水环境中耐热大肠菌群的抗生素耐药性与质粒谱研究

用滤膜法、mFC培养基从5种水体中分离出疑似耐热大肠菌162株,用API微生物分析系统鉴定到种,以Kirby-Bauer法分析其对人畜常用10种抗生素的耐药性,碱裂解法小量制备各菌株质粒DNA做质粒谱分析.结果表明,埃希氏大肠杆菌为优势菌,占分离菌总数的96.3%.除分离自泉水的3株外,其它菌株都对3种及3种以上抗生素耐药,多重耐药率为98.1%.不同水体分离菌株对氧氟沙星、诺氟沙星、庆大霉素、丁胺卡那霉素、链霉素的耐药率有显著性差异(P<0.005).92株菌(56.8%)提取到大小为0.90～158.83kb、数量为1～6个的质粒,有81种质粒谱型.70株(43.2%)未提取到质粒的细菌中有67株为多重耐药.具有相同质粒谱型的菌株耐药谱都不相同.未发现质粒谱与抗生素耐药性间有明显相关性.图1表3参15     

不同补给水源的城市人工湖中异养菌耐药性及耐药菌种属分布研究

为了阐明不同补给水源的城市人工湖中异养菌耐药状况和耐药菌种属分布特征,选取分别以地表水和再生水为补水来源的XQ湖和FQ湖为代表进行研究。从2018年4—11月逐月采集水样,考察了各水样中对氨苄西林(AMP)、磺胺甲恶唑(SMZ)和四环素(TET)这3种不同种类抗生素具有耐药性的异养菌含量,并对分离菌株的耐药表型、耐药菌株的种属分布以及水质理化指标进行了分析。结果表明,2处人工湖中的AMP耐药菌和SMZ耐药菌含量约为10~2～10~4CFU·(100 m L)~(-1),而TET耐药菌含量则约10~1～10~3CFU·(100 m L)~(-1)。分离出的84株耐药菌归属于19个种,其中蜡状芽孢杆菌、大肠埃希氏菌、维氏气单胞菌、豚鼠气单胞菌和鸟氨酸拉乌尔菌为2个湖共有耐药菌。71.4%的耐药菌都是对AMP单一耐药,以蜡状芽孢杆菌为主。由于具有固有耐药性的细菌在分离出的耐药菌中占比很低,获得性耐药很可能在城市人工湖中异养菌耐药性的发展上发挥了主要贡献作用。地表水补水的XQ湖和再生水补水的FQ湖在总异养菌含量、耐药菌含量和检出率上均无显著差异。分离来源对16株耐药性气单胞菌的聚类无明显影响。     

Gastrointestinal Bacterial Transmission among Humans,Mountain Gorillas,and Livestock in Bwindi Impenetrable National Park,Uganda

Abstract: Habitat overlap can increase the risks of anthroponotic and zoonotic pathogen transmission between humans, livestock, and wild apes. We collected Escherichia coli bacteria from humans, livestock, and mountain gorillas (Gorilla gorilla beringei) in Bwindi Impenetrable National Park, Uganda, from May to August 2005 to examine whether habitat overlap influences rates and patterns of pathogen transmission between humans and apes and whether livestock might facilitate transmission. We genotyped 496 E. coli isolates with repetitive extragenic palindromic polymerase chain reaction fingerprinting and measured susceptibility to 11 antibiotics with the disc‐diffusion method. We conducted population genetic analyses to examine genetic differences among populations of bacteria from different hosts and locations. Gorilla populations that overlapped in their use of habitat at high rates with people and livestock harbored E. coli that were genetically similar to E. coli from those people and livestock, whereas E. coli from gorillas that did not overlap in their use of habitats with people and livestock were more distantly related to human or livestock bacteria. Thirty‐five percent of isolates from humans, 27% of isolates from livestock, and 17% of isolates from gorillas were clinically resistant to at least one antibiotic used by local people, and the proportion of individual gorillas harboring resistant isolates declined across populations in proportion to decreasing degrees of habitat overlap with humans. These patterns of genetic similarity and antibiotic resistance among E. coli from populations of apes, humans, and livestock indicate that habitat overlap between species affects the dynamics of gastrointestinal bacterial transmission, perhaps through domestic animal intermediates and the physical environment. Limiting such transmission would benefit human and domestic animal health and ape conservation.     

