Antimicrobial resistant genes associated with Salmonella from retail meats and street foods

We examined the antimicrobial resistance of Salmonella isolates from 300 meat products (raw beef, chicken meat and street foods). A total of 88 non-duplicate Salmonella from 66 (22.0%) retail meat and 22 (7.5%) street food samples were recovered and 11 serovars were identified. Among the 88 Salmonella isolates, the highest resistance was to tetracycline (73.8%), followed by sulfonamide (63.6%), streptomycin (57.9%), nalidixic acid (44.3%), trimethoprim–sulfamethoxazole (19.3%), ampicillin (17.0%), chloramphenicol (10.2%), cephalotin (8.0%), kanamycin (6.8%), ciprofloxacin (2.2%) gentamycin (2.2%), cefoxitin (2.2%), amoxicillin–clavulanate (1.0%) and amikacin (1.0%). Sixty-seven percent of the isolates (59/88) were multidrug resistant (MDR). Ten out of 17 resistance genes (bla_TEM-1, strA, strB, aadA, sulI, sulII, tetA, tetB, floR, cmlA) were detected. Twelve of the 59 MDR Salmonella isolates from serovars Typhimurium (6), Newport (3), Agona (1), Albany (1) and Weltevreden (1) had class 1 integrons. The gene cassettes identified were dfrA1, dfrV, dfrA12, aadA2, sul1 genes and an open reading frame orfC of unknown function. Four integron-positive isolates could transfer resistance phenotypes to the recipient strain, E. coli J53 via conjugation. These data revealed that the Salmonella isolates recovered from the retail meats and cooked street foods were resistant to multiple antimicrobials, which can be transmitted to humans through food products. The occurrence of mobile genetic elements such as integrons reiterates the roles of food of animal origins as a reservoir of MDR Salmonella.     

烟台地区食源性疾病中副溶血性弧菌的病原特征及溯源分析

目的了解烟台地区引发食物中毒的副溶血性弧菌分离菌株的主要血清型、抗生素耐药情况、致病力的强弱以及传播流行趋势。方法对2017～2019年11起食物中毒爆发事件中分离的14株副溶血性弧菌进行血清分型;采用微量肉汤稀释法进行药敏试验;采用PCR技术检测毒力基因不耐热溶血素(tld)、耐药直接溶血素(tdh)和溶血相关溶血素(trh);采用脉冲场电泳(pulse-field gel electrophoresis,PFGE)进行分子分型的溯源分析。结果引发烟台地区食物中毒爆发的副溶血性弧菌血清型多样,但以O3:K6型为主(28.6%),分离菌株中11株(78.6%)表现为tdh阳性和trh阴性,3株(21.4%)未携带tdh和trh基因,对头孢唑啉(71.4%)、多粘菌素E(57.1%)、美罗培南和阿莫西林/克拉维酸(7.1%)均出现耐药情况,对氨苄西林(21.4%)、多粘菌素B(14.3%)为中度敏感,耐药谱显示没有出现对3种以上同时耐药的情况,但是42.9%的分离菌株对2种同时耐药情况。结论烟台地区引发食物中毒爆发的副溶血性弧菌以O3:K6型为主,主要携带tdh,对头孢唑啉耐药和多粘菌素耐药普遍存在,具有聚集性爆发的风险,因此加强腹泻病的监测,充分利用国家致病菌识别网集中进行本地区分离菌株的病原特征研究是防控食物中毒爆发的有效手段。     

High prevalence of antimicrobial‐resistant Escherichia coli from animals at slaughter: a food safety risk

