四川泡豇豆原料表皮附生乳酸菌抗生素抗性及抗性基因污染分析

以鲜豇豆为研究对象,利用生理生化特征、16S r RNA、抗生素抗性和抗性基因检测分析其表皮附生乳酸菌对氯霉素(CHL)、四环素(TET)和氨苄青霉素(AMP)的抗性与抗性基因。结果表明:从鲜豇豆表皮共分离得到182株乳酸菌,分别属于Weissella confuse(50.0%)、Weissella cibaria(15.4%)、Enterococcus sulfurous(7.1%)和Lactococcus lactis subsp.lactis(27.5%)。32株(17.6%)乳酸菌对CHL和TET有抗性,其中,6株W.confuse(3.3%)和4株L.lactis subsp.Lactis(2.1%)对CHL单一抗性,4株W.confuse(2.1%)和5株L.lactis subsp.Lactis(2.7%)对TET单一抗性,6株W.confuse(3.2%)、2株W.cibaria(1.1%)、1株E.sulfurous(0.5%)和4株L.lactis subsp.Lactis(2.1%)对CHL和TET二重抗性;编码外排泵基因efr A、efr B、nor B、nor C、nor E和sug E中,efr B的检出率最高,达31.3%,efr B和sug E的检出率最低,为3.1%。TET被检抗性基因tet(A)、tet(B)、tet(C)、tet(D)、tet(H)和tet(L)中,tet(C)的检出率最高,达90.9%,tet(B)的检出率最低,为4.5%。      

四川泡豇豆原料表皮附生乳酸菌抗生素抗性及抗性基因污染分析

以鲜豇豆为研究对象,利用生理生化特征、16S r RNA、抗生素抗性和抗性基因检测分析其表皮附生乳酸菌对氯霉素(CHL)、四环素(TET)和氨苄青霉素(AMP)的抗性与抗性基因。结果表明:从鲜豇豆表皮共分离得到182株乳酸菌,分别属于Weissella confuse(50.0%)、Weissella cibaria(15.4%)、Enterococcus sulfurous(7.1%)和Lactococcus lactis subsp.lactis(27.5%)。32株(17.6%)乳酸菌对CHL和TET有抗性,其中,6株W.confuse(3.3%)和4株L.lactis subsp.Lactis(2.1%)对CHL单一抗性,4株W.confuse(2.1%)和5株L.lactis subsp.Lactis(2.7%)对TET单一抗性,6株W.confuse(3.2%)、2株W.cibaria(1.1%)、1株E.sulfurous(0.5%)和4株L.lactis subsp.Lactis(2.1%)对CHL和TET二重抗性;编码外排泵基因efr A、efr B、nor B、nor C、nor E和sug E中,efr B的检出率最高,达31.3%,efr B和sug E的检出率最低,为3.1%。TET被检抗性基因tet(A)、tet(B)、tet(C)、tet(D)、tet(H)和tet(L)中,tet(C)的检出率最高,达90.9%,tet(B)的检出率最低,为4.5%。     

健康人体及保健品中乳酸菌和双歧杆菌的抗药性分析

目的:研究常用抗生素对来自健康人体和保健品的乳酸菌的影响.为构建乳酸菌和双歧杆菌食品级基因克隆和表达系统,为研制益生菌食品级微生态制剂而筛选无耐药性受体菌株和安全菌株.为乳酸菌的抗性筛选提供科学依据.方法:采用标准药敏纸片琼脂扩散法(K-B法),以金黄色葡萄球菌ATCC 25923为标准对照菌株,研究15株乳酸球菌,14株乳酸杆菌,5株双歧杆菌对8大类19种常用抗生素的敏感性.结果:15株乳酸球菌对氨苄西林、青霉素G、头孢霉素、头孢曲松、卡那霉素、四环素、红霉素、氯霉素均敏感或中度敏感:对磺胺、多黏菌素B均耐药;对其它抗生素表现出不同药敏性.14株乳杆菌都对氨苄西林、青霉素G、头孢霉素、卡那霉素、红霉素、四环素、氯霉素敏感或中度敏感;对磺胺、甲氧嘧啶、多黏菌素B均耐药:对其它表现出不同药敏性.5株双歧杆菌均对氨苄西林、青霉素G、头孢霉素、卡那霉素、甲氧苄氨嘧啶、红霉素、四环素、氯霉素敏感或中度敏感;对庆大霉素、阿米卡星、链霉素、磺胺、杆菌肽、环丙沙星均耐药;对其它抗生素表现出不同药敏性.     

