Gut Bacterial and Lactobacilli Communities of Weaning Piglets in Response to Mannan Oligosaccharide and Sugar Beet Pulp In vitro Fermentation

doi:10.1016/S1671-2927(00)8516

Journal of Integrative Agriculture ›› 2012, Vol. 12 ›› Issue (1): 122-133.DOI: 10.1016/S1671-2927(00)8516

Gut Bacterial and Lactobacilli Communities of Weaning Piglets in Response to Mannan Oligosaccharide and Sugar Beet Pulp In vitro Fermentation

HANG Su-qin , ZHU Wei-yun

1.Laboratory of Gastrointestinal Microbiology, Nanjing Agricultural University, Nanjing 210095, P.R.China

收稿日期:2010-11-18 出版日期:2012-01-01 发布日期:2012-01-13
通讯作者: Correspondence ZHU Wei-yun, Tel: +86-25-84395523, Fax: +86-25-84395314, E-mail:zhuweiyun@njau.edu.cn
作者简介:HANG Su-qin, Mobile: 13512532889, E-mail: suqinhang69@yahoo.com.cn
基金资助:
This work was funded by the National Basic Research Program of China (973 Program, 2004CB117500).

Gut Bacterial and Lactobacilli Communities of Weaning Piglets in Response to Mannan Oligosaccharide and Sugar Beet Pulp In vitro Fermentation

HANG Su-qin , ZHU Wei-yun

1.Laboratory of Gastrointestinal Microbiology, Nanjing Agricultural University, Nanjing 210095, P.R.China

Received:2010-11-18 Online:2012-01-01 Published:2012-01-13
Contact: Correspondence ZHU Wei-yun, Tel: +86-25-84395523, Fax: +86-25-84395314, E-mail:zhuweiyun@njau.edu.cn
About author:HANG Su-qin, Mobile: 13512532889, E-mail: suqinhang69@yahoo.com.cn
Supported by:
This work was funded by the National Basic Research Program of China (973 Program, 2004CB117500).

摘要/Abstract

摘要： Microbiota in the gastrointestinal tract (GIT) of piglets during weaning transition can experience a sharp change which could result in growth reduction and diarrhea of weaned piglets. Dietary manipulations can play an important role in attenuating such changes caused by weaning stress. Therefore, ileal and colonic contents of weaned piglets were used as inocula, mannan oligosaccharide (MOS) or sugar beet pulp (SBP) was supplied as single energy sources to investigate effects of MOS or SBP on the shifts of gastro-intestinal microflora and lactobacilli populations. The universal bacteriaand lactobacilli-specific PCR/denaturing gradient gel electrophoresis (DGGE), cloning and sequencing techniques were used. DGGE profiles of the universal bacteria showed that great changes were found in the position, numbers and intensity of dominant bands after fermentation. The similarity of bacterial community between ileum and colon was increased to 85-97% by MOS or SBP treatment after fermentation from the similarity with 20% before fermentation. MOS treatment significantly increased the bacterial diversity and band number in both ileal and colonic fermentation (P<0.05). SBP treatment significantly increased the bacterial diversity and band number in colon (P<0.05). It implies that some species were enriched by the addition of MOS or SBP to increase the similarity and diversity of bacterial community in weaned piglets. Five specific bands appearing in MOS or SBP treatment group after fermentation were cloned and sequenced, the changes of species related to Prevotella and Ruminococcus were observed. Two bands related to uncultured bacterium with 98% similarity were detected by MOS or SBP treatment. However, there were no effects on the similarity, diversity index and lactobacilli species revealed by MOS or SBP treatment. These results imply that MOS or SBP could have beneficial effects on the weaning piglets by stablizing microbiota in the GIT microflora.

关键词: mannan oligosaccharide (MOS), sugar beet pulp (SBP), bacterial communities, lactobacilli, weaning piglets