BACKGROUND: There has been concern about the increase of antimicrobial resistant bacteria and protection of animal and public health, along with food safety. In the present study, we evaluate the incidence of antimicrobial resistance among 192 strains of Escherichia coli isolated from faecal samples of healthy food‐producing animals at slaughter in Portugal. RESULTS: Ninety‐seven % of the pig isolates, 74% from sheep and 55% from cattle were resistant to one or more antimicrobial agents, with the resistances to ampicillin, streptomycin, tetracycline and trimethoprim–sulfamethoxazole the most common phenotype detected. Genes encoding resistance to antimicrobial agents were detected in most of the resistant isolates. Ninety‐three % of the resistant isolates were included in the A or B1 phylogenetic groups, and the virulence gene fimA (alone or in association with papC or aer genes) was detected in 137 of the resistant isolates. Five isolates from pigs belonging to phylogroup B2 and D were resistant to five different antimicrobial agents. CONCLUSION: Our data shows a high percentage of antibiotic resistance in E. coli isolates from food animals, and raises important questions in the potential impact of antibiotic use in animals and the possible transmission of resistant bacteria to humans through the food chain. © 2012 Society of Chemical Industry     

广州市白云口岸航空食品中金黄色葡萄球菌凝固酶基因分型及肠毒素基因研究

目的对2013—2015年从广州市白云口岸航空食品中分离的金黄色葡萄球菌进行基因分型研究,为食源性金黄色葡萄球菌分子溯源提供基础数据。方法以血浆凝固酶和肠毒素为目标基因,采用聚合酶链式反应(PCR)方法对9株金黄色葡萄球菌进行基因分型,其中6株为航空食品分离株,1株为配餐车间大门手拭分离株,2株为标准菌株。肠毒素基因检测包括5种传统肠毒素基因(sea、seb、sec、sed、see)和6种新型肠毒素基因(ser、seg、seh、sei、sej、sep)。结果 6株航空食品分离株的血浆凝固酶基因扩增分型结果为2个PCR型,酶切后得3种亚型;肠毒素基因检测结果显示有2株航空食品分离株含有肠毒素基因,检出率为33.3%(2/6),检出的基因为2种传统肠毒素基因(sec、sed)和4种新型肠毒素基因(ser、seg、sei、sej),均同时携带2种以上肠毒素基因。结论血浆凝固酶基因扩增分型结果显示,不同时间、不同采集地点存在相同的基因型,提示金黄色葡萄球菌存在交叉污染的可能性;航空食品分离株共检出6种肠毒素基因,提示金黄色葡萄球菌基因型多样性,应加强其他新型肠毒素基因检测。     

酿酒酵母的基因改良

酿酒酵母的发酵性能直接影响到酒的质量与生产成本。利用基因工程改良酿酒酵母可提高其生产性能。基因改良酿酒酵母的研究和应用有：①增加酿酒酵母发酵性能的基因改良，如：构建含α-乙酰乳酸脱羧酶基因的低双乙酰工程酵母；含乙醇乙酰酶基因的高生香工程酵母；含糖化酶、葡聚糖酶等基因的高发酵度工程酵母；高絮凝性工程酵母和嗜杀酵母。②增强或缺失酵母自身基因的菌种改良，如构建高级醇低生成量的工程酵母和构建双乙酰低生成量的工程酵母。     

食源性单核细胞增生李斯特菌四环素、红霉素耐药基因研究

研究了食源性单核细胞增生李斯特菌四环素、红霉素耐药基因的分布状况及和耐药表型的关系。采用微量肉汤稀释法对2005~2007年河北省疾病预防控制中心分离到的食源性单核细胞增生李斯特菌株进行四环素、红霉素药敏实验;应用PCR方法对实验菌株进行四环素耐药基因tet(M)、tet(S)、tet(L)、tet(K)、tet(B)、及与tet(M)基因关系密切的转座子Tn916、红霉素核糖体甲基化酶基因ermB、ermC、及与ermB基因关系密切的转座子Tn917检测,对阳性样本序列进行鉴定分析;应用血清学分型、脉冲场凝胶电泳(PFGE)、及脂肪酸聚类分析方法分析四环素耐药菌株之间的相关性,确定基因型和多态性。结果表明,91株单核细胞增生李斯特菌四环素敏感77株、耐药14株;红霉素敏感89株、耐药2株,其中包含1株菌同时交叉耐药四环素和红霉素;14株四环素耐药株中含tet(M)基因的13株,在13株tet(M)基因阳性菌中,tet(M)位于Tn916转座子上的9株;1株同时交叉耐药四环素、红霉素菌同时携带tet(S)、ermB基因;ermC基因、转座子Tn917均为阴性;四环素、红霉素敏感株中未检测到上述任何耐药基因。14株四环素耐药菌株血清型分布以1/2a型为主(n=12),部分菌株PFGE、脂肪酸分型完全一致。食源性单核细胞增生李斯特菌获得tet(M)基因是耐四环素的主要机制之一,具有水平传播耐药基因能力的接合型转座子Tn916与该菌四环素耐药播散有直接关系;ermB基因介导的核糖体靶位点改变存在食源性单核细胞增生李斯特菌红霉素耐药株中;PFGE基因型结合脂肪酸聚类分析能够用来分析菌株之间的相关性。     

