Enterococcus: review of its physiology, pathogenesis, diseases and the challenges it poses for clinical microbiology

The genus Enterococcus is composed of 38 species, the most important of which are Enterococcus faecalis and Enterococcus faecium—both human intestinal colonizers. Hospitals within the United States and around the world commonly isolate these bacteria because they are a cause of bacteremia, urinary tract infections (UTIs), endocarditis, wound infections, meningitis, intraabdominal and pelvic infections, and nosocomial and iatrogenic infections. Given the ubiquity of enterococci within the human population, it is important for laboratories to be able to distinguish these agents within hospitalized patients from other bacterial genera and also differentiate different species within the Enterococcus genus as well as different strains within each species. Unfortunately, the enterococci are emerging as serious pathogens in both the developed world, where surveillance needs to be improved and speciation procedures are inadequate or cumbersome, and in developing nations, which lack the trained hospital personnel or funding to sufficiently identify enterococci to the genus or species level. This review explores the Enterococcus genus and highlights some of the concerns for national and international clinical microbiology laboratories.     

Sequence of the 50-kb Conjugative Multiresistance Plasmid pRE25 from Enterococcus faecalis RE25

The complete 50,237-bp DNA sequence of the conjugative and mobilizing multiresistance plasmid pRE25 from Enterococcus faecalis RE25 was determined. The plasmid had 58 putative open reading frames, 5 of which encode resistance to 12 antimicrobials. Chloramphenicol acetyltransferase and the 23S RNA methylase are identical to gene products of the broad-host-range plasmid pIP501 from Streptococcus agalactiae. In addition, a 30.5-kb segment is almost identical to pIP501. Genes encoding an aminoglycoside 6-adenylyltransferase, a streptothricin acetyltransferase, and an aminoglycoside phosphotransferase are arranged in tandem on a 7.4-kb fragment as previously reported in Tn5405 from Staphylococcus aureus and in pJH1 from E. faecalis. One interrupted and five complete IS elements as well as three replication genes were also identified. pRE25 was transferred by conjugation to E. faecalis, Listeria innocua, and Lactococcus lactis by means of a transfer region that appears similar to that of pIP501. It is concluded that pRE25 may contribute to the further spread of antibiotic-resistant microorganisms via food into the human community.     

基于全基因组关联分析研究粪肠球菌发酵食品分离株的耐药性

【目的】研究15株分离自自然发酵食品的粪肠球菌(Enterococcus faecalis)和1株粪肠球菌模式株对15种(1种β-内酰胺类和14种非β-内酰胺类)抗生素的耐药性,并通过菌株耐药表型与基因型的关联分析找到粪肠球菌潜在的耐药基因。【方法】利用微量肉汤稀释法检测试验菌株对15种抗生素的耐药性,运用Scoary软件进行表型与基因型的关联分析。【结果】16株粪肠球菌对卡那霉素、万古霉素、利奈唑胺和红霉素100%敏感;对其余11种抗生素均表现出不同程度的耐药,其中对克林霉素为100%耐药。基因组关联分析发现了9个功能基因与5种抗生素(氯霉素、环丙沙星、甲氧苄啶、新霉素和四环素)存在显著相关关系,其中基因FAM000296和FAM005768与氯霉素、甲氧苄啶和环丙沙星的耐药性有关。进一步分析发现基因FAM000296和FAM005768同被注释为Sec G,但基因FAM005768与基因FAM000296相比在3′端丢失了21个碱基。【结论】分离自自然发酵食品的粪肠球菌对多种抗生素具有耐药性,其基因组中携带有潜在的耐药基因。因此,对分离自自然发酵食品的粪肠球菌需要进行全面的安全性评估后方可考虑应用。     

