粪肠球菌生长曲线的测定及其对小鼠脑组织的影响     

王蒙蒙  张莉  李慧  汪骁轩  魏殿华  高静雯  高建鹏  张华雷  齐亚银 《中国畜牧兽医》2018,45(4):1041-1049

为测定致羔羊脑炎粪肠球菌（Enterococcus faecalis）的生长曲线，寻求一种快速而准确的方法测定不同生长时期粪肠球菌数量，并客观评价其毒性强弱及其对小鼠脑组织的影响，试验采用平板菌落计数法和OD-Monitor振荡比浊法（D_λ值法）测定粪肠球菌的生长曲线，探究该菌在合适时间段内的吸光值（D_{600 nm}）与平板菌落计数法测定的活菌数（CFU）的关系。用粪肠球菌感染小鼠，观察记录小鼠的死亡情况，最后采用Karber法计算粪肠球菌感染小鼠的半数致死量（LD₅₀）。用LD₅₀的剂量感染小鼠，及时采集死亡小鼠脑组织，未死亡的小鼠72 h后全部剖杀取脑组织，一部分做涂片染色，制作病理切片，观察病理变化；一部分进行培养，用于PCR方法进行细菌的回收鉴定。结果显示，用两种方法测定此株粪肠球菌的生长曲线基本一致，在2~8 h生长迅速，为对数生长期，8~14 h生长缓慢，为稳定期，14 h之后死亡数增加，进入衰亡期；对12 h粪肠球菌D_{600 nm}与CFU的关系进行探讨，成功建立回归方程：y=20.769x-1.3422，R²=0.997；其感染小鼠的LD₅₀为7.77×10¹¹个活菌。以此剂量感染小鼠，脑组织涂片染色和培养染色，均能看到革兰氏阳性球菌；PCR结果显示，均出现了大小为112 bp的条带。对脑组织进行病理学观察发现该菌可导致脑组织充血、出血、形成微血栓，脑膜充血。通过生长曲线和其D_{600 nm}与CFU关系的建立，可实时监测粪肠球菌数量，为后期更深入研究粪肠球菌穿越血脑屏障的机制奠定重要的理论基础。  相似文献   

粪肠球菌的生物学特性及其在养殖业中的应用   总被引：1，自引：0，他引：1  

安玉亭  胡浩  王智刚  周新秀 《安徽农业科学》2015,(27):115-117

粪肠球菌(Enterococcus faecalis)是乳酸菌的一类,属于链球菌科(Streptococcaceae)肠球菌属(Enterococcus),细胞呈球形,链状排列,无芽孢,兼性厌氧,为革兰氏阳性菌,是人类和动物肠道中重要菌群之一。粪肠球菌能够调节动物肠道菌群平衡,增强机体免疫力,促进肠道上皮细胞生长与修复,提高饲料转化率,促进动物体生长。该菌容易在动物肠道中定植并发挥作用,具有广阔的应用前景。  相似文献   

