血标本检出420株菌的菌谱分布及药敏分析

本文报告中山医科大学附一院从１９９５年１０月至１９９７年８月应用Ｂａｃｔ／Ａｌｅｒｔ血培养仪检测３６００份血液培养结果,阳性标本４１４份,其中６份为复数菌,菌株数４２０,其中革兰阳性球菌（Ｇ＋Ｃ）２３６株,革兰阴性杆菌（Ｇ－Ｂ）发酵菌７２株,Ｇ－Ｂ非发酵菌３４株,真菌５３株,革兰阳性杆菌（Ｇ＋Ｂ）１３株,革兰阴性球菌（Ｇ－Ｃ）２株,厌氧菌１０株。主要的细菌分离率由高到低为凝固酶阴性葡萄球菌（ＳＣＯＮ）１７８６％,金黄色葡萄球菌（ＳＡ）１２８６％,α—溶血性链球菌９５２％,肠球菌属９０５％,大肠埃希氏菌８１％,微球菌属６４３％,铜绿假单胞菌４２９％和沙门氏菌３１％。药敏结果显示Ｇ＋Ｃ中万古霉素的耐药率最低,Ｇ－Ｂ却对各种药物表现出不同的耐药率,其中对泰能和头胞噻甲肟的耐药率较低。     

弗劳地枸椽酸盐杆菌复合群基因种的生化鉴定

用Ｂｒｅｎｎｅｒ等的方法对从小儿腹泻大便中分离的４０株弗劳地枸椽酸盐杆菌进行基因种的鉴定和分析。７５％的细菌可被准确鉴定,共鉴定出４个基因种。弗劳地枸椽酸盐杆菌（Ｃ．ｆｒｅｕｎｄｉ）居首位（５６．７％）,依次为布拉基枸椽酸盐杆菌（Ｃ．ｂｒａａｋｉ）（２０．０％）,杨格枸椽酸盐杆菌（Ｃ．ｙｏｕｎｇａｅ）（１３．３）和沃克马尼枸椽酸盐杆菌（Ｃ．ｗｅｒｋｍａｎｉ）（１０．０％）     

医院空气中溶血性微生物微生态分布及耐药谱分析

在医院空气中,有１１．２４％的细菌具有溶血能力。溶血性微生物超过５００ｃｆｕ／ｍ３者占４３．４４％。在２０９１株溶血性细菌中,Ｇｒａｍ’ｓ阴性菌占９３．６％,主要为葡萄球菌（１３３３株）．Ｇｒａｍ’ｓ阳性菌占３．３１％,均为不发酵杆菌。葡萄球菌分布于７个种,以中间型葡萄球菌（２７．９８％）和金葡菌（２４．７６％）为主,同时,还检出耐甲氧西林葡萄球菌９１８株。药敏结果的提示,葡萄球菌耐药率高,耐药谱宽。８重以上耐药株占５２．２９％。     

慢性前列腺炎患者前列腺菌群的调查与分析

本文对１６９例慢性前列腺炎患者前列腺进行了微生物的分离与药物敏感试验,证实慢性前列腺炎患者前列腺内可有多种微生物混合感染,绝大多数为条件致病性或非病原性的耐药性或多重耐药性微生物。其中细菌２１８株占８０４％,含革兰氏阳性细菌９２２％株,革兰氏阴性细菌７８％株,真菌１１株占４１％;支原体４０株占１４８％;衣原体２株占０７％。我们又根据所检出的各种病原体的药物敏感性选用不同的抗菌药物以常规途径给药（口服、肌肉或静脉注射）治疗,获得了良好的治愈效果。因此认为,机体抵抗力降低或抗菌药物的滥用是导致前列腺微生态紊乱而发生前列腺炎症状的主要原因,前列腺复杂的菌群及其耐药性则是导致前列腺炎难以治愈的的关键因素。     

