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1.
【目的】 通过对瘤胃液中分离所得的菌株进行研究,为益生菌制剂的制备提供基础数据。【方法】 采集10头健康荷斯坦奶牛的瘤胃液,通过涂布、特性培养、纯化等步骤获得单一菌株,提取细菌DNA,进行16S rDNA的PCR扩增。通过16S rDNA基因测序鉴定后进行序列比对并构建系统发育树,确定菌株种类。对不同种类的细菌进行0~48 h的培养测定其生长特性,并进行耐酸碱和耐胆盐试验。【结果】 通过涂布分离菌株得到20株分离菌,经PCR扩增后测序发现20株菌属于7种不同的菌,大致确定L7为解淀粉芽孢杆菌、K7为枯草芽孢杆菌、S7为嗜麦芽窄食单胞菌、C2为地衣芽孢杆菌、M7为马链球菌、F7为弗氏酸柠檬杆菌、T7为吉氏库特菌。通过生长曲线可以看出,7株菌分别在24~36 h达到最快生长期。M7和K7对酸有较强的耐受性,而S7、C2和F7对酸性较为敏感,C2对碱的耐受性最强,而T7对胆盐的耐受性最强,S7对胆盐最不耐受。【结论】 本研究从瘤胃中分离得到的7个菌株对酸碱和胆盐具有不同程度的耐受性,以及最适生长周期。  相似文献   

2.
The degree of piliation of 29 haemolytic and 4 non-haemolytic Australian strains of Moraxella bovis representing 7 different pilus antigen groups was determined. The infectivity and virulence for the eye was measured in steroid-treated mice and in cattle. Non-piliated strains failed to infect the murine eye. Most moderately or heavily piliated strains reproducibly produced the highest infectivity and virulence scores in mice when compared with lightly or very lightly piliated strains (p less than 0.05). Non-haemolytic, piliated strains were infective and in one instance virulent for mice. Almost similar levels of infectivity and virulence were observed for 7 representative haemolytic strains tested in both cattle and mice. The relative molecular weight of pilin sub-units was compared using sodium dodecyl-sulphate polyacrylamide gel electrophoresis. Three classes of pili, alpha, beta and gamma of ascending sub-unit size were identified among the 7 pilus antigen serogroups. Pilin sub-unit size bore no relationship to the degree of piliation but most strains that were highly virulent in mice and cattle expressed alpha and gamma sub-units. Some strains appeared capable of switching from alpha to beta or form beta to gamma sub-unit production.  相似文献   

3.
Randomly amplified polymorphic DNA (RAPD) analysis was used to differentiate 7 strains of Mycoplasma gallisepticum. Six commercially available primers or primer combinations were screened for their ability to differentiate vaccine and type strains. Although major and minor bands were produced with each primer, many of the primers were unsuitable for strain differentiation. The use of primer 6 and combined primers 3 and 4 resulted in complementary RAPD banding patterns for each M. gallisepticum strain. Eleven different isolates representing 7 different strains were segregated into 7 different patterns, corresponding to the 7 strains.  相似文献   

4.
试验旨在对分离自天津地区发病猪场的7株猪链球菌3型菌株(Streptococcus suis type 3,SS 3)进行致病性和耐药特性研究。应用剂量为1.0×107、1.0×108和1.0×109 CFU/只的细菌对小鼠进行致病力研究,用PCR方法检测7株SS 3型菌株的毒力基因mrp、ef、sly、gdh、gapdh、fbpsorf2,并对这7株SS 3型菌株进行药物敏感试验。致病力试验结果显示,接种剂量为1.0×109和1.0×108 CFU/只时分别有6和3株SS 3型菌株可使小鼠100.0%(5/5)发病死亡,接种剂量1.0×107 CFU时仍有1株SS 3型菌株可使80.0%(4/5)小鼠发病死亡,这7株SS 3型菌株对小鼠的致病力依次为R15056 > R15042=S15030 > Y12024=Y09011 > Y13125 > Y13164。毒力基因检测结果表明,共有3种毒力基因型,R12024、Y13164、R15042和S15030株为gdh、gapdh、fbpsorf2毒力基因阳性,Y13125和R15056株为sly、gdh、gapdh、fbpsorf2毒力基因阳性,Y09011株为gdh、fbpsorf2毒力基因阳性。药物敏感性试验结果显示,7株SS 3型对头孢喹肟相对最敏感,其次为阿莫西林、环丙沙星和磺胺间甲氧嘧啶钠,但对强力霉素100.0%耐药,且所有菌株均呈现3重以上的耐药性。本研究为进一步开展天津地区SS 3型流行特点及致病机理研究奠定了基础,可为天津地区SS 3型菌株的综合防控提供理论指导。  相似文献   

