细菌的消毒剂耐药性

细菌的消毒剂耐药性指细菌与消毒剂多次接触后,使该类消毒剂的最小抑菌浓度或最小杀菌浓度升高的现象。细菌的消毒剂耐药性普遍存在,多种细菌可对一种消毒剂耐药,一种细菌可对多种消毒剂耐药。消毒剂选择件压力是耐药性产生的外在原因,产生的机制包括生化结构、遗传学途径和酶学途径。消毒剂耐药与抗生素耐药之间存在一定的关系。应加强研究,制定统一标准,加强监测,并制定合理应用消毒荆规范,以减少消毒剂耐药性的发生。     

生牛奶中分离大肠杆菌对季铵盐类消毒剂耐药性研究

目的为调查牛奶中大肠杆菌Escherichia coli的污染状况和对常见季铵盐类消毒剂的耐药情况,为大型养殖场季铵盐类消毒剂的合理使用提供指导。方法对从74份生牛奶样品中分离的45株大肠杆菌采用琼脂稀释法测定4种季铵盐类消毒剂[十六烷基三甲基溴化铵(CTAB)、苯扎氯铵(BC)、双十烷基二甲基氯化铵(DDAC)、氯代十六烷基吡啶(CPC)]的最小抑菌浓度(MIC),采用PCR法检测10种消毒剂耐药基因,并分析其MIC值与耐药基因之间的相关性。结果大肠杆菌对4种消毒剂表现出不同程度的抗性,所检测的4种消毒剂对大肠杆菌的MIC值均大于标准菌株,且比例为BC=DDACCTABCPC。大肠杆菌季铵盐类消毒剂的耐药基因检出率为ydgF(84.44%)ydgE(80.00%)sugE(c)(66.67%)emrE(40.00%)mdfA(37.78%)qacEΔ1(33.33%)qacE(17.78%)qacF(13.33%)qacG(11.11%)sugE(p)(4.44%)。分析大肠杆菌消毒剂基因检出情况与MIC值之间的关系发现,qacF基因的检出率与128 mg·L~(-1)的CPC之间差异有统计学意义。结论由此可见,牛奶中存在大肠杆菌污染严重的现象对季铵盐类消毒剂耐药情况不容乐观,需进一步规范季铵盐类消毒剂的使用。     

鹌鹑源鼠伤寒沙门氏菌的分离鉴定与耐药性分析

【背景】在鹌鹑养殖过程中,抗菌药物和消毒剂的不规范使用加剧了耐药菌株在动物、场所和食品之间的相互传播,因此,掌握致病菌株在养殖动物中的耐药状况至关重要。【目的】检测北京周边地区鹌鹑蛋源致病菌株的耐药特征和耐药基因的流行情况。【方法】在天津市武清区部分鹌鹑养殖场采集鹌鹑泄殖腔粪便、鹌鹑蛋表、养殖环境和鹌鹑饮水的样品,通过细菌分离培养、菌落形态观察、染色镜检、生化鉴定、血清分型、沙门氏菌inv A基因序列测定等方法对分离菌株进行鉴定。同时进行小鼠攻毒试验,测定小鼠半数致死量(median lethal dose, LD50)。再通过药敏试验和PCR方法对分离菌的耐药表型、耐药基因及毒力基因进行检测。【结果】分离菌株菌落颜色、镜检形态和生化试验结果符合沙门氏菌特性,沙门氏菌inv A基因序列测定与鼠伤寒沙门氏菌参考株相似度为99.44%,鉴定为鼠伤寒沙门氏菌,血清型为1,4,[5],[12]:i:l,2。该菌株对小鼠有致病作用,小鼠LD50为2.10×107 CFU/mL;药敏试验结果显示该菌株对氨苄西林、阿莫西林/克拉维酸、头孢噻呋、链霉素、磺胺甲啞唑、磺胺异啞唑、诺氟沙星、环丙沙星表现耐...     

