky, R. M. Smith, H. R. Spires and C. L. Davis, “Measurement of Volatile Fatty Acid Production Rates in the Cecum of the Pony,” Journal of Animal Science, Vol. 42, No. 6, 1976, pp. 1465-1470.
[14]	K. L. Anderson, T. G. Nagaraja, J. L. Morrill, T. B. Avery, S. J. Galitzer and J. E. Boyer, “Ruminal Microbial Development in Conventionally or Early Weaned Calves,” Journal of Animal Science, Vol. 64, No. 4, 1987, pp. 1215-1226.
[15]	G. Fonty, P. Gouet, J. P. Jouany and J. Senaud, “Establishment of the Microflora and Anaerobic Fungi in the Rumen of Lambs,” Journal of General Microbiology, Vol. 133, No. 7, 1984, pp. 1835-1843.
[16]	P. Gouet, J. M. Nebout, G. Fonty and J. P. Jouany, “Cellulolytic Bacteria Establishment and Rumen Digestion in Lambs Isolated after Birth,” Canadian Journal of Animal Science, Vol. 64, No. 5, 1984, 163-16423. doi:10.4141/cjas84-205
[17]	H. Minato, M. Otsuka, S. Shirasaka, H. Itabashi and M. Mitsumori, “Colonization of Microorganisms in the Rumen of Young Calves,” Journal of General and Applied Microbiology, Vol. 38, No. 5, 1992, pp. 447-456. doi:10.2323/jgam.38.447
[18]	F. W. Lengemann and N. N. Allen, “Development of Rumen Function in the Dairy Calf. II. Effect of Diet upon Characteristics of the Rumen Flora and Fauna of Young Calves,” Journal of Dairy Science, Vol. 42, No. 7, 1959, pp. 1171-1181. doi:10.3168/jds.S0022-0302(59)90709-X
[19]	R. E. Mueller, E. L. Iannotti and J. M. Asplund, “Isolation and Identification of Adherent Epimural Bacteria during Succession in Young Lambs,” Applied and Environmental Microbiology, Vol. 47, No. 4, 1984, pp. 724-730.
[20]	A. Zoilecki and C. A. E. Briggs, “The Microflora of the Rumen of the Young Calf: II. Source, Nature, and Development,” Applied and Environmental Microbiology, Vol. 24, No. 2, 1961, pp. 148-163.
[21]	Y. Sakaitani, N. Yuki, F. Nakajima, S. Nakanishi, H. Tanaka, R. Tanaka and M. Morotomi, “Colonization of Intestinal Microflora in Newborn Foals,” Journal of Intestinal Microbiology, Vol. 13, No. 1, 1999, pp. 9-14.
[22]	V. Julliand, A. De Vaux, L. Villard and Y. Richard, “Preliminary Studies on the Bacterial Flora of Faeces Taken from Foals, from Birth to Twelve Weeks. Effect of the Oral Administration of a Commercial Colostrum Replacer,” Pferdeheilkunde, Vol. 12, No. 3, 1996, pp. 209-212.
[23]	N. R. Pace, “A Molecular View of Microbial Diversity and the Biosphere,” Science, Vol. 276, No. 5313, 1997, pp. 734-740. doi:10.1126/science.276.5313.734
[24]	E. O. Casamayor, H. Sch?fer, L. Ba?eras, C. Pedrós-Alió and G. Muyzer, “Identification of and Spatio-Temporal Differences between Microbial Assemblages from Two Neighboring Sulfurous Lakes: Comparison by Microscopy and Denaturing Gradient Gel Electrophoresis,” Applied and Environmental Microbiology, Vol. 66, No. 2, 2000, pp. 499-508. doi:10.1128/AEM.66.2.499-508.2000
[25]	L. R. Dice, “Measures of the Amount of Ecologic Association between Species,” Ecology, Vol. 26, No. 3, 1945, pp. 297-302. doi:10.2307/1932409
[26]	S. Crowell-Davis, “Normal Behavior and Behavioral Problems,” In: C. Kobluk, T. Ames, R. Geor, Eds., The Horse: Diseases and Clinical Management, W.B. Saunders, Philadelphia, 1995, pp. 1-20.
[27]	S. L. Crowell-Davis, K. Houpt and J. Carnevale, “Feeding and Drinking Behavior of Mares and Foals with Free Access to Pasture and Water,” Journal of Animal Science, Vol. 60, No. 4, 1985, pp. 883-889.
[28]	J. H. Connell and R. O. Slayter, “Mechanisms of Succession in Natural Communities and Their Role in Community Stability and Organization,” American Naturalist, Vol. 111, No. 982, 1977, pp. 1119-1144. doi:10.1086/283241
[29]	C. E. Shannon, “A Mathematical Theory of Communication,” Bell System Technical Journal, Vol. 27, 1948, pp. 379-423.
[30]	C. J. Krebs, “Ecological Methodology,” Addison Wesley Longman, Menlo Park, 1999.
[31]	J. C. Frey, E. R. Angert and A. N. Pell, “Assessment of Biases Associated with Profiling Simple, Model Communities Using Terminal-Restriction Fragment Length Polymorphism-Based Analyses,” Journal of Microbiological Methods, Vol. 67, No. 1, 2006, pp. 9-19. doi:10.1016/j.mimet.2006.02.011
[32]	G. Muyzer, E. C. De Waal and A. G. Uitterlinden, “Profiling of Complex Microbial Populations by Denaturing Gradient Gel Electrophoresis Analysis of Polymerase Chain Reaction-Amplified Genes Coding for 16S rRNA,” Applied and Environmental Microbiology, Vol. 59, No. 3, 1993, pp. 695-700.
[33]	S. Hayes, H. Werner and L. M. Lawrence, “In Vitro Assessment of Fiber Digestion Capacity in Foals,” Proceedings of the 18th Equine Nutrition and Physiology Symposium, East Lansing, 4-7 June 2003, pp. 273-274.
[34]	H. Yoshioka, K. Iseki and K. Fujita, “Development and Differences of Intestinal Flora in the Neonatal Period in Breast-Fed and Bottle-Fed Infants,” Pediatrics, Vol. 72, No. 3, 1983, pp. 317-321.