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Journal of Integrative Agriculture  2016, Vol. 15 Issue (7): 1575-1583    DOI: 10.1016/S2095-3119(15)61236-9
Animal Science · Veterinary Science Advanced Online Publication | Current Issue | Archive | Adv Search |
Fermentation characteristics of Megasphaera elsdenii J6 derived from pig feces on different lactate isomers
JIANG Xiao-lin, SU Yong, ZHU Wei-yun
Jiangsu Key Laboratory of Gastrointestinal Nutrition and Animal Health, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, P.R.China
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Abstract      D-Lactate-utilizing bacteria play important roles in maintaining the balance of gut lactate; however, studies on gut D-lactate-utilizing bacteria have been limited. This study aimed to isolate and identify D-lactate-utilizing bacteria from pig gut using the Hungate roll-tube method, and to investigate their metabolic characteristics in vitro. Six different anaerobes were isolated from pig feces, which were identified as related to Bacteroides fragilis, Bacteroides acidifaciens, Veillonella denticariosi, Veillonella caviae, Bacteroides uniformis, and Megasphaera elsdenii based on the 16S rRNA gene sequences. All strains had a significant ability to utilize D-lactate, which was concluded after in vitro fermentation with 25 mmol L–1 D-lactate as the primary carbon source. Of all 6 strains, M. elsdenii J6 showed the highest efficiency of D-lactate utilization and produced a higher ratio of butyrate in total short chain fatty acids (SCFAs). Thus, the in vitro fermentation characteristics of this strain in D-, L-, and DL-lactate mixtures (D-lactate:L-lactate=1:1 or 1:2) were further studied. The results showed that M. elsdenii J6 preferred utilizing D-lactate, and produced more SCFA when using D-lactate as the primary carbon source. The findings suggest that the administration of D-lactate-utilizing bacteria such as M. elsdenii J6 may have a potential advantage in the alleviation of D-lactic acidosis in the animal gut.
Keywords:  D-lactate-utilizing bacteria       Megasphaera elsdenii       butyrate        pig        gut  
Received: 07 April 2015   Accepted:
Fund: 

This research has received funding from the National Basic Research Program of China (2013CB127603 and 2012CB124705), the National Natural Science Foundation of China (31572414 and 30901036), and the Natural Science Foundation of Jiangsu Province, China (BK20130058).

Corresponding Authors:  SU Yong, Tel: +86-25-84395523, Fax: +86-25-84395314, E-mail: yong.su@njau.edu.cn   

Cite this article: 

JIANG Xiao-lin, SU Yong, ZHU Wei-yun. 2016. Fermentation characteristics of Megasphaera elsdenii J6 derived from pig feces on different lactate isomers. Journal of Integrative Agriculture, 15(7): 1575-1583.

Allison M J. 1978. Production for branched-chain volatile fatty acids by certain anaerobic bacteria. Applied and Environmental Microbiology, 35, 872.

Barcenilla A, Pryde S E, Martin J C, Duncan S H, Stewart C S, Henderson C, Flint H J. 2000. Phylogenetic relationships of butyrate-producing bacteria from the human gut. Applied and Environmental Microbiology, 66, 1654–1661.

Biddle A S, Black S J, Blanchard J L. 2013. An in vitro model of the horse gut microbiome enables identification of lactate-utilizing bacteria that differentially respond to starch induction. PloS ONE, 8, e77599.

Black F, Einarsson K, Lidbeck A, Orrhage K, Nord C E. 1991. Effect of lactic acid producing bacteria on the human intestinal microflora during ampicillin treatment. Scandinavian Journal of Infectious Diseases, 23, 247–254.

Bongaerts G, Tolboom J, Naber A, Sperl W, Severijnen R, Bakkeren J, Willems J L. 1997. Role of bacteria in the pathogenesis of short bowel syndrome-associated D-lactic acidemia. Microbial Pathogenesis, 22, 285–293.

Bourriaud C, Akoka S, Goupry S, Robins R, Cherbut C, Michel C. 2002. Butyrate production from lactate by human colonic microflora. Reproduction Nutrition Devlopment, 42, S55.

Bourriaud C, Robins R, Martin L, Kozlowski F, Tenailleau E, Cherbut C, Michel C. 2005. Lactate is mainly fermented to butyrate by human intestinal microfloras but inter-individual variation is evident. Journal of Applied Microbiology, 99, 201–212.

