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1.
利用BIOLOG自动微生物分析系统和传统鉴定方法对1株猪源粪肠球菌进行了鉴定，并对其生物学特性进行了系统研究。结果显示，2种鉴定方法的结果一致，与传统的鉴定方法相比，BIOLOG自动微生物分析系统更快捷、方便、高效，可快速为临床诊断和用药提供依据。生物学试验表明，本株粪肠球菌在液体和固体培养基内均可有效抑制大肠杆菌生长，从而维持肠道平衡；另外，该菌株可使培养液的pH值降低至3．69。上述结果为该菌株的进一步开发利用提供了理论支持。 相似文献
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为研究猪繁殖与呼吸综合征病毒(PRRSV)与细菌混合感染的情况,本研究采用RT-PCR方法对不同猪场采集的样品进行PRRSV检测,并进一步对PRRSV阳性样品进行细菌培养分离,经BIOLOG细菌鉴定仪鉴定,分离培养的细菌为猪霍乱沙门氏菌(S.cholersuis).利用分离获得的2株PRRSV和一株S.cholersuis进行动物回归试验,结果表明所有人工感染猪在感染后的第5d出现病毒血症;2份PRRSV分离株单独人工感染的发病率均为100％,死亡率分别为55.6％和100％;PRRSV与S.cholersuis共感染的动物死亡率均为100％.与单独的PRRSV感染组相比,PRRSV与S.cholersuis共感染后实验动物的发病和死亡时间大大缩短,进一步表明其混合感染具有协同致病性,导致发病率和死亡率的上升. 相似文献
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为查明贵阳市花溪区麦坪镇某猪场仔猪发生呼吸道疾病的病因,对送检的2头病猪采集病料进行细菌分离培养、染色镜检、生化试验、PCR扩增及测序、药敏试验。结果:从病料样本中分离得到1株细菌,根据形态学和生化试验初步鉴定为副猪嗜血杆菌;应用细菌16S rRNA序列分析技术从分子水平对分离细菌进行分型鉴定,运用DNAStar软件与不同血清型副猪嗜血杆菌基因序列进行比对,发现分离菌与不同血清型副猪嗜血杆菌菌株16S rRNA序列同源且相似性为97.4%～100%,其中与血清5型相似性最高;系统进化分析显示,分离菌株与血清5型副猪嗜血杆菌进化关系最近;分离菌株对利福平、头孢氨苄、阿米卡星、环丙沙星、万古霉素敏感。结论:综合分离细菌传统鉴定方法和分子生物学鉴定方法的实验结果,确定分离菌株属于血清5型副猪嗜血杆菌。 相似文献
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据《Scientia Horticulturae》(2012.10.18)的一篇研究报道,来自南昌大学食品科技国家重点实验室的研究人员对生物控制柑桔青霉菌的拮抗细菌进行了筛选和鉴定。研究人员从宽皮柑桔、苹果、柠檬和金柑表面分离出81种细菌,从中筛选与青霉菌拮抗的细菌用于控制采后青霉病的发生。通过BIOLOG自动微生物鉴定系 相似文献
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禽大肠杆菌的分离与16S rRNA的鉴定 总被引：9，自引：4，他引：5
从疑似患有大肠杆菌病的病死鸡群中采取粪便样品,分离病原进行生化鉴定,从8份样品中分离鉴定出6株大肠杆菌。根据细菌16S rRNA 基因的高度保守性,设计合成大肠杆菌的共同引物,对随机选取的1株细菌进行PCR扩增,并与GenBank中的E.coli 16S rRNA进行序列比对,确定这株细菌与大肠杆菌的同源性达99%以上。本方法特异性好,为实验室鉴定大肠杆菌提供了一种简单、容易操作的手段。 相似文献
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袋鼠源铜绿假单胞菌分离鉴定 总被引：1，自引：0，他引：1
为确定袋鼠死亡原因,本研究采用Biolog快速鉴定系统、16SrRNA序列分析以及传统细菌鉴定方法对2株分离自北京动物园袋鼠肺脏的菌株进行形态学、培养特性、生化特性、小鼠致病性、系统发育分析等生物特性的鉴定和分析,结果表明2菌株均为铜绿假单胞菌,对昆明小鼠有强致病性。系统发育分析结果表明2株袋鼠源铜绿假单胞菌16SrRNA序列与ATCC10145模式株差异很小,同源性分别为99.9%和100%,并且位于系统发育树的同一分支。 相似文献
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本试验通过对不同地区3个奶牛场临床型乳房炎细菌病原的分离鉴定,分析其种类和感染情况,并结合牧场环境状况和管理方式,提出控制乳房炎的建议.从中国福建、新疆、重庆3个不同规模化奶牛场采集61份临床型乳房炎乳样,采用传统微生物鉴定方法与PCR技术进行细菌分离鉴定.结果表明,61份临床型乳房炎乳样中单一感染44份(72.13%),混合感染15份(24.59%);共分离到细菌80株,经传统微生物鉴定方法与PCR技术鉴定主要为大肠杆菌、葡萄球菌和链球菌,其中传染性微生物13株(16.25%)、环境性微生物46株(57.50%)、机会性微生物11株(13.75%)、其他微生物10株(12.50%);3个奶牛场临床型乳房炎仍以单一感染为主,混合感染占有一定的比例,传染性致病菌和环境性致病菌为主要致病菌.在实际生产中,规模化牧场可以通过对乳房炎病原菌的分离鉴定来摸清牧场奶牛乳房炎的感染情况和致病菌种类,结合生产实际有助于了解牛群管理情况,发现问题并加以改善,更好的控制乳房炎的发生. 相似文献
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猪霍乱沙门氏菌的快速分离鉴定 总被引：3，自引：0，他引：3
猪霍乱沙门氏菌是引起仔猪副伤寒的主要病原菌,给养猪业造成重大危害。本文旨在建立一套细菌快速分离鉴定方法,为该病流行病学调查提供有效手段。从临床初诊为仔猪副伤寒的病猪采集粪便,接种亚硒酸盐亮绿增菌液增菌后,划线于SS琼脂培养基,挑无色透明菌落纯化。纯化菌落用美国B IOLOG自动细菌鉴定仪GN2肠杆菌鉴定板鉴定,同时用常规生化鉴定管鉴定,结果符合猪霍乱沙门氏菌的生化特征。PCR扩增invA基因,测序鉴定PCR产物,结果与猪霍乱沙门氏菌参考序列同源性达99%以上。分离菌株接种小白鼠,证明其对小白鼠有致病性。用沙门氏菌属标准血清检测证实其抗原式为6,7∶c∶1,5,符合猪霍乱沙门氏菌的抗原特征。应用以上方法从全国各地病料中分离鉴定了40多株猪霍乱沙门氏菌,为猪霍乱沙门氏菌病的流行病学调查提供了依据,同时证明该方法切实可行。 相似文献