畜禽养殖场排放物病原微生物危险性调查

以粪大肠菌群和沙门菌为指标,采集江苏省内10家不同养殖类型畜禽养殖场的排放物及其周边水、土样共37个进行观察与计数,同时分析排放污水中大肠杆菌的耐药性以及禽畜排泄物对土壤和水体耐药细菌数的影响.结果表明:调查的10家养殖场中有9家粪便未经无害化处理直接排放到水体或施于农田,且排放物的粪大肠菌群数全部严重超标,沙门菌检出率达19%,施新鲜粪肥的土壤中粪大肠菌群数(MPN,最大可能数)在105 g-1以上,水体中分离出的大肠杆菌表现出多重耐药性,抗生素抗性细菌总数也远高于未施新鲜粪肥的土壤及水体.     

土壤环境中四环素类抗生素残留及抗性基因污染的研究进展(Tetracycline Residues and Tetracycline Resistance Gene Pollution in Soil: A Review)

近年来,致病菌耐药性的增加和扩散已成为全世界关注的热点问题,而人类医疗和畜禽养殖业抗生素的滥用正不断加剧这一问题.众多研究表明土壤环境作为一个巨大的抗性基因储存库,在抗性微生物和抗性基因的传播中起重要的作用,具有潜在的生态与健康风险.在总结国内外最新研究基础上,对土壤环境中典型抗生素-四环素类抗生素的主要污染源以及其在土壤中的基本环境行为等进行了分析,并探讨了土壤中四环素类抗性基因的来源、迁移和扩散及分子检测手段等问题.我国作为抗生素的生产和消费大国,抗生素污染问题较其他国家更为严重,而国内相关研究才刚刚起步,迫切需要开展有关环境中抗生素和抗生素抗性基因污染的系统研究.     

磺胺和四环素类抗生素对大肠杆菌联合突变效应的研究

抗生素的滥用造成的环境安全问题已不可忽视,关于抗生素联合毒性效应研究较多,但联合突变效应研究较少。因此,本文以大肠杆菌为受试生物,研究了2种磺胺类抗生素(磺胺氯哒嗪(SCP)、磺胺二甲嘧啶(SMZ))和3种四环素类抗生素(二甲胺四环素(MH)、盐酸四环素(TH)和盐酸强力霉素(DH))单一及联合暴露时对大肠杆菌的突变效应。结果表明:在单一暴露下,磺胺类抗生素会促进大肠杆菌的突变效应,四环素类抗生素没有明显的促进作用;联合暴露下,磺胺类抗生素对大肠杆菌突变效应为相加,磺胺和四环素类抗生素对大肠杆菌突变效应为拮抗。本研究初步探索了抗生素对大肠杆菌的联合致突变风险,为今后环境中抗生素混合暴露的生态风险评价和抗生素污染控制标准制定提供一种理论支撑。     

莱州湾近岸海域中典型抗生素与抗性细菌分布特征及其内在相关性

为了揭示海陆衔接区环境中抗生素与抗性细菌分布特征及其内在相关性,以莱州湾及其主要入海河流为研究区域,利用HPLC-MS/MS分析样品中15种磺胺类抗生素(SAs)和6种喹诺酮类抗生素(QNs)的浓度,并通过改良的Method 1604(US EPA)评估海水与沉积物中2种典型水传病原微生物大肠杆菌(E.coli)与金黄色葡萄球菌(S.aureus)的抗生素抗性水平,进而探讨该区域水体中抗性菌株的分布特点以及微生物抗性率与相应抗生素浓度的相关性。结果显示,莱州湾水体与沉积物中普遍存在磺胺与喹诺酮类抗生素残留及抗性污染问题。两大类抗生素在水体中平均残留浓度分别为3.89 ng·L~(-1)(SAs)和234.68ng·L~(-1)(QNs),在沉积物中分别为0.91 ng·g~(-1)(SAs)和49.37 ng·g~(-1)(QNs),且分布特征基本呈现自河流向海洋逐渐递减的趋势,说明河流输入是莱州湾抗生素污染的主要来源。在水体中,具有磺胺类抗性的E.coli和S.aureus平均检出量分别达到2 018和4 683 CFU·L~(-1),抗性率范围分别在0%~37.3%和10.6%~45.8%之间;而2种喹诺酮类抗性病原微生物的平均检出量则相对较低,分别为1 315 CFU·L~(-1)(E.coli)和1 461 CFU·L~(-1)(S.aureus),抗性率分别为0%~50.0%和0%~20.8%;此外,相比于E.coli,S.aureus为沉积物中的主要抗性病原微生物,磺胺与喹诺酮类抗性S.aureus检出率均高于80%,平均检出量分别为24CFU·g~(-1)和18 CFU·g~(-1)。相关性分析表明,莱州湾近岸海域水体中磺胺类抗生素浓度与磺胺类抗性微生物总量之间具有良好的线性关系,然而其与微生物抗性率之间并未表现出相似的规律,说明近岸海洋环境中抗生素的残留量不是影响抗性菌株丰度的唯一因素。     