高加索酸奶中乳酸菌的分离与鉴定

从自然发酵的5份酸奶样品中,通过平板划线等方法分离筛选乳酸菌。经形态特征,生理生化特性及糖发酵试验等,筛选到12株乳酸菌,分别为:乳杆菌7株,其中:3株德氏乳杆菌保加利亚亚种(Lactobacillus delbrueckii subsp.bulgaricus),3株瑞士乳杆菌(Lactobacillus hel-veticus),1株罗伊氏乳杆菌(Lactobacillus reuteri);乳酸球菌5株,包括3株嗜热链球菌(Streptococcus thermophilus),2株乳酸乳球菌乳脂亚种(Lactococcus lactis subsp.Cremoris)。     

乳酸菌对酸乳中霉菌的抑制作用研究

以酸乳中的腐败霉菌为指示菌,采用双层平板拮抗法对28株乳酸菌进行了抗霉菌筛选,得到了9株抗性菌株,分别属于戊糖乳杆菌(Lactobacillus pentosus)、乳酸乳球菌乳酸亚种(Lactococcus lactis subsp)以及植物乳杆菌(Lactobacillus plantarum)。对戊糖乳杆菌(Lactobacillus pentosus)ZJ28在不同培养条件下的抑菌能力进行了研究。结果表明,L.pentosus ZJ28在37℃下,恒温培养26~50 h,pH值为4.5或5.5时抗霉菌活性最好。     

食源性乳酸菌安全性的评价

从不同来源的样品中分离得到36 株乳酸菌,对其进行常见抗生素如氯霉素、四环素、红霉素抗药性分析,结果表明,26 株菌具有抗药性,其中4 株携带抗生素抗性基因,并且菌株Enterococcus faecium KN9所携带的四环素抗性基因tet（M）和tet（L）能通过接合作用,转移到受体菌Lactococcus lactis MG1614中。对这些乳酸菌产生有害代谢产物分析结果表明,部分菌株可以产生生物胺,所有的菌株都不产生硝基还原酶和偶氮还原酶。所以,本实验分离到的大部分乳酸菌是安全的,只有携带有抗四环素基因的E. faecium KN9具有潜在的安全隐患。因此,需要在食用前对乳酸菌进行安全性评价,以减少可能的安全隐患。     

内蒙古传统发酵乳制品中乳酸菌的分离及其耐药性研究

以内蒙古通辽地区自制的奶豆腐和酸性奶油（乌日莫）为材料,采用稀释涂布法从中分离乳酸菌,通过分子生物学技术对其进行菌种鉴定,并对其耐药性进行研究。结果表明,共分离纯化得到80株乳酸菌,经鉴定,归属于13个种,分别为短乳杆菌（Lactobacillus brevis）、粪肠球菌（Enterococcus faecalis）、乳酸片球菌（Lactobacillus acidilacticii）、乳酸乳球菌（Lactococcus lactis）、瑞士乳杆菌（Lactobacillus helveticus）、屎肠球菌（Enterococcus faecium）、戊糖片球菌（Pediococcus pentosaceus）、副干酪乳杆菌（Lactobacillus paracasei）、食二酸乳杆菌（Lactobacillus digitalis）、奥塔基乳杆菌（Lactobacillus ottaki）、耐久肠球菌（Enterococcus durans）、植物乳杆菌（Lactobacillus plantarum）、开菲尔乳杆菌（Lactobacillus kefir）,表现出高的生物多样性。80株乳酸菌对萘啶酸的耐药率为100%;对链霉素、环丙沙星、万古霉素的耐药率较高,分别为95.00%、80.00%、72.50%;对利福平和四环素的耐药率中等,分别为51.25%,45.00%;对红霉素、氯霉素的耐药率较低,分别为26.25%、23.75%。此外,76株（95%）的乳酸菌还具有多重耐药性。     