Bioinformatics of Differentially Expressed Genes in Phorbol 12-Myristate 13-Acetate-Induced Megakaryocytic Differentiation of K562 Cells by Microarray Analysis

Megakaryocytes are large hematopoietic cells present in the bone marrow cavity, comprising less than 0.1% of all bone marrow cells. Despite their small number, megakaryocytes play important roles in blood coagulation, inflammatory responses, and platelet production. However, little is known about changes in gene expression during megakaryocyte maturation. Here we identified the genes whose expression was changed during K562 leukemia cell differentiation into megakaryocytes using an Affymetrix GeneChip microarray to determine the multifunctionality of megakaryocytes. K562 cells were differentiated into mature megakaryocytes by treatment for 7 days with phorbol 12-myristate 13-acetate, and a microarray was performed using RNA obtained from both types of cells. The expression of 44,629 genes was compared between K562 cells and mature megakaryocytes, and 954 differentially expressed genes (DEGs) were selected based on a p-value < 0.05 and a fold change >2. The DEGs was further functionally classified using five major megakaryocyte function-associated clusters—inflammatory response, angiogenesis, cell migration, extracellular matrix, and secretion. Furthermore, interaction analysis based on the STRING database was used to generate interactions between the proteins translated from the DEGs. This study provides information on the bioinformatics of the DEGs in mature megakaryocytes after K562 cell differentiation.     

花生miRNA与其靶基因的生物信息学预测

本研究利用生物信息学方法预测花生中的miRNA及其靶基因。将miRBase注册的已知植物miRNA与花生EST和GSS数据库进行比对搜索,筛选得到13条miRNA,进一步预测得到20个靶基因。分析结果显示大多数靶基因属于转录因子,调控植物的生长发育等多种生物过程。     

Characterization of Antibiotic Resistance Genes from Lactobacillus Isolated from Traditional Dairy Products

Lactobacilli are widely used as starter cultures or probiotics in yoghurt, cheese, beer, wine, pickles, preserved food, and silage. They are generally recognized as safe (GRAS). However, recent studies have shown that some lactic acid bacteria (LAB) strains carry antibiotic resistance genes and are resistant to antibiotics. Some of them may even transfer their intrinsic antibiotic resistance genes to other LAB or pathogens via horizontal gene transfer, thus threatening human health. A total of 33 Lactobacillus strains was isolated from fermented milk collected from different areas of China. We analyzed (1) their levels of antibiotic resistance using a standardized dilution method, (2) their antibiotic resistance gene profiles by polymerase chain reaction (PCR) using gene‐specific primers, and (3) the transferability of some of the detected resistance markers by a filter mating assay. All Lactobacillus strains were found to be resistant to vancomycin, but susceptible to gentamicin, linezolid, neomycin, erythromycin, and clindamycin. Their susceptibilities to tetracycline, kanamycin, ciprofloxacin, streptomycin, quinupristin/dalfopristin, trimethoprim, ampicillin, rifampicin, and chloramphenicol was different. Results from our PCR analysis revealed 19 vancomycin, 10 ciprofloxacin, and 1 tetracycline‐resistant bacteria that carried the van(X), van(E), gyr(A), and tet(M) genes, respectively. Finally, no transferal of the monitored antibiotic resistance genes was observed in the filter mating assay. Taken together, our study generated the antibiotic resistance profiles of some milk‐originated lactobacilli isolates and preliminarily assessed their risk of transferring antibiotic gene to other bacteria. The study may provide important data concerning the safe use of LAB.