临床分离耐甲氧西林溶血性葡萄球菌的SCCmec分型及同源性分析

目的了解临床分离耐甲氧西林溶血性葡萄球菌（MRSH）的SCCmec基因型别及相同SCCmec型别菌株的同源性。方法多重PCR进行SCCmec分型,ERIC-PCR法对相同SCCmec型别菌株进行同源性分析。结果83株临床分离MRSH菌株中,SCCmecI型有23株（27．7％）,SCCmecⅡ型有10株（12．1％）,SCCmecm型有24株（28．9％）,SCCmecIV型有1株（1．2％）,I、Ⅱ混合型有8株（9．6％）,I、Ⅲ混合型有6株（7．2％）,Ⅱ、11混合型有5株（6．0％）,I、Ⅱ、Ⅲ混合型有3株（3．6％）,未分型3株（3．6％）。ERIC—PCR结果显示,23株SCCmecI型分为11型,其中A型5株,B型5株,C型3株,其余8株各为1型,2株未分型;10株SCCmecⅡ型分为6型,其中D型4株,E型2株,3株各为1型,1株未分型;24株SCCmecm型分为9型,其中F型11株,G型2株,H型2株,I型2株,5株各为1型,2株未分型。结论临床分离MRSH中,SCCmecI、Ⅲ型为多,部分菌株呈混合型别;相同SCCmec型别的部分菌株之间可能存在克隆传播。     

The nodular microsymbionts of Gymnostoma spp. are Elaeagnus-infective Frankia strains.

The phylogenetic relationships of Frankia strains infective on Gymnostoma with other Frankia strains was analyzed. Partial sequencing of the 16S rDNA and use of specific primers showed that the Frankia strains present in Gymnostoma are phylogenetically close to Elaeagnus-infective strains. This finding was confirmed by using the sequences of the hypervariable nifDK intergenic spacer. The strains present in Gymnostoma nodules were close to one another. Clustered with Elaeagnus-infective strains, and distantly related to Casuarina and Alnus-infective strains. Morphological observations of strains and cross-inoculation trials showed that Gymnostoma-infective strains are indistinguishable from Elaeagnus-infective strains. Results of both phenotypic and genotypic approaches indicate that Gymnostoma-infective strains are Elaeagnus infective and not Casuarina infective.     

替加环素与多粘菌素B对泛耐药鲍曼不动杆菌体外研究

目的为治愈耐舒普深的泛耐药鲍曼不动杆菌（PDR-Ab）提供新药。方法替加环素采用二倍琼脂稀释法,多粘菌素B用E-test条法测分离目标菌株100株的最低抑菌浓度（M IC）,并用WHONET 5.4软件分析数据。结果多粘菌素B对耐舒普深的泛耐药鲍曼不动杆菌株的M IC值分布情况为：1.5 mg/L为2株,1.0 mg/L为9株,0.75 mg/L为9株,0.5 mg/L为38株,0.38 mg/L为34株,0.25 mg/L为8株,均为敏感;替加环素对耐舒普深的泛耐药鲍曼不动杆菌株的M IC值分布情况为：≥32 mg/L为0株、16 mg/L为2株、8 mg/L为3株、4 mg/L为4株、2 mg/L为6株、1 mg/L为9株、0.5 mg/L为30株、0.25 mg/L为33株、0.125 mg/L为9株、0.06 mg/L为2株、0.03 mg/L为2株,敏感率为95.0%,中敏率为3.0%,耐药率为2.0%。结论替加环素或多粘菌素B是目前对耐舒普深的泛耐药鲍曼不动杆菌最有效药物之一。     

Properties of poliovirus strains isolated over the period 1969-1978 from the main municipal sewerage system of the city of Prague