甘肃肃南牧区传统发酵乳中乳酸菌的分离鉴定

以5份甘肃肃南牧区传统发酵乳制品为原料进行乳酸菌的分离和鉴定,共分离出25株乳酸菌。对其中23株产乳酸的菌株利用形态、生理生化分析和碳水化合物发酵产酸实验进行鉴定。经鉴定得出23株分属为4个属,11个种。其中乳杆菌属[Lactobacillus(L.)]11株,乳球菌属(Lactococcus)6株,明串珠菌属(Leuconostoc)3株,肠球菌属(Enterococcus)3株。具体来说,L.plantarum6株、L.fermentum2株、L.brevis1株、L.minor2株、L.lactis subsp.lactis3株、L.lactis subsp.cermoris3株、L.mesenteroides1株、L.lactis2株、E.faecium1株、E.faecalis1株、E.durans1株。L.plantarum占分离菌株的26.1%,是优势菌株。      

四川泡菜发酵原料-灯笼辣椒附生乳酸菌的抗生素耐药性评估与耐药基因分析（英文）

以四川泡菜蔬菜原料——新鲜灯笼辣椒为对象,分析其表面附生乳酸菌Enterococcus mundtii(5株)、Enterococcus faecalis(2株)、Enterococcus hirae(5株)、Lactococcus lactis(7株)、Leuconostoc mesenteroides(2株)、Leuconostoc holzapfelii(3株)和Weissella cibaria(79株)对青霉素(penicillin,PEN)、红霉素(erythromycin,ERY)、四环素(tetracycline,TET)、链霉素(streptomycin,STR)和氯霉素(chloramphenicol,CHL)的抗生素耐药性和耐药基因分布,为制定合理的食品安全防控措施提供科学依据。研究表明:所有分离菌株均无PEN和ERY耐药性,其他种属部分菌株对TET、STR和CHL表现出单一、二重或三重耐药性。除E.hirae、E.faecalis和L.holzapfelii部分菌株对STR表现出单一耐药性外,所有L.mesenteroide菌株只表现出了STR单一耐药性;STR和TET、STR和CHL二重耐药菌株在E.faecalis、E.hirae、L.lactis和W.cibaria分离菌株中都有发现,但是STR、TET、CHL三重耐药菌株仅在W.cibaria中发现。聚合酶链式反应检测发现:除基因nor A、sep A、tet(A)、tet(O)和aac(6’)-aph(2’)未被检出外,其他耐药菌株都有相应1个或多个耐药基因被检出。多重耐药外排泵基因efr A、tol C、nor C、sug E和mdf A较核糖体蛋白质保护和酶修饰基因检出率高,分别达到了49%、41%、48%、41%和47%。虽然辣椒表面附生乳酸菌的抗生素耐药基因在四川泡菜发酵过程中的扩散行为需要进一步研究,但根据食品加工过程安全规范标准,也应关注其表面附生的乳酸菌抗生素耐药性存在的潜在食品安全问题。     

益生乳杆菌质粒抗生素抗性基因与其耐药性的相关性探讨

采用K-B药敏纸片法检测了5株具有潜在益生乳杆菌的耐药性,通过质粒消除,分析了菌株质粒与耐药性之间的联系,应用PCR确定了质粒决定的耐药基因。5株乳杆菌对万古霉素、多粘菌素B以及链霉素等7种抗生素普遍表现出抗性,但主要对四环素敏感。采用SDS与SDS-高温两种方法消除戊糖乳杆菌CH8的质粒后,菌株CH8表现出头孢噻吩和氯霉素敏感性。设计β-内酰胺类抗性基因blr、ECP-1569和nps-1以及氯霉素抗性基因cmlA、cat和cmlA1的引物进行PCR,与目的产物测序比对表明,戊糖乳杆菌CH8的质粒上含有β-内酰胺类抗性基因blr,该基因与其头孢噻吩抗性有关。该研究为探讨乳酸菌的抗药基因转移性提供了前期基础,有助于益生乳杆菌安全性评价体系的建立与完善。      

四株乳酸菌的加工性能

选用红霉素（Erythromycin）、氯霉素（Chloramphenicol）、四环素（Tetracycline）等常见的抗生素,用于测定4株乳酸菌在不同抗生素中的最高耐受浓度,并比较4株乳酸菌体外模拟胃肠道环境的耐受性。实验结果表明:4株乳酸菌均有不同程度的抗生素抗性作用。肠膜明串珠菌肠膜亚种（Leuconostoc mesenteroides subsp.mesenteroides）BD1710在四环素和氯霉素中的最高耐受浓度分别为1、2μg/m L;植物乳杆菌ST-Ⅲ（Lactobacillus plantarum ST-Ⅲ）在氯霉素中的最高耐受浓度为1μg/m L;干酪乳杆菌BD0054（Lactobacillus casei BD0054）在四环素和氯霉素中的最高耐受浓度都为1μg/m L;干酪乳杆菌LC2W（Lactobacillus casei LC2W）对氯霉素的最高耐受浓度为1μg/m L,4株乳酸菌都不耐红霉素。模拟胃肠道环境实验表明,在胃液环境下,ST-Ⅲ、LC2W的生长不受影响,但BD0054受到影响较大;在肠液环境下,ST-Ⅲ、BD1710生长不受影响,LC2W肠道不耐。高温处理过程,EDTA会加重菌体的损伤程度。冷冻干燥处理,存活率在90%⁹5%之间。      