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根据现有细菌分类鉴定方法,采用血清学、Biolog、16S rRNA序列分析和全基因组序列分析等方法对羊败血性链球菌病疫苗生产检验用菌种CVCC 553、55001和55002进行系统鉴定。结果显示,菌种CVCC 553、55001和55002血清群为兰氏C群;Biolog鉴定为马链球菌反刍亚种;16S rRNA序列分析与马链球菌反刍亚种模式菌株CECT 5772的同源性最高(均为99.72%),且在系统发育树中位于同一分支;与马链球菌反刍亚种模式菌株CCUG 47520的DNA-DNA的杂交值大于70%,分别为80.5%、80.2%、80.5%,与马链球菌反刍亚种模式菌株CECT 5772的平均核苷酸同源性分别为97.73%、97.64%、97.77%。因此本研究认为羊败血性链球菌病疫苗生产用菌种CVCC 553、55001和55002应为马链球菌反刍亚种。 相似文献
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百脉根根际高效溶磷菌LC15的特性研究及菌株鉴定 总被引：2，自引：0，他引：2
研究了分离自百脉根(Lotus corniculatus)根际的溶磷菌LC15,通过对其进行生理生化特性及16S rDNA序列分析表明,其菌株具有较强的溶解无机磷和有机磷的能力,且对无机磷的溶解能力更强,D/d比值均超过3.00;振荡培养7 d后其可溶性磷含量高达400.49 mg/L,分泌总有机酸量30.67 mmol/L,且菌悬液pH 4.39呈酸性;革兰氏染色呈阴性,在厌氧生长、V-P、柠檬酸盐利用、ONPG测定、精氨酸双水解酶等生理生化试验中均呈阳性;经BIOLOG测定,可利用碳源占95种供试碳源种类的54%;结合菌株16S rDNA序列同源性比对分析,初步鉴定为柠檬酸杆菌属(Citrobcter sp.)。 相似文献
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An evaluation of the API ZYM system as a means of classifying spirochaetes associated with swine dysentery. 总被引：4，自引：0，他引：4
Thirty-one strains of spirochaetes from pigs and two strains from poultry were tested in the API ZYM enzyme system. The spectrum of enzymatic reactions provided a means of differentiating between strains related to swine dysentery and strains not so related from pigs. The numerical coding system simplified the interpretation of the results. 相似文献
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Salmonella Livingstone is occasionaly isolated from humans, animals and feedstuffs in Sweden. To follow the spread of infection and trace the source of isolates, adequate typing methods are needed. We have developed an automated typing system based on biochemical fingerprinting of bacteria (the PhP system) for typing of different Salmonella serotypes. The system measures the kinetics of various biochemical reactions of bacteria grown in liquid medium in microtiter plates and uses numerical techniques to identify biochemical phenotypes (BPTs) among the tested strains. In the present study we used a set of 16 highly discriminatory tests to differentiate strains of Salmonella of serotype Livingstone and evaluated the system for its discriminatory ability using a collection of 34 unrelated human isolates of S. Livingstone. We also used the system to investigate BPTs of 45 Livingstone strains isolated from animals and feedstuffs in Sweden between 1987 and 1991. Altogether 19 different BPTs were found among human isolate giving a diversity index (Di) of 0.930. In contrast, most strains isolated from animals and feedstuffs in Sweden belonged to 2 dominating BPTs (Di = 0.704). One of these contained 17 strains mainly isolated during 1992 whereas the other contained 18 strains isolated between 1987 and 1991. None of the Swedish human isolates were identical to those of animals and feedstuffs. These findings suggest that 2 different BPTs of Salmonella Livingstone strains are particularly common among animals and feedstuffs in Sweden and that they are not related to human cases of enteritis in this country. We also conclude that biochemical fingerprinting with the PhP system is a reliable and highly discriminatory method for detecting epidemic strains of Salmonella Livingstone. 相似文献