广东省养禽场肠球菌耐药性及毒力因子流行分布特征     

章婧  蔡萍  贺奕卓  陈伟涛  黄宇晨  蒋红霞 《畜牧兽医学报》2021,52(9):2650-2659

旨在调查和分析广东省养禽场肠球菌的亚型屎肠球菌和粪肠球菌耐药性及其毒力因子流行分布特征,为控制禽源肠球菌耐药性传播、保障公共卫生安全提供理论依据。作者于2018年从广东省4个养禽场采集肠道样品493份,进行屎肠球菌和粪肠球菌的分离鉴定;采用琼脂二倍稀释法测定肠球菌的最小抑菌浓度（MIC）;PCR方法检测肠球菌的耐药基因和毒力基因。结果显示：1）共分离到125株肠球菌,其中粪肠球菌84株（鸡源66株,鸭源18株）;屎肠球菌41株,均来自鸡肠道样本。2）菌株对四环素、多西环素、红霉素几乎全部耐药,对氟苯尼考和氯霉素的耐药率高达89.60%和74.40%。屎肠球菌耐药率普遍高于粪肠球菌,而粪肠球菌对环丙沙星和利奈唑胺的耐药率高于屎肠球菌;鸭源粪肠球菌对利奈唑胺的耐药率（94%）显著高于鸡源粪肠球菌（39.4%）,屎肠球菌对利奈唑胺均敏感。从鸡分离的1株粪肠球菌对万古霉素耐药。3）耐药基因在屎肠球菌中的检出率高于粪肠球菌,鸭源分离株检出率高于鸡源。耐药基因tetL、fexA、ermB最为流行,检出率均高于90%。其次是optrA基因,检出率为73.60%,poxtA和fexB的检出率均低于20%。在3株鸭源粪肠球菌中检测出cfr基因。4）已检测的毒力基因中efaA的携带率最高,为63.04%（58/92）,其他依次为gelE（54.35%,50/92）、ace（47.83%,44/92）、asa1（44.57%,41/92）。对环丙沙星及高浓度氨基糖苷类耐药的菌株及携带cfr基因的菌株,大多携带agg、asal、gelE或ace。本研究显示养殖场禽源肠球菌耐药严重,鸭源肠球菌对利奈唑胺耐药率高,耐药基因和毒力基因流行且多样,且检测出人医临床重要抗生素耐药基因,应加强对养禽场肠球菌耐药性监测。  相似文献   

Isolation and Identification of Enterococcus faecalis from Fur-bearing Animals     

CI Yang  SHI Xiaojie  SONG Peng  DING Qi  LI Wenli 《中国畜牧兽医》2007,47(9):2997-3005

In order to obtain Enterococcus faecalis from fur animals and evaluate its prebiotic properties,in this study,Enterococcus faecalis was isolated from the feces of healthy adult fur-bearing animals (mink,fox,raccoon dog),identified by morphological observation,biochemical test and 16S rRNA sequence analysis.The growth curve,acid production capacity and antibiotic sensitivity of the Enterococcus faecalis isolates were measured to evaluate their probiotic properties.Some strains were selected to determine their tolerance to temperature,artificial gastric juice and artificial bile salt.The results showed that five strains were Gram-positive,and their biochemical characteristics were basically consistent with the standard strains of Enterococcus faecalis,and they were identified as Enterococcus faecalis by 16S rRNA sequence analysis.The five strains all entered the logarithmic phase at 2 h after culture,and entered the stable phase at 8-10 h,and had weak acid production capacity.The resistance rate of the isolates to tetracycline and levofloxacin was 100%,followed by penicillin (80%),erythromycin (80%),gentamicin (80%) and chloramphenicol (40%).All the isolates were sensitive to ampicillin and vancomycin.Enterococcus faecalis from mink,fox and raccoon dog had strong tolerance to temperature below 60 ℃,artificial gastric juice with pH>3.0 and 0.3%-0.5% concentration of bile salt,but poor tolerance to temperature above 70 ℃,and artificial gastric juice with pH<3.0.In conclusion,five strains of Enterococcus faecalis from fur animals (mink,fox,raccoon dog) were obtained in this study.The isolated strains propagated rapidly,which were suitable for colonization and played a prebiotic role in fur animals' intestines,and had good prebiotic characteristics and stress resistance.They could be used as candidate strains for animal microbiological agents for further study.  相似文献   

Cloning,Expression and Bioinformatics Analysis of Enterococcus faecalis from Northeast Black Bear     

QIN Xiao-dong  CHU Xi-tong  LU Cheng  SUN Fu-liang  SUN Xing-zhong 《中国畜牧兽医》2017,44(8):2248-2254

In order to detect Enterococcus faecalis endocarditis antigen (EfaA) of bear that was used to disclose the infected Northeast Black bear immediately, a pair of PCR primers was synthesized by EfaA gene of Enterococcus faecalis that was recorded in GenBank (accession number:U03756.1) and the target fragment was 689 bp. The PCR product was cloned by pMD19-T vector, then was transferred to Escherichia coli DH5α competent cells and the positive clones were filtered. Recombinant plasmid (pMD19-T-EfaA) was extracted. Identification of PCR and digestion, sequencing, the structure prediction of proteins were operated. The results showed that EfaA gene was cloned successfully, and the gene homology with ATCC 29212 was 100.0%. pMD19-T-EfaA was constructed successfully, structure of proteins was predicted. This study provided theoretical basis for diagnostic methods of Enterococcus faecalis and laid basis for prevention and control of Northeast Black bear disease.  相似文献   

