嗜吡啶红球菌Rp3生长及降解粪臭素特性研究

为获得适宜嗜吡啶红球菌（Rhodococcus pyridinivorans）Rp3生长的碳氮源并明确不同因素对Rp3菌株降解粪臭素的影响,采用单因素和正交试验优化Rp3菌株的发酵培养基,并采用高效液相色谱法测定培养液中粪臭素含量,分析最适碳氮源、金属离子、温度和传代对Rp3菌株生长和降解粪臭素效率的影响。结果表明,Rp3菌株能够以粪臭素为唯一碳源生长。碳氮源对Rp3菌株生长的影响排序依次为酵母浸粉>牛肉膏>蔗糖,适宜碳氮源用量为蔗糖16 g·L^-1、牛肉膏14 g·L^-1、酵母浸粉6 g·L^-1。培养液中不添加Mg²⁺时,粪臭素不能被降解,Mg²⁺被Fe²⁺、Mn²⁺、Zn²⁺替代后,Rp3菌株对粪臭素的降解率显著下降。Rp3菌株对粪臭素的降解率随温度的升高而增加,28~40 ℃,培养24 h,降解效果可达到82.9%~95.5%。碳氮源和传代不影响Rp3菌株对粪臭素的降解。研究结果表明,嗜吡啶红球菌Rp3是易培养、不易退化的粪臭素高效降解菌株,具有一定的开发潜力。     

日粮中纳豆芽孢杆菌对体外猪粪发酵液中臭气及微生物的影响

为了减少舍外粪便污染,研究日粮中纳豆芽孢杆菌对体外猪粪发酵液中氨气、硫化氢、粪臭素及菌群的影响。试验组生猪饲喂纳豆芽孢杆菌日粮,对照组无纳豆芽孢杆菌。结果表明,试验组氨气、硫化氢、色氨酸、吲哚-3-乙酸、粪臭素浓度极显著低于对照组(P0.01),吲哚显著低于对照组(P0.05)。对照组猪粪发酵液优势菌以芽孢杆菌(CWBI B1434)为主,含量为26%;试验组以非解乳糖链球菌(AF201899)为主,含量为36%,其次为噬淀粉乳酸杆菌,含量为16%。结果表明纳豆芽孢杆菌通过调整猪粪菌群降低臭味物质浓度。为了减少舍外粪便污染,建议猪日粮中添加纳豆芽孢杆菌。     

应用反相高效液相色谱法测定绵羊瘤胃液、肉鸡粪中吲哚和粪臭素含量

[目的]建立适于绵羊瘤胃液和肉鸡粪中痕量吲哚、粪臭素测定的反相高效液相色谱法.[方法]肉鸡日粮中分别添加饲用抗生素、葡萄糖酸和乳果糖,收集13～15、20～22、33～35和40 ～41日龄肉鸡的粪;采集绵羊饲喂前(记为0h),饲喂后1.5、3、6和9h的瘤胃内容物.[结果]色谱条件,流动相:乙腈:水,流速1mL/min,柱温30℃,激发波长270nm,发射波长350nm,梯度洗脱;方法具有分离时间短(10 min以内),样品处理简单、分离度好、灵敏度高(吲哚142.8 ng/mL,绝对量2.142 ng/15 μL;粪臭素1 ng/mL,绝对量0.015ng/15 μL)的特点.葡萄糖酸组、乳果糖组肉鸡粪中吲哚含量在肉鸡34～36日龄时达到高峰值,抗生素组肉鸡粪中吲哚含量随日龄呈现上升趋势;从粪臭素含量来看,40 ～41日龄时肉鸡粪中含量有所增加,其中,饲用抗生素组、乳果糖组增加最明显,葡萄糖酸组增加最少.绵羊瘤胃液中吲哚含量在饲喂后1.5h达到最高值,饲喂800 g精料日粮时含量极显著高于饲喂400 g精料日粮,其余各时间点两种日粮之间差异不显著;而瘤胃液中粪臭素含量在饲喂后3h达到最高值,且饲喂800 g精料日粮时含量都有高于饲喂400 g精料日粮时.[结论]肉鸡粪中的吲哚、粪臭素含量不仅与日粮有关,还与肉鸡的日龄有关.饲喂两种水平的精料补充料时绵羊瘤胃液中吲哚、粪臭素含量的动态变化规律基本相似;高精料饲喂后1.5 ～9 h内,瘤胃液中的粪臭素含量普遍升高.     