我国部分地区即食食品和蔬菜中金黄色葡萄球菌污染分布及耐药和基因分型情况

【目的】系统调查了我国15个代表性城市在即食食品(卤肉、烤肉、凉拌菜和巴氏奶)和蔬菜样品中金黄色葡萄球菌(Staphylococcus aureus)的污染分布情况,并对分离株进行耐药性分析及多位点序列分型(Multilocus sequence typing,MLST)研究,为食源性金黄色葡萄球菌的风险识别和分子溯源提供基础数据。【方法】依据GB 4789.10-2010对540份即食食品和蔬菜样品进行定性和最大可能数(Most probable number,MPN)分析;采用K-B纸片扩散法检测金黄色葡萄球菌分离株的耐药特征并通过PCR检测mecA基因;应用MLST方法确证金黄色葡萄球菌的主要ST型。【结果】结果发现9.3%(50/540)的样品中检出金黄色葡萄球菌,其中卤肉污染率最高,为16.3%(30/184),其次为烤肉(9.2%,6/65),蔬菜(6/150,4.0%)污染率最低。定量分析发现62.0%的阳性样品污染水平处于0.3–1.0 MPN/g,其中3份阳性样品污染水平≥110 MPN/g。对50株分离株进行24种抗生素耐药性检测,发现82.0%的分离株对氨苄西林和青霉素耐药,64.0%的分离株为多重耐药株。对mecA阳性分离株进行SCCmec分型,发现均为SCCmecⅣa。所有分离株进行MLST分型,共检出14种型别,其中ST3595和ST3847为新ST型。【结论】普遍的多重耐药性表明我国食源性金黄色葡萄球菌的耐药状况已较为严重,对消费者的安全健康存在潜在威胁。ST型与耐药存在一定的关联性,这为进一步了解该菌在我国食品中的流行趋势和风险评估提供了科学的数据支撑。     

Toxic and non-toxic Nodularia strains can be distinguished from each other and from eukaryotic algae with chlorophyll fluorescence fingerprinting

Chlorophyll fluorescence induction curves of toxic and non-toxic strains of the cyanobacterium Nodularia were measured and compared with fluorescence curves measured from four species of eukaryotic algae. Both cyanobacteria and algae were isolated from the Baltic Sea. The results show that Nodularia strains can be distinguished from the eukaryotes by applying a pattern recognition procedure to the fluorescence induction curves, suggesting that the fluorescence fingerprinting technique might be useful in environmental monitoring of marine algae. The six studied Nodularia strains could not be distinguished from each other from their fluorescence induction kinetics. However, their fluorescence curves fell into two clear categories, the toxic and the non-toxic Nodularia. Emission spectroscopy and differences in the fluorescence induction curves showed that the ratio of the intensity of the Photosystem I emission peak to the Photosystem II peak is higher in non-toxic Nodularia than in the toxic strains, suggesting that the toxicity affects the structure of the photosynthesis machinery. The effect on photosynthesis may be related to the ability of the microcystins to chelate iron.     

不同抗生素对金发草愈伤组织生长分化的影响

该研究以金发草愈伤组织为材料,通过分析比较不同抗生素种类(卡那霉素、潮霉素、头孢噻呋钠和氨苄青霉素)和浓度对金发草愈伤组织生长分化的影响,来确定适用于金发草遗传转化体系中的抗性筛选剂和抑菌剂。结果表明：(1)金发草愈伤组织对卡那霉素很敏感,且其分化率随着卡那霉素浓度的增加显著减少( P＝0.01)。当卡那霉素浓度为10 mg·L-1时,金发草愈伤组织的生长分化受到明显抑制,且有大量的白化苗形成,但分化率仍有36.56％;当卡那霉素浓度为15 mg·L-1时,金发草愈伤组织的分化率为11.94％,只有很少部分的愈伤分化出绿色的丛生苗;当卡那霉素浓度为20 mg·L-1时,金发草愈伤组织基本褐化死亡,分化率仅为2.26％。因此,浓度为15 mg·L-1的卡那霉素适合作为金发草遗传转化体系中的抗性筛选剂。(2)金发草愈伤组织对潮霉素的敏感性要比卡那霉素弱,且潮霉素对金发草愈伤组织分化率的影响小,但毒害作用大。因此,潮霉素不适合作为金发草遗传转化体系中的抗性筛选剂。(3)300 mg·L-1的头孢霉素和氨苄青霉素对金发草愈伤组织生长分化影响很小且能有效抑制杂菌的生长,较高浓度的氨苄青霉素对金发草愈伤组织的抑制作用不太明显。因此,300 mg·L-1的头孢霉素和较高浓度的氨苄青霉素均可作为金发草遗传转化体系中的抑菌剂。该研究确定了适用于农杆菌介导的金发草遗传转化体系中的抗性筛选剂和抑菌剂,为金发草的遗传改良及功能性基因的研究奠定了基础。     

耐碳青霉烯鲍曼不动杆菌的临床分布及基因组流行病学分析

[背景]鲍曼不动杆菌是造成临床感染的重要病原菌之一,其对碳青霉烯类抗生素的耐药形势日益严重,利用基因组测序技术解析其临床分布特征和流行病学规律有助于临床感染的有效防治.[目的]研究沧州市中心医院2018年检出的200株耐碳青霉烯鲍曼不动杆菌(carbapenem-resistant Acinetobacter baum...     

