植物乳杆菌BS22对黄曲霉毒素B1的吸附效果 及对肉鸡消化道菌群的影响

为探明植物乳杆菌BS22对黄曲霉毒素的解毒作用及对肉鸡肠道菌群的影响,设饲料添加不同量的BS22试验(不饲喂)和饲料添加BS22饲喂肉鸡试验。在含有50μg黄曲霉毒素B1(AFB1)的1 kg饲料中加入106、107、108 CFU/g植物乳杆菌BS22,于加入0、15、30 d检测AFB1含量。检测结果:加入BS22后,各组饲料中的AFB1含量均显著下降;同一添加量下,随饲料贮存时间的增加,AFB1含量均有所提高。肉鸡饲喂设3个组:I组(对照组),饲喂基础日粮;II组,饲喂基础日粮+50μg/kg黄曲霉毒素B1;III组,饲喂基础日粮+50μg/kg黄曲霉毒素B1+1.0×108 CFU/g植物乳杆菌BS22。试验期为28 d。采用RCR–DGGE比较4周龄肉鸡嗉囊、腺胃、十二指肠、空肠、回肠及盲肠内容物及黏膜细菌群落的结构。对消化道内容物而言,除回肠外,消化道其他部位的内容物的细菌多样性指数、均匀度和丰富度均是III组的高于II组和对照组;聚类图显示,除腺胃和盲肠外,其他部位均是II组与对照组聚在一起,而III组单独存在。对消化道黏膜而言,除空肠和盲肠外,其他部位的黏膜的细菌多样性指数和丰富度均是II组的均高于III组和对照组,且III组和对照组的细菌多样性指数相近或相同;除回肠和盲肠外,其他部位III组和对照组聚在一起,而试验II组单独存在。综合分析,黄曲霉毒素B1对各肠段内容物菌群的作用不明显,对黏膜菌群有较大影响,尤其是对十二指肠的黏膜菌群影响更明显,植物乳杆菌BS22能调节由于黄曲霉毒素B1引起的肠道黏膜菌群失衡。     

仔猪源罗伊氏乳酸杆菌生物学特性的研究

 研究了罗伊氏乳酸杆菌的生物学特性包括生长曲线、耐热存活率、耐酸存活率、贮藏存活率以及发酵参数。采用均匀设计的试验设计对影响乳酸杆菌发酵较大的6个因素如碳源、氮源和时间进行优化。试验结果表明,罗伊氏乳酸杆菌的浓度在开始时有所下降,经过2 h之后从1.0×10³数量级迅速上升,到第22 h接近1.0×10¹⁰数量级,到40 h细菌的数量级开始缓慢下降。75 ℃处理15 min之后存活率为31.3%,经过1个月贮藏存活率为85.5%,pH 2.0处理6 h的存活率为72%. 优化的发酵参数为:时间20 h;葡萄糖10 g/L;蔗糖60 g/L;胰蛋白胨30 g/L;酵母浸粉5 g/L;柠檬酸铵12 g/L. 试验表明了罗伊氏乳酸杆菌具有良好的抗逆性,生长繁殖迅速,可以用作益生菌生产菌种。     

藏猪源唾液乳酸杆菌的分离鉴定及生物学特性研究

【目的】对藏猪粪便中乳酸杆菌进行分离、鉴定及生物学特性研究。【方法】采集不同生长阶段藏猪（仔猪、保育猪、母猪）粪便,通过MRS选择性培养基培养、形态学观察、产酸性能测定、细菌生化鉴定及16S rDNA基因测序技术等方法从中分离、鉴定乳酸杆菌,检测藏猪源乳酸杆菌的生长曲线、耐酸耐胆盐特性、溶血活性、体外抑菌效果、益生基因,并对其进行安全性评估。【结果】从藏猪粪便中分离出48株乳酸杆菌,经16S rDNA基因测序和生物学特性分析,最终得到1株藏猪源唾液乳酸杆菌（Lactobacillus salivarius）,该菌株具有较好的产酸性能,产酸量可达（0.070±0.005） mol/L;对高质量浓度胆盐和低pH具有良好的耐受能力;该菌株中检测到肠菌素P2基因（Ent P2）,无溶血活性;体外抑菌结果显示,藏猪源唾液乳酸杆菌对金黄色葡萄球菌（CMCC 26003）和大肠杆菌（CMCC 44102）具有良好的抑菌效果;15 d小鼠灌胃试验结果显示,小鼠的体质量变化和脏器指数与生理盐水组无显著性差异,未观察到菌血症和细菌移位现象。【结论】筛选得到1株安全无毒副作用的藏猪源唾液乳酸杆菌,该菌具有较好的生物学特性,可作为发酵饲料和防治细菌性肠道疾病的潜在益生菌。     