四川泡菜发酵原料-灯笼辣椒附生乳酸菌的抗生素耐药性评估与耐药基因分析

以四川泡菜蔬菜原料——新鲜灯笼辣椒为对象,分析其表面附生乳酸菌Enterococcus mundtii（5 株）、Enterococcus faecalis（2 株）、Enterococcus hirae（5 株）、Lactococcus lactis（7 株）、Leuconostocmesenteroides（2 株）、Leuconostoc holzapfelii（3 株）和Weissella cibaria（79 株）对青霉素（penicillin,PEN）、红霉素（erythromycin,ERY）、四环素（tetracycline,TET）、链霉素（streptomycin,STR）和氯霉素（chloramphenicol,CHL）的抗生素耐药性和耐药基因分布,为制定合理的食品安全防控措施提供科学依据。研究表明：所有分离菌株均无PEN和ERY耐药性,其他种属部分菌株对TET、STR和CHL表现出单一、二重或三重耐药性。除E. hirae、E. faecalis和L. holzapfelii部分菌株对STR表现出单一耐药性外,所有L. mesenteroide菌株只表现出了STR单一耐药性;STR和TET、STR和CHL二重耐药菌株在E. faecalis、E. hirae、L. lactis和W. cibaria分离菌株中都有发现,但是STR、TET、CHL三重耐药菌株仅在W. cibaria中发现。聚合酶链式反应检测发现：除基因norA、sepA、tet(A)、tet(O)和aac(6’)-aph(2’)未被检出外,其他耐药菌株都有相应1 个或多个耐药基因被检出。多重耐药外排泵基因efrA、tolC、norC、sugE和mdfA较核糖体蛋白质保护和酶修饰基因检出率高,分别达到了49%、41%、48%、41%和47%。虽然辣椒表面附生乳酸菌的抗生素耐药基因在四川泡菜发酵过程中的扩散行为需要进一步研究,但根据食品加工过程安全规范标准,也应关注其表面附生的乳酸菌抗生素耐药性存在的潜在食品安全问题。     

传统发酵食品中乳酸菌的安全性评估

本文以传统发酵食品中分离的38株乳酸菌(lactic acid bacteria,LAB)为研究对象,利用溶血试验、明胶液化试验和吲哚试验测试受试菌株相关产毒代谢产物,采用K-B纸片法检测菌株对25种抗生素的耐药表型,运用聚合酶链式反应检测菌株携带的耐药基因.结果表明:38株乳酸菌的溶血试验、明胶液化试验和吲哚试验均为...     

Antibiotic susceptibility profiles of new probiotic Lactobacillus and Bifidobacterium strains

The antimicrobial susceptibilities and presence of plasmids in four new probiotic lactic acid bacteria (LAB) strains, Lactobacillus rhamnosus HN001 (DR20) HN067, Lactobacillus acidophilus HN017 and Bifidobacterium lactis HN019 (DR10), were determined. Resistance to 18 commonly used antibiotics was assessed by disk diffusion. The three Lactobacillus strains had similar antibiotic susceptibility profiles to those of Lactobacillus plantarum strain HN045 and two commercial probiotic Lactobacillus strains, GG and LA-1. The B. lactis strain HN019 had a similar profile to three commercial probiotic B. lactis strains (Bb12, HN049 and HN098). All 10 strains were sensitive to the Gram-positive spectrum antibiotics erythromycin and novobiocin, the broad-spectrum antibiotics rifampicin, spectinomycin, tetracycline and chloramphenicol and the beta-lactam antibiotics penicillin, ampicillin and cephalothin. By contrast, most strains were resistant to the Gram-negative spectrum antibiotics fusidic acid, nalidixic acid and polymyxin B and the aminoglycosides neomycin, gentamicin, kanamycin and streptomycin. All three L. rhamnosus strains (HN001, HN067 and GG) were resistant to vancomycin and several strains were also resistant to cloxacillin. Of the four new probiotic strains, only L. rhamnosus HN001 contained plasmids; however, a plasmid-free derivative of HN001 had the same antibiotic susceptibility profile as the parent strain.     