模拟增温对青藏高原多年冻土区小嵩草和藏嵩草生长与抗氧化特征的影响

采用开顶式增温小室(OTCs)方法模拟气候变暖,分别选取青藏高原腹地风火山地区高寒小嵩草草甸和高寒藏嵩草沼泽草甸优势物种小嵩草和藏嵩草为研究对象,对比分析增温处理下两种优势物种叶片的形态与生理特征变化,从而探索高寒植物对气候变暖的内在响应机理.结果表明: 增温显著增加了小嵩草叶片长度(40.0%)和叶片数量(72.7%),也显著增加了藏嵩草株高(11.9%)和叶片长度(19.3%),促进了两种优势植物的形态生长和地上生物量增加.增温处理下小嵩草和藏嵩草叶片的膜透性(电导率),活性氧(过氧化氢和超氧阴离子自由基),超氧化物歧化酶、过氧化物酶、抗坏血酸过氧化物酶和过氧化氢酶活性,丙二醛含量均没有显著变化.但抗坏血酸和游离脯氨酸含量在藏嵩草叶片内分别显著增加了29.8%和53.8%,而在小嵩草叶片内没有明显变化.可见,增温下小嵩草和藏嵩草均能够维持正常的抗氧化水平,以维持该区域优势植物生长;但藏嵩草生理过程对增温更加敏感.     

木麻黄浸提液对榄仁树幼苗生长及生理生化的影响

该研究试图将榄仁树引入木麻黄海防林以期形成混交林型的海防林。以15 d榄仁树种子苗为材料,分别采用浓度为125 g·L~-1的木麻黄根、木麻黄凋落物和林下表层土壤的水浸提液胁迫处理榄仁树幼苗60 d,研究木麻黄化感作用对榄仁树幼苗生长、光合作用、膜系统伤害、抗氧化系统酶活性及游离Pro含量的影响。结果表明:木麻黄根、木麻黄凋落物及林下表层土壤的水浸提液都能降低榄仁树幼苗的存活率,影响幼苗株高、叶和根生物量,可以推断不同浸提液中对影响植物生长成分的浓度有所不同。3种浸提液均显著降低处理15~45 d期间幼苗的净光合速率(Pn)、气孔导度(Ccond)、胞间CO_2浓度(Ci)、蒸腾速率(Tr),且显著增加幼苗水分利用率(WUE)。第60天时3种木麻黄浸提液能显著增加幼苗叶片、根的Pro含量,使得幼苗根系组织细胞膜通透性增加,而木麻黄根浸提液能显著增加幼苗Ccond、Ci、叶SOD、根CAT含量,木麻黄凋落物浸提液能显著增加幼苗Ccond和Tr,同时凋落物浸提液会显著降低幼苗根POD含量。榄仁树幼苗通过调整自身的生理生化能适应木麻黄化感胁迫的影响,为此可考虑榄仁树作为沿海防护林建设的树种之一。     

Antagonistic activity of Lactobacillus acidophilus RY2 isolated from healthy infancy feces on the growth and adhesion characteristics of enteroaggregative Escherichia coli

Enteroaggregative Escherichia coli (EAggEC) infection is an important cause of acute diarrhea, affecting children in developing countries and travelers visiting tropical or subtropical areas. Three probiotics can exert bacteriostatic or bactericidal effects on human and animal intestinal pathogens, the efficiency of probiotics on EAggEC infection remains unclear. In this study, the antagonistic activity of probiotic bacteria isolated from infant faeces was examined against several EAggEC stains. While three isolates, Lactobacillus acidophilus RY2, Lactobacillus salivarius MM1 and Lactobacillus paracasei En4 were shown to significantly inhibit the growth of EAggEC. In addition, the antagonistic activity of the Lactobacillus species was maintained despite heating (100 °C, 15 min) of cell free culture supernatant (CFCS). The antagonistic activity of the CFCS however, could be reduced following lactate dehydrogenase treatment and at pH 7.2. Furthermore, in an adhesion–inhibition assay, L. acidophilus RY2 was shown to be more effective than L. salivarius MM1 and L. paracasei EN4. This study suggests that L. acidophilus RY2 could be used as a probiotic organism against EAggEC.     

