Effect of laboratory‐isolated Lactobacillus plantarum LGFCP4 from gastrointestinal tract of guinea fowl on growth performance,carcass traits,intestinal histomorphometry and gastrointestinal microflora population in broiler chicken

The study aimed to investigate the effect of feed supplements, viz Lactobacillus plantarum LGFCP4 (laboratory isolate from GIT of Guinea fowl), Lactobacillus acidophilus (NCDC, Karnal) and in‐feed antibiotic bacitracin methylene disalicylate (BMD) on growth performance, FCR, carcass traits and immune organs weight, intestinal histomorphometry and gastrointestinal microflora population in broiler chickens. In a completely randomized design, CARIBRO‐Dhanraja broiler chicks (n = 160) were used with four treatment groups. During the entire experimental duration of 35 days, treatment groups were provided with different dietary treatments (T1 – basal diet (negative control), T2 – antibiotic growth promoter BMD 20 g/100 kg feed (positive control), T3 – 1 × 10⁸ cfu of L. acidophilus/gm‐fermented feed +MOS 1 g/kg feed and T4 – 1 × 10⁸ cfu of laboratory‐isolated L. plantarum LGFCP4/gm‐fermented feed+ MOS 1 g/kg feed. After 35 days of experimental period, no significant results have been observed in different growth performance traits among treatment groups. Cut‐up parts and edible organs' weight remained unaffected by dietary supplementation, whereas weight of immune organs were significantly higher (p < 0.05) in L. plantarum LGFCP4‐supplemented group. At the end of feeding trial, significantly (p < 0.05) lower E. coli count was observed in crop of T4 birds, while in ileum, T2 and T3 showed lower count. In caeca, T2 group showed lowest E. coli count. Salmonella count in crop and ileum was significantly (p < 0.05) low in T3 and T4, while in caeca, T2 group showed lowest count. In terms of histomorphometry, duodenal villous height (VH), crypt depth (CD) and VH:CD ratio were higher for T3 and T4 and lowest values were obtained for T2 group. The results of the study showed that L. plantarum LGFCP4 isolated from GIT of guinea fowl can effectively replace in‐feed antibiotic growth promoters in broiler diets by altering intestinal villi morphology and improving the gut health by reducing the pathogenic microbial load.     

Influence of pH, Heat and Enzymatic Treatments on the Activity of Antibacterial Substance in MRS and Milk Media Produced by Lactobacillus fermentum F6

This study aimed to investigate the influence of pH,heat and enzymatic treatments on the activity of antibacterial substance in MRS,skim soy milk and bovine milk media fermented by a potential probiotic Lactobacillus fermentum F6.The antibacterial activity of the culture supernatant of L.fermentum F6 was tested against a wide range of Gram-positive and Gram-negative pathogenic bacteria including Staphylococcus aureus,Escherichia coli,Listeria monocytogenes,Salmonella typhimurium,and Shigella flexneri.Different antibacterial activities were detected in MRS and milk but not in soy milk.We presumed that three kinds of probable components including organic acids,heat-stable and heat-labile proteinaceous compounds were involved in antibacterial activity of fermented MRS and milk.The influence of acids on antibacterial activity was pH-dependent and this effect multiplied with thermal treatments seemed complex.Data analyses showed various significant differences of antibacterial activity among five pathogens were observed in pH and heat treatments.The untreated fermented milk showed higher inhibitory activity to Gram-positive pathogenic bacteria than Gram-negative bacteria （P 〈 0.05）,indicating antibacterial substances of fermented milk are greatly different from fermented MRS.An accurate stimulated gastrointestinal transit showed antibacterial substances would have no influence on intestinal flora.Acidic range of antibacterial substances from pH 2.0 to 6.0 can be potentially used as food biopreservatives and alternative therapeutics.     

抗猪链球菌戊糖乳杆菌素的纯化及特性研究

对分离自健康未断奶仔猪粪便的抗猪链球菌的戊糖乳杆菌LPL1—5所产细菌素进行提取、纯化,以及理化和生物学特性研究。戊糖乳杆菌LPL15所产细菌素经饱和度为80％的硫酸铵盐析、葡聚糖凝胶树脂G-10层析和SPSepharoseFastFlow阳离子树脂交换层析可得到纯化的细菌素,其比活力为1732．16AU／mg,纯度较发酵液提高了26．37倍;Tricine—SDS-PAGE凝胶电泳及活性检测试验结果显示,3313～5856u范围内存在1条活性蛋白带;纯化后细菌素特性分析发现,该细菌素具有良好的热稳定性、酸耐受性及溶解性,并基本确定其等电点为8．7;胃蛋白酶、胰蛋白酶、中性蛋白酶和蛋白酶K处理可使细菌素完全失活;该细菌素抑菌谱较广,不仅对猪链球菌（Sreptococcussuis）、单核细胞增生李斯特氏菌（Listeriamonocytogenes）、金黄色葡萄球菌（sfnphylococcus aereu）等革兰氏阳性菌有明显抑制作用,同时对大肠杆菌（Escherichiacoli）及假单胞菌（Pseudomonasspp．）等革兰氏阴性菌也表现出抑菌活性,具有作为食品防腐剂良好的应用前景。     

黑豆酸奶发酵微生物鉴定与发酵特性研究

利用形态学观察、生理生化方法对黑豆酸奶发酵菌株FJAT-7926和FJAT-7928进行分类鉴定.2株菌的菌体均为杆状,无芽孢,革兰氏阳性,过氧化氢酶阴性.菌株FJAT-7926的生理生化特征与植物乳杆菌Lactobacillus plantarum一致,菌株FJAT-7928的生理生化特征与干酪乳杆菌Lactobac...     