Abstract: Microbiota in the gastrointestinal tract (GIT) of piglets during weaning transition can experience a sharp change which could result in growth reduction and diarrhea of weaned piglets. Dietary manipulations can play an important role in attenuating such changes caused by weaning stress. Therefore, ileal and colonic contents of weaned piglets were used as inocula, mannan oligosaccharide (MOS) or sugar beet pulp (SBP) was supplied as single energy sources to investigate effects of MOS or SBP on the shifts of gastro-intestinal microflora and lactobacilli populations. The universal bacteriaand lactobacilli-specific PCR/denaturing gradient gel electrophoresis (DGGE), cloning and sequencing techniques were used. DGGE profiles of the universal bacteria showed that great changes were found in the position, numbers and intensity of dominant bands after fermentation. The similarity of bacterial community between ileum and colon was increased to 85-97% by MOS or SBP treatment after fermentation from the similarity with 20% before fermentation. MOS treatment significantly increased the bacterial diversity and band number in both ileal and colonic fermentation (P<0.05). SBP treatment significantly increased the bacterial diversity and band number in colon (P<0.05). It implies that some species were enriched by the addition of MOS or SBP to increase the similarity and diversity of bacterial community in weaned piglets. Five specific bands appearing in MOS or SBP treatment group after fermentation were cloned and sequenced, the changes of species related to Prevotella and Ruminococcus were observed. Two bands related to uncultured bacterium with 98% similarity were detected by MOS or SBP treatment. However, there were no effects on the similarity, diversity index and lactobacilli species revealed by MOS or SBP treatment. These results imply that MOS or SBP could have beneficial effects on the weaning piglets by stablizing microbiota in the GIT microflora.

Key words: mannan oligosaccharide (MOS), sugar beet pulp (SBP), bacterial communities, lactobacilli, weaning piglets

HANG Su-qin , ZHU Wei-yun. Gut Bacterial and Lactobacilli Communities of Weaning Piglets in Response to Mannan Oligosaccharide and Sugar Beet Pulp In vitro Fermentation[J]. Journal of Integrative Agriculture, 2012, 12(1): 122-133.

参考文献

[1]Araya-Kojima T, Yaeshima T, Ishibashi N, Shimamura S, Hayasawa H. 1995. Inhibitory effects of Bifidobacterium longum BB536 on harmful intestinal bacteria. Bifidobacteria Microflora, 14, 59-66.

[2]Bauer E, Williams B A, Voigt C, Mosenthin R, Verstegen M W A. 2001. Microbial activities of faeces from unweaned and adult pigs, in relation to selected fermentable carbohydrates. Animal Science, 73, 313-322.

[3]van den Bogaard A E. 1997. Antimicrobial resistancerelation to human and animal exposure to antibiotics. Journal of Antimicrobial Chemotherapy, 40, 453-454.

[4]Cromwell G L. 2002. Why and how antibiotics are used in swine production. Animal Biotechnology, 13, 7-27.

[5]Crosby L D, Criddle C S. 2003. Understanding bias in microbial community analysis techniques due to rrn operon copy number heterogeneity. BioTechniques, 34, 790-802.

[6]Davies D R, Theodorou M K, Baughan J, Brooks A E, Newbold J R. 1995. An automated pressure evaluation system (APES) for determining the fermentation characteristics of ruminant feeds. Annual Zootechnology, 44, 36-36.

[7]Edwards S A. 1996. A new look on the role of fibre in the diet of pigs. In: Proceedings of the 6th European Society of Veterinary Internal Medicine. Organizing Committee of the 6th ESVIM Congress, Veldhoven, Netherlands. pp. 90-91.

[8]Gibson G R, Roberfroid M B. 1995. Dietary modulation of the human colonic microbiota: introducing the concept of prebiotics. Journal of Nutrition, 125, 1401-1412.

[9]Hang S Q, Dai Z L, Zhu W Y. 2009. Effects of mannanoligosaccharide on the growth of Lactobacillus pure culture and co-culture. Microbiology, 36, 51-56. (in Chinese)

[10]Hang S Q, Mao S Y, Yu Z T, Zhu W Y. 2007. Effect of mannanoligosaccharide and sugar beet pulp on intestinal microbial fermentation in vitro. Journal of Nanjing Agricultural University, 30, 79-83. (in Chinese)

[11]Harmsen H J M, Elfferich P S F, Welling G W. 1999. A 16S rRNA-targeted probe for detection of Lactobacilli and Enterococci in faecal samples by fluorescent in situ hybridization. Microbial Ecology in Health and Disease, 11, 3-12.

[12]Hayakawa K, Mizutani J, Wada K, Masai T, Yoshihara I, Mitsuoka T. 1990. Effects of soybean oligosaccharides on human faecal microflora. Microbial Ecology in Health and Disease, 3, 293-303.

[13]Heilig H G, Zoetendal E G, Vaughan E E, Marteau P, Akkermans A D L, de Vos W M. 2002. Molecular diversity of Lactobacillus spp. and other lactic acid bacteria in the human intestine as determined by specific amplification of 16S ribosomal DNA. Applied Environmental Microbiology, 68, 114-123.