Characterization of the lactic acid bacteria in ewe's milk and cheese from northwest Argentina

Indigenous lactic acid bacteria in ewe's milk and artisanal cheese were studied in four samples of fresh raw milk and four 1-month-old cheeses from the provinces of northwest Argentina. Mean growth counts on M17, MRS, and MSE agar media did not show significant differences (P < 0.05) in raw milk and cheeses. Isolates of lactic acid bacteria from milk were identified as Enterococcus (48%), lactococci (14%), leuconostocs (8%), and lactobacilli (30%). All lactococci were identified as Lactococcus lactis (subsp. lactis and subsp. cremoris). Lactobacilli were identified as Lactobacillus plantarum (92%) and Lactobacillus acidophilus (8%). Enterococci (59%) and lactobacilli (41%) were isolated from cheeses. L. plantarum (93%), L. acidophilus (5%), and Lactobacillus casei (2%) were most frequently isolated. L. lactis subsp. lactis biovar diacetylactis strains were considered as fast acid producers. L. lactis subsp. cremoris strains were slow acid producers. L. plantarum and L. casei strains identified from the cheeses showed slow acid production. The majority of the lactobacilli and Lactococcus lactis strains utilized citrate and produced diacetyl and acetoin in milk. Enzyme activities (API-ZYM tests) of lactococci were low, but activities of L. plantarum strains were considerably higher. The predominance of L. plantarum in artisanal cheese is probably important in the ripening of these cheeses due to their physiological and biochemical characteristics.     

Antimicrobial potential of immobilized Lactococcus lactis subsp. lactis ATCC 11454 against selected bacteria

Immobilization of living cells of lactic acid bacteria could be an alternative or complementary method of immobilizing organic acids and bacteriocins and inhibit undesirable bacteria in foods. This study evaluated the inhibition potential of immobilized Lactococcus lactis subsp. lactis ATCC 11454 on selected bacteria by a modified method of the agar spot test. L. lactis was immobilized in calcium alginate (1 to 2%)-whey protein concentrate (0 and 1%) beads. The antimicrobial potential of immobilized L. lactis was evaluated in microbiological media against pathogenic bacteria (Escherichia coli, Salmonella, and Staphylococcus aureus) or Pseudomonas putida, a natural meat contaminant, and against seven gram-positive bacteria used as indicator strains. Results obtained in this study indicated that immobilized L. lactis inhibited the growth of S. aureus, Enterococcus faecalis, Enterococcus faecium, Lactobacillus curvatus, Lactobacillus sakei, Kocuria varians, and Pediococcus acidilactici. Only 4 h of incubation at 35 degrees C resulted in a clear inhibition zone around the beads that increased with time. With the addition of 10 mM of a chelating agent (EDTA) to the media, results showed growth inhibition of E. coli; however, P. putida and Salmonella Typhi were unaffected by this treatment. These results indicate that immobilized lactic acid bacteria strains can be successfully used to produce nisin and inhibit bacterial growth in semisolid synthetic media.     

酸和盐胁迫对乳酸菌活性的影响

该研究以13株食品常见乳酸菌为研究对象,考察酸和盐胁迫对其活性的影响,探究这13株乳酸菌的最适酸、盐生长条件及耐酸、耐盐能力。结果表明,适当的低盐浓度（1%、2%）对部分乳酸菌的生长有促进作用,植物乳杆菌和副干酪乳杆菌的最适盐浓度为1%,保加利亚乳杆菌和嗜酸乳杆菌的最适盐浓度为2%;盐浓度进一步增大,13株乳酸菌的生长受到抑制;盐浓度为10%时,除戊糖片球菌、乳酸乳球菌乳酸亚种、嗜热链球菌和乳酸片球菌外,其余9株菌的生长抑制率都＞95%。当pH值为5～7时,13株乳酸菌都生长良好,pH=6时,7株乳酸菌活性均最高;随着酸胁迫的增强,13株乳酸菌生长受抑制程度增大;pH为1～3时,13株乳酸菌的生长抑制率都＞90%,高盐及高酸对9株杆菌的抑制作用强于4株球菌。