Bryant M. 1972. Commentary on the Hungate technique for culture of anaerobic bacteria. American Journal of Clinical Nutrition, 25, 1324–1328.

Buchanan R E, Gibbons N E. 1984. Bergey’s Manual of Systematic Bacteriology. Science Press, Beijing. pp. 535–536.

Chan L, Slater J, Hasbargen J, Herndon D N, Veech R L, Wolf S. 1994. Neurocardiac toxicity of racemicd, L-lactate fluids. Integrative Physiological and Behavioral Science, 29, 383–394.

Connor H, Woods H, Ledingham J. 1983. Comparison of the kinetics and utilisation of D (–)- and L (+)-sodium lactate in normal man. Annals of Nutrition and Metabolism, 27, 481–487.

Counotte G, Lankhorst A, Prins R. 1983. Role of DL-lactic acid as an intermediate in rumen metabolism. Journal of Animal Science, 56, 1222–1235.

Counotte G, Prins R, Janssen R. 1981. Role of Megasphaera elsdenii in the fermentation of DL-[2-13C] lactate in the rumen of dairy cattle. Applied and Environmental Microbiology, 42, 649–655.

Duncan S H, Louis P, Flint H J. 2004. Lactate-utilizing bacteria, isolated from human feces, that produce butyrate as a major fermentation product. Applied and Environmental Microbiology, 70, 5810–5817.

Duncan S H, Scott K P, Ramsay A G, Harmsen H J M, Welling G W, Stewart C S, Flint H J. 2003. Effects of alternative dietary substrates on competition between human colonic bacteria in an anaerobic fermentor system. Applied and Environmental Microbiology, 69, 1136–1142.

Ewaschuk J B, Naylor J M, Zello G A. 2005. D-Lactate in human and ruminant metabolism. Journal of Nutrition, 135, 1619–1625.

Hashizume K, Tsukahara T, Yamada K, Koyama H, Ushida K. 2003. Megasphaera elsdenii JCM1772T normalizes hyperlactate production in the large intestine of fructooligosaccharide-fed rats by stimulating butyrate production. Journal of Nutrition, 133, 3187–3190.

Hino T, Kuroda S. 1993. Presence of lactate dehydrogenase and lactate racemase in Megasphaera elsdenii grown on glucose or lactate. Applied and Environmental Microbiology, 59, 255–259.

Hino T, Shimada K, Maruyama T. 1994. Substrate preference in a strain of Megasphaera elsdenii, a ruminal bacterium, and its implications in propionate production and growth competition. Applied and Environmental Microbiology, 60, 1827–1831.

Hove H, Mortensen P B. 1995. Colonic lactate metabolism and D-lactic acidosis. Digestive Diseases and Sciences, 40, 320–330.

Hungate R E. 1979. Evolution of a microbial ecologist. Annual Review of Microbiology, 33, 1–21.

Kaneko T, Bando Y, Kurihara H, Satomi K, Nonoyama K, Matsuura N. 1997. Fecal microflora in a patient with short-bowel syndrome and identification of dominant Lactobacilli. Journal of Clinical Microbiology, 35, 3181–3185.

Kung L, Hession A. 1995. Preventing in vitro lactate accumulation in ruminal fermentations by inoculation with Megasphaera elsdenii. Journal of Animal Science, 73, 250–256.

Lane D J. 1991. 16S/23S rRNA sequencing. In: Stackebrandt E, Goodfellow M, eds., Nucleic Acid Techniques in Bacterial Systematics. John Wiley & Sons, Chichester, United Kingdom. pp. 115–175.

Liu W, Zhu W Y, Yao W, Mao S Y. 2007. Isolation and identification of a lactate-utilizing, butyrate-producing bacterium and its primary metabolic characteristics. Acta Microbiologica Sinica, 47, 435–440. (in Chinese)

Long L M, Mao S Y, Su Y, Zhu W Y. 2008. Isolation and in vitro metabolic characterization of a lactate-utilizing bacterium from goat rumen. Acta Microbiologica Sinica, 48, 1571–1577. (in Chinese)

Mackie R I, Heath S. 1979. Enumeration and isolation of lactate-utilizing bacteria from the rumen of sheep. Applied and Environmental Microbiology, 38, 416–421.

