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1.
《现代畜牧兽医》2021,(9)
试验以55株乳源金黄色葡萄球菌为研究对象,通过结晶紫染色法分析其生物被膜形成能力,应用PCR技术检测乳源金黄色葡萄球菌的所携带的耐药基因和生物被膜形成相关基因。结果表明,55株金黄色葡萄球菌中34株生物被膜形成能力较强,17株生物被膜形成能力较弱,4株无生物被膜形成能力;55株细菌中17株(31%)携带mec A耐药基因,17株金黄色葡萄球菌分为9个ST型,均属于已发现的ST型,其中ST97和ST398为优势株。上述来源相近的金黄色葡萄球菌分型后,基本位于ST97和ST98两个大簇,说明该地区引起奶牛乳房炎的细菌型别相对较集中。生物被膜相关基因Ica A、Ica D、Ica R、Eno、Atl、aap、Bap和sig B基因的检出率分别为44%、36%、27%、89%、60%、42%、18%和25%。研究表明,乳源金黄色葡萄球菌的生物被膜形成能力较强,生物被膜形成能力较强的菌株均携带多个生物被膜形成相关基因,说明生物被膜的形成过程可能是受多个基因共同调节。 相似文献
2.
金黄色葡萄球菌是引起奶牛细菌性乳腺炎的主要原因，生物被膜的形成是金黄色葡萄球菌在不利环境条件下持久性存在的关键因素。探索同一株菌在生物被膜态与浮游态生长状态下的耐药性与其生长状态的相关性，可为进一步探究金黄色葡萄球菌的耐药性机制奠定基础。本研究培养了金黄色葡萄球菌的生物被膜，使用光学显微镜和扫描电镜观察其形成过程。测定并比较了9种抗菌药物对32株金黄色葡萄球菌在生物被膜态和浮游态的最小抑菌浓度，并对两种状态下的金黄色葡萄球菌进行转录组学测序，筛选出具有显著性差异的细胞信号通路和表达基因，同时对主要差异表达的基因进行RT-qPCR验证。结果发现，在生物被膜形成前期，随着培养时间的延长，显微镜下观察到的生物被膜态菌聚集面积越来越大，结构也越来越紧密，培养至72 h后，生物被膜逐渐开始分散。MIC测定结果显示浮游态菌的抑菌浓度低于生物被膜态菌。转录组结果显示两种状态菌的差异表达基因共1 512个，其中，生物被膜态菌中上调基因760个，下调752个。GO与KEGG富集分析显示，相比于浮游态菌，生物被膜态菌中与代谢相关的通路显著富集，其次为氨基酸的生物合成和ABC转运蛋白通路。与生物被膜形成相关的基因，如编码ABC转运蛋白的基因表达上调，而与代谢途径相关的基因下调。RT-qPCR验证了10个主要差异基因，其表达差异趋势与转录组测序结果一致。这些差异可能对金黄色葡萄球菌生物被膜态的高耐药性和细菌毒力的研究有所帮助。 相似文献
3.
研究鱼腥草水提物对金黄色葡萄球菌生物被膜的干预作用。采用微量肉汤稀释法测定鱼腥草水提物对金黄色葡萄球菌CMCC(B)26003的最低抑菌浓度;采用结晶紫染色法、扫描电镜及激光共聚焦显微镜评估鱼腥草水提物对CMCC(B)26003生物被膜形成的影响;最后采用实时荧光定量PCR方法测定鱼腥草水提物对O-乙酰丝氨酸硫化氢裂解酶B(O-acetylserine sulfhydrylase-B)cysM基因转录的影响。结果显示,鱼腥草水提物在12.500 mg/mL、6.250 mg/mL、3.125 mg/mL质量浓度下均可以有效抑制金黄色葡萄球菌生物被膜的形成,在12.500 mg/mL质量浓度下可下调cysM基因的表达。鱼腥草可能通过下调cysM基因的表达而抑制金黄色葡萄球菌生物被膜的形成。 相似文献
4.