粪肠球菌替代抗生素对断奶仔猪生长性能、腹泻率、血液生化指标和免疫器官的影响   总被引：6，自引：0，他引：6  

史自涛  姚焰础  江山  肖融  刘作华  杨飞云  董国忠  郭致君 《动物营养学报》2015,(6)

本试验在断奶仔猪饲粮中添加不同水平的粪肠球菌,研究其替代抗生素对断奶仔猪生长性能、腹泻指数和腹泻率、血液生化指标以及免疫器官的影响。选取120头(28±2)日龄平均体重为(7.60±0.81)kg的"杜×长×大"断奶仔猪,随机分为5组,每组6个重复(公母各占1/2),每个重复4头,每个重复为1圈。对照组饲喂基础饲粮,抗生素组在基础饲粮中添加100mg/kg硫酸黏菌素和400 mg/kg杆菌肽锌,3个粪肠球菌组分别在基础饲粮中添加40、200和1 000 mg/kg粪肠球菌,试验期31 d。试验结果显示:1)在1~14 d,饲粮中添加抗生素和粪肠球菌对断奶仔猪的平均日采食量(ADFI)、平均日增重(ADG)以及料重比(F/G)无显著影响(P0.05)。在15~31 d,与对照组和抗生素组相比,200和1 000 mg/kg组可极显著提高ADFI和ADG(P0.01)。在1~31 d,与对照组相比,200和1 000 mg/kg组的ADFI均极显著提高(P0.01),而ADG分别显著和极显著增加(P0.05和P0.01);与抗生素组相比,1 000 mg/kg组可极显著提高ADFI(P0.01),而200和1 000 mg/kg组的ADG分别显著和极显著增加(P0.05和P0.01)。2)在1~14 d,与对照组相比,40 mg/kg组的腹泻指数和腹泻率显著降低(P0.05)。在15~31 d和1~31 d,与对照组和抗生素组相比,40和1 000 mg/kg组均可在一定程度上降低腹泻率(P0.05)。3)与对照组相比,粪肠球菌可以显著提高血液中总蛋白含量(P0.05),其中200和1 000 mg/kg组达到极显著水平(P0.01),且这2组分别可以显著和极显著提高球蛋白含量(P0.05和P0.01)。与抗生素组相比,200和1 000 mg/kg组可以显著提高总蛋白含量(P0.05)。与对照组相比,抗生素组和1 000 mg/kg组的白球比显著降低(P0.05),另外,抗生素组血液中谷丙转氨酶和谷草转氨酶活性显著降低(P0.05),200 mg/kg组的谷丙转氨酶活性显著降低(P0.05)。4)与对照组相比,1 000 mg/kg组的肝脏重量和脾脏指数有一定程度的提高(P0.05),与抗生素组相比,200和1 000 mg/kg组可显著提高肝脏重量(P0.05)。结果表明,饲粮中添加粪肠球菌可改善断奶仔猪的生长性能(饲喂15~31 d),降低腹泻率,提高仔猪免疫力,其中添加量为1 000 mg/kg时效果最好。  相似文献   

3种不同微生态制剂两两组合对池塘水质的影响     

杜兴  韩庆  吴有富  杨茂兴  罗玉双  邹万生 《农业科学与技术》2016,(9):1985-1989

研究了3种微生态制剂枯草芽孢杆菌(A)、粪链球菌(B)、光合细菌(C)两两配伍,以105 CFU/ml的浓度按9∶1、4∶1、1∶1、1∶4、1∶9混合制成15种制剂,对池塘水质中COD、氨态氮、亚硝酸盐氮、硫化物的影响.结果表明,以1:4混合制成的枯草芽孢杆菌和光合细菌的混合制剂对COD的处理效果最好(P＜0.05),以4∶1混合制成的枯草芽孢杆菌和粪链球菌的混合制剂对氨态氮的处理效果最好,对亚硝酸盐氮处理效果最好的是以4∶1配制的粪链球菌和光合细菌的混合制剂(P＜0.05),对硫化物的处理效果最好是以9∶1混合制成的粪链球菌和光合细菌的混合制剂(P＜0.05).  相似文献   