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The Vitek Gram-positive identification system (GPI, Vitek Systems, Inc., Hazelwood, MO) and the API Rapid Strep system (Analytab Products, Plainview, NY) were evaluated for species identification of streptococci isolated from bovine mammary glands and compared to conventional biochemical methods. A total of 144 strains including Streptococcus uberis (60), S. dysgalactiae (32), S. agalactiae (15), S. bovis (15), Enterococcus faecium (10) and Ent. faecalis (12) were evaluated. All reference strains were identified correctly by both systems. Vitek GPI card system identified 94.4% of strains, including 95% of S. uberis, 93.8% of S. dygalactiae, 93.3% of S. agalactiae and S. bovis II, 90% of Ent. faecium and 100% of Ent. faecalis. Majority of strains were identified with a 90-99% level of confidence, with an average of 8 h needed for identification. The API Rapid Strep system identified 96.5% of strains correctly, including 95% of S. 96.9% of S. dysgalactiae, 93.3% of S. agalactiae, and 100% of S. bovis II, Ent. faecium, and Ent. faecalis. Majority of strains were identified with excellent level of identification. With the exception of S. uberis, most strains were identified at 4 h of incubation. 相似文献
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P A Grieve D A Dionysius A C Vos 《Zentralblatt für Veterin?rmedizin. Reihe B. Journal of veterinary medicine. Series B》1992,39(7):537-545
The lactoperoxidase-thiocyanate-hydrogen peroxide (LP) system inhibited the growth of enterotoxigenic Escherichia coli strains responsible for scouring in neonatal and post-weaning piglets. An enzymatic system for hydrogen peroxide generation (glucose oxidase, GO; 0.1 U/ml) and a chemical source (sodium carbonate peroxyhydrate, SCP; 90 mg/l) were used in the LP system to test 19 strains in a 6-h growth assay at 37 degrees C. Only three strains were highly sensitive to the LP/GO system, while all exhibited significant growth inhibition with the LP/SCP system. Hydrogen peroxide alone had less effect than the complete system. The bactericidal activity of the LP/GO system towards a previously resistant strain was greatly increased by increasing the level of glucose oxidase in the system by three- or five-fold. 相似文献
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Canine distemper (CD) is a contagious disease, which can damage the immune system, respiratory system, digestive system, and even nervous system, leading to systemic pathological changes, and has a huge threat to pet dogs, fur animals, etc. At present, the commonly used colloidal gold test cannot effectively distinguish vaccine immunity from animal natural infection. To establish an efficient and accurate detection method for identifying wild strains and vaccine strains of canine distemper virus(CDV), the whole genome of six canine distemper wild strains isolated from dogs