研究简报 Short Communications

外源L-色氨酸、果聚糖、酪蛋白对猪粪发酵液粪臭素浓度的影响

展开
  • 山东省农科院畜牧兽医研究所, 山东省畜禽疫病防治与繁育重点实验室, 济南 250100
盛清凯(1971-),男,山东陵县人,研究员,博士,研究方向为单胃动物营养与环境科学。E-mail:qksheng@163.com

收稿日期: 2014-04-03

  网络出版日期: 2014-09-10

基金资助

国家自然科学基金项目(31172245)

Effects of Additional L-Tryptophan, Fructan and Casein on Skatole Concentration in Pig Manure Fermentation Broths

Expand
  • Shandong Provincial Key Laboratory of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan 250100, China

Received date: 2014-04-03

  Online published: 2014-09-10

摘要

为了减少粪臭素在猪体外的污染,本试验旨在研究不同剂量的外源L-色氨酸、果聚糖和酪蛋白对猪粪发酵液中粪臭素及相关合成物浓度的影响。将80头体重为(50.0±0.5) kg的“杜×长×大”三元杂交猪随机分为对照组和试验组,每组4个重复,每重复10头猪,分别饲喂基础饲粮和添加0.2%枯草芽孢杆菌的试验饲粮。6周后,每组随机选取8头猪,采集其新鲜粪便,配制粪水厌氧发酵液用于发酵试验。发酵试验采用3×3正交设计,发酵液中外源L-色氨酸、酪蛋白、果聚糖的添加剂量分别为0、0.05%、0.10%,0、0.125%、0.250%,0、0.75%、1.50%。色谱法测定发酵液中色氨酸、吲哚、吲哚-3-乙酸、粪臭素浓度,荧光标记单链构象多态性及片段长度多态性技术与毛细管电泳技术结合测定空白对照组和空白试验组发酵液中微生物的种群结构。结果表明:1)对照组色氨酸、吲哚、吲哚-3-乙酸、粪臭素浓度显著高于试验组(P<0.05)。2)外源L-色氨酸为发酵液中色氨酸、粪臭素的主要影响因素,酪蛋白为对照组中吲哚的主要影响因素,果聚糖为发酵液中吲哚-3-乙酸的主要影响因素。3)外源L-色氨酸、果聚糖与饲粮中的枯草芽孢杆菌无互作(P>0.05)。4)空白对照组猪粪发酵液优势菌以芽孢杆菌(CWBIB1434)为主,含量为26%;空白试验组发酵液以非解乳链球菌(AF201899)为主,含量为36%。 由此得出,降低猪粪中色氨酸的含量及在饲粮中添加枯草芽孢杆菌有助于减少猪粪发酵液中的粪臭素浓度。

本文引用格式

盛清凯, 成建国, 赵红波, 窦红艳, 宣玉娟, 武英 . 外源L-色氨酸、果聚糖、酪蛋白对猪粪发酵液粪臭素浓度的影响[J]. 动物营养学报, 2014 , 26(9) : 2797 -2804 . DOI: 10.3969/j.issn.1006-267x.2014.09.043

Abstract

In order to reduce the pollution of pig skatole in vitro, this study investigated the effects of different doses of additional L-tryptophan (L-Trp), fructan and casein on skatole and its related compounds concentration in pig manure fermentation broths. Eighty Duroc×Landrace×Yorkshire pigs with (50.0±0.5) kg body weight were randomly allotted into control group and test group with 4 replicates per group and 10 pigs per replicate. Pigs were accordingly fed a basal diet and an adding 0.2% Bacillus subtilis test diet. After 6 weeks, 8 pigs in each group were randomly selected; their fresh manures were collected and prepared manure anaerobic broths for fermentation experiment. A 3×3 orthogonal design was used in the fermentation experiment. The additional doses of L-Trp, casein, fructan in fermentation broths were 0, 0.05% and 0.10%; 0, 0.125% and 0.250%; 0, 0.75% and 1.50%, respectively. The concentrations of Trp, indole-3-acetic acid (IAA), indole and skatole were detected by high performance liquid chromatography (HPLC) and microbial communities in fermented broths in blank control and blank test groups were detected by fluorescnce-based PCR-SSCP-FLP (F-PCR-SSCP-FLP) and capillary electrophoresis. The results showed as follows: 1) the concentrations of Trp, indole, IAA and skatole in test group were significantly decreased compared with the control group (P<0.05). 2) L-Trp was the main influence factor on Trp and skatole concentration in fermentation broths, respectively, casein was the main influence factor on indole concentration in the control group, and fructan was the main factor on IAA concentration in fermentation broths. 3) Additional L-Trp or fructan and Bacillus subtilis did not interact (P>0.05). 4) The bacteria Bacillus (CWBIB1434) was predominant and its content was 26% in the blank control group, and the Lactose Streptococcus (AF201899) was predominant and its content was 36% in the blank test group. The conclusion is that the reduction of Trp in pig manure and additional Bacillus subtilis in diet are beneficial to the abatement of skatole pollution in vitro.