奶牛乳房炎金黄色葡萄球菌生物被膜的形成受许多物质及其同一种物质不同浓度添加的影响。本研究以乳房炎金黄色葡萄球菌分离株N_2为研究对象,采用XTT法研究不同浓度的羊血、鸡血和驴血对金黄色葡萄球菌分离株N_2生物被膜的影响。结果表明,羊血对生物被膜形成的促进作用最大,鸡血次之,驴血反而抑制生物被膜的形成;羊血、鸡血的浓度梯度范围在3%~5%内对金黄色葡萄球菌分离株N_2的生物被膜形成的促进作用最强,在低于该浓度范围时对生物被膜的形成呈抑制作用。综上所述,不同种类动物的血液以及同一种动物的不同浓度血液都会对金黄色葡萄球菌分离株N_2生物被膜的形成产生较大影响。 相似文献
5.
奶牛乳房炎是造成奶牛养殖业巨大经济损失的疾病,金黄色葡萄球菌是导致奶牛乳房炎感染的主要致病菌之一。以生物被膜的群落形式存在的金黄色葡萄球菌,更易逃避宿主免疫系统、降低药物敏感性,从而导致奶牛乳房炎反复发作,最终形成顽固性乳房炎。研究显示,天然抗生物被膜物质对金黄色葡萄球菌生物被膜的抑制作用明显,可作为新的抗菌药物应用于奶牛乳房炎的防治。对金黄色葡萄球菌生物被膜耐药机制和天然抗金黄色葡萄球菌生物被膜物质的最新研究进展进行了概述,以期对防治金黄色葡萄球菌导致的奶牛乳房炎提供参考。 相似文献
6.
[目的]分离鉴定新疆维吾尔自治区阿拉尔市猪乳源金黄色葡萄球菌,了解其致病性和耐药性。[方法]从阿拉尔市某规模化养殖场收集6份新鲜猪乳样本,采用甘露醇高盐琼脂（MSA）培养基分离乳样中的金黄色葡萄球菌,利用表型鉴定方法及细菌16S rDNA序列分析法对分离到的菌株进行鉴定,检测分离菌株的生物被膜形成能力,利用K-B纸片扩散法进行药物敏感性试验,利用PCR法检测毒力基因和耐药基因。[结果]经分离培养及分子生物学鉴定,从6份乳样中获得2株金黄色葡萄球菌,分离率为33.33%(2/6);2株金黄色葡萄球菌均具有强生物被膜形成能力（+++）;2株金黄色葡萄球菌均对利福平敏感,均对青霉素、头孢西丁、四环素、红霉素、替米考星、克林霉素和磺胺甲噁唑耐药;在检测的5种耐药基因中,2株金黄色葡萄球菌均携带ant(4)基因和mecA基因;在检测的20种毒力基因中,共检测到6种毒力基因,其中,2株金黄色葡萄球菌均携带seg、sei、hlb、fnbA、clfA基因,有1株携带hla基因。[结论]该养殖场猪乳源金黄色葡萄球菌生物被膜形成能力强,携带多种毒力基因和耐药基因,表现为多重耐药性。 相似文献
7.
《动物医学进展》2015,(12)
为了展示金黄色葡萄球菌生物被膜领域的整体研究状况和研究热点,基于CitespaceⅢ研究了1995年-2015年Web of Science~(TM)核心合集数据库所收录的有关金黄色葡萄球菌生物被膜的3 582条科技文献,分析了年度发文量,绘制了国家、机构、作者、被引期刊、被引文献及关键词的知识图谱。金黄色葡萄球菌生物被膜研究领域的发文量从1995年-2014年持续上升,2015年有延续的趋势;研究金黄色葡萄球菌生物被膜的国家和机构及刊登该领域文献的期刊主要分布在北美洲和欧洲,大学是该研究领域的主要研究机构;金黄色葡萄球菌生物被膜的研究热点集中于生物被膜的形成机制和抑制药物的研发两个方面,未来一段时间的研究方向将侧重于阻碍生物被膜形成和促进已形成生物被膜瓦解的药物研发。 相似文献
8.