海南特种野公猪不同世代膻味物质含量及相关性分析

【目的】通过比较不同世代杂交海南特种野公猪F1、F2、F3膻味物质雄烯酮、粪臭素和吲哚含量,及血清性激素水平的差异,揭示公猪膻味物质的变化规律及性激素的变化对膻味物质含量的影响,为选育低膻味性状的公猪提供理论基础。【方法】以12月龄F1、F2、F3不同世代杂交公猪(野猪♂×屯昌猪♀)各18头为研究对象。试验猪空腹12 h,于前腔静脉采取血液样本并提取血清样品,采血后将试验猪屠宰,按照商品猪分割标准从第三和第四颈椎之间的颈部区采集脂肪和肌肉样品。利用(ELISA)试剂盒检测血清样品雄烯酮和性激素水平及脂肪和肌肉样本的雄烯酮含量,采用高效液相色谱法检测血清、脂肪和肌肉样本的粪臭素和吲哚含量。【结果】海南特种野公猪膻味物质雄烯酮、粪臭素和吲哚在血清、脂肪和肌肉组织中非均匀分布。脂肪和肌肉组织中的粪臭素和雄烯酮含量均与血清中的粪臭素和雄烯酮含量存在显著或极显著相关性(P0.05或P0.01),脂肪中粪臭素的含量与血清中粪臭素含量的相关系数最大为0.94。公猪肉品膻味物质粪臭素、雄烯酮和吲哚的沉积量在不同杂交代公猪间存在明显差异,其中雄烯酮的沉积量随着野猪血统的增加而增加,粪臭素的沉积量则随着野猪血统的增加而降低,而吲哚的沉积量不存在随野猪血统的变化规律。性激素水平在不同杂交代野猪间存在明显差异,其中雄激素睾酮水平随着野猪血统的增加而极显著增加(P0.01),而F1公猪的雌二醇水平极显著低于F2和F3公猪(P0.01)。雄烯酮含量均与血清睾酮水平呈极显著正相关(P0.01),而粪臭素含量均与血清睾酮水平呈显著或极显著负相关(P0.01),血清雄烯酮含量与睾酮水平的相关系数最大为0.95。【结论】随着海南特种野公猪血统的增加,睾酮水平和雄烯酮的沉积量显著增加,而粪臭素的沉积量显著降低。因此,海南特种野公猪睾酮水平的变化影响粪臭素和雄烯酮含量的变化。     

In vitro effects of inulin and soya bean oligosaccharide on skatole production and the intestinal microbiota in broilers

The experiment was conducted to investigate the in vitro effects of inulin and soya bean oligosaccharide (SBO) on the metabolism of L‐tryptophan (L‐try) to skatole production, and the intestinal microbiota in broilers. Treatments were as follows: caecal microbiota control (Cc), Cc + inulin, Cc + SBO, rectal microbiota control (Rc), Rc + inulin and Rc + SBO. Microbial suspensions were anaerobically incubated at 38°C for 24 hr. The results showed that concentrations of skatole and acetic acid were significantly lower in caecal microbiota fermentation broth (MFB) than those in rectal MFB (p < .05). Addition of inulin or SBO significantly decreased the concentrations of indole and skatole and rate of L‐try degradation (p < .05). Inulin groups had lower indole than SBO groups (p < .05). PCR‐DGGE analysis revealed that addition of inulin or SBO decreased the microbiota richness (p < .05), but no significant differences in Shannon index (p > .05). Four distinct bands were detected in inulin and SBO groups, which were related to two of Bacteroides, one of Firmicutes and Bifidobacteria. Six bands were detected only in control groups, which represented uncultured Rikenellaceae, Roseburia, Escherichia/Shigella dysenteriae, Bacteroides uniformis (T), Parabacteroides distasonis and Enterobacter aerogenes. Populations of Lactobacilli, Bifidobacteria and total bacteria in inulin groups were higher than those in control groups (p < .05). For SBO groups, only population of total bacteria increased (p < .05). However, there were no significant differences in Escherichia coli population among treatments (p > .05). These results suggest that reduced concentrations of skatole and indole in the presence of inulin and SBO may be caused by decrease in L‐try degradation rate, which were caused by change in microbial ecosystem and pH value. Uncultured B. uniformis (T) and E. aerogenes may be responsible for degradation of L‐try to skatole.     