沙门氏菌对季铵盐类消毒剂的适应性耐受机制研究进展

沙门氏菌（Salmonella）是一种极其重要的食源性致病菌,具有耐高温、抗逆性强的特点,能在食品加工环境中长久存活。季铵盐类消毒剂在食品工业中的广泛应用使得沙门氏菌对其产生了适应性耐受和交叉耐受,为食品健康带来了极大的安全隐患。文章简要介绍了季铵盐类消毒剂的主要种类及其作用机理,分析并归纳了沙门氏菌对季铵盐类消毒剂的适应性耐受和交叉耐受现状,进一步从细胞膜生理特征、生物被膜形成、外排泵活性等角度,重点阐述了沙门氏菌对季铵盐类消毒剂的适应性耐受机制进展及新发现,并对未来研究方向进行了展望,旨在为季铵盐类消毒剂的合理使用和沙门氏菌的防控提供重要参考。     

大熊猫源大肠杆菌及肺炎克雷伯氏菌对消毒剂耐药性研究

本实验对大熊猫肠道分离的88株大肠杆菌、32株肺炎克雷伯氏菌对季铵盐类消毒剂BC、CTPC、CTAB及DDAC的最小抑菌浓度(MIC)值进行测定,并扩增了消毒剂的耐药基因。结果显示,大肠杆菌对季铵盐类消毒剂MIC值为:BC的MIC值介于8~128 mg/L;CTPC的MIC值在32~256 mg/L之间;CTAB的MIC值为64~512 mg/L;DDAC的MIC值介于8~128 mg/L。肺炎克雷伯氏菌对季铵盐类消毒剂的耐药情况为:BC的MIC值介于16~512 mg/L;CTPC的MIC值在64~256 mg/L之间;CTAB的MIC值介于128~512 mg/L;DDAC的MIC值介于8~64 mg/L。可见,肺炎克雷伯氏菌对季铵盐类消毒剂的MIC值要大于大肠杆菌对季铵盐类消毒剂的MIC值。耐药基因检测结果表明,大肠杆菌季铵盐类消毒剂的染色体型耐药基因扩增率为68.18%~98.86%,最高为sug E(98.86%),emr E最低(68.18%),没有检测出qac E、qac F、qac G,检出率最高的可移动遗传元件介导耐药基因为qac EΔ1(19.31%)。肺炎克雷伯氏菌的染色体型耐药基因检出率为13.64%~28.41%,ydg E最高(28.14%),emr E检出率最低(13.64%),可移动遗传元件介导耐药基因sug E(p)检出率最高(6.82%),qac EΔ1、qac F、qac G基因未检出。测定大熊猫源大肠杆菌及肺炎克雷伯氏菌对消毒剂的耐药性,对圈养大熊猫消毒剂的规范使用,防控大熊猫细菌性疾病以及细菌对消毒剂耐药性有着重要意义。     

醋酸钙不动杆菌对22种抗生素的耐药性研究

目的：检测102株醋酸钙不动杆菌对22种抗生素的耐药性。方法：AMS法,结果：耐药率IMI,AMI,CIP和TIA为5．9％－9．8％,CFT,CFZ,TOB和MEZ为10．8－19．6％,TIC,PIP,CAR,CEN和TET为21．6％－39．2％,CFP,CFS,CFU,AZT和AMP为52．0－59．8％,CFX,CFA和NIT为82．4％－85．3％,CEP为92．2％,在痰液,伤口及其他标本分离醋酸钙不动杆菌的测定结果之间,有多种抗生素的敏感性差异有显著性（P＜0．050－0．001）,结论：结果与文献报道一致或接近。     

四川省猪源沙门氏菌及耐药性变迁调查

为了解四川地区近几年猪源沙门氏菌的耐药性变迁情况,本研究于2009—2014年间采集了四川省主要猪养殖场的肛拭子样本2660份,分离鉴定获得沙门氏菌151株,总分离率为5.68%,占主导的血清型为德尔卑沙门氏菌(60.26%)。采用微量肉汤稀释法测定分离菌株对13种抗菌药物的敏感性。药敏结果显示,沙门氏菌对氨苄西林、阿莫西林/克拉维酸、大观霉素、四环素、磺胺异噁唑、复方新诺明和恩诺沙星7种药物的耐药率最高,分别为94.7%、92.7%、93.4%、95.4%、91.4%、90.1%和86.8%,对头孢噻呋和多粘菌素E相对敏感。分析猪源沙门氏菌对不同抗菌药物的耐药性随年份的变迁情况,结果表明:2009—2014年猪源沙门氏菌对氨苄西林、阿莫西林/克拉维酸、大观霉素、四环素、磺胺异噁唑和复方新诺明6种药物的耐药率维持较高的平稳动态趋势,对头孢噻呋、庆大霉素、多西环素、氟苯尼考、恩诺沙星、氧氟沙星和多粘菌素E这7种药物的耐药率呈上升趋势,尤其对头孢噻呋、多西环素、氟苯尼考和氧氟沙星的耐药率上升显著。6年间多重耐药菌株比例维持在较高水平(>60%)。研究表明,四川地区猪源沙门氏菌的耐药情况十分严重,亟需加强养殖业抗菌药物的合理应用以控制耐药性发展。     