[14]Hidaka H, Eida T, Takizawa T, Tokunaga T, Tashiro Y. 1986. Effects of fructooligosaccharides on intestinal flora and human health. Bifidobacteria Microflora, 5, 37-50.

[15]Houdijk J G, Bosch M W, Tamminga S, Verstegen M W, Berenpas E B, Knoop H. 1999. Apparent ileal and totaltract nutrient digestion by pigs as affected by dietary nondigestible oligosaccharides. Journal of Animal Science, 77, 148-158.

[16]Jensen B B, Jorgensen H. 1994. Effect of dietary fibre on microbial activity and microbial gas production in various regions of the gastrointestinal tract of pigs. Applied Environmental Microbiology, 60, 1897-1904.

[17]Kane M D, Poulsen L K, Stahl D. 1993. Monitoring the enrichment and isolation of sulfate reducing bacteria by using oligonucleotide hybridization probes designed from environmentally derived 16S rRNA sequences. Applied Environmental Microbiology, 59, 682-686.

[18]Konstantinov S R, Awati A A, Williams B A, Miller B G, Jones P, Stokes C R, Akkermans A D L, Smidt H, de Vos W M. 2006a. Post-natal development of the porcine microbiota composition and activities. Environmental Microbiology, 8, 1191-1199.

[19]Konstantinov S R, Poznanski E, Fuentes S, Akkermans A D L, Smidt H, de Vos W M. 2006b. Lactobacillus sobrius sp. nov., a novel isolate abundant in the intestine of weaning piglets. International Journal of Systematic and Evolutionary Microbiology, 56, 29-32.

[20]Konstantinov S R, Smidt H, de Vos W M. 2005. Representative difference analysis and real-time PCR for strain-specific quantification of Lactobacillus sobrius sp. nov. Applied Environmental Microbiology, 71, 7578-7581.

[21]Konstantinov S R, Zhu W Y, Williams B A, Tamminga S, de Vos W M, Akkermans A D L. 2003. Effect of fermentable carbohydrates on piglet faecal bacterial communities as revealed by denaturing gradient gel electrophoresis analysis of 16S ribosomal DNA. FEMS Microbiology Ecology, 43, 225-235.

[22]Lane D J. 1991. 16S/23S rRNA sequencing. In: Stackebrandt E, Goodfellow M, eds., Nucleic Acid Techniques in Bacterial Systematics. John Wiley & Sons, UK. pp. 115-175.

[23]Loddi M M, Nakaghi L S O, Edens F, Tucci F M, Hannas M I, Moraes V M B, Ariki J. 2002. Manna noligosacharide and organic acids on intestinal morphology integrity of broilers evaluated by scanning electron microscopy. In: Proceedings of the 11th European Poultry Science Conference. Bremen, Germany. p. 121.

[24]Lowe S E, Theodorou M K, Trinci A P J, Hespell R B. 1985. Growth of anaerobic rumen fungi on defined and semidefined media lacking rumen fluid. Journal of General and Applied Microbiology, 131, 2225-2229.

[25]Mathew A G, Upchurch W G, Chattin S E. 1998. Incidence of antibiotic resistance in faecal Escherichia coli isolated from commercial swine farms. Jouranl of Animal Science, 76, 429-434.

[26]Miguel J C, Rodrogies-Zas S L, Pettigrew J E. 2002. Practical effects of Bio-Mos in nursery diets: Ameta-analysis. In: Lyons T P, Jacques K A, eds., Biotechnology in the Feed and Food Industry (Proceedings of the 13th Annual Symposium). Nottingham University Press, Nottingham, UK. pp. 425-433.

[27]Mikkelsen L L, Knudsen K E B, Jensen B B. 2004. In vitro fermentation of fructo-oligosaccharides and transgalacto-oligosaccharides by adapted and unadapted bacterial populations from the gastrointestinal tract of piglets. Animal Feed Science and Technology, 116, 225-238.

[28]Moore W E, Moore L V, Cato E P, Wilkins T D, Kornegay E T. 1987. Effect of high-fiber and high-oil diets on the faecal flora of swine. Applied Environmental Microbiology, 53, 1638-1644.

[29]Murray A E, Hollibaugh J T, Orrego G. 1996. Phylogenetic composition of bacterioplankton from two California estuaries compared by denaturing gradient gel electrophoresis of 16S rDNA fragments. Applied Environmental Microbiology, 62, 2676-2680.

[30]Nübel U, Engelen B, Felske A, Snaidr J, Wieshuber A, Amann R I, Ludwig W, Backhaus H. 1996. Sequence heterogeneities of genes encoding 16S rRNAs in Paenibacillus polymyxa detected by temperature gradient gel electrophoresis. Journal of Bacteriology, 178, 5636-5643.

