一株布氏菌新种的综合鉴定

从国外得到一株布氏菌新种,用常规以外的DNA同源性、多种R型血清凝集和对分群噬菌体裂解试验系列检查结果:G+C mol%值为57.1,Tm为78.7℃,与抗IV型猪种血清、抗牛种45/20菌血清、抗羊种B115菌血清的效价均为1∶3200,与抗犬种RM6/66菌血清和阴性兔血清均为(一),与M血清为(+),与A血清为(-),能被Wb、BK₂噬菌体裂解,对Tb、Fi、R/C和8j-309噬菌体不敏感。其生长不需CO₂、不产生H_2S、对硫堇染料不敏感、对碱性复红染     

布氏菌噬菌体SA对各种布氏菌的裂解活性

布氏菌噬菌体SA是从猪种布氏菌培养物中分离到的。其裂解活性当菌液浓度为10^-4时,对光滑型猪种菌1、3型,牛种菌1、3、6型和沙林鼠种菌可产生混合性裂解,产生噬菌斑直径3—4mm。噬菌体SA原液或稀释浓度为10^-1、10^-2时,对羊种菌1、3型不裂解。对羊种菌2型、粗糙型牛种菌45/20、犬种菌和绵羊附睾菌产生混合性裂解或噬菌斑,噬菌斑为1—2mm或更小。     

国内分离的犬布氏菌的电镜观察

本文报告了用电镜对国内分离的{株犬布氏菌和W．H. O.的4种对照菌(包括犬布氏菌、猪布氏菌、羊布氏菌和牛布氏茵)进行的电镜观察结果。所有上述菌株的形状和大小基本一致,均为球杆状,其球状体直径为0.70—0．73μm;杆状体长1.0一1.46μm,宽0.5--0.7μm。细胞壁均由外膜和肽聚糖两部分组成,具有革兰氏阴性细胞壁的特征。国内株犬布氏菌的细胞表面,经常产生乳头状突起,它包有外膜,内含肽聚糖,实际上是细胞壁的一部分。外胰主要由脂多糖(LPS)组成,因此乳头状突起有可能是细菌释放内毒素(Endotoxin)的一种形式。这一现象在W．H．O．菌株中比较少见。细胞分又生长,在布氏菌中比较普遍,尤以W．H．O．菌株更为多见。超薄切片的结果表明,细胞分又是由杆状细菌的侧分裂形成的。但 是,一般报道多认为细菌的繁殖方式是横分裂。     

溶藻弧菌噬菌体的分离

从海产品中检出29株溶藻弧菌噬菌体,从中选出4株进行鉴定。据噬菌斑特征可分为两类：一类是透明,一类是不透明。但大小各有差异,直径均在0．5—3.0mm。电镜可见形态也可分成两种类型：即头部呈长轴六角形,尾部细长,结构简单;另一类头部呈等轴六角形,但棱角不甚明显,尾部很短。这些噬菌体的增殖效价均可达108-9pfu／ml,对溶藻弧菌的综合裂解率为72.22％,单株平均裂解率在9.72—44．4％。4株噬苗体的特异性高,原液与612株属外常见菌作交叉裂解试验,均未出现交叉裂解现象;与697株属内弧菌I昊I试,也仅对副溶血弧菌有39.0％的交叉裂解,但将原液稀释至10RTD时,大部分交叉现象消失,表明两菌间有明显的亲缘关系。     

应用肠杆菌科四个属的诊断噬菌体快速诊断沙门氏菌

本文报告应用弗氏柠檬酸细萄噬菌体3组,大肠埃希氏菌噬菌体4组,阴沟肠杆菌噬菌体1组和沙门氏菌O-I噬菌体快速诊断沙门氏菌的结果。沙门氏菌0-I噬菌体可裂解沙门氏菌属地方株1393株中的1351株(97％)。柠檬酸细菌属噬菌体和共可裂解柠橡酸细菌属地方株381株中的362株(95％)。阴沟肠杆菌噬菌体Ent可裂解阴淘肠杆菌地方株l 50株中的133株(84．2％)。埃希氏菌属噬菌体E—1、E一2、E-3和E-4共可裂解埃希氏菌属地方株683株中的567株(83％)。由于E一1和E一2噬菌体的联合使用,可使o I噬菌体对埃希氏菌属地方株的误诊率从6．3％下降到0．6％。E一4噬菌体对沙门氏菌属地方株的误诊率可因与。一I噬菌体的联合使用而从0．36％下降到0．07％。     