成都平原不同种植方式折耳根表面附生细菌群落结构与抗生素抗性基因分析

【目的】分析有机、化肥和野生折耳根表面的附生细菌群落结构和抗生素抗性基因(ARGs),揭示细菌群落结构与ARGs相互关系。【方法】高通量测定16SrRNAV3-V4可变区序列分析样品表面附生细菌群落结构;PCR和qPCR扩增29种ARGs基因分析样品表面ARGs污染情况;冗余分析(RDA)探讨细菌群落结构与ARGs的相互关系。【结果】折耳根表面检测到35个属的细菌,其中有机折耳根表面附生细菌多样性低于化肥和野生折耳根(P0.05);29种被检的ARGs中,有14种在折耳根中被检出,其中有机折耳根含有全部被检出的ARGs,化肥和野生折耳根则含有部分被检出的ARGs。折耳根表面ARGs污染的多样性和丰度显著受到样品表面的菌群结构影响,其中Lactococcus、 Escherichia、Fluviicola、Enterococcus、Sanguibacter和Acidovorax是影响ARGs最主要的菌群。【结论】有机种植极大地改变了折耳根表面附生细菌的群落结构,增加了ARGs的多样性和丰度,对有机折耳根的食品安全带来了潜在威胁。因此,有必要将ARGs污染监测纳入到有机折耳根的食品安全考核范围内。     

不同隔盐措施对滨海盐碱地土壤水盐运移及刺槐光合特性的影响

土地盐碱化是限制土地资源利用的一个主要障碍,在耕地面积逐渐减少的今天,改良利用盐碱地早已提上了科学日程。在天津滨海盐碱地区,通过田间完全随机区组实验设计,对比分析了4种土壤盐分隔离措施(对照-不设隔盐处理,CK;沸石隔盐处理,FS;陶粒隔盐处理,TL;河沙隔盐处理,HS)对0—80 cm土层水盐运移及刺槐(Robinia pseudoacacia)光合特性的影响,目的是通过评估不同隔盐材料的控盐改土效果,为滨海地区盐渍土改良和沿海防护林营造等林业生态工程建设提供理论依据。研究结果表明:(1)FS和TL可以显著提高土壤相对水分含量,为刺槐生长创造更加适宜的水分环境。FS导致0—80 cm土体内盐分含量和土壤盐溶质浓度显著降低,控盐效果显著。TL仅在40 cm以下土层有显著降盐效果,控盐效果仅次于FS。与对照相比,HS对0—80 cm土体内盐分含量和土壤盐溶质浓度无显著影响,降盐效果最差。(2)FS处理能显著提高刺槐的净光合速率(Pn)、蒸腾速率(Tr)、气孔导度(Gs)及叶片水分利用效率(LWUE),降低胞间CO2浓度(Ci)。TL可显著提高刺槐叶片的Pn、Tr,但改善效果不及FS,并且其对Gs、Ci和LWUE没有显著影响。与对照相比,HS虽然能显著增加刺槐叶片Pn和Tr,但却导致LWUE显著降低。综上所述,在滨海地区采用沸石作为隔盐材料比采用传统材料河沙更能有效保水降盐,促进植物光合及生长,可以作为滨海盐碱地区隔盐材料的优先选择。     

乌头产吲哚乙酸内生细菌遗传多样性、抗逆性及其对水稻幼苗生长的影响

[目的]探究乌头产吲咮乙酸(IAA)内生细菌的遗传多样性、溶磷解钾能力、抗逆能力及其对水稻幼苗生长的影响,为道地产区乌头产业可持续发展提供科技支撑.[方法]从健康乌头植株分离可培养内生细菌,采用Salkowski比色法测定内生细菌产IAA能力,16S rDNA限制性片段长度多样性(16S rDNA-RFLP)及16S ...     

龙须菜(Gracilaria lemaneiformis)的生长、光合作用及其对扇贝排泄氮磷的吸收

研究了不同温度、光照条件下大型藻类龙须菜 (Gracilaria lemaneiformis)的生长和光合作用特性,及其对扇贝排泄氮、磷的吸收作用。结果表明温度与龙须菜的生长和光合作用显著相关,在本实验条件下,15～25℃都适宜龙须菜生长,其中20℃龙须菜具有较高的生长率(SGR),为2.8%/d;光合作用速率随温度升高和光照的增加而升高,30℃和120 μmol/(m^２•s)时最高,最大光合作用速率（Pmax）为5.0 mg O₂/(g dw•h)。龙须菜对扇贝排泄氮、磷有较强的吸收作用,其吸收率和去除效率与放养密度和养殖时间有关。对NH4-N和PO4-P的最大去除效率分别为83.7%和70.4%,最大吸收率分别为9.9 μmol/(g ww•h)和4.3 μmol/(g ww•h)。实验证明龙须菜生长温度范围和光照范围适合中国北方海区养殖,并且能有效吸收和去除扇贝排泄氮、磷,可以作为生物滤器与贝类及其他养殖动物进行综合养殖。