本实验以中药提取物小檗碱为研究对象,探究其对金黄色葡萄球菌生物被膜形成的抑制作用。实验采用常量肉汤稀释法测定小檗碱对金黄色葡萄球菌的最小抑菌浓度(MIC);随后分别选取2 MIC、1 MIC、0.5 MIC浓度小檗碱,采用结晶紫染色法测定其对金黄色葡萄球菌生物被膜生长的抑制率,同时采用实时荧光定量PCR测定2 MIC、1 MIC、0.5 MIC小檗碱对金黄色葡萄球菌Nuc基因的表达情况。结果显示,小檗碱对金黄色葡萄球菌的最小抑菌浓度(MIC)为62.5μg/mL,2 MIC、1 MIC、0.5 MIC浓度小檗碱对金黄色葡萄球菌生物被膜的抑制率分别为77.14%、66.91%和55.55%,荧光定量PCR测得各组金葡菌Nuc基因的Ct值大小为Ct值(0.5 MIC)> Ct值(1.0 MIC)>Ct值(2 MIC),说明随着小檗碱浓度的增大,对金黄色葡萄球菌生物被膜的抑制作用增强,呈现明显的浓度依赖性。本研究为小檗碱对金黄色葡萄球菌感染的防治提供数据支持。 相似文献
9.
试验旨在探究苦参、连翘、白头翁、蒲公英、黄柏和五倍子等6种中药水提物对金黄色葡萄球菌生物被膜形成的影响。通过银染法和结晶紫染色法检测6种中药水提物对金黄色葡萄球菌生物被膜的黏附和抑制作用。结果显示,中药水提物减少了金黄色葡萄球菌生物被膜的黏附;与对照组相比,在6种中药水提物作用下,金黄色葡萄球菌生物被膜洗脱液的OD590 nm值均极显著降低（P<0.01）,其中五倍子水提物的抑制效果最优,抑制率为74.9%。研究表明,6种中药水提物均能够抑制金黄色葡萄球菌生物被膜的形成,其中五倍子水提物效果最好。 相似文献
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11.
The implication of biofilm in chronic bacterial infection in many species has triggered an increasing interest in the characterization of genes involved in biofilm formation. The bap gene is a newly identified gene that encodes the biofilm-associated protein, BAP, which is involved in biofilm formation in Staphylococcus aureus. So far the bap gene has only been found in a small proportion of S. aureus strains from bovine mastitis in Spain. In order to study the presence of the bap gene in S. aureus isolates obtained from other species and various locations, a collection of 262 isolates was tested by PCR, using published primers and dot-blot. The results indicated that none of the isolates carried the bap gene suggesting that the prevalence of this gene among S. aureus isolates should be very low. 相似文献
12.
Phenotypic and genotypic characterization of bovine mastitis isolates of Staphylococcus aureus for biofilm formation 总被引：1，自引：0，他引：1
Staphylococcus aureus is one of the most common pathogens responsible for contagious mastitis in ruminants. The ability of S. aureus to form biofilm in vivo is considered to be a major virulence factor influencing its pathogenesis in mastitis. The objectives of the study were to examine in vitro slime production, biofilm formation, and the presence of the ica gene locus and icaA and icaD genes in S. aureus isolates from bovine mastitis. Thirty-two of the 35 isolates tested produced slime on Congo red agar, whereas only 24 of the isolates were found to produce biofilm in vitro. However, all the 35 isolates possessed the ica locus, icaA and icaD genes. This study indicates a high prevalence of the ica genes among S. aureus mastitis isolates, and their presence is not always associated with in vitro formation of slime or biofilm. A combination of phenotypic and genotypic tests is recommended for investigating biofilm formation in S. aureus. 相似文献
13.
The purpose of this study was to observe and analyze the ability of biofilm formation of the Enterococcus faecalis (E.faecalis) XJ05 and to detect if the E.faecalis which could induce lamb encephalitis carried the biofilm-associated genes. The E.faecalis XJ05 was used as the sample. The biofilm model was established by the 96-well microtiter plate and read by microplate reader in the D595 nm at the same time observed by the inverted microscope. The primers were designed to examine if the 11 strains of E.faecalis carried the biofilm related genes and then the amplified products were sequenced and analyzed for homology. The results showed that the amount of BF produce in XJ05 strain was significantly different from blank control group at different time (P<0.05), and the highest D595 nm value was at 24 h and significant difference compared with other time (P<0.05). The biofilm of 6 h was scattered when observed under the microscope and the time between 12 to 24 h biofilm to be more denser, after 24 h the biofilm became to degradate and loose gradually. It turned out that the 11 strains of bacteria carried different biofilm-associated genes, 8 biofilm-associated genes were found in 6 strains of bacteria, 7 biofilm-associated genes were find in 1 strain of bacterium, 6 biofilm-associated genes were found in 1 strain of bacterium, 2 biofilm-associated genes were found in 1 strain of bacterium, 2 strains of bacteria could not text out anyone of the detected genes.The homology of all detected genes were all between 93.3% to 100.0%. It concluded that the E.faecalis XJ05 could form the completed biofilm and there were 9 kinds carried the biofilm-associated genes in 11 strains of E.faecalis. 相似文献
14.