Biological properties (rct marker and antigenic relatedness) were compared in vaccine prototype strains and in 62 poliovirus strains isolated during a period 1969-1978 from the main municipal sewerage system of the City of Prague. None of the strains isolated from the municipal sewerage showed biologic properties that would fully differ from those observed in vaccine-derived strains. The strains detected very late postvaccination (after about a year) showed a lesser extent of changes than strains isolated earlier after vaccination. The most frequent changes were recorded in type 2 strains, less frequent in type 3 strains and the least frequency of changes was found in type 1 strains. To facilitate comparisons of these changes in dependence on time of postvaccination virus excretion a supporting evaluation criterion has been developed to help express the dynamics of changes in the isolated poliovirus strains. The recorded degree of the dynamics of changes was highest in type 2 strains, lower in type 3 strains and lowest in type 1 strains. The dynamics of changes detected in strains of various types was not always constant in the course of years: in a given year (or in a period of several years) changes occurred always in strains of the same serologic type, whereas strains of the other two types changed only insignificantly during the respective period.     

The application of PCR fingerprinting to the differentiation of Yersinia enterocolitica strains isolated from humans and pigs

The distribution of different genotypes of Yersinia enterocolitica strains recovered from humans and from healthy pigs was investigated using PCR fingerprinting. The thirty six strains of Y. enterocolitica from humans, thirty five strains from pigs and Y. enterocolitica ATCC 9610 strain were included in this study. The tested strains of Y. enterocolitica belonged to O3 and O9 serogroups. The PCR fingerprinting using EAE5 primer (5' CTT AAT CTC AGT AAT GCT GGC CTT GG) made it possible to form five groups among the tested Y. enterocolitica strains. Two groups were very numerously represented by the tested strains. The thirty of Y. enterocolitica O3 strains from humans (thirty one of tested) and eighteen of Y. enterocolitica O3 strains from pigs (twenty of tested) belonged to one group. This group also included Y. enterocolitica ATCC9610 strain and four Y. enterocolitica O9 strains from pigs. All investigated Y. enterocolitica O9 strains from humans and the majority of Y. enterocolitica O9 strains isolated from pigs created a second, numerous group. The third genotype was created by two strains O9 from pigs, and the remaining two strains, isolated from pigs, belonging to O3 and O9 serogroups showed different binding patterns revealed by gel electrophoresis and created two other genotypes. The tested Y. enterocolitica strains which were isolated from humans formed only two groups but Y. enterocolitica strains isolated from pigs were found in five groups but such as the Y. enterocolitica strains from humans, the majority of strains from pigs were in first and second group. The Y. enterocolitica O3 strains regardless of their origin mostly represented the same PCR fingerprinting profile. The tested Y. enterocolitica O9 strains were more genetically diverse and represented four PCR fingerprinting profiles.     

Characteristics of E.coli O157 strains isolated in Poland from clinical material and food samples

E. coli belonging to the O157 serological group are among the organisms isolated most frequently out of all the so called entero-hemorrhagic E. coli strains (EHEC). Since several years they have been isolated also in Poland. The purpose of the present study was determination on selected phenotypic and genotypic properties of E. coli O157 strains isolated in our country from clinical material samples and from food. The serotype of the strains was determined, together with the following properties regarded as pathogenicity markers of verotoxic E. coli strains such as absence of beta-glucuronidase activity and sorbitol fermentation ability, as well as production of verotoxins SLT I and/or SLT II and entero-hemolysin. Besides that, by the PCR method the fragments of the genes coding for verotoxins, intimin and enterohaemolysin were amplified. The products of PCR were analysed by the restriction enzyme analysis (RFLP). All verotoxic E. coli O157 strains isolated in Poland were analysed by the pulsed field gel electrophoresis of genomic DNA (PFGE). The studied group comprised E. coli O157 strains, among them 40 strains were isolated from human faeces and 5 from food. The remaining strains were the reference E. coli O157:H7 EDL 933 and G 5244 strains and strains from NIH collection. The obtained results showed that the tested strains were a very varying population. 21 of them (all isolated from food, 11 from faeces and 5 reference strains) belonged to serotype O157:H7, five were not peritrichous O157:NM and the remaining ones had other ciliary antigen than H7. All strains isolated from food, reference strains and only 3 O157:NM strains isolated from humans were verotoxic. The strains from food and two reference strains produced only SLT II, 2 of 3 strains isolated from humans and one reference strain also produced only SLT II and the other produced both verotoxins. Apart from these 13 verotoxic strains all remaining strains caused sorbitol fermentation.     