益生菌对16种抗生素的耐药性分析

目的评估市场常见益生菌对临床常用抗生素的体外耐药情况,为临床预防抗生素相关性肠道菌群紊乱和抗生素相关性腹泻提供参考。方法分离培养9种益生菌制剂中的20株益生菌,采用浓度梯度试条法(E-test)测定益生菌对16种抗生素的最低抑菌浓度(MIC)。结果细菌类益生菌对一些口服抗生素敏感,其中枯草芽胞杆菌、保加利亚乳杆菌、长双歧杆菌、婴儿双歧杆菌和嗜热链球菌对12种以上的抗生素敏感;屎肠球菌只对5种抗生素敏感,而真菌类益生菌如布拉酵母CNCM I-745则对16种抗生素均具有耐药性。结论细菌类益生菌和真菌类益生菌表现出不同的抗生素敏感性,在临床预防和治疗抗生素相关性腹泻时应注意益生菌与抗生素的合理使用。     

广州地区淋球菌抗生素耐药性及质粒图谱研究

目的：了解广州地区淋球菌对抗生素的耐药性及质粒图谱。方法：用琼脂稀释法测定5种抗生素对167株菌的最低抑菌浓度（MIC）及用破裂解法分析160株菌的质粒图谱。结果：167株淋球菌检出PPN9株（5．4％）,TRNG16株（9．6％）,环丙沙星耐药率达78．4％,头孢三嗪,壮观霉素未发现耐药菌株。160株菌其质粒谱型2．6Md 4.5Md(24.4%),2.6Md 4.5Md 24.5Md(20.6%),2.6Md 24.5Md(20.6%),4.5Md检出率（45％）24.5Md检出率（41．3％）。结论：表明了广州地区淋球菌抗生素的耐药性和质粒图谱,有助于淋病的治疗和防治。     

香芹酚联合头孢曲松对耐药性沙门氏菌的体外抗菌作

【背景】由于抗生素的滥用,使得细菌耐药性问题严峻,寻找解决耐药性细菌感染的治疗策略迫在眉睫。临床上,中药与抗菌药物联用在抗耐药性细菌感染方面效果显著。【目的】研究香芹酚联合头孢曲松对耐药性沙门氏菌(SJ2)的协同效应及机制。【方法】通过二倍肉汤稀释法测定香芹酚和头孢曲松对SJ2的最小抑菌浓度(minimal inhibitory concentration, MIC);通过棋盘法和生长曲线测定探究香芹酚和头孢曲松联合抗SJ2活性;通过膜电位检测、胞外碱性磷酸酶(alkaline phosphatase, AKP)含量测定、菌体内抗生素蓄积分析、细菌生存活力测定及扫描电镜研究香芹酚联合头孢曲松对SJ2细胞壁、细胞膜的影响。【结果】香芹酚和头孢曲松对SJ2的MIC分别为256μg/mL和2 048μg/mL;香芹酚联合头孢曲松对SJ2具有协同作用,其分级抑制浓度指数(fractional inhibitory concentration index, FICI)为0.375;香芹酚可协同头孢曲松使SJ2细胞膜出现去极化,显著增加AKP泄漏至胞外的量(P<0.05),显著增加菌体胞内抗...     

Antibiotic resistance and genotyping of clinical group B Salmonella isolated in Accra,Ghana

AIMS: The purpose of this study was to investigate the antibiotic resistance and clonal lineage of serogroup B Salmonella isolated from patients suspected of suffering from enteric fever in Accra, Ghana. METHODS AND RESULTS: Serogroup B Salmonella were isolated from blood (n=28), cerebral spinal fluid (CSF) (n=1), or urine (n=2), and identified based on standard biochemical testing and agglutinating antisera. Isolates were examined for their susceptibility to ampicillin, chloramphenicol, tetracycline and trimethoprim-sulfamethoxazole. Most of the isolates could be classified as multiple-drug resistant. Furthermore, the genetic location of resistance genes was shown to be on conjugative plasmids. Genetic fingerprinting by plasmid profiling, enterobacterial repetitive intergenic consensus (ERIC)-PCR, and repetitive element (REP)-PCR were performed to determine the diversity among the isolates. Plasmid profiling discriminated five unique groupings, while ERIC-PCR and REP-PCR resulted in two and three groupings, respectively. CONCLUSIONS: A high rate of antibiotic resistance was associated with the Salmonella isolates and the genes responsible for the resistance are located on conjugative plasmids. Also, there appears to be minimal diversity associated with the isolates. SIGNIFICANCE AND IMPACT OF THE STUDY: As a result of the increasing antibiotic resistance among bacteria of all genera, surveys to monitor microbial populations are critical to determine the extent of the problem. The inability to treat many infectious diseases with current antibiotic regimens should prompt the medical community to be more prudent with its antibiotic use.     