鸡、猪粪便源大肠杆菌对4种抗生素耐药性的比较

以琼脂稀释法对67株鸡源大肠杆菌及61株猪源大肠杆菌进行最低抑菌浓度(MIC)测定,并以SPSS分别进行卡方检验和Probit模型估算,分析不同来源大肠杆菌耐药差异显著性及不同抗生素的半数抑菌浓度(MIC50),以期对不同畜禽粪便来源的大肠杆菌耐药差异进行详细准确的探讨。结果显示,鸡源大肠杆菌对头孢噻肟、诺氟沙星、环丙沙星和四环素耐药率分别为98.51%、68.66%、56.72%和100%,猪源大肠杆菌对头孢噻肟耐药率为88.52%,对诺氟沙星、环丙沙星和四环素则100%耐药。除四环素,鸡源、猪源大肠杆菌对诺氟沙星、环丙沙星、头孢噻肟的耐受差异显著(P0.05),51%的鸡源大肠杆菌和89%的猪源大肠杆菌均呈4重耐药。SPSS分析结果表明,Probit模型估算结果优于当前MIC50常规计算方法,头孢噻肟、诺氟沙星、环丙沙星和四环素对鸡源大肠杆菌MIC50分别为40.031μg·m L~(-1)、40.020μg·m L~(-1)、2.683μg·m L~(-1)和101.418μg·m L~(-1),对猪源大肠杆菌MIC50分别为8.724μg·m L~(-1)、56.044μg·m L~(-1)、31.214μg·m L~(-1)和130.915μg·m L~(-1)。回归方程显示环丙沙星对鸡源大肠杆菌抑制作用最强;高于40.031μg·m L~(-1)时,诺氟沙星抗菌作用弱于头孢噻肟,低于120.23μg·m L~(-1)时,诺氟沙星抗菌作用强于四环素;头孢噻肟、环丙沙星、诺氟沙星和四环素对猪源大肠杆菌的抑菌作用则呈递减趋势。同时验证了诺氟沙星和环丙沙星属同种作用机制,基于Probit模型计算更简单、快速、直观。研究结果可为畜禽养殖中大肠杆菌的耐药差异监控提供一定的数据基础。     

抗生素最小抑制浓度法(MICs)评估环境大肠杆菌抗生素抗性

2015年世界卫生组织将抗生素的滥用列为21世纪最大的挑战之一,全球范围内抗生素抗药性的散播已严重威胁人类的健康。如何检测环境中的细菌抗药性,并有效评估抗药性感染的风险,是环境微生物研究的一项重要课题。本文通过改进的培养基微量稀释(broth micro-dilution)法,确定了2个地区(中国成都和美国夏威夷)不同环境来源(天然水系和市政污水)的大肠杆菌的抗生素最小抑制浓度(minimum inhibitory concentrations,MICs)。计算所得的MIC分位数(MIC50和MIC90)、菌体抗性百分比及多抗药性指数(multiple antibiotic resistance indexes,MARIs)显示两地不同环境区划的抗生素抗性存在明显的差异。天然水系(成都锦江)中的抗生素抗性是随时间可变的,与当地的降雨事件相关。环境菌株的抗药性模式通过聚类和非度量多维测度(non-metric multidimensional scaling,NMDS)进行分析。广谱β-内酰胺酶基因筛查显示出抗性基因与抗性表型之间的正相关性。结合现有的两地抗生素的使用数据讨论了两地环境抗生素抗性与当地人类活动及抗生素的使用实践之间的紧密联系。采集环境菌株抗生素MIC数据的实验及数据分析方法实现了环境抗药性的跨时空对比,为规范抗生素的区域性使用提供了指导作用。     