参考文献

[1] WESOLY R,WEILER U.Influences of feed and feed additives on the skatole formation and accretion in adipose tissue of pigs[J].Züchtungskunde,2012,84(5):412-426.  

[2] ZAMARATSKAIA G,SQUIRES E J.Biochemical,nutritional and genetic effects on boar taint in entire male pigs[J].Animal,2009,3(11):1508-1521.  

[3] 杨锋.枯草芽孢杆菌的抗逆特性及其对仔猪生化指标和氨气排放的影响[D].硕士学位论文.杭州:浙江工商大学,2011.

[4] 李彩燕.饲粮纤维对猪体粪臭素沉积的调控及分子机理研究[D].博士学位论文.杭州:浙江大学,2009.

[5] 夏枚生,胡彩虹,许梓荣.果寡糖对猪粪便细菌群作用下L-色氨酸代谢的影响[J].中国畜牧杂志,2004,40(1):11-13.

[6] 刘毅,徐维家,李妍.人工神经原网络处理PCR数据在细菌鉴定中的应用[J].中国微生态学杂志,2009,21(5):468-473.

[7] WEEMS J M,CUTLER N S,MOORE C,et al.3-methylindole is mutagenic and a possible pulmonary carcinogen[J].Toxicological Sciences,2009,112(1):59-67.  

[8] TERENCE R W,NEIL P P,HAROID L D,et al.Catabolic pathway for the production of skatole and indoleacetic acid by the acetogen Clostridium drakei,Clostridium scatologenes,and swine manure[J].Applied and Environmental Microbiology,2008,74(6):1950-1953.  

[9] LI C Y,LIU J X,WANG Y Z,et al.Influence of differing carbohydrate sources on L-tryptophan metabolism by porcine fecal microbiota studied in vitro[J].Livestock Science,2009,120(1/2):43-50.

[10] DOERNER K C,COOK K L,MASON B P.3-methylindole production is regulated in Clostridium scatologenes ATCC 25775[J].Letters in Applied Microbiology,2009,48(1):125-132.  

[11] YOKOYAMA M T,JOHNSON K A,CARLSON J R.Factors influencing the production of p-cresol and skatole by Lactobacillus isolated from the rumen and pig feces//Proceedings of 17 conference on rumen function.Chicag:USA,1983:19-20.

[12] JENSEN B B.Prevention of boar taint in pig production.Factors affecting the level of skatole[J].Acta Veterinaria Scandinavica,2006,48(Suppl.1):S6.

[13] PEDERSEN J K,MORTENSEN A B,MADSEN A,et al.Foderets indflydelse pa ornelugt i svinekod statens hysdyrbrugsforsog[J].Meddelse,1986(638):1-7.

[14] RAFFAEL W,ULRIKE W.Nutritional influences on skatole formation and skatole metabolism in the pig[J].Animals,2012,2(2):221-242.

[15] LIN R S,ORCUTT M W,ALLRICH R D,et al.Effect of dietary crude protein content on skatole concentration in boar serum[J].Meat Science,1992,31(4):473-479.  

[16] LUNDSTRÖMA K,MALMFORSB B,STERNB S,et al.Skatole levels in pigs selected for high lean tissue growth rate on different dietary protein levels[J].Livestock Production Science,1994,38(2):125-132.  

[17] AN C,KUDA T,YAZAKI T,et al.Caecal fermentation,putrefaction and microbiotas in rats fed milk casein,soy protein or fish meal[J].Applied Microbiology and Biotechnology,2014,98(6):2779-2787.  

[18] RUSSELL W R,DUNCAN S H,SCOBBIE L,et al.Major phenylpropanoid-derived metabolites in the human gut can arise from microbial fermentation of protein[J].Molecular Nutrition & Food Research,2013,57(3):523-535.  