Salmonella serotypes isolated from pet chews in New Zealand

AIMS: To survey the prevalence of Salmonella in imported and domestic pet chews for assessing their potential in introducing novel pathogenic and antimicrobial resistant Salmonella serotype clones into New Zealand, and as vehicles of salmonellosis in the domestic home environment. METHODS AND RESULTS: Three hundred samples, each of imported and domestic pet chews, were examined bacteriologically for the presence of Salmonella. Salmonella cells in the pre-enrichment culture were concentrated by using Dynabeads, and then selective enrichment and plating were performed by a method described in the Bacteriological and Analytical Manual, USFDA. Salmonella was isolated from 16 (5.3%) of the imported and 20 (6.7%) of the domestic pet chews, but the prevalences of Salmonella in imported and domestic products were not significantly different. All Salmonella isolates were serotyped and genotyped by pulsed-field gel electrophoresis and antimicrobial susceptibility determined by the Clinical and Laboratory Standards Institute disc diffusion methods. Salmonella Borreze has never been recorded earlier in New Zealand and was detected from Australian raw hide. Three isolates of Salmonella London were resistant to ampicillin and gentamicin, and two isolates of Salmonella Infantis were resistant to nalidixic acid, one of which was also resistant to streptomycin. CONCLUSIONS: Novel pathogenic and antimicrobial-resistant Salmonella are being introduced into New Zealand through the import of pet chews. This indicates that pet chews are a potential source of exposure to Salmonella in the domestic home environment. SIGNIFICANCE AND IMPACT OF THE STUDY: Contaminated pet chews are potential sources of Salmonella infection for domestic pets, and humans are at risk of exposure either directly by contact through handling or inadvertently by cross-contamination of food or food-contact surfaces in home environments.     

Molecular characterization of antibiotic resistance in clinical Salmonella typhi isolated in Ghana

Fifty-eight clinical Salmonella typhi strains isolated from patients suspected of suffering from typhoid fever were obtained at the Korle-Bu Teaching Hospital and the Noguchi Memorial Institute for Medical Research, both located in Ghana, Africa. Each isolate was examined for susceptibility to ampicillin, chloramphenicol, streptomycin, tetracycline, and trimethoprim/sulfamethoxazole by the disk diffusion assay. Five of the isolates were resistant to all five antibiotics while 10 isolates were resistant to ampicillin, chloramphenicol, and trimethoprim/sulfamethoxazole, which are considered 'first line' antibiotics in the treatment of typhoid fever. Thirty-four isolates were resistant to at least one of the antibiotics tested and 62% of these isolates possessed conjugable plasmids belonging to incompatibility group IncHI. Ninety percent of the conjugable plasmids conferred a multiple drug-resistant phenotype on the strains harboring them. Additionally, 14 strains contained plasmids that were transformable and six of them encoded multiple drug resistance. Our findings indicate that multiple drug resistance to the 'first line' antibiotics in S. typhi may be more prevalent in Africa than previously thought.     

Prevalence of the multiple antibiotic resistance operon (marRAB) in the genus Salmonella

The multiple antibiotic resistance operon (marRAB) is a member of the multidrug resistance (mdr) systems. Similar to other mdr systems, this operon when induced encodes resistance to structurally and functionally unrelated antibiotics. marRAB has been shown to be conserved in the family Enterobacteriaceae, but within the genus Salmonella certain species appeared to be lacking this operon. To investigate how conserved the marRAB operon was in Salmonella, 30 veterinary isolates were examined by PCR, Southern blot, and dot blot analysis. Using DNA primers based on the marRAB operon of S. typhimurium, a predicted 2.3-kb amplicon resulted after PCR in 16 of the 30 organisms. The 2.3-kb DNA band from S. enteritidis was cloned and sequenced and shown to possess 99% sequence homology to marRAB from S. typhimurium. Using a labeled marRAB gene probe from S. enteritidis, Southern blot and dot blot analysis confirmed the presence of the operon in all 30 Salmonella species examined. Furthermore, when these isolates were induced with low levels of either tetracycline or chloramphenicol, increased antimicrobial resistance was observed to structurally and functionally unrelated antibiotics. Thus, the widespread occurrence of the marRAB locus in this genus prescribes judicious use of antimicrobials to avoid induction of a mdr phenotype.     