5.
本试验对7株A型产气荚膜梭菌菌株在不同培养基、不同培养温度和不同培养时间的产毒力进行了试验。结果表明,培养基的种类和培养时间对菌株的产毒素能力影响较大,而培养温度则影响较小。所试菌株中C57-1菌株的产毒素能力最强,是制备疫苗的最佳菌株。  相似文献   

6.
Streptococcus suis is an important agent of swine and human meningitis. Sequence type (ST) 7 emerged in China and was responsible for the human epidemic caused by S. suis in 2005. The virulence of S. suis ST7 is greater than the wild type pathogenic S. suis, ST1; however, the mechanisms for this increased pathogenicity are unknown. The aim of this study was to determine the role of different toll-like receptors (TLRs) involved in regulating the host response to the S. suis infection and to speculate on differing mechanisms used by ST7 strains to induce disease. Here we compared two ST7 strains isolated in the 2005 Sichuan outbreak to two ST1 strains. Our data show TLR2, 6 and 9 are involved in the recognition of heat-killed S. suis independent of the ST type. We found the TLR-dependent cytokine production differed between the two types of strains using whole cell lysate proteins. TLR6 played a greater role in cytokine production induced by the whole cell lysate proteins from the ST7 strain than in that induced by the ST1 strain lysates. The data suggest that mechanisms of inflammation induced by S. suis strains differ where this will be useful in designing efficient strategies in combating streptococcal toxic shock-like syndrome caused by the S. suis ST7 strains.  相似文献   

7.
A PCR assay for simultaneous species identification and separation of Actinobacillus pleuropneumoniae serovars 1, 7 and 12 was developed. Primers specific for genes involved in biosynthesis of the capsular polysaccharides (cps genes) of serovars 1, 7, and 12 were combined with a species-specific PCR test based on the omlA gene. The PCR test was evaluated with the serovar reference strains of A. pleuropneumoniae as well as 183 Danish field isolates. For all typable strains, a complete correspondence was found between results obtained with the multiplex PCR test and results from the traditional serotyping methods. Among eight serologically cross-reacting strains designated K1:O7, seven isolates produced amplicons of similar sizes as serovar 1 and one isolate produced amplicons of similar sizes as serovar 7. The species specificity of the assay was evaluated using a collection of 126 strains representing 25 different species within the family Pasteurellaceae including 45 field strains of the phylogenetically affiliated species Actinobacillus lignieresii. All these isolates tested negative for the cps genes by the multiplex PCR test except for 6 isolates of A. lignieresii. Five of these isolates produced an amplicon identical to the cps gene of serovar 7, whereas one isolate produced an amplicon identical to the cps gene of serovar 1. In addition, four isolates of Actinobacillus genomospecies 1 tested positive for the omlA gene but negative for the cps genes. The test represents a convenient and specific method for serotyping A. pleuropneumoniae in diagnostic laboratories.  相似文献   

8.
为了解内江市猪源大肠埃希菌对抗菌药与消毒剂的交叉耐药情况及交叉耐药机制。本试验以大肠埃希菌标准菌株(ATCC 25922)为质控菌,检测从内江市分离的66株猪源大肠埃希菌的耐药性,采用微量肉汤稀释法测定常用抗菌药恩诺沙星、土霉素和消毒剂苯扎溴铵、戊二醛对受试菌株的最小抑菌浓度(MIC),进行耐药性检测,并筛选出对抗菌药和消毒剂均耐药的交叉耐药菌;PCR扩增标准菌株和交叉耐药菌的qacE△1、acrA、acrB和tolC基因,采用琼脂糖凝胶电泳检测基因携带和扩增情况,并对acrA、acrB和tolC基因进行测序,以标准菌株的acrA、acrB和tolC基因为参照,比对交叉耐药菌相应基因突变情况。结果显示,从内江市分离的66株猪源大肠埃希菌对恩诺沙星和土霉素完全耐药,对苯扎溴铵耐药的有3株,而所有菌株均对戊二醛敏感,即对抗菌药和消毒剂均耐药的交叉耐药菌有3株,编号分别为5、6、7号。在检测的3株交叉耐药菌中,仅6号菌检出qacE△1基因;基因序列比对结果显示,与标准菌株相比,交叉耐药菌的acrA、acrB和tolC基因碱基和氨基酸均有突变发生,碱基突变率最高的是7号菌的tolC基因(2.10%),突变率最低的是7号菌的acrA基因(0.97%);氨基酸突变率最高的为7号菌的acrB基因(2.39%),突变率最低的是tolC基因,且3株菌突变率相同(均为0.22%);3个基因有部分碱基或氨基酸有一致突变。所检测的内江市猪源大肠埃希菌对常用抗菌药恩诺沙星和土霉素耐药性严重,已有对抗菌药和消毒剂交叉耐药的菌株,且其交叉耐药的机制与外排泵AcrAB-TolC有相关性。  相似文献   