应用肠杆菌科诊断噬菌体检测志贺氏菌的评价

应用肠杆菌科诊断噬菌体对2280株疑似志贺氏菌进行了检测,同时进行了常规鉴定。结果表明,志贺氏菌属Sh噬菌体10³RTD对属内裂解率为100%,1RTD为99.9%;65株与志贺氏菌分型血清呈现凝集的非志贺氏菌,10³RTD裂解率为12.3%,1RTD为4.6%。裂解模式的测定表明,2215株志贺氏菌分属于7个裂解模式,仅模式3中3株鲍氏5型为文献[2,3]所未列入,余者完全一致。Sh10³RTD裂解的非志贺氏菌均可用1RTD和裂解模式排除     

鲍曼不动杆菌裂解性噬菌体LZ35的分离及全基因组分析

目的 通过分离鉴定鲍曼不动杆菌噬菌体并进行遗传信息分析,为今后噬菌体用于治疗鲍曼不动杆菌引起的感染提供依据。方法 以鲍曼不动杆菌临床分离株为宿主菌,从医院污水中分离鲍曼不动杆菌噬菌体并进行纯化、电镜观察形态特征、提取噬菌体DNA,进行全基因组测序,分析全基因组的结构特征,比较基因组分析其进化关系。结果 分离到鲍曼不动杆菌裂解性噬菌体LZ35,电镜观察显示,该噬菌体属于有尾噬菌体目肌尾病毒科。基因组全长44 885 bp,G＋C含量为37.95%,含有83个开放阅读框,其中22个编码序列可预测其功能,61个编码序列为未知基因。噬菌体LZ35的基因组与鲍曼不动杆菌噬菌体IME-AB2和YMC-13-01-C62具有很高的同源性（分别为97%和99%）,与鲍曼不动杆菌噬菌体YMC11/12/R1215的进化关系最近。结论 以鲍曼不动杆菌临床分离株为宿主菌,分离到鲍曼不动杆菌裂解性噬菌体LZ35,明确了其形态和基因组特征,为防治噬菌体疗法奠定基础。     

应用噬菌体GH15和K治疗金黄色葡萄球菌感染

[目的]研究金黄色葡萄球菌噬菌体GH15和K的生物学特性及联合用于治疗金黄色葡萄球菌感染的潜力.[方法]通过透射电镜观察噬菌体GH15和K的形态;测定二者的裂解谱和一步生长曲线;通过体外裂解实验和体内治疗实验分别测定单独使用GH15、K及二者混合使用时的裂解能力和对菌血症小鼠的保护效果.[结果]通过电镜观察发现两个噬菌体的外形相似,但GH15的尾部比K的长.GH15裂解谱较宽,可以裂解28株金葡菌,而K仅能裂解7株金葡菌.通过对两株噬菌体感染7株共同宿主菌形成的一步生长曲线进行拟合曲线分析,表明二者对不同宿主菌的增殖趋势是不同的.在体外,混合噬菌体与单个噬菌体的抑菌活性无明显差别;在体内实验中,混合噬菌体表现出优于单个噬菌体的治疗效果,用较低的剂量即可以达到高剂量单个噬菌体的治疗效果.[结论]GH15和K所形成的混合噬菌体在治疗金黄色葡萄球菌感染具有更大的应用潜力.     

金黄色葡萄球菌荚膜分型血清研制及初步应用

为了解中国金葡菌荚膜流行型,用5型和8型国际标准菌株的菌悬液免疫家兔,吸收除去交叉凝集素研制出金葡菌5型和8型荚膜分型血清,并应用于333株临床菌株荚膜分型。该血清效价为1：1280和1：640,与本菌及其它同型菌株玻片凝集反应呈强凝集,与其它型菌株不凝集,且与美国标准血清同时对333株临床分离菌株进行分型比较的符合率为100％。333株金葡地方菌株荚膜分型结果显示,8型菌株占绝对优势,所占百分率为70．9％;5型占6．3％,5型和8型菌株共占77．2％。这与国外金葡菌荚膜5型和8型占70％-80％的报道相似。试验为金葡菌疫苗株选择提供了流行病学依据。     