快速评估近岸海洋水体与沉积物中E.coli与S.aureus抗生素抗性水平的新方法

随着抗生素抗性污染日益严重,快速评估环境中典型病原菌与条件性致病菌的抗生素抗性水平,对掌握区域环境抗生素抗性污染状况、揭示抗性污染传播规律至关重要。通过以最低抑菌浓度浸入抗生素改进MI、VJ培养基,并结合滤膜法,建立了针对近岸海洋环境中指示性病原微生物大肠杆菌(Escherichia coli,E.coli)与金黄色葡萄球菌(Staphylococcus aureus,S.aureus)的抗生素抗性监测方法。水体和沉积物样品抗生素抗性水平评估实验结果显示,该方法具有较好重现性(水体和沉积物中E.coli和S.aureus抗生素抗性水平的相对标准偏差分别为11%、8%)与准确度(水体和沉积物中E.coli和S.aureus的平均回收率分别为83.5%、68.4%;相对于CLSI药敏试验的偏离度为±0.1)。且与CLSI药敏实验相比,该方法过程简便、耗时短(36 h/84 h),能最大限度节约经济和人员成本提高抗性评价效率。应用该方法评估辽河口与莱州湾环境中2种病原微生物磺胺类抗生素抗性水平,结果显示辽河口水体中E.coli和S.aureus磺胺二甲嘧啶的平均抗性率分别为27.0%、28.4%,沉积物中分别为35.5%、34.6%;莱州湾水体中E.coli和S.aureus磺胺二甲嘧啶的平均抗性率分别为26.0%、14.5%,沉积物中分别为12.0%、32.9%。该方法适用于河口、近岸海洋及入海排污口水体与沉积物样品中E.coli与S.aureus的快速分析及抗生素抗性水平评估。     

Influence of lignin, pentachlorophenol and heavy metal on antibiotic resistance of pathogenic bacteria isolated from pulp paper mill effluent contaminated river water

Pulp paper mill pollutants are the major source of aquatic contamination having metals, lignin and chlorophenols. Study was conducted to see the effect of these contaminants on antibiotic resistance pattern of isolated bacteria. Pulp paper effluents were evaluated for its physico-chemical properties i.e, BOD 72143 +/- 164.81 to 22.32 +/- 2.48, COD 213136 +/- 583.59 to 60.40 +/- 6.34, total phenol 386 +/- 71.24 to 0.43 +/- 0.0, lignin 26312 +/- 258.59 to 73.67 +/- 31.81and microbial quality i.e. K. pneumonae, S. typhi, S. faecalis, P. aeruginosa, E. coli, Ent. faecalis, A. hydrophila, B. subtilis, S. aureus, Y enterolitica and V vulrificus. Antibiotic sensitivity (10-30 microg), heavy metal resistance (100-1000 microg ml(-1)), lignin (1000-10,000 ppm) and pentachlorophenol (100-1000 ppm) tolerance of bacterial strains were assessed by seven classes of antibiotics. Eleven bacterial isolates were found multidrug resistant towards antibiotics, heavy metal, lignin and PCP. Out of 11 isolates, 90.9% were found resistant against eleven antibiotics which acquired 100% resistant in presence of heavy metal, lignin and chlorophenols. Results also revealed that concentration of lignin (50-350 ppm) and PCP (5-30 ppm) induced maximum growth (273-8050 cfu ml(-1)) of pathogenic bacteria in river water.     

Effect of ultraviolet irradiation and chlorination on ampicillin-resistant <Emphasis Type="Italic">Escherichia coli</Emphasis> and its ampicillin resistance gene