9.
【目的】了解河南地区副猪嗜血杆菌(Haemophilus parasuis,Hps)血清7型流行菌株的毒力及耐药情况。【方法】以Hps血清7型参考菌株为对照,对6株Hps血清7型临床分离菌株进行16S rRNA基因扩增、序列分析、毒力基因检测、致病性试验和药敏试验研究。【结果】16S rRNA基因测序及相似性比对结果显示,菌株1436与参考菌株0007相似性为100%,菌株1565、1624与参考菌株0007相似性均为98.6%。系统发育树分析表明,菌株1436与参考菌株0007亲缘关系最近;菌株1624与参考菌株0007亲缘关系最远。6株临床菌株与参考菌株表现为2种毒力基因型,菌株均携带有vta1、vta2、vta3、wzananHcdtAcdtBcdtCespP2毒力基因,其中5株临床菌株还携带ompP2毒力基因。豚鼠致病性试验结果显示,临床菌株毒力较参考菌株表现出不同程度的增强,参考菌株不能感染豚鼠发病,未表现任何临床症状;临床菌株可感染豚鼠发病,表现出一定的临床症状,发病率为20%~40%,死亡率为0。参考菌株和临床菌株均对头孢他啶和头孢噻呋敏感,对强力霉素、氟苯尼考中介,对新霉素、卡那霉素、阿米卡星、红霉素、替米考星等耐药,且均表现出多重耐药现象。【结论】Hps血清7型临床菌株有一定的致病性,但毒力较弱,对头孢类药物敏感,有明显的多重耐药现象,临床上应注意对该血清型的防控。本研究为Hps血清7型流行病学、致病机制研究奠定了基础,为Hps血清7型的临床防控提供了参考依据。  相似文献   

10.
从已分离的多个狗牙根(Cynodon dactylon)内生固氮菌株中筛选到两株固氮酶活性较高的菌株7D和BM13,为明确固氮菌株对环境因子的适应性,研究了在不同的生长环境条件下菌株的生长量。采用Ashby无氮培养基,测定了温度、pH、渗透压以及不同氮源和碳源等环境条件对固氮菌生长的影响。结果表明,在10~40℃的温度范围内、pH值3.92~9.64的酸碱环境和30g·L~(-1) NaCl的高渗透压下,固氮菌7D和BM13均能正常生长;菌株能耐受0~10mmol·L~(-1)的NH_4~+和NO_3~-,而且能利用蔗糖、甘油、麦芽糖、甘露醇和葡萄糖等多种碳源生长。说明狗牙根内生固氮菌株7D和BM13对环境因子有很强的抗逆性和适应性,但两菌株之间存在一定差异,可望进一步研发成为优良的固氮微生物肥料生产菌种。  相似文献   

11.
为从岷山红三叶根际土壤中筛选出大量高效溶磷菌株,本研究采用Pikovaskaia’s、PKOC1和PKOC2三种溶磷培养基进行溶磷菌株的初步筛选和优良溶磷菌株16S rRNA基因序列鉴定,并对分离筛选效果较好的PKOC2培养基进行响应面优化。结果表明:采用Pikovaskaia’s、PKOC1和PKOC2三种溶磷培养基,从红三叶根际分离出26株溶磷菌株;经16S rRNA基因序列对7株优良溶磷菌株进行初步比对鉴定,初步鉴定结果为:菌株MHS4为蜡样芽胞杆菌(Bacillus cereus);菌株MHS7和MHS19为枯草芽胞杆菌(Bacillus subtilis);菌株MHS27为苏云金杆菌(Bacillus thuringiensis);菌株MHS30为荧光假单胞菌(Pseudomonas fluorescens);菌株MHS31为盖氏假单胞菌(Pseudomonas gessardii);菌株MHS49为产酸克雷伯菌(Klebsiella oxytoca)。PKOC2溶磷培养基优化配方为:葡萄糖18.19 g·L-1,Ca3(PO4)2 5 g·L-1,MgCl2·6H2O 3.21 g·L-1,MgSO4·7H2O 0.25 g·L-1,KCl 0.2 g·L-1,(NH4)2SO4 0.08 g·L-1。红三叶根际溶磷菌资源丰富,优化后的溶磷培养基为高效溶磷菌资源筛选提供资源支持。  相似文献   