MRSA和MRCNS院内分布调查

对１６３７ 人（ 件） 次医、护人员手及护理工作台和病房床头桌进行了金黄色葡萄球菌（ＳＡ） 、凝固酶阴性葡萄球菌（ＣＮＳ） 、耐甲氧苯青霉素金黄色葡萄球菌（ ＭＲＳＡ） 及耐甲氧苯青霉素凝固酶阴性葡萄球菌（ ＭＲＣＮＳ） 的分布调查。结果表明,四种标本上ＳＡ 携带率分别为４．５ ％ 、３．１ ％ 、３．０ ％ 和６．１ ％ ;ＣＮＳ 携带率分别是３１．３ ％ 、３７．７ ％ 、３７．６ ％ 和４３．４ ％ 。而四种标本上ＭＲＳＡ 占检出ＳＡ 的百分率分别是２６．２ ％ 、２４．３ ％ 、２３．０ ％ 、３１．８ ％ ;ＭＲＣＮＳ占检出ＣＮＳ的百分率分别是２８．１ ％ 、２７．６ ％ 、２２．０ ％ 、３４．８ ％ 。所检出的ＭＲＳＡ 和ＭＲＣＮＳ对１４ 种抗生素的耐药性较ＳＡ 和ＣＮＳ都高。鉴于ＭＲＳＡ 和ＭＲＣＮＳ在医院环境中的分布,注意医护人员手及工作环境的清洁和消毒,是切断该类菌交叉感染的有效途径。     

115株来自泌尿系感染病原菌的细菌学及其药敏分析

作者对泌尿系统感患者中段尿分离出的１１５株细菌进行分析。结果：来自住院患者男女比例为１∶３,共检出２０个菌种,其中革兰氏阴性杆菌占７８．８％,以大肠埃希氏菌检出率最高,占５１．８％;革兰氏阳性球菌以表皮葡萄球菌和粪肠球菌为主,占１１．７％;药敏试验结果表明：革兰氏阴性杆菌对丁胺卡那、头孢三嗪、亚胺硫霉素、头孢噻肟、呋喃妥因较敏感;革兰氏阳性球菌除对万古霉素、环丙氟哌酸、利福平较敏感外,对其它抗菌素都有相当高的耐药性。来自门诊患者男女之比为１∶９,共检出１８个菌种,其中革兰氏阴性杆菌占６６．７％,以大肠埃希氏菌为主,占３０．０％,其次为变形杆菌（６．６％）;革兰氏阳性球菌以粪肠球菌为主,占总检出菌的１３．３％,药物敏感率较住院患者高,细菌分布更广。     

961份血培养的细菌及其药敏分析

对９６１份血培养的细菌和药敏结果进行了分析。结果：有１２３份培养阳性,阳性率为１２．８％,共分离出１２６株细菌。其中革兰氏阳性球菌９１（Ｇ~＋Ｃ）株,占７２．２％,以表皮葡萄球菌．金黄色葡萄球菌和粪肠球菌为主,占所有检出菌的５６．３％;革兰氏阴性杆菌（Ｇ~－Ｂ）２６株,真菌７株,提示革兰氏阳性球菌为本地区引起菌血症的主要病原菌,真菌感染有增多趋势。药敏结果提示：针对Ｇ~＋Ｃ的首选药物为万古霉素,对其它抗菌素都有普遍的不同程度的耐药性,应引起临床注意。     

细菌L型败血症324例临床分析

自１９８６年６月至１９９４年２月住院收治细菌Ｌ型败血症３２４例。年龄４天－７８天。病原多为金黄色葡萄球菌。其次为表皮葡萄球菌、大肠杆菌、链球菌、枸椽酸杆菌、鼠伤寒沙门菌、腊样芽胞杆菌、级脓杆菌变形直菌等。做细菌Ｌ型培养同时应用光镜、电镜、酶联免疫法快速诊断,可提高诊断率,使患者早日得到治疗。     

泌尿系感染常见病原菌分析及耐药性测定

１８５６例中段尿细菌培养和药敏结果,共检出致病菌６０８株;２２个种类细菌。革兰阴性杆菌３７０株,占６０．９％;革兰阳性球菌１７７株,占２９％;真菌６１株,占１０．１％。革兰阴性杆菌中最多的是大肠埃希菌（占５２％）和普通有杆菌（占１０．９％）;革兰阳性球菌中最多的是粪球菌（占４０．１）和金黄色葡萄球菌（占２６．１）。本文还分析了常见病原菌的耐药性。     