Antibiotic resistance is a serious public health risk that may spread via potable and reclaimed water. Effective disinfection is important for inactivation of antibiotic-resistant bacteria and disruption of antibiotic resistance genes. Ampicillin is a widely prescribed antibiotic but its effectiveness is increasingly undermined by resistance. In this study, changes in ampicillin resistance for Escherichia coli (E. coli) CGMCC 1.1595 were analyzed after exposure to different doses of ultraviolet (UV) or chlorine, and damage incurred by the plasmid encoding ampicillin resistance gene bla _TEM-1 was assessed. We reported a greater stability in ampicillinresistant E. coli CGMCC 1.1595 after UV irradiation or chlorination when compared with previously published data for other E. coli strains. UV irradiation and chlorination led to a shift in the mortality frequency distributions of ampicillin-resistant E. coli when subsequently exposed to ampicillin. The ampicillin hemiinhibitory concentration (IC₅₀) without disinfection was 3800 mg·L^–1, and an increment was observed after UV irradiation or chlorination. The IC₅₀ of ampicillin-resistant E. coli was 1.5-fold higher at a UV dose of 40 mJ·cm^–2, and was 1.4-fold higher when exposed to 2.0 mg·L^–1 chlorine. These results indicate that UV irradiation and chlorination can potentially increase the risk of selection for E. coli strains with high ampicillin resistance. There was no evident damage to bla _TEM-1 after 1–10 mg Cl₂·L^–1 chlorination, while a UV dose of 80 mJ·cm^–2 yielded a damage ratio for bla _TEM-1 of approximately 1.2-log. Therefore, high UV doses are required for effective disruption of antibiotic resistance genes in bacteria.     

离子液体[BMIm][PF6]增强抗性质粒RP4介导的接合转移基因mRNA的表达水平

环境中广泛存在的抗生素和抗生素抗性基因会导致很严重的人类健康风险。在我们前期研究中发现,离子液体1-丁基-3-甲基咪唑六氟磷酸盐([BMIm][PF6])作为环境选择性压力可以促进抗生素抗性基因的水平转移。本研究以E.coli DH5α(RP4)和同属的E.coli HB101,以及E.coli DH5α(RP4)和跨属的Salmonella enterica之间的纯菌接合转移体系为考察对象,从mRNA基因表达调控水平角度阐明离子液体[BMIm][PF6](0.001~2.5 g·L-1)影响质粒RP4接合转移的机理。结果表明,离子液体[BMIm][PF6]通过抑制基因kor A,kor B和trb A的mRNA表达水平来提高接合和跨膜转运基因trb Bp和trf Ap的表达;增强水平转移基因tra F的mRNA表达水平,并通过增强负调控基因kil A和kil B的mRNA表达水平抑制质粒的垂直传递过程;通过抑制基因trb K的mRNA表达水平降低接合转移体系的排斥效果,从而促进质粒RP4的接合转移。该结果有助于为抗生素抗性基因在环境中水平转移扩散的机理研究提供理论依据。     

群体感应信号分子存在下磺胺对大肠杆菌生长、突变及R388质粒接合转移的影响

抗生素的滥用使细菌耐药性问题日益突出,给许多疾病的预防与控制增加了难度。基因突变和质粒接合转移是细菌获得抗生素抗性基因的主要方式,许多研究围绕抗性基因来展开,但是关于群体感应对于抗性基因产生和传播的影响鲜有报道。本文以大肠杆菌(Escherichia coli)为模式生物,群体感应信号分子N-(β-酮己酰)-L-高丝氨酸内酯(3-oxo-C6-HSL,C6)和3种磺胺类抗生素(磺胺嘧啶、磺胺甲恶唑、磺胺氯哒嗪)为研究对象,测定了其对大肠杆菌生长效应、突变效应及接合转移效应的影响。结果表明:C6不影响磺胺对大肠杆菌的生长抑制率,但能够削弱磺胺对大肠杆菌突变的促进作用,并且能增强磺胺对大肠杆菌R388质粒接合转移的抑制作用。本文为从群体感应角度研究大肠杆菌耐药性的产生与传播提供新思路。     

利用GFP和抗性双类型标记监测联合固氮菌在玉米根际的定殖

将来自质粒ｐＫＲＰ１０、ｐＫＲＰ１１和ｐＫＲＰ１２的氯霉素、卡那霉素和四环素抗性基因分别插入质粒ＧＦＰｍｕｔ２中ｇｆｐ基因下游的ＰｓｔＩ位点，得到ｇｆｐ和不同抗性基因共存的重组质粒，转化Ｅｎｔｅｒｏｂａｃｔｅｒ ｇｅｒｇｏｖｉａｅ ５７－７野生型菌株和耐铵工程菌Ｅ７后，得到既有抗生素抗性又在蓝光下呈现亮绿荧光的菌株。用它们接种玉米后，利用这两种选择标记双重筛选重新分离到的细菌确定了接种菌在玉米幼苗     