粘质沙雷菌裂解性噬菌体ФSM9-3Y的分离和鉴定

目的从医院污水中分离粘质沙雷菌噬菌体,并分析其生物学特性,为进一步研究针对耐药性粘质沙雷菌的噬菌体制剂提供依据。方法采用双层琼脂平板法分离纯化针对粘质沙雷菌的裂解性噬菌体,观察噬菌体对宿主菌的裂解特异性,通过负染法电镜观察噬菌体的形态结构,提取噬菌体核酸进行酶切电泳,测定噬菌体的最佳感染复数和一步生长曲线,SDS-PAGE电泳初步分析噬菌体的结构蛋白和非结构蛋白。结果从医院污水分离出7株可裂解粘质沙雷菌的噬菌体,对其中一株噬菌体(命名为ФSM9-3Y)的生物学特征进行了初步研究。电镜显示噬菌体呈蝌蚪状,头部为20面体立体对称、直径约70 nm;尾部长约50 nm。ФSM9-3Y的最佳感染复数为1。一步生长曲线表明;ФSM9-3Y的潜伏期约30 min,暴发时间70 min,暴发量为629 PFU/cell。凝胶电泳显示噬菌体基因组为双链DNA、大小约54 kb。SDS-PAGE呈现至少包括13种蛋白,相对分子质量范围在25～130 kD,其中主要蛋白的相对分子质量约为48 kD。结论此次分离的噬菌体ФSM9-3Y为裂解性噬菌体,根据形态和结构特征,粘质沙雷菌噬菌体ФSM9-3Y属于有尾病毒目,肌尾噬菌体科。     

Genomic island 2 is an unstable genetic element contributing to Brucella lipopolysaccharide spontaneous smooth-to-rough dissociation

Brucella is a Gram-negative bacterium that causes a worldwide-distributed zoonosis. The genus includes smooth (S) and rough (R) species that differ in the presence or absence, respectively, of the O-polysaccharide of lipopolysaccharide. In S brucellae, the O-polysaccharide is a critical diagnostic antigen and a virulence determinant. However, S brucellae spontaneously dissociate into R forms, a problem in antigen and S vaccine production. Spontaneous R mutants of Brucella abortus, Brucella melitensis, and Brucella suis carried the chromosomal scar corresponding to genomic island 2 (GI-2) excision, an event causing the loss of the wboA and wboB O-polysaccharide genes, and the predicted excised circular intermediate was identified in B. abortus, B. melitensis, and B. suis cultures. Moreover, disruption of a putative phage integrase gene in B. abortus GI-2 caused a reduction in O-polysaccharide loss rates under conditions promoting S-R dissociation. However, spontaneous R mutants not carrying the GI-2 scar were also detected. These results demonstrate that the phage integrase-related GI-2 excision is a cause of S-R brucella dissociation and that other undescribed mechanisms must also be involved. In the R Brucella species, previous works have shown that Brucella ovis but not Brucella canis lacks GI-2, and a chromosomal scar identical to those in R mutants was observed. These results suggest that the phage integrase-promoted GI-2 excision played a role in B. ovis speciation and are consistent with other evidence, suggesting that this species and B. canis have emerged as two independent lineages.     

Intraspecies biodiversity of the genetically homologous species Brucella microti

Brucellosis is one of the major bacterial zoonoses worldwide. In the past decade, an increasing number of atypical Brucella strains and species have been described. Brucella microti in particular has attracted attention, because this species not only infects mammalian hosts but also persists in soil. An environmental reservoir may pose a new public health risk, leading to the reemergence of brucellosis. In a polyphasic approach, comprising conventional microbiological techniques and extensive biochemical and molecular techniques, all currently available Brucella microti strains were characterized. While differing in their natural habitats and host preferences, B. microti isolates were found to possess identical 16S rRNA, recA, omp2a, and omp2b gene sequences and identical multilocus sequence analysis (MLSA) profiles at 21 different genomic loci. Only highly variable microsatellite markers of multiple-locus variable-number tandem repeat (VNTR) analysis comprising 16 loci (MLVA-16) showed intraspecies discriminatory power. In contrast, biotyping demonstrated striking differences within the genetically homologous species. The majority of the mammalian isolates agglutinated only with monospecific anti-M serum, whereas soil isolates agglutinated with anti-A, anti-M, and anti-R sera. Bacteria isolated from animal sources were lysed by phages F1, F25, Tb, BK2, Iz, and Wb, whereas soil isolates usually were not. Rough strains of environmental origin were lysed only by phage R/C. B. microti exhibited high metabolic activities similar to those of closely related soil organisms, such as Ochrobactrum spp. Each strain was tested with 93 different substrates and showed an individual metabolic profile. In summary, the adaptation of Brucella microti to a specific habitat or host seems to be a matter of gene regulation rather than a matter of gene configuration.     