乳杆菌乳糖酶的基因异源表达及酶学性质分析

采用简并引物PCR和TAIL-PCR方法,从乳杆菌Lactobacillus sp. B164中克隆得到一个乳糖酶基因bg42-164。基因全长2 031 bp,编码676个氨基酸和一个终止密码子,预测分子量76 kDa,无信号肽序列。将bg42-164连接pET-30a(+)载体并转入大肠杆菌BL21(DE3),经IPTG诱导表达后可检测到乳糖酶活力。SDS-PAGE分析乳糖酶BG42-164的蛋白分子量为76 kDa,使用组氨酸标签亲和层析方法进行蛋白纯化,并对纯化的乳糖酶BG42-164进行了酶学性质分析。该酶最适反应温度为50℃,经50℃处理30 min后,剩余酶活力保留80%以上,pH 6.0时该酶的水解活力最高。牛奶水解试验表明,乳糖酶BG42-164对乳糖的水解效果良好,5 mL牛奶中添加250 U酶蛋白,在50℃条件下2 h乳糖水解率为100%。该酶温度范围宽,乳糖水解效果好,为其在低乳糖奶生产中应用奠定了理论基础。     

单一菌种与复合菌种益生菌制剂对蛋鸡生产效果研究

为了探讨单一菌种与复合菌种益生菌制剂对蛋鸡生产效果的影响。本研究以鸡源枯草芽孢杆菌、乳酸杆菌为益生菌株,利用发酵培养制备乳酸杆菌益生菌制剂、枯草芽孢杆菌益生菌制剂和乳酸杆菌—枯草芽孢杆菌复合益生菌制剂;在140日龄罗曼褐商品蛋鸡基础日粮上,按照质量比0.1%比例分别添加不同益生菌制剂,进行蛋鸡生产效果研究。结果表明,益生菌制剂可以显著提高平均蛋重、日产蛋量、产蛋率,降低料蛋比,提高蛋品质。与乳酸杆菌益生菌制剂、枯草芽孢杆菌益生菌制剂相比,乳酸杆菌—枯草芽孢杆菌复合益生菌制剂可以使平均蛋重分别显著提高1.87%、1.91%;平均日产蛋量分别提高了3.34%、2.77%;产蛋率分别提高1.56%、1.00%;料蛋比分别降低1.99%、1.49%; A级蛋比例分别提高9.16%、5.91% ;蛋黄比率分别提高7.71%、4.14%;每只鸡毛利润分别提高10.84%、7.73%;而在降低不合格蛋率、死淘率、平均日耗料量、蛋形指数和提高蛋比重、蛋壳厚度等方面,各益生菌制剂组差异不显著。因此,乳酸杆菌—枯草芽孢杆菌复合益生菌制剂对蛋鸡生产效果优于乳酸杆菌益生菌制剂、枯草芽孢杆菌益生菌制剂。     

以嗜酸乳杆菌发酵虾头虾壳回收蛋白质和甲壳素的研究

利用嗜酸乳杆菌发酵虾头、虾壳,结合虾头、虾壳中内源蛋白酶的自溶作用,回收蛋白质和甲壳素。研究了影响发酵的因素并优化了发酵条件,分析了用稀盐酸脱除粗甲壳素(即发酵残渣)中的矿物质,制备食用级甲壳素的条件。研究结果表明,添加15%葡萄糖,固液比1∶2.5,接种量15%,初始pH值6.5,于40℃发酵48 h,得到的虾头、虾壳发酵液pH值低至3.79,蛋白质水解度达到21.9%;蛋白质回收率达到95.2%,发酵残渣(粗甲壳素)中仅残留2.71%矿物质,达到工业级甲壳素的纯度要求;以稀盐酸浸泡处理粗甲壳素,当浸泡温度为50℃,浸泡时间为1.5 h,粗甲壳素脱矿物质的效果最好,甲壳素中仅残留0.2%矿物质,达到食用级甲壳素的纯度要求。     

质粒pUC19-CM-D的构建及应用

[目的]用克隆载体pUC19构建乳杆菌自杀质粒及乳杆菌基因缺失工程菌[方法]在质粒pUC19的多克隆位点插入氯霉素抗性基因构建pUC19-CM载体;在pUC19-CM载体氯霉素抗性基因的两侧均添加1个用于同源重组的同源臂,再构建成自杀质粒pUC19-CM-D。将自杀质粒pUC19-CM-D转化乳杆菌进行抗性筛选即可得到目标基因被抗性基因替换的突变株。[结论]pUC19-CM-D质粒的构建及应用为乳杆菌基因缺失工程菌的构建提供了一个快速有效的手段,也为乳杆菌基因功能研究奠定了基础。     

动物乳杆菌发酵液的体外抑菌试验

采用牛津杯法测定了动物乳杆菌发酵液及除酸、除过氧化氢后的发酵液对各种指示菌的抑制作用。研究结果显示：动物乳杆菌原始发酵液的抑菌效果最好;除酸后的发酵液的抑菌效果有一定变化,抑菌谱减小;而除过氧化氢后的发酵液的抑菌谱与除酸后的没有变化。     

嗜酸乳杆菌的亚油酸异构酶基因克隆及其pMG36e表达载体的构建

以嗜酸乳杆菌基因组为模板,对亚油酸异构酶基因进行PCR,PCR产物克隆到pMD-19T质粒中,经菌落PCR、酶切分析和DNA测序鉴定克隆成功后,亚克隆入乳酸菌表达质粒pMG36e,构建乳酸菌表达载体pMG36e-LAI并转化到大肠杆菌DH5α中,经菌落PCR和酶切分析初步证明表达载体pMG36e-LAI构建成功。该研究为利用工程菌工业化生产高产量、高活性亚油酸异构酶制剂来生产CLA奠定基础。