and three widely used CDV vaccine strains collected from Wuhan area were sequenced. After comparing and analyzing the amino acid and the base sequences, the H gene was determined as the target gene for AS-PCR primer design. By genotyping the H gene, it was found that the prevalent CDVs in Wuhan were all Asia-Ⅰ, while the vaccine strain Y2 was America-I, and the vaccine strains Y1 and Y3 were both America-Ⅱ. Comparing the CDV-H gene sequences of 179 Asia-Ⅰtypes (6 wild-type strain samples + 173 GenBank Asia-Ⅰtype stains) and 3 vaccine strains, using AS-PCR technology (3'mismatch) to design a pair of primers. It can effectively distinguish CDV Asia-I wild strain and vaccine strain. The upstream primer sequence is 5'-TTAATATAATAATGACAGTG-3', and the downstream primer sequence is 5'-CCTCAAGGGGCACA-3'. The results showed that the primer had a strong specificity and the wild strains could amplify an 894 bp fragment, while the vaccine strains could not. There are 9 more regular bases (amino acids) variation sites in H gene of wild strain, and the 277th amino acids of all vaccine strains are Asparagine. These mutations may lead to an increase in N-glycosylation sites, which will have an impact on the virulence of canine distemper virus vaccine strains. The AS-PCR method established in this study can effectively distinguish the canine distemper vaccine and Asia-Ⅰwild strains. 相似文献
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Sasaki Y Yamamoto K Kojima A Tetsuka Y Norimatsu M Tamura Y 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2000,62(12):1275-1281
Clostridium chauvoei causes blackleg, which is difficult to distinguish from the causative clostridia of malignant edema. Therefore, a single-step PCR system was developed for specific detection of C. chauvoei DNA using primers derived from the 16S-23S rDNA spacer region and partial 23S rDNA sequences. The specificity of the single-step PCR system was demonstrated by testing 37 strains of clostridia and 3 strains of other genera. A 509 bp PCR product, which is a C. choauvoei-specific PCR product, could be amplified from all of the C. chauvoei strains tested, but not from the other strains. Moreover, this single-step PCR system specifically detected C. chauvoei DNA in samples of muscle from mice 24 hr after inoculation with 100 spores of C. chauvoei, and in clinical materials from a cow affected with blackleg. These results suggest that our single-step PCR system may be useful for direct detection of C. chauvoei in culture and in clinical materials from animals affected with blackleg. 相似文献
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主动外排系统介导的大肠杆菌多重耐药性的确证 总被引：9，自引：1，他引：8
为检测和证实主动外排系统(外输泵)在细菌多重耐药机制中的作用，用四环素和水扬酸钠对大肠杆菌质控株进行了体外人工诱导，筛选出多重耐药株，再用能量抑制剂CCCP(Carbongl cyanide m-chloropheyhydrazone)时耐药株和质控株进行了环丙沙星摄取抑制试验。结果：在耐药株，其菌体内环丙沙星稳态浓度明显低于质控株；多重耐药株在有CCCP存在时，菌体内环丙沙星的浓度随时间的延长而递增，而无CCCP的抑制时，菌体内环丙沙星的浓度递增缓慢；在质控株，不论有无CCCP存在，菌体内环丙沙星的浓度都随时间的延长而呈递增趋势，且保持较耐药株明显高的水平。结论：体外诱导多重耐药大肠杆菌多重耐药性的产生，可能主要依赖于菌体外输泵的激活。 相似文献