为观察致羔羊脑炎粪肠球菌XJ05株生物被膜（BF）的形成能力并检测致羔羊脑炎粪肠球菌BF形成相关基因的携带情况,本研究以XJ05为研究对象,运用96孔板建立不同时间BF模型,染色后酶标仪测定D595 nm值,并用倒置显微镜观察BF形态。同时检测11株菌中与BF形成相关基因（gelE、esp、ace、asa1、asa373、efaA、ef0591、ef3314、ahrc、eep）的携带情况,并对相关基因片段做同源性分析。结果显示,在不同时间段XJ05株BF的生成量与空白对照组相比差异显著（P<0.05）,24 h时D595 nm值最高且与其他各个时间段相比均差异显著（P<0.05）。显微镜下观察6 h时BF初具规模,可见散落的网状结构,12~24 h矩阵网格结构明显,24 h时形成高度有序的网格结构,24 h后网状结构逐渐脱落,BF开始降解。11株菌中10种相关基因的携带情况不同,有6株携带8种基因,1株携带7种基因,1株携带6种基因,1株携带2种基因,有2株未检测到10种基因的任何一种。检出的基因片段同源性均在93.3%~100.0%之间。结果表明,XJ05株能形成完整的BF,11株分离的致羔羊脑炎粪肠球菌中有9株携带部分与BF形成相关的基因。 相似文献
15.
Melchior MB Fink-Gremmels J Gaastra W 《Journal of veterinary medicine. B, Infectious diseases and veterinary public health》2006,53(7):326-332
Mastitis is one of the most important diseases in dairy cattle of which Staphylococcus aureus is a major pathogen. Despite an apparently good antimicrobial susceptibility in vitro, the cure of diseased animals from this bacteriological infection is often disappointing, which results in cases of recurrent clinical- and chronic subclinical infections. It has been suggested that these recurrent and chronic Staphylococcus infections can be attributed to the growth of bacteria in biofilm. The objective of this study was to compare the susceptibility for antimicrobial agents of S. aureus isolates obtained from bovine mastitis growing under different conditions. These conditions include a conventional conventional microbroth dilution assay in which minimal inhibitory concentration values are determined, the MBEC assay which measures both the susceptibility in biofilm and the susceptibility of sequester cells released from the biofilm. A comparison of the susceptibility for antimicrobial agents of a number of representative S. aureus isolates grown in broth (representing in vitro growth conditions) or milk (representing in vivo growth conditions) is also made. The results indicate that S. aureus isolates obtained from bovine mastitis are highly resistant to antimicrobial agents when growing in biofilms. 相似文献
16.
Extended antimicrobial susceptibility assay for Staphylococcus aureus isolates from bovine mastitis growing in biofilms 总被引：1，自引：0，他引：1
Staphylococcus aureus is one of the most prevalent causes of bovine mastitis. The antimicrobial treatment of this disease is currently based on antimicrobial susceptibility tests according to CLSI standards. However, various studies have shown that there is a discrepancy between the results of this standard susceptibility test and the actual cure rate of the applied antimicrobial treatment. Increasing evidence suggests that biofilm formation by S. aureus is associated with this problem. The currently available antimicrobial susceptibility assays for bacteria growing in biofilms, are not considered reliable enough for routine application. Therefore, the objective of this study was to further develop a susceptibility test for bacteria growing in biofilm, suitable for routine testing of the antimicrobial susceptibility of S. aureus. With the expansion of the available MBEC assay to an extended biofilm susceptibility test, that comprises 2 and 4 consecutive days of antimicrobial challenge, the antimicrobial susceptibility for S. aureus growing in biofilm was further analysed. The results showed clear differences between strains and various antimicrobial agents with respect to the effect of longer duration of the antimicrobial challenge on the eradication of S. aureus growing in biofilm. The extended biofilm susceptibility test also indicates that each bacterial strain requires a specific duration of antimicrobial therapy, which cannot be derived from a standard susceptibility test or from a 24-h biofilm susceptibility test. 相似文献
17.