Identification of enterotoxigenic staphylococci from sheep and sheep cheese

The total of 127 Staphylococcus aureus strains obtained from sheep and sheep cheese were examined for their biochemical activities, biotypes, phage patterns, and ability to produce enterotoxins. Of the 83 staphylococcal strains isolated from animals 77 (93%) were classified as the C biotype. Of this group of sheep-adapted strains, 61 (79%) were sensitive to phage 78, and 46 (60%) produced enterotoxin C exclusively. The three isolated belonging to the A biotype produced enterotoxin D, and two of the three unclassifiable strains produced enterotoxin A. Of the 44 staphylococcal strains isolated from sheep cheese, there were 37 (84%) identified as the C biotype. From this series, 31 (84%) strains were lysed with phage 78, 6 (16%) strains produced enterotoxin C, and 1 strain produced enterotoxin A. One of the six strains determined as the A biotype produced enterotoxin D. C biotype strains, especially of ovine origin, are an exception among animal staphylococci, because a large number of them are enterotoixgenic. The C antigenic type is the most usual of the known enterotoxins in staphylococci of animal provenance.     

METABOLIC CHARACTERIZATION OF THE GENUS BRUCELLA IV. 3: Correlation of Oxidative Metabolic Patterns and Susceptibility to Brucella Bacteriophage, Type abortus, Strain.

Meyer, Margaret E. (University of California, Davis). Metabolic characterization of the genus Brucella. IV. Correlation of oxidative metabolic patterns and susceptibility to Brucella bacteriophage, type abortus, strain 3. J. Bacteriol. 82:950-953. 1961.-A total of 212 strains of brucellae that had been identified as Brucella melitensis, B. abortus, B. suis, or B. neotomae by their oxidative metabolism were tested for their susceptibility to Brucella bacteriophage, type abortus, strain 3. It was demonstrated that only those organisms that displayed the oxidative metabolic pattern that is singular for B. abortus were susceptible to this strain of phage, irrespective of their identity by the conventional methods usually employed for differentiating members of this genus. Strains of organisms that display the features of B. melitensis by the conventional determinative methods, but display the metabolic characteristics of B. abortus, are susceptible to lysis by this phage. These organisms are in fact B. abortus. Strains of organisms that display the features of B. melitensis by the classical methods, and display the metabolic pattern of B. melitensis, are not lysed by this phage. These organisms are B. melitensis. The conclusions then were drawn that B. abortus is the only species that can serve as host for this strain of phage, that oxidative metabolic patterns accurately identify the species in this genus, and that by the conventional methods of differentiation, many strains of B. abortus are misidentified as B. melitensis.     

Phage types of Staphylococcus aureus strains and their antibiotic resistance in carriers of medical students population

The phage types of 78 S. aureus strains isolated from nose swabs obtained from a medical students in 2005 -2006 was determined and antibiotic resistance of the phage types was analysed. 680 students were tested in order to obtain the strains and 11.5% of them were carriers of S. aureus. Phage typing was performed using basic set of23 phages and 3 additional phages: 88, 89 and 187. Drug resistance was determined by the disc-diffusion method. The most frequent in studied population were the group III (21.8%) and strains lysed by phages belonging to varied groups (21.8%). Highly different phage patterns were observed among strains belonging to each of the group. Strains belonging to the group III as the strains lysed by phages from varied groups were most frequently resistant only to penicillin (52,9% respectively). Resistance to penicillin was also most often observed in the strains belonging to another groups and phage types. Usefulness of the additional phages 88,89 and 187 was in the investigations as no more than 51% of strains was lysed by this phages.     

Evaluation of Micro Complement Fixation Tests for Antibodies Against Group A Streptococcal M and M-Associated Antigens in Rabbit and Human Sera

The microslide gel-diffusion and macro-tube agglutination techniques to detect Brucella canis antibodies in dogs were compared. Sera from dogs experimentally infected with B. canis and a random sample of dog sera with unknown histories of exposure to this organism were examined. The results of the gel-diffusion method employing specific rough Brucella saline-extract antigens of B. canis and Brucella ovis were comparable to those obtained by the tube agglutination test. The easily