Enzymatic characterization of Vibrionaceae strains isolated from environment and cold-blooded animals.

Enzymatic profiles were determined by the API ZYM system for 15 strains of non 01 Vibrio cholerae, 4 strains of V. metschnikovii, 9 strains of V. anguillarum, 6 strains of Plesiomonas shigelloides and 115 strains motile Aeromonas sp. All of the tested strains produced alkaline phosphatase, leucine aminopeptidase and did not possess alpha-fucosidase and alpha-mannosidase. Some differences in enzymatic activities among the tested Vibrionaceae strains were noted. The strains of non 01 V. cholerae, V. metschnikovii, V. anguillarum and P. shigelloides did not produce trypsin, whereas all of the tested Vibrio sp. strains appeared to be positive for this enzyme. Only the strains of P. shigelloides produced BI-Phospho-hydrolase. The lack of acid phosphatase activity was observed among the strains of V. anguillarum.     

The interchangeability of siderophores in the Enterococcus genus

The functional interchangeability of siderophores was tested among 62 strais belonging to 12 species of genus Enterococcus. Most investigated strains were from E. faecalis and E. faecium species. The majority of examined enterococcal strains appeared highly resistant to EDDA (ethylene-di-amine-di-ortho-hydroxyphenylacetic acid), therefore the group of sensitive strains involved only 11 used as indicator strains. The determination of interchangeability of siderophores within enterococcal strains was performed using EDDA-agar media into which the indicator strains were included. Test colonies (donor strains) were applied to the surface of the media to determine whether the indicator organisms could obtain the required iron for growth by utilizing chelators from the test colony. Only two strains: E. solitarius DSM 5634 and E. pseudoavium DSM 5632 did not demonstrate the ability to utilize siderophores synthesized by all investigated strains. The other tested indicator strains appeared to be recipients of siderophores from 20-52 donor enterococcal strains. The ability to exchange siderophores in enterococci was found as the feature characterizing individual strains.     

Lactic acid bacteria isolated from soy sauce mash in Thailand

Fourteen sphere-shaped and 30 rod-shaped lactic acid bacteria were isolated from soy sauce mash of two factories in Thailand. These strains were separated into two groups, Group A and Group B, by cell shape and DNA-DNA similarity. Group A contained 14 tetrad-forming strains, and these strains were identified as Tetragenococcus halophilus by DNA similarity. Group B contained 30 rod-shaped bacteria, and they were further divided into four Subgroups, B1, B2, B3, and B4, and three ungrouped strains by phenotypic characteristics and DNA similarity. Subgroup B1 contained 16 strains, and these strains were identified as Lactobacillus acidipiscis by DNA similarity. Subgroup B2 included two strains, and the strains were identified as Lactobacillus farciminis by DNA similarity. Subgroup B3 contained five strains. The strains had meso-diaminopimelic acid in the cell wall, and were identified as Lactobacillus pentosus by DNA similarity. The strains tested produced DL-lactic acid from D-glucose. Subgroup B4 contained four strains. The strains had meso-diaminopimelic acid in the cell wall, and they were identified as Lactobacillus plantarum by DNA similarity. Two ungrouped strains were homofermentative, and one was heterofermentative. They showed a low degree of DNA similarity with the type strains tested, and were left unnamed. The distribution of lactic acid bacteria in soy sauce mash in Thailand is discussed.     

Extracellular Enzymic Activity of Poultry Spoilage Bacteria

The extracellular enzymic activity has been studied of 224 strains of bacteria isolated mainly at 1° from spoiling chickens and turkeys and from poultry processing plants. The isolates comprised 44 strains of pigmented Pseudomonas , 57 strains of nonpigmented Pseudomonas , 29 strains of Ps. putrefaciens , 50 strains of oxidase positive Acinetobacter and 44 strains of oxidase negative Acinetobacter. None of the strains showed any significant activity against dextrin, starch, glycogen, inulin, dextran, xylan or pectin. Proteolytic activity was found mainly amongst 2 groups of pigmented pseudo-monads, and Ps. putrefaciens. Nuclease activity was found particularly amongst strains of Ps. putrefaciens and the oxidase negative Acinetobacter strains isolated from spoiling poultry. Almost all of the strains showed lipolytic activity when tested with tributyrin and a proportion of strains could also attack chicken fat. This latter property was particularly evident amongst the nonpigmented Pseudomonas strains.     