大豆低聚糖及其功能组分对体外条件下肉仔鸡盲肠内容物粪臭素产量及菌群组成的影响

本试验旨在研究大豆低聚糖(SBO)及其功能组分对体外条件下肉仔鸡盲肠内容物粪臭素产量及菌群组成的影响。试验采用单因素完全随机设计,以42日龄肉仔鸡盲肠内容物为菌源,将厌氧培养液分装于无菌培养瓶中,分成5个组,每组3个重复。对照组添加250μmol/L的L-色氨酸,蔗糖(SUC)、水苏糖(STA)、棉籽糖(RAF)和SBO组分别在对照组的基础上添加1%的SUC、STA、RAF和SBO。另外,每组均设1个不加L-色氨酸但其他成分都相同的空白对照。利用ANKOM RFS体外产气系统,39℃厌氧培养24 h。采用PCR-变性梯度胶凝电泳(DGGE)技术研究了发酵液菌群组成的差异。结果表明:1)SUC、STA、SBO组24 h发酵液累积产气量极显著高于对照组和RAF组(P0.01);SBO、SUC和STA组吲哚浓度分别比对照组降低了98.15%、97.72%和94.17%(P0.01),粪臭素浓度分别比对照组降低了79.04%、71.88%和70.28%(P0.05);SUC组乳酸浓度极显著高于其他各组(P0.01);对照组p H极显著高于除RAF组之外的其他各组(P0.01)。2)SBO组菌群均匀度显著低于对照组(P0.05);SUC、STA、RAF、SBO组菌群丰富度显著高于对照组(P0.05);SBO、STA、RAF组菌群相似性高于对照组;试验组有3条特异性条带,相似菌分别为延长布劳特氏菌(Blautia producta)、迪氏副拟杆菌(Parabacteroides distasonis)、路氏乳杆菌(Lactobacillus reuteri)。综上,在本试验条件下,添加1%的SUC、STA和SBO显著降低了肉仔鸡体外盲肠内容物培养液中L-色氨酸代谢生成吲哚和粪臭素的浓度,提高了菌群丰富度,并促进了特异性菌的增殖。降低粪臭素效果优劣依次为SBO、SUC、STA。     

日粮纤维调控猪体粪臭素水平的研究进展

粪臭素(3-甲基吲哚)是猪大肠内微生物降解L-色氨酸的产物,是引起公猪膻味的主要物质,易在脂肪组织中沉积,对猪肉风味有负面作用。本文综述了粪臭素的生物合成和代谢途径,以及日粮纤维对猪肠道、血浆、粪便和脂肪组织中粪臭素水平影响的研究进展;并从粪臭素的产生、吸收和代谢等方面,详细讨论了纤维物质影响猪体粪臭素沉积的可能机制。     

组成型雄烷受体与公猪膻味物质代谢关系的研究进展

饲养不去势公猪较去势公猪而言具有很多优势,但导致公猪膻味物质雄烯酮和粪臭素一直是限制其饲养的主要瓶颈。作者综述了组成型雄烷受体对膻味物质雄烯酮和粪臭素代谢关系的研究,为寻求一种较合适的方法减少公猪膻味提供参。     

Spatial variations in intestinal skatole production and microbial composition in broilers

Spatial variations in intestinal skatole production and microbial composition in broilers were evaluated. Fifteen 42‐day‐old broilers were slaughtered. Samples were taken from the broilers’ ileum, cecum, and rectum and analyzed for skatole levels. Denaturing gradient gel electrophoresis (DGGE) technique was used to analyze the microbial community from the intestinal digesta. The skatole levels could be arranged in decreasing order: cecum > rectum > ileum. Cecal lactate and acetate levels were higher than those of ileum and rectum (p < 0.01). Cecal microbial diversity and richness were higher than those of ileum (p < 0.05). One specific DGGE band was found in cecal sample and is closely related to Bacteroides uniformis. Cecum and rectum samples consisted of three coexistence bands, the related bacteria included Lactobacillus vaginalis and two members of Candidatus Arthromitus. The total bacterial population in cecum was higher than that in ileum and rectum (p < 0.05). Skatole levels were positively correlated with microbial Shannon‐Wiener index, richness, total bacteria (p < 0.01) and Lactobacilli and Bifidobacterium (p < 0.05) populations. These results suggest that the variations in fermentation patterns are more likely to explain differences in intestinal skatole level. Bacteroides uniformis may play a role in the production of skatole.     

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

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(CWBIB 1434)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.