12.
The genetic relatedness of 7 Korean type O field strains of foot-and-mouth disease virus (FMDV) in clinical specimens collected from 5 different geographic locations in 2000 was investigated. The sequence of 162 nucleotides (nt 478-639) at the 3' end of the 1D (VP1) genes was determined from amplified cDNA fragments, and subjected to the analysis for the sequence identity/divergence and phylogenetic relationship. The overall nucleotide sequence divergence among the 7 field strains was 0 to 3.8%, suggesting that they are closely related to each other. Phylogenetic analysis with the known Middle East-South Asia (ME-SA) topotype strains showed that the 7 Korean field strains formed two distinct clusters within the same lineage of the ME-SA topotype strains. Cluster 1 consisted of the strains of the primary foci of infection (Paju and Hongseong), and closely related to the strains prevailed in the Far East. Cluster 2 comprised those of subsequently affected regions (Boryeong, Yongin, and Chungju), and was further diverged from the Cluster 1. The result of phylogenetic analysis indicated that the Korean strains may have evolved from a common ancestor of the Pan Asia strains, and that at least 2 phylogenetically clustered variants within the same lineage were prevalent during the epidemic. The potential origin and sources of the virus introduction to Korea were discussed.  相似文献   

13.
Escherichia (E.) coli serotype O157:H7 is a globally distributed human enteropathogen and is comprised of microorganisms with closely related genotypes. The main reservoir for this group is bovine bowels, and infection mainly occurs after ingestion of contaminated water and food. Virulence genetic markers of 28 O157:H7 strains were investigated and multilocus enzyme electrophoresis (MLEE) was used to evaluate the clonal structure. O157:H7 strains from several countries were isolated from food, human and bovine feces. According to MLEE, O157:H7 strains clustered into two main clonal groups designated A and B. Subcluster A1 included 82% of the O157:H7 strains exhibiting identical MLEE pattern. Most enterohemorrhagic E. coli (EHEC) O157:H7 strains from Brazil and Argentina were in the same MLEE subgroup. Bovine and food strains carried virulence genes associated with EHEC pathogenicity in humans.  相似文献   

14.
冯世文  李军  曾芸  杨威  陈泽祥  潘艳  彭昊 《中国畜牧兽医》2015,42(12):3315-3322
为初步研究猪源大肠杆菌O157:H7 (E.coli O157:H7)对氟苯尼考耐药性的产生和消除机制,本研究采用亚抑菌浓度体外耐药诱导的方法将两株猪源大肠杆菌O157:H7诱导成氟苯尼考高度耐药菌株,采用无氟苯尼考压力下连续传代培养的方法将获得的氟苯尼考耐药菌株的氟苯尼考耐药性消除,检测耐药诱导菌和耐药消除菌对抗菌药物的敏感性,并检测菌株质粒携带的耐药基因。结果显示,经氟苯尼考耐药诱导,猪源大肠杆菌O157:H7对氟苯尼考、阿莫西林、头孢唑啉、头孢拉定和头孢噻吩由敏感变为耐药,对头孢噻肟的敏感性由敏感变为中介,对氧氟沙星、环丙沙星和阿奇霉素由中介变为耐药;而经耐药消除后,菌株恢复对上述药物的敏感性;在菌株的质粒中检测到氟苯尼考耐药基因、喹诺酮类耐药基因和β-内酰胺酶基因,与耐药表型相符。结果表明,在氟苯尼考压力的长期存在下,猪源大肠杆菌O157:H7对氟苯尼考产生耐药,且对青霉素类、头孢类和喹诺酮类药物产生交叉耐药,在去除氟苯尼考压力下连续培养,可消除菌株的部分耐药性。  相似文献   