Relationship between O-serogroup and presence of pathogenic factor genes in Escherichia coli

A total of 383 isolates of serogroup-based enteropathogenic and enteroinvasive Escherichia coli (310 strains of EPEC and 73 strains of EIEC) were examined for the presence of corresponding pathogenic genes. The serogroup-based EPEC consisted of 232 strains isolated from diarrhea patients and of 78 strains from healthy carriers. The gene encoding intimin, eaeA, was detected in 42 of the 232 EPEC strains from patients (18.1%) and 9 of the 78 strains from carriers (11.5%). The difference was not significant. The bfp gene on the EAF plasmid was detected in 7 of the 42 eaeA-positive EPEC strains from patients but was not detected in the 9 strains from carriers. In serogroup-based EIEC, a chromosomal ipaH gene encoding one of the invasive plasmid antigens was detected in 4 of the 60 strains from patients (6%) but not in the 13 strains from carriers. The 4 ipaH-positive strains possessed the invasive plasmid. These results suggested that the serogroup-based diagnosis of EPEC and EIEC is not sufficient for identifying strains carrying the eaeA or ipaH gene.     

迟缓爱德华菌的外膜蛋白图谱及耐药性分析

分析 2 7株不同来源的迟缓爱德华菌 (Et)的外膜蛋白 (OMP) ,可分为A～H 8个型。 2 1株致病株与 6株非致病株有明显不同的图谱。致病株OMP条带多而深浓 ,并且相同来源的菌株有几乎一致的图谱。其中国内致病株以E型为主 ,与ATCC参考株相似。非致病株OMP条带则浅而稀疏 ,来源虽不同 ,但图谱极类似。另外 ,6株非致病株对磺胺、庆大、四环素等抗生素普遍耐药 ,而致病株除少数几株外 ,均对抗生素有不同程度的敏感     

Properties of poliovirus strains isolated from waste waters of nurseries in Prague in 1969-1982

The rct marker and antigenic marker were studied in poliovirus strains of all the three types isolated from waste waters of selected communities of children in Prague in 1969-1982. Only three isolated strains (two of type 2 and one of type 3) i.e. 1.38% of strains from nursery waste waters and 0.99% of strains from the total Prague municipal sewerage system, differed in both markers from prototype vaccine strains. The rct marker was most frequently altered in strains of type 1; antigenic marker varied most often in strains of type 3, less conspicuously in strains of type 2 and minimum changes were found in type 1 strains. The dependence of changes in both studied markers of isolated strains on the interval between vaccination and isolation from waste waters was reflected in the dynamics of changes. The highest dynamics of changes in rct marker and antigenic marker was recorded in type 3 and type 2 strains, minimal dynamics being found in type 1 strains. Study of type 2 strains isolated for a long time in some years showed that the longer the interval between vaccination and isolation, the higher the similarity between the isolated strain and the prototype vaccine strain.     

297例浅部真菌病临床及病原菌菌种分析

目的 探讨皮肤浅部真菌病致病真菌菌种的构成.方法 对297例真菌涂片阳性和培养阳性的浅部真菌病患者,取标本进行分离培养及菌种鉴定,培养阳性标本在形态学上不能准确鉴定的,进行梅里埃API 20C AUX酵母菌鉴定试剂盒或核糖体DNA （rDNA） ITS区序列测定,确切鉴定菌种.使用SPSS 17.0统计软件对于结果进行统计分析.结果 共分离培养出致病菌13种,其中红色毛癣菌86株（29.0％）,须癣毛癣菌68株（22.9％）,念珠菌属59株（19.9％）,暗色真菌属13株（4.4％）,曲霉菌属13株（4.4％）,红酵母菌12株（4.0％）,青霉菌属9株（3.0％）,毛霉菌9株（3.0％）,犬小孢子菌5株（1.7％）,浅白隐球菌3株（1.0％）,毛孢子菌属2株（0.7％）,絮状表皮癣菌1株（0.3％）,混合感染17株（5.7％）.结论 本地区浅部真菌病以甲癣为主,主要致病真菌是红色毛癣菌,但其他种类真菌感染尤其是念珠菌属有明显上升趋势.     