Association between heavy metals and antibiotic-resistant human pathogens in environmental reservoirs: A review

Heavy metals can act as co-selecting agents and promote antibiotic resistance. Most frequent resistances to heavy metals are observed for zinc and cadmium. P. aeruginosa and E. coli are commonly resistant to heavy metals and antibiotics. Heavy metals proliferate antibiotic resistance through co- and cross-resistance. Heavy metal and antibiotic resistances are common near anthropogenic activities. Antibiotic resistance in human pathogens can proliferate under selective pressures. Heavy metals in environmental reservoirs may contribute to selecting antibiotic-resistant strains. To determine the associations between heavy metals and antibiotic resistance, a literature review was conducted to systematically collect and categorize evidence for co-occurrence of resistance to heavy metals and antibiotics within human pathogenic bacteria in water, wastewater, and soil. In total, 42 publications adhered to inclusion criteria. Across the reservoirs, zinc and cadmium were the most commonly observed heavy metals associated with resistance to antibiotics. Pseudomonas aeruginosa and Escherichia coli were the most commonly studied bacteria with reported co-occurrence of resistance to several heavy metals and antibiotic classes. As co-selecting agents, prevalence of heavy metals in the environment can proliferate resistance to heavy metals and antibiotics through co-resistance and cross-resistance mechanisms. In comparing different reservoirs, soils and sediments harbor higher heavy metal and antibiotic resistances compared to water environments. Additionally, abiotic factors such as pH can affect the solubility and hence, the availability of heavy metals to bacterial pathogens. Overall, our review demonstrates heavy metals act as co-selecting agents in the proliferation of antibiotic resistance in human pathogens in multiple environmental reservoirs. More studies that include statistical data are needed to further describe the exposure-response relationships between heavy metals and antibiotic resistance in different environmental media. Moreover, integration of culture-based and molecular-based methods in future studies are recommended to better inform our understanding of bacterial co- and cross-resistance mechanisms to heavy metals and antibiotics.     

Antimicrobial resistance among enteric bacteria, isolated from runoff of the Gangotri glacier, western Himalaya India

An attempt has been made to study antimicrobial resistance among the coliforms, faecal coliforms and faecal streptococci isolated from the runoff of the Gangotri glacier. The number of multiple antibiotic resistances (MAR) coliform isolates increases from upper stretch (33.33%) to lower stretch (83.33%). All faecal coliforms (100%) and faecal streptococci (100%) of lower stretch of study area showed multiple antibiotic resistances. Among coliforms, the value of Antibiotic resistance index (ARI) was found maximum in lower stretch (0.34) followed by middle stretch (0.29) and upper stretch (0.25).     

Plasmid mediated antibiotic resistance in marine bacteria

This research work was conducted in Uppanar estuary to ascertain the role of plasmids in the antibiotic resistance of bacteria. Water and sediment samples were collected for a period of three months. When tested against 20 antibiotics 22 MAR strains were isolated from the samples, which were found resistant to 5-13 antibiotics. They belong to 7 genera and 10 species. Gram-negative bacteria namely Neisseria mucosa, N. sicca, Branhamella catarrhalis, Klebsiella ozaenae, Citrobacterintermedius, Pseudomonas fluorescens and Enterobacter aerogenes were isolated. Gram-positive bacteria were of Bacillus subtilis, B. megaterium and Micrococcus luteus. When plasmid curing was done using acredine orange, the resistance against penicillin-G, ampicillin, tetracycline, amoxycillin, kanamycin, and chloramphenicol were totally lost in all strains, which confirmed the role of plasmid in these strains against antibiotics. Ten strains belong to different species were selected for the plasmid isolation and electrophoresis was done. Presence of plasmids in all strains was confirmed and the molecular weight was in the range of 2850 to 3170 bp. The study revealed that MAR strains are common in Uppanar estuary and they are plasmid mediated. This environment is seemed to be deteriorating at an alarming rate.     

牛至挥发油对肠炎常见菌的体外抗菌作用

不同浓度牛至挥发油对常见的致肠炎沙门氏菌属、大肠杆菌属和葡萄球菌属等31株菌的抗菌作用研究结果表明，该挥发油对上述31株菌均有不同程度的杀菌和抑菌作用．