1 276例新生儿临床血液培养的分析

目的研究新生儿血液感染的细菌种类和培养方法（需氧、厌氧培养）。方法血标本在mini VITAL全自动血培养仪中培养,采用VITEK32自动微生物分析仪进行细菌鉴定。结果1276例新生儿血中共分离出细菌95株,阳性检出率为7．37％。95株菌分属37个菌种。其中革兰阳性球菌71株,占74．74％,革兰阴性杆菌20株,占21．05％,专性厌氧菌1株,占1．05％,感染率最高的前3位细菌是凝固酶阴性葡萄球菌、金黄色葡萄球菌和大肠埃希菌。1149例新生儿中需氧和厌氧配对培养结果,阳性83例,阳性检出率为7．24％,其中需氧和厌氧均生长者34例,占40．96％;仅需氧生长者21例,占25．30％;仅厌氧生长者28例,占33．73％。结论快速血培养技术是血液感染病原学诊断必要手段,同时做需氧、厌氧培养对培养结果至关重要。     

母婴同室新生儿皮肤菌群分析

目的了解母婴同室新生儿皮肤菌群及皮肤感染致病菌情况,为医院感染管理提供依据。方法选取我院母婴同室区2017年3月至7月出生的新生儿,对新生儿出生5min内、出生24h后第1次沐浴前、出生48h后第2次沐浴前、出生72h后第3次沐浴前及皮肤感染部位使用琼脂直接接触培养(RODCA)法采样并将采样平皿送入检验科微生物室进行培养观察。结果本研究共纳入157例新生儿,其中23例新生儿发生皮肤感染。随着出生时间的增加,检出的细菌菌落数显著增加,差异有统计学意义(P0.05)。新生儿出生5min内无细菌生长,出生5min之后主要检出表皮葡萄球菌(320株)、微球菌(118株)和枯草芽胞杆菌(96株)。阴道分娩新生儿皮肤感染发生率低于剖宫产新生儿(P0.05),其感染部位致病微生物为表皮葡萄球菌(18株)。结论我院母婴同室区无外来致病菌,不会造成医院感染的暴发。新生儿出生后72h内是皮肤感染的高危时间段,医院感染管理者应高度重视。     

Initial development and structure of biofilms on microbial fuel cell anodes

Background  

Microbial fuel cells (MFCs) rely on electrochemically active bacteria to capture the chemical energy contained in organics and convert it to electrical energy. Bacteria develop biofilms on the MFC electrodes, allowing considerable conversion capacity and opportunities for extracellular electron transfer (EET). The present knowledge on EET is centred around two Gram-negative models, i.e. Shewanella and Geobacter species, as it is believed that Gram-positives cannot perform EET by themselves as the Gram-negatives can. To understand how bacteria form biofilms within MFCs and how their development, structure and viability affects electron transfer, we performed pure and co-culture experiments.     

Study of the inactivation of antibiotics by bacterial colonies

A procedure of bacteria application to disks from the colonies was used for determining antibiotic inactivation in the disks by the bacteria colonies after the disk direct contact with the colonies. Changes in the antibiotic activity in the disks were registered after incubation at 37 degrees C for 2 hours. It was shown that ampicillin resistant strains of E. coli K12 carrying R plasmids and strains of S. typhimurium and S. aureus inactivated the antibiotics in the disks and their population were homogenous in this respect. It is advisable to use the procedure in assaying drug resistance of bacterial populations.     

Numerical analysis of electrophoretic protein patterns of 'Achromobacter'group B, E and F strains from human blood

Thirty-two clinical strains representing ' Achromobacter 'groups B, E and F were characterized by one-dimensional SDS-PAGE of cellular proteins. All the strains were isolated from blood samples from hospital patients in the United Kingdom. The protein patterns, which contained 40 to 45 discrete bands, were highly reproducible and were used as the basis for a numerical analysis which included all the protein bands. The 32 ' Achromobacter ' strains formed two clusters at the 77% S level. The first, phenon 1, included the 28 group B and the two group E strains and the second, phenon 2, contained the two strains of group F. The strains in each phenon were characterized by a clearly distinct pattern of protein bands. Phenon 1 could be further divided at the 87% S level into three subphenons which correlated with differences in the principal bands found between 40.0 and 42.5 kD. Strains of group E clustered with group B strains from which they could not be distinguished by protein patterns. We conclude that high resolution PAGE combined with computerized analysis of protein patterns provides a useful method for the classification of this group of bacteria. Reference strains of each of the PAGE types identified are available from NCTC for inclusion in future studies.     