粪肠球菌F71的增殖培养基优化研究     

周英  高紫兰  袁凌南  周家春 《现代农业科技》2012,(3):35-36,39

为达到增殖菌体的目的,对粪肠球菌F71进行了培养基优化正交试验。结果表明,最佳培养基的组成为:胰蛋白胨10 g/L,酵母粉10 g/L,葡萄糖10 g/L,无水氯化钙0.04 g/L,七水合硫酸镁0.019 2 g/L,磷酸氢二钾0.04 g/L,磷酸二氢钾0.04 g/L,磷酸氢钠0.4 g/L,氯化钠0.08 g/L,土豆汁50 g/L,碳酸钙1 g/L,乙酸钠10 g/L,碳酸铵2 g/L,乙酸铵1 g/L,半胱氨酸盐酸盐0.5 g/L,pH值为9,最适培养温度为35℃。用此培养基进行增殖,F71的最高活菌数达到2.78×108 cfu/mL,比基础培养基提高了54.6%。  相似文献   

Enterococcus faecalis of human and poultry origin share virulence genes supporting the zoonotic potential of E. faecalis     

Olsen RH  Schønheyder HC  Christensen H  Bisgaard M 《Zoonoses and public health》2012,59(4):256-263

Enterococcus faecalis is a major cause of nosocomial infections in humans and has been linked to severe extra‐intestinal infections in poultry. A zoonotic potential has been suggested and the aim of the present study was to investigate similarities in virulence gene profiles of E. faecalis originating from infections in humans and poultry respectively. A total of 106 isolates of E. faecalis [26 human clinical isolates, 60 poultry clinical isolates (including two small‐colony variants (SCVs) and 20 poultry cloacal isolates] were investigated for presence of seven virulence‐associated genes: ace, asa1, cylA, efaA, EF0591, esp and gelE. For each gene, the PCR‐amplification product was sequenced from one isolate in each group to explore intragenic variations between genes of human and poultry origin. Haemolytic and protease activities were assessed and isolates were assigned a sequence type (ST). Three of the seven genes investigated (ace, efaA and gelE) were present in all isolates. The asa1 was detected in 63/80 and 13/26 isolates of poultry and human origin respectively. For cylA, the numbers were 46/80 and 14/26 respectively. Among poultry isolates, esp and EF0591 were the least frequently observed genes (1/80 and 20/80 respectively); the prevalences among human isolates were 1/26 and 18/26 respectively. A high degree of similarity between genes in human and poultry isolates were confirmed by sequencing of amplification products. None of the cylA‐positive isolates demonstrated haemolytic activity, while the phenotypic expression of gelatinase varied. The ST16 was the only ST shared by human and poultry isolates. The SCV isolates did not show a unique virulence profile or phylogeny. In conclusion, regardless of the distinct phylogenetic background of most E. faecalis isolates of human and poultry origin, we found major similarities in virulence gene profile and gene sequences in isolates from the two sources, supporting the zoonotic risk associated with this organism.  相似文献   

粪产碱菌耐铵工程菌与水稻联合共生固氮作用   总被引：8，自引：1，他引：7  

陈明  张维  林敏 《核农学报》1999,13(6):373-376

粪产碱菌（ Ａａｅｃａｌｉｓｆａｅｃａｌｉｓ） 在ＬＷ 培养基下能产生植物激素ＩＡＡ。盆栽结果表明,接种Ａ．ｆａｅｃａｌｉｓ 显著提高了水稻的生长,接种Ａ１５０１ 和Ａ１５１３ 的水稻稻谷产量分别比未接种提高了８５％ 和１０３ ％ ,用１５ Ｎ 同位素稀释法估测水稻地上部总氮中来源联合固氮的百分率达９００ ％ 和１１５ ％ ,其结果与水稻地上部（ 茎叶及稻谷） 总氮量的增加幅度（８６ ％ 和１１６ ％ ）基本一致。研究还表明Ａ１５１３ 耐铵工程菌的产ＩＡＡ作用以及对水稻的生长和产量、联合固氮作用均优于原始菌株Ａ１５０１ 。  相似文献