Marounek M, Bartos S. 1987. Interactions between rumen amylolytic and lactate-utilizing bacteria in growth on starch. Journal of Applied Bacteriology, 63, 233–238.

Mizock B A. 1989. Lactic acidosis. Disease-A-Month, 35, 237–300.

Moazeni F, Zhang G, Sun H J. 2010. Imperfect asymmetry of life: earth microbial communities prefer D-lactate but can use L-lactate also. Astrobiology, 10, 397–402.

Munoz-Tamayo R, Laroche B, Walter E, Dore J, Duncan S H, Flint H J, Leclerc M. 2011. Kinetic modelling of lactate utilization and butyrate production by key human colonic bacterial species. FEMS Microbiology Ecology, 76, 615–624.

Ng S K, Hamilton I R. 1971. Lactate metabolism by Veillonella parvula. Journal of Applied Bacteriology, 105, 999–1005.

Schultz J, Breznak J A. 1979. Cross-feeding of lactate between Streptococcus lactis and Bacteroides sp. isolated from termite hindguts. Applied and Environmental Microbiology, 37, 1206–1210.

Oh M S, Uribarri J, Alveranga D, Lazar I, Bazilinski N, Carroll H J. 1985. Metabolic utilization and renal handling of D-lactate in men. Metabolism, 34, 621–625.

Prabhu R, Altman E, Eiteman M A. 2012. Lactate and acrylate metabolism by Megasphaera elsdenii under batch and steady-state conditions. Applied and Environmental Microbiology, 78, 8564–8570.

Pryde S E, Duncan S H, Hold G L, Stewart C S, Flint H J. 2002. The microbiology of butyrate formation in the human colon. FEMS Microbiology Letter, 217, 133–139.

Rowe J B, Lees M J, Pethick D W. 1994. Prevention of acidosis and laminitis associated with grain feeding in horses. Journal of Nutrition, 124, 2742S–2744S.

Stanton T B, Humphrey S B. 2011. Persistence of antibiotic resistance: Evaluation of a probiotic approach using antibiotic-sensitive Megasphaera elsdenii strains to prevent colonization of swine by antibiotic-resistant strains. Applied and Environmental Microbiology, 77, 7158–7166.

Stolberg L, Rolfe R, Gitlin N, Merritt J, Mann Jr L, Linder J, Finegold S. 1982. D-Lactic acidosis due to abnormal gut flora: Diagnosis and treatment of two cases. New England Journal of Medicine, 306, 1344.

Su Y, Li B, Zhu W Y. 2013. Fecal microbiota of piglets prefer utilizing dl-lactate mixture as compared to D-lactate and L-lactate in vitro. Anaerobe, 19, 27–33.

Tubbs P. 1965. The metabolism of D-α-hydroxy acids in animal tissues. Annals of the New York Academy of Sciences, 119, 920–926.

Tsukahara T, Koyama H, Okada M, Ushida K. 2002. Stimulation of butyrate production by gluconic acid in batch culture of pig cecal digesta and identification of butyrate-producing bacteria. Journal of Nutrition, 132, 2229–2234.

Ushida K, Hoshi S, Ajisaka K. 2002. 13C-NMR Studies on lactate metabolism in a porcine gut microbial ecosystem. Microbial Ecology in Health and Disease, 14, 242–247.

Vernia P, Caprilli R, Latella G, Barbetti F, Magliocca F M, Cittadini M. 1988. Fecal lactate and ulcerative colitis. Gastroenterology, 95, 1564–1568.

Yang C J, Mao S Y, Long L M, Zhu W Y. 2012. Effect of disodium fumarate on microbial abundance, ruminal fermentation and methane emission in goats under different forage: Concentrate ratios. Animal, 6, 1788–1794.

Zhang D L, Jiang Z W, Jiang J, Cao B, Li J S. 2003. D-Lactic acidosis secondary to short bowel syndrome. Postgraduate Medical Journal, 79, 110–112.

Zoetendal E G, Akkermans A D, De Vos W M. 1998. Temperature gradient gel electrophoresis analysis of 16S rRNA from human fecal samples reveals stable and host-specific communities of active bacteria. Applied and Environmental Microbiology, 64, 3854–3859.
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