Antibiotic resistant Escherichia coli and Salmonella in Russian rooks (Corvus frugilegus) wintering in the Czech Republic

AIMS: To characterize antibiotic resistant Escherichia coli and Salmonella isolates in rooks wintering in the Czech Republic. METHODS AND RESULTS: Three hundred and sixty-three faeces samples from rooks were examined for antibiotic resistant Escherichia coli and Salmonella. Altogether 13.7%E. coli isolates were resistant to antimicrobial agents tested. The dominant type of resistance was to tetracycline. Resistant E. coli isolates were examined for antibiotic resistance genes and class 1 integrons. Five of 29 antibiotic resistant isolates possessed the int1 gene. Nine Salmonella isolates (2.5%) were found in rook faeces. All the isolates belonged to serotype Salmonella enterica serovar Enteritidis phage type PT8 and PT23. CONCLUSIONS: The study suggests that rooks can be infected by antibiotic resistant E. coli and Salmonella isolates, probably reflecting the presence of such isolates in their sources of food and/or water in the environment. SIGNIFICANCE AND IMPACT OF THE STUDY: Rooks can serve as reservoirs and vectors of antibiotic resistant E. coli and Salmonella isolates and potentially transmit these isolates over long distances.     

Competitive exclusion against Salmonella Enteritidis in layer chickens by yoghurt microbiota: impact on egg production, protection and yolk-antibody and cholesterol levels

AIMS: This work aims at studying the impact of competitive exclusion of Salmonella serotype Enteritidis infection in layer chickens, by microbiota of fresh and dried-modified yoghurt, on egg production and weight, protection against infection, and on yolk-antibody and cholesterol levels. METHODS AND RESULTS: Four groups of 27-week-old layer chickens were included in this study. After an initial enrofloxacin treatment, groups 1 and 2 were administered fresh or dry yoghurt (respectively) for 14 days. Groups 1, 2 and 3 were challenged intraoesophageally with Salm. Enteritidis, on the sixth day of yoghurt administration, while group 4 was left unchallenged and without yoghurt administration. No significant difference in percent infectivity of visceral organs with Salm. Enteritidis was observed between the groups. The yoghurt administered groups showed an early significant antibody response in their yolk on the seventh day postchallenge (P < 0.05) and highest egg production and weight. Finally, the egg yolk cholesterol concentration was higher in Salm. Enteritidis-challenged groups than that observed in the unchallenged group. CONCLUSIONS: The results point to the possible involvement of yoghurt administration in immunopotentiation and improvement of egg production and weight. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings warrant further research that could improve immunity and production in layer chickens infected with Salm. Enteritidis.     

Identification and characterization of class 1 integron resistance gene cassettes among Salmonella strains isolated from healthy humans in China

Twenty-three strains of Salmonella spp. isolated from healthy humans in Guangdong, China, were examined for their susceptibility to ten common antibiotics and the presence of antibiotic resistance integrons. All the strains were resistant to at least one antibiotic, and 4 strains were positive for the intI1 gene. Polymerase chain reaction using in-F and in-B primers showed the existence of amplicons of 1,009 bp in two, 1,664 bp in one, and 1,009 bp and 1,664 bp in one of the intI1 -positive isolates, respectively. Sequence analysis revealed that the 1,009-bp amplicon harbored gene cassette aadA2, conferring resistance to spectinomycin, and the 1,664-bp amplicon harbored genes aadA5 and dfr17, conferring resistance to spectinomycin, streptomycin and trimethoprim. Meanwhile the experiments of plasmid conjugation and Southern hybridization with intI1 as the DNA probe indicated that all the integrons found in these strains were chromosomal. Because the strains carrying class 1 integrons were isolated from healthy humans, it suggests the need for all-round surveillance of the antibiotic resistance of pathogens.