[31]Pieper R, Janczyk P, Zeyner A, Smidt H, Guiard V, Souffrant W B. 2008. Ecophysiology of the developing total bacterial and Lactobacillus communities in the terminal small intestine of weaning piglets. Microbial Ecology, 56, 474-483.

[32]Sanguinetti C J, Dias N E, Simpson A J. 1994. Rapid silver staining and recovery of PCR products separated on polyacrylamide gels. Biotechniques, 17, 914-921.

[33]Scipioni R, Martelli G. 2001. Consequences of the use of ensiled sugar beet-pulp in the diet of heavy pigs on performances,carcass characteristics and nitrogen balance: a review. Animal Feed Science and Technology, 90, 81-91.

[34]Simpson J M, McCracken V J, Gaskins H R, Mackie R I. 2000. Denaturing gradient gel electrophoresis analysis of 16S ribosomal DNA amplicons to monitor changes in fecal bacterial populations of weaning pigs after introduction of Lactobacillus reuteri strain MM53. Applied Environmental Microbiology, 66, 4705-4714.

[35]Sims M D, Dawson K A, Newman K E, Spring P, Hooge D M. 2004. Effects of dietary mannan oligosaccharide, bacitracin methylene disalicylate, or both on the live performance and intestinal microbiology of Turkeys. Poultry Science, 83, 1148-1154.

[36]Spring P, Privulescu M. 1998. Mannan-oligosaccharide: its logical role as a natural feed additive for piglets. In: Lyons T P, Jacques K A, eds., Biotechnology in the Feed Industry (Proceedings of the 4th Annual Symposium of Alltech). Nottingham University Press, Nottingham, UK. pp. 553-561.

[37]© 2012, CAAS. All rights reserved. Published by Elsevier Ltd. Strickling J A, Harmon D L, Dawson K A, Gross K L. 2000. Evaluation of oligosaccharide addition to dog diets: influences on nutrient digestion and microbial populations. Animal Feed Science and Technology, 6, 205-219.

[38]Su Y, Yao W, Perez-Gutierrez Odette N, Smidt H, Zhu W Y. 2008. Changes inabundance of Lactobacillus spp. and Streptococcus suis in the stomach, jejunumand ileum of piglets after weaning. FEMS Microbiology Ecology, 66, 546-555.

[39]Sutton A L, Patterson J A. 1996. Effects of dietary carbohydrates and organic acid additions on pathogenic Escherichia coli and other microorganisms in the weanling pig. In: Proceedings of the 5th International Symposium on Animal Nutrition. Pannon Agricultural University, Kaposvar, Hungary. pp. 31-61.

[40]Verstegen M W, Williams B A. 2002. Alternatives to the use of antibiotics as growth promotors for monogastric animals. Animal Biotechnology, 13, 113-127.

[41]Williams B A, van der Poel A F B, Boer H, Tamminga S. 1995. The use of cumulative gas production to determine the effect of steam explosion on the fermentability of two substrates with different cell wall quality. Journal of the Science of Food and Agriculture, 69, 33-39.

[42]Williams B A, Verstegen M W A, Tamminga S. 2001. Fermentation in the large intestine of single-stomached animals and its relationship to animal health. Nutrition Research Reviews, 14, 207-227.

[43]Yu T, Morrison M. 2004. Comparisons of different hypervariable regions of rrs genes for use in fingerprinting of microbial communities by PCRdenaturing gradient gel electrophoresis. Applied Environmental Microbiology, 70, 4800-4806.

[44]Zhu W Y, Williams B A, Konstantinov S R, Tamminga S, de Vosa Willem M, Akkermans A D L. 2003. Analysis of 16S rDNA reveals bacterial shift during in vitro fermentation of fermentable carbohydrate using piglet faeces as inoculums. Anaerobe, 9, 175-180.

[45]Zoetendal E G, Akkermans A D L, de Vos W M. 1998. Temperature gradient gel electrophoresis analysis from human fecal samples reveals stable and host specific communities of active bacteria. Applied Environmental Microbiology, 64, 3854-3859.

Gut Bacterial and Lactobacilli Communities of Weaning Piglets in Response to Mannan Oligosaccharide and Sugar Beet Pulp In vitro Fermentation

Gut Bacterial and Lactobacilli Communities of Weaning Piglets in Response to Mannan Oligosaccharide and Sugar Beet Pulp In vitro Fermentation

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