Starch gel electrophoresis: an effective method for separation of pathogenic and nonpathogenic Naegleria strains

Isoenzyme electrophoresis of 7 different enzyme systems was used to compare 24 strains of Naegleria fowleri and 6 strains of N. gruberi. The 30 strains could be grouped into 4 distinct categories based upon zymogram patterns. No interstrain band variation in all enzyme systems was demonstrated in pathogenic strains of N. fowleri. Three nonpathogenic high temperature-tolerant strains of Naegleria had similar zymograms. Four of the 5 remaining nonpathogenic Naegleria strains had no interstrain band variation. Based upon zymograms, the 22 pathogenic strains constitute a homogenous species. Similarly the high temperature-tolerant nonpathogenic strains formed a cohesive group. The remaining nonpathogenic strains could be separated into 2 groups.     

Characterisation of persistent and sporadic Listeria monocytogenes strains by pulsed-field gel electrophoresis (PFGE) and amplified fragment length polymorphism (AFLP)

This study was set up to evaluate the genetic similarity or dissimilarity of persistent and sporadic Listeria monocytogenes strains existing in eleven food processing facilities, including fish, dairy, meat and poultry processing plants. In each plant persistent and sporadic strains were selected on the basis of PFGE typing results. A total of 17 strains representing persistent strains and 38 sporadic strains originating from eleven food processing plants were included in the study. PFGE macrorestriction patterns of persistent and sporadic strains from different processing plants were compared and the strains were further studied by amplified fragment length polymorphism (AFLP), being a characterisation method giving more whole genome based information. The 17 persistent and 38 sporadic strains showed 14 and 35 pulsotypes, 14 and 28 AFLP types, respectively. The combination of PFGE and AFLP typing results yielded a total of 48 genotypes. Thirteen of 15 genotypes presented by persistent strains were only associated with persistent strains and similarly 94% (33/35) of genotypes showed by sporadic strains were recovered among sporadic strains only. Our results showed that L. monocytogenes strains causing persistent contamination differ from sporadic strains. In AFLP analysis persistent strains did not, however, form any specific clusters and neither was there any difference between the known two genomic groups. These results indicate that even though persistent strains differ from sporadic strains there seems not to be any specific evolutional lineage of persistent strains.     

Reciprocal effect of Klebsiella pneumoniae and Lactobacillus on their cytadhesion

Reciprocal effect of 4 strains of Klebsiella pneumoniae and 6 Lactobacillus strains on their cytadhesion in mixed populations was studied on a model of formalinized human erythrocytes. The Lactobacillus strains included 2 strains of Lactobacillus casei subsp. casei, 2 strains of L. plantarum and 2 strains of L. fermentum. It was shown that adhesion of both the Klebsiella and the Lactobacillus strains changed under their reciprocal effect. The changes were characterized by the strain differences and depended on the quantitative ratio of the microorganisms.     