Biofilm production by 221 Staphylococcus aureus isolates from 45 dairy herds was evaluated. Isolates were from composite milk of 117 cows, from teat skin of 70 cows, and from 34 milking machine unit liners. Of S. aureus from milk samples, 41.4% were biofilm producers, as compared to 24.7 and 14.7% of the isolates collected from skin and liners. Pulsed field gel electrophoresis (PFGE) best categorized S. aureus biofilm producers as compared to phage typing and binary typing. PFGE types that were significantly associated with isolation from milk as opposed to teat skin or liners, had isolates that were more likely to produce biofilm than PFGE types that were isolated from milk, skin and liners at similar frequencies. By contrast, PFGE type A was significantly associated with isolation from teat skin and had few biofilm producers. PFGE type Q, which is exclusively a milk, isolate produced more biofilm as evidenced by absorbance values. Given S. aureus that are associated with milk are more likely to produce biofilm as compared to extramammary sources (teat skin and milking unit liners), suggests that biofilm production is a risk factor for infection. 相似文献
18.
Oliveira M Bexiga R Nunes SF Carneiro C Cavaco LM Bernardo F Vilela CL 《Veterinary microbiology》2006,118(1-2):133-140
Biofilm-forming ability has been increasingly recognized as an important virulence factor in Staphylococci, facilitating their persistence in the host, evading its defences and allowing bacterial survival at high antimicrobial concentrations. Staphylococcus aureus remains a major pathogen of chronic mastitis, but in the last years Staphylococcus epidermidis has emerged as a relevant mastitis pathogen. The present work aimed at the evaluation of the biofilm-forming ability of Staphylococci field isolates from bovine subclinical mastitis and at the development of a fluorescent in situ hybridisation (FISH) protocol that would allow the direct observation of biofilm formation in milk samples. The analysis of phenotypic expression in Congo Red Agar (CRA) and by FISH, showed that 37.5% of the S. aureus isolates produced biofilm, while by optical density measurement only 18.75% isolates revealed this phenotype. The results showed a fair agreement according to the kappa coefficient test (kappa = 0.259). Regarding S. epidermidis mastitis isolates, 37.5% revealed the ability to produce biofilm, but only four isolates were positive by all methods. This agreement was moderate (kappa = 0.467). The application of FISH to artificially contaminated milk samples allowed the direct observation of biofilm production by 37.5% isolates, showing total agreement with the CRA results. This method better mimics the in vivo conditions, especially in terms of the presence of calcium and iron, which in high concentrations, respectively, are known to inhibit or induce biofilm production. 相似文献
19.
In this study, Staphylococcus aureus strains (n = 110) isolated from seven ewe flocks in Sanliurfa, Turkey were screened for antibiotic resistance and biofilmforming ability as well as for genes associated with antibiotic resistance and biofilm-forming ability. All isolates were found to be susceptible to oxacillin, gentamicin, clindamycin, cefoxitin, tetracycline, vancomycin, amoxicillin-clavulanic acid, ciprofloxacin and sulphamethoxazole-trimethoprim. The percent proportions of strains resistant to penicillin G, ampicillin and erythromycin were 27.2% (n = 30), 25.4% (n = 28) and 6.3% (n = 7), respectively. Regarding the antibiotic resistance genes, 32 (29%) isolates carried the blaZ and 8 (7.2%) the ermC gene. Other resistance genes were not detected in the isolates. All isolates showed biofilm-forming ability on Congo red agar (CRA), while 108 (98.18%) and 101 (91.81%) of them were identified as biofilm producers by the use of standard tube (ST) and microplate (MP) methods, respectively. All isolates carried the icaA and icaD genes but none of them harboured the bap gene. The results demonstrated that S. aureus isolates from gangrenous mastitis were mainly resistant to penicillins (which are susceptible to the staphylococcal beta-lactamase enzyme), and less frequently to erythromycin. Furthermore, all of the S. aureus isolates produced biofilm which was considered a potential virulence factor in the pathogenesis of staphylococcal mastitis. 相似文献