[19] XU X,HU C,WANG M.Effects of fructooligosaccharide on conversion of L-tryptophan to skatole and indole by mixed populations of pig fecal bacteria[J].The Journal of General and Applied Microbiology,2002,48(2):83-89.  

[20] VHILE S G,KJOS N P,SØ RUM H,et al.Feeding Jerusalem artichoke reduced skatole level and changed intestinal microbiota in the gut of entire male pigs[J].Animal,2012,6(5):807-814.  

[21] LI C Y,WU C,LIU J X,et al.Spatial variation of intestinal skatole production and microbial community in Jinhua and Landrace pigs[J].Journal of the Science of Food and Agriculture,2009,89(4):639-644.  

[22] PASCAL P,HUBERT B,ANNE-MARIE P,et al.Dynamics of a pig slurry microbial community during anaerobic storage and management[J].Applied and Environmental Microbiology,2006,72(5):3578-3585.  

[23] OLSON A B,SIBLEY C D,SCHMIDT L,et al.Development of real-time PCR assays for detection of the Streptococcus milleri group from cystic fibrosis clinical specimens by targeting the cpn60 and 16S rRNA genes[J].Journal of Clinical Microbiology,2010,48(4):1150-1160.  

[24] RINKINEN M L,KOORT J M,OUWEHAND A C,et al.Streptococcus alactolyticus is the dominating culturable lactic acid bacterium species in canine jejunum and feces of four fistulated dogs[J].FEMS Microbiology Letters,2004,230(1):35-39.  

[25] VANDAMME P,DEVRIESE L A,HAESEBROUCK F,et al.Streptococcus intestinalis Robinson et al.,1988 and Streptococcus alactolyticus Farrow et al.,1984 are phenotypically indistinguishable[J].International Journal of Systematic Bacteriology,1999(49):737–741.

[26] Ø VERLANDA M,KJOSB N P,BORGC M,et al.Organic acids in diets for entire male pigs:effect on skatole level,microbiota in digesta,and growth performance[J].Livestock Science,2008,115(2/3):169-178.

[27] NOWAK A,LIBUDZISZ Z.Ability of probiotic Lactobacillus casei DN 114001 to bind or/and metabolise heterocyclic aromatic amines in vitro[J].European Journal of Nutrition,2009,48(7):419-427.  

[28] 李凤.公猪膻味物质粪臭素的乳酸菌降解研究[D].博士学位论文.重庆:西南大学,2011,141-142.

[29] KONSTANTINOV S R,AWATI A,SMIDT H,et al.Specific response of a novel and abundant Lactobacillus amylovorus-like phylotype to dietary prebiotics in the guts of weaning piglets[J].Applied and Environmental Microbiology,2004,70(7):3821-3830.  

[30] KIM P I,JUNG M Y,CHANG Y H,et al.Probiotic properties of Lactobacillus and Bifidobacterium strains isolated from porcine gastrointestinal tract[J].Applied Microbiology and Biotechnology,2007,74(5):1103-1111.  

[31] MARTINEZ R C,AYNAOU A E,ALBRECHT S,et al.In vitro evaluation of gastrointestinal survival of Lactobacillus amylovorus DSM 16698 alone and combined with galactooligosaccharides,milk and/or Bifidobacterium animalis subsp.lactis Bb-12[J].International Journal of Food Microbiology,2011,149(2):152-158.  

[32] FINAMORE A,ROSELLI M,IMBINTO A,et al.Lactobacillus amylovorus inhibits the TLR4 inflammatory signaling triggered by enterotoxigenic Escherichia coli via modulation of the negative regulators and involvement of TLR2 in intestinal Caco-2 cells and pig explants[J].PLoS One,2014,9(4):e94891.

[33] EOM H J,MOON J S,SEO Y,et al.Heterologous expression and secretion of Lactobacillus amylovorus alpha-amylase in Leuconostoc citreum[J].Biotechnology Letters,2009,31(11):1783-1788.  

[34] MARTINEZ R C,CARDARELLI H R,BORST W,et al.Effect of galactooligosaccharides and Bifidobacterium animalis Bb-12 on growth of Lactobacillus amylovorus DSM 16698,microbial community structure,and metabolite production in an in vitro colonic model set up with human or pig microbiota[J].FEMS Microbiology Ecology,2013,84(1):110-123.  

[35] MENG X,HE Z F,LI H J.Purification and characterization of a novel skatole-degrading protease from Lactobacillus brevis 1.12[J].Food Science and Biotechnology,2013,22(5):1367-1373.
文章导航

/