Numerical analysis of electrophoretic protein patterns of 'Achromobacter' group B, E and F strains from human blood

Thirty-two clinical strains representing 'Achromobacter' groups B, E and F were characterized by one-dimensional SDS-PAGE of cellular proteins. All the strains were isolated from blood samples from hospital patients in the United Kingdom. The protein patterns, which contained 40 to 45 discrete bands, were highly reproducible and were used as the basis for a numerical analysis which included all the protein bands. The 32 'Achromobacter' strains formed two clusters at the 77% S level. The first, phenon 1, included the 28 group B and the two group E strains and the second, phenon 2, contained the two strains of group F. The strains in each phenon were characterized by a clearly distinct pattern of protein bands. Phenon 1 could be further divided at the 87% S level into three subphenons which correlated with differences in the principal bands found between 40.0 and 42.5 kD. Strains of group E clustered with group B strains from which they could not be distinguished by protein patterns. We conclude that high resolution PAGE combined with computerized analysis of protein patterns provides a useful method for the classification of this group of bacteria. Reference strains of each of the PAGE types identified are available from NCTC for inclusion in future studies.     

外源性磷酸肌酸对重度窒息新生儿血清S—100B蛋白及NSE水平的影响

目的：探讨外源性磷酸肌酸对重度窒息新生儿血清S-100B蛋白和特异性神经元烯醇化酶含量（NSE）的影响。方法：重度窒息的新生儿40例,随机分为两组,常规治疗组21例,给予一般治疗（氧疗、支持、对症）和胞二磷胆碱治疗。磷酸肌酸治疗组19例,在常规治疗基础上,生后12h内给予磷酸肌酸治疗lg／d）,另外同期住院的新生儿湿肺和黄疸患儿14例为正常对照组。均与生后48h和生后10天取血检测血清S-100B蛋白和NSE含量。并于生后第14天进行新生儿行为神经测定（NBNA评分）。结果：磷酸肌酸治疗组和常规治疗组患儿生后48h血清s．100B蛋白和NsE含量无显著差异（※P〉0．05,※P〉0．05）,与正常对照组比较差异具有显著性（△P〈0．05,AP〈0．05）,生后10天血清S．100B和NSE含量在常规治疗组患儿和磷酸肌酸组相比具有显著差异,磷酸肌酸治疗组两者明显下降（※P〈0．05,※P〈0．05）。生后三周的行为神经评估（NBNA评分）〈35分者所占百分比磷酸肌酸治疗组27％与常规治疗组组53％比较,差异均具有显著性意义（x2=6．112,※P〈0．05）。结论：磷酸肌酸用于治疗新生儿缺氧缺血性脑病能够改善脑的能量代谢,降低脑损伤的程度,改善神经行为,降低致残率。     

Investigation of antibiotic and antibacterial agent cross-resistance in target bacteria from homes of antibacterial product users and nonusers

AIM: To describe the relationship between antibiotic and antibacterial resistance in environmental and clinical bacteria from home environments across geographical locations, relative to the use or nonuse of antibacterial products, with a focus on target organisms recognized as potential human pathogens. METHODS AND RESULTS: In a randomized study, environmental and clinical samples were collected from the homes of antibacterial product users (n=30) and nonusers (n=30) for the isolation of target bacteria for antibiotic and antibacterial testing in three geographical areas (in USA and UK). Isolates were tested for antibiotic susceptibility, with selected antibiotic-resistant and antibiotic-susceptible isolates tested against four common antibacterial agents (triclosan, para-chloro-meta-xylenol, pine oil and quaternary ammonium compound). Prequalified users and nonusers at each location were randomly selected after meeting exclusionary criteria. Of 1238 isolates, more target bacteria were recovered from nonuser than user homes. Of Staphylococcus aureus isolates (n=33), none showed resistance to oxacillin or vancomycin; for Enterococcus sp. (n=149), none were resistant to ampicillin or vancomycin; and for Klebsiella pneumoniae (n=54)and Escherichia coli (n=24), none were resistant to third generation cephalosporins. Antibiotic resistance to one or more of the standard test panel drugs for Gram-positive and Gram-negative target bacteria was comparable between nonuser and user homes for both environmental and clinical isolates [e.g. resistance of environmental coagulase-negative (CN) Staphylococcus sp. was 73.8% (124/168) from nonuser homes and 73.0% (111/152) from user homes, and Enterobacteriaceae other than E. coli, 75.9% (186/245) from nonuser homes compared with 78.0% from user homes]. Of 524 Gram-negatives tested against preferred/alternative drugs, 97.1% (509/524) were susceptible to all antibiotics, across both groups. Isolates of S. aureus, Enterococcus sp. and CN Staphylococcus sp. susceptible to all preferred treatment drugs showed comparable antibacterial minimum inhibitory concentration (MIC) results between nonuser and user home isolates. For Gram-positives resistant to one or more preferred drugs, greatest resistance to antibacterial active ingredients was found in the nonuser group. For Gram-negatives, the antibacterial MIC data were comparable for isolates that were fully susceptible and resistant to one or more preferred/alternative treatment antibiotics. CONCLUSIONS: The results showed a lack of antibiotic and antibacterial agent cross-resistance in target bacteria from the homes of antibacterial product users and nonusers, as well as increased prevalence of potential pathogens in nonuser homes. SIGNIFICANCE AND IMPACT OF THE STUDY: It refutes widely publicized, yet unsupported, hypotheses that use of antibacterial products facilitates the development of antibiotic resistance in bacteria from the home environment.     

Use of the Gram stain in microbiology

The Gram stain differentiates bacteria into two fundamental varieties of cells. Bacteria that retain the initial crystal violet stain (purple) are said to be 'Gram-positive,' whereas those that are decolorized and stain red with carbol fuchsin (or safranin) are said to be 'Gram-negative.' This staining response is based on the chemical and structural makeup of the cell walls of both varieties of bacteria. Gram-positives have a thick, relatively impermeable wall that resists decolorization and is composed of peptidoglycan and secondary polymers. Gram-negatives have a thin peptidoglycan layer plus an overlying lipid-protein bilayer known as the outer membrane, which can be disrupted by decolorization. Some bacteria have walls of intermediate structure and, although they are officially classified as Gram-positives because of their linage, they stain in a variable manner. One prokaryote domain, the Archaea, have such variability of wall structure that the Gram stain is not a useful differentiating tool.     

Transformation of group A streptococci by electroporation

Abstract The introduction, via electroporation, of free plasmid DNA into three strains of Streptococcus pyogenes is described. The method is very simple and rapid and efficiencies vary from 1 × 10³ to 4 × 10⁴ per μg of DNA. The method was also used to introduce an integrative plasmid and transformants were obtained, albeit at a somewhat lower frequency (2 × 10²). Some of the plasmids used in this study are derivatives of the Lactococcus lactis subsp. cremoris Wg2 plasmid pWV01. These broad host range vectors replicate in Gram-positives as well as Gram-negatives (viz. Escherichia coli ). Here we show that they also replicate in S. pyogenes and S. sanguis .     

12株猪瘟病毒E2基因主要抗原区域的序列差异分析

用RTPCR扩增了12个不同时期分离的HCV毒株E2基因主要抗原区域的cDNA片段并对其进行了序列测定。应用DNAstar序列分析软件对所测的12个HCV毒株与国内外已知的6个毒株Alfort株、Ald株、Brescia株、Gpe株、C株、CW株及早期已测定的HCLV株、HCVSM株和北京顺义株(BJSY2/96)3个毒株的相应片段进行了同源性比较分析。E2基因主要区域长度均为224 bp,包括从HCV 2485到2708位的E2基因B、C区域。所测的疫苗株HCLV与国外测得的疫苗株C株核苷酸及氨基酸同源性分别为991%和100%,表明目前应用的疫苗株是稳定的;用目前我国流行的部分野毒株对HCLV株免疫猪的攻击试验表明,HCLV对野毒株均具有很好的免疫力,这与序列分析结果相吻合。根据系统树分析,可将HCV分为两大群,5株90年代的野毒株及1株80年代的野毒株(其中北京3株、河南2株、广东1株)均与国内外C株、标准株属同一群(即第一群),其核苷酸及氨基酸的同源性分别为857%～100%和838%～100%;与Alfort株同属第二群的有6个野毒株(广西北海、辽宁、河北黄骅、吉林、深圳光明、四川成都),其中80年代与90年代的野毒株各有三株,核苷酸及氨基酸的同源性分别为843%～100%和851%～100%;21株HCV的核苷酸及氨基酸的同源性分别为781%～100%和784%～100%。两群之间的特征性差异表现在713和729位氨基酸位点的不同,经分析发现猪瘟野毒株具有复杂性与多样性。