15.
通过分离青海高寒草地针茅根际土壤细菌,以番茄早疫病菌、马铃薯枯萎病菌、马铃薯坏疽病菌和马铃薯炭疽病菌为指示菌,采用平板对峙法对其拮抗功能进行评价,并利用16SrDNA序列分析法进行鉴定。结果表明,从针茅根际土壤中共分离获得23株土壤细菌,17株可转代培养,根际土壤细菌数量介于6.4×10^6~36×10^6cfu·g^-1,其中4T1、5T1、10T1和10T3至少对1种病原真菌有抑菌作用,占细菌总数的17.39%,抑菌率介于46.68%~75.71%;菌株10T1的抑菌效果最好,对番茄早疫病菌、马铃薯枯萎病菌、马铃薯坏疽病菌和马铃薯炭疽病菌的抑菌率分别为67.14%、68.57%、61.43%和75.71%;23株土壤细菌均呈革兰氏阳性,杆状,大小介于3.79μm×0.85μm^10.50μm×2.37μm。结合培养形状、形态特征和16SrDNA序列分析将17株土壤细菌鉴定为7属12种,其中芽孢杆菌属8株,占鉴定总数的47.05%,为优势属。通过对青海高寒极端生境针茅根际土壤细菌的拮抗功能评价,为生防菌剂的开发提供了菌株资源。  相似文献   

16.
Sixty-two E. insidiosa strains isolated from joints or regional lymph nodes of pigs were examined from the point of view of morphology, cultural aspects, biochemical activity and virulence. All the strains consisted of gram-positive, short rods, which were similar on solid and fluid media. All strains formed H2S. Otherwise the biochemical activity was rather low except in 1 strain (no. 18), which was very active. One strain (no. 36) was rather inactive, since it showed no other activity than H2S formation. This latter strain was the only one that was avirulent for mice. The rest of the strains (61) were strongly virulent for mice (LD50 0.5 × 10−4.17 to 0.5 × 10−8.5).Of 7 strains examined for virulence for pigs by intracutaneous injection of 0.1 ml broth culture, 6 were virulent. The 7th, which was avirulent, was the one that was also avirulent for mice.  相似文献   

17.
为筛选适宜湖州地区利用桑枝屑栽培的黑木耳品种,本文通过对6个黑木耳菌株在PDA培养基的菌丝生长情况、桑枝培养基上的菌丝生长情况及子实体产量3个方面进行了比较试验,结果显示,在PDA培养基上,黑木耳916、神农7号菌丝粗壮有力,菌丝生长快;在桑枝屑培养基中,黑木耳916菌丝生长最快,神农7号次之;在子实体产量上,黑木耳9...  相似文献   

18.
Long polar fimbriae (Lpf) are recently discovered adhesins and increasingly important genetic markers of pathogenic Escherichia coli strains. The presence and genotype diversity of Lpf operons was screened in a collection of 97 Escherichia coli O157 strains representing different pathotypes, isolated from healthy cattle (n = 43) and human patients (n = 54) in several countries. Individual structural genes of Lpf were scanned by PCR, and allelic variants were detected with a recently developed typing scheme. Ninety-five strains carried at least one whole Lpf operon (genes lpfABCD and/or lpfABCDE). The 64 enterohaemorrhagic (EHEC) and 24 enteropathogenic (EPEC) strains all carried two Lpf operons, allele 3 of lpfA1 and allele 2 of lpfA2, a combination characteristic of the O157:H7/NM serotype. Out of the 9 bovine atypical (AT; stx-, eae-) strains, 7 carried one complete Lpf operon, allele 1 of lpfA2. The atypical strains belonged to main phylogenetic groups A and B1, while the EHEC and EPEC strains were from group D. Lpf variants carried by the 72 strains of the Escherichia coli Reference Collection (ECOR) were determined with the same typing scheme. Alleles were detected in 25 strains, of which 6 were found negative for the respective Lpf operons in earlier studies. The marker value of the Lpf allelic combination for the O157:H7/NM serotype was confirmed, and further evidence was given for the presence of at least two different genetic lineages of atypical bovine E. coli O157 strains.  相似文献   

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为快速检测临床样品中鸭疫里默氏杆菌(RA),建立检测RA的免疫胶体金方法。以血清1型和2型RA分别免疫BALB/c小鼠,通过细胞融合技术制备单克隆抗体。经ELISA筛选获得4株单克隆抗体2F7、3G9、5G7和2E6,5G7可与1、2、3、11型RA交叉反应,而其他3株与1、2、3型RA有交叉反应。选取单克隆抗体5G7进行纯化和胶体金标记,制备免疫胶体金试纸条,可检测1、2、3、11型RA,而与禽巴氏杆菌、大肠杆菌、沙门菌无反应。对RA模拟样品最低检出剂量为6×10^3 CFU细菌,与细菌分离方法相比,免疫胶体金试纸条对临床病料中RA检测的敏感性为100%,符合率为88.9%。  相似文献   

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