阴道中益生特性乳杆菌的分离与功能评价

目的 分离健康女性阴道中的乳杆菌并鉴定其益生特性,为开发治疗妇科疾病的复方益生菌制剂提供新型菌株。方法 采集健康女性阴道分泌物并分离筛选乳杆菌,通过16S rDNA序列分析鉴定乳杆菌分离株,并对其产酸性能、产H2O2能力、抑菌能力、产生物膜能力进行检测。结果 从50名健康女性阴道内共分离出179株乳杆菌,其中卷曲乳杆菌101株、詹氏乳杆菌42株、格氏乳杆菌26株、植物乳杆菌5株、唾液乳杆菌3株以及干酪乳杆菌2株。179株乳杆菌中有146株具有产酸能力,发酵液pH值的最低的5株菌分别为卷曲乳杆菌J3、卷曲乳杆菌J8、詹氏乳杆菌J87,植物乳杆菌J75以及格氏乳杆菌J35,其pH分别为4.20、4.23、4.24、4.26及4.36;产H2O2弱阳性菌株有87株、阳性有37株、强阳性有9株,这9株菌分别为卷曲乳杆菌J3、卷曲乳杆菌J8、卷曲乳杆菌J20、詹氏乳杆菌J87,詹氏乳杆菌J90、詹氏乳杆菌J15、格氏乳杆菌J11、植物乳杆菌J75、植物乳杆菌J69以及植物乳杆菌J40;能拮抗大肠埃希菌的菌株有115株、拮抗金黄色葡萄球菌的有84株、拮抗白假丝酵母的有52株;经统计,对三者同时有拮抗作用且作用最强的只有6株,分别为卷曲乳杆菌J3、卷曲乳杆菌J50、卷曲乳杆菌J62、詹氏乳杆菌J87、詹氏乳杆菌J16和格氏乳杆菌J66;不同乳杆菌产生物膜能力数值范围在1.0~5.4,卷曲乳杆菌、詹氏乳杆菌、干酪乳杆菌的生物被膜形成能力显著高于其他三种菌（P<0.05）。在全部179株菌中,卷曲乳杆菌J3和詹氏乳杆菌J87既具有强的产酸能力和产过氧化氢能力,又有较强抑菌活性,同时产生物膜能力也最强。结论 卷曲乳杆菌J3和詹氏乳杆菌J87具有优良的生物学特性,有望成为用于治疗妇科疾病微生态制剂的备选菌株。     

6个杏鲍菇菌株生理生化研究

目的:对收集的6个杏鲍菇菌种菌丝进行生理、生化研究,酯酶同工酶法分析菌株间的差异与关系。方法:利用杏鲍菇菌丝阶段的生长速度测定、诱变剂抗性试验和酯酶同工酶比较,对杏鲍菇菌株营养生长阶段的理化指标进行研究,对酯酶同工酶谱进行相似系数和遗传距离分析,确立各菌株的异同。结果:菌株间菌丝生长速度差异不显著;抗诱变剂实验结果显示,2、5、6号菌株对诱变剂抗性较强;酯酶同工酶分析表明,杏鲍菇6个菌株同工酶酶带存在的差异很大。结论:通过试验表明,所选6株杏鲍菇菌种为各自独立菌株。     

SSR标记在毛木耳种质资源遗传多样性研究中的应用

本研究利用基于毛木耳全基因组开发的SSR标记对27份毛木耳菌株（野生14株、栽培13株）的遗传多样性进行分析。首先随机选取3个菌株（2个野生菌株、1个栽培菌株）的DNA为模板,从144对SSR引物中筛选出扩增条带清晰、稳定性强、多态性丰富的引物24对。24对SSR引物共检测到116个多态性SSR片段,每对引物的多态性片段有3-7个,引物平均检测效率为4.83个,Shannon’s遗传多样性指数范围是0.866-1.885,多态性位点比率100%。供试菌株遗传相似系数范围是0.618-0.971,说明毛木耳种质资源具有丰富的遗传多样性。野生菌株与栽培菌株间平均遗传相似系数分别为0.746、0.779,说明毛木耳野生菌株遗传多样性更为丰富。经聚类分析,在遗传相似系数为0.680时,可将供试菌株分为无色（白色）类群Ⅰ和有色（浅黄色到红褐色）类群Ⅱ。遗传相似系数为0.704时,可将供试菌株中栽培菌株和野生菌株明显区分（14株野生菌株均在类群Ⅱ-2中,13株栽培菌株分别在类群Ⅰ和Ⅱ-1中）。本研究表明基于全基因组的SSR标记能从分子水平上揭示各菌株间的遗传差异,丰富毛木耳遗传多样性的研究手段,并为进一步进行毛木耳的品种选育、遗传学研究等提供有力手段。