干酪乳杆菌T1生物特性及保藏方法研究

干酪乳杆菌是一种具有广泛研究价值和良好研究前景的有益微生物。从青藏高原牧区的牦牛酸奶中分离出一株产细菌素的干酪乳杆菌T1(Lactobacillus casei T1)。通过探究菌株基本性质,研究能替代MRS的低成本培养基,筛选冻干保护剂等提高候选生产菌株T1的潜在生产价值。结果显示干酪乳杆菌T1的生长温度区间为20~40℃,耐受的p H值范围为p H2~pH 10。T1菌株对供试革兰阳性细菌和革兰阴性细菌均有较好的抑制作用,对部分供试真菌也有抑制作用,豆粕MRS培养基适应性好。以存活率为指标,进行冻干保护研究的结果表明,供试的4种保护剂的保护效果由大到小依次为乳糖、甘油、甘露醇和抗坏血酸。对候选生产菌株T1生物特性及菌种的保藏研究为该菌的开发打下坚实的理论基础。     

蜡样芽胞杆菌纤维素酶基因的克隆与其在乳酸菌中的表达

全球每年可产生的纤维素数量庞大。为了探索纤维素资源,更为科学合理的利用纤维素类饲料提供理论依据,根据蜡样芽胞杆菌纤维素酶基因序列设计1对特异性引物,扩增纤维素酶(Cell)基因,获得片段大小约为1 269bp的目的基因。该基因与Bacillus cereus B4264、Bacillus cereus G9842纤维素酶基因序列同源性分别达96%和95%。将Cell基因与乳酸乳球菌表达载体pNZ8149进行连接,电转化入乳酸菌NZ3900中,构建纤维素高效分解基因工程乳酸菌,经乳链菌肽(Nisin)诱导表达,SDS-PAGE分析得到分子量大小约为42kDa的目的片段。刚果红平板验证该表达蛋白具有纤维分解活性。FPA法测定表达产物酶活力为0.57U/mL。该基因的成功表达,对纤维素基因工程菌的相关研究具有指导意义。     

Effect of formic acid and a bacterial inoculant on the amino acid composition of grass silage and on animal performance

An Italian ryegrass and hybrid ryegrass sward was harvested on 11 May 1994. The mean dry‐matter (DM) content of the herbage was 197 g kg^–1 fresh matter (FM), and mean nitrogen and water‐soluble carbohydrate contents were 20 and 272 g kg^–1 DM respectively. Approximately 72% of total nitrogen (TN) was in the form of protein‐nitrogen. The herbage was treated with either no additive, formic acid (3·3 l t^–1) (Add‐F, BP) or inoculant (2·3 l t^–1) (Live‐system, Genus) and ensiled in 100 t silos. Changes in effluent composition with time showed that silage fermentation and protein breakdown were delayed by treatment with formic acid. Formic acid and inoculant treatments also inhibited amino acid catabolism during ensilage. All silages were well fermented at opening with pH values < 4·0 and ammonia‐N concentrations of ≤ 50 g kg^–1 TN after 120 d ensilage. Treatment had an effect on protein breakdown as measured by free amino acid concentration, with values of 21·5, 18·2 and 13·2 mol kg^–1 N at opening (191 d) for untreated, formic acid‐treated and inoculated silages respectively. Amino acid catabolism occurred to the greatest extent in untreated silages with significant decreases in glutamic acid, lysine and arginine, and increases in gamma amino butyric acid and ornithine. The silages were offered ad libitum without concentrate supplementation to thirty‐six Charolais beef steers for a period of 69 d (mean live weight 401 kg). Silage dry‐matter intakes and liveweight gains were significantly (P < 0·05) higher on the treated silages. Silage dry‐matter intakes were 7·42, 8·41 and 8·23 kg d^–1 (s.e.d. 0·27) with liveweight gains of 0·66, 0·94 and 0·89 kg d^–1 (s.e.d. 0·058) for untreated, formic acid‐treated and inoculated silage‐fed cattle respectively. In conclusion, additives increased the intake of silage and liveweight gain by the beef steers, and it is suggested that this may be caused in part by the amino acid balance in these silages.     

Effects of synbiotic containing Lactobacillus plantarum 7–40 and galactooligosaccharide on the growth performance of white shrimp,Litopenaeus vannamei

This study aimed to develop a synbiotic combination with probiotic, Lactobacillus plantarum 7–40 and one of three prebiotics, fructooligosaccharide (FOS), galactooligosaccharide (GOS) and mannan oligosaccharide (MOS). The best in vitro growth was observed when probiotic was cultured in the medium containing either FOS or GOS as the sole of carbon source. The analysis of enzyme activity revealed that GOS induced the highest activities of protease and β‐galactosidase of probiotic. Based on the findings, probiotic + GOS were selected as synbiotic to evaluate if it could promote the growth of white shrimp, Litopenaeus vannamei. For this, four diets, including a basal diet with no GOS or probiotic (control), 0.4% GOS (PRE), 10⁸ CFU probiotic kg^?1 (PRO) and 0.4% GOS in combination with 10⁸ CFU probiotic kg^?1 (SYN), were fed to shrimp for 60 days, and then the growth performance, intestinal microbiota (including total Vibrio counts, VBCs; and lactic acid bacteria, LAB) and digestive enzyme (including protease, leu‐aminopeptidase and β‐galactosidase) were evaluated. The weight gain (WG) of shrimp fed the PRO did not significantly differ from those of control (p > .05). Shrimp fed the SYN had significantly higher WG compared with the other treatments (p < .05). In addition, the SYN‐fed shrimp had significantly higher LAB and protease, leu‐aminopeptidase and β‐galactosidase activity (p < .05). The lowest presumptive Vibrio count (VBC) was also observed in intestines of SYN‐fed shrimp. Therefore, we suggested that Lac. plantarum 7–40+ GOS can be used as a synergistic synbiotic for shrimp culture.     

植物乳杆菌的生理功能和组学研究进展

植物乳杆菌是乳酸杆菌的一种,在自然界中广泛存在,尤其是各类发酵食品中。植物乳杆菌是人体肠道微生物的一员,由其生产的发酵食品的发酵作用与其对人体的益生作用使得植物乳杆菌逐渐成为基因组学、蛋白质组学、转录组学和代谢组学研究的模式微生物,阐明微生物环境适应性及发挥有益功能等的分子机制。本文综述了近年来植物乳杆菌的生理功能和组学研究进展,以期为研究者提供借鉴。     

乳清蛋白适度水解与其致敏性及益生菌生长的相关性

牛乳过敏问题是影响婴儿健康的重要问题,乳清蛋白中的主要过敏原为α-乳白蛋白和β-乳球蛋白,将乳清蛋白进行适度水解可以显著降低其致敏性。但是目前对于乳清蛋白的最适水解度还没有一个明确的评价标准。本研究从降低致敏性和促进益生菌（嗜酸乳杆菌08001）增殖角度建立乳清蛋白最适水解度的标准。结果表明：水解度为14.27%的乳清蛋白水解物符合本研究所要寻找的最适水解度乳清蛋白衡量标准,在此水解度下,乳清蛋白的致敏性降低较多,促嗜酸乳杆菌08001生长效果也较好。     

异常汉逊酵母和植物乳杆菌菌液浓度与光密度值的关系研究

[目的]探索一种快速测定异常汉逊酵母和植物乳杆菌菌液浓度的方法。[方法]采用分光光度法测定菌液的光密度值(OD值),平板菌落计数法测定异常汉逊酵母和植物乳杆菌的菌液浓度,并研究两者的相关关系。[结果]异常汉逊酵母在YEPD培养基中指数期的菌液浓度与光密度值呈现极显著的线性相关关系,菌液浓度(y)与光密度值(x)线性回归方程:y=2.246x-0.030(R2=0.998 6)。植物乳杆菌在MRS肉汤培养基中指数期菌液浓度的对数值与光密度值呈现极显著的线性相关关系,菌液浓度的对数值(y)与光密度值(x)线性回归方程:y=0.172 4x+8.432 1(R~2=0.996 6)。[结论]分光光度法可用于快速测定异常汉逊酵母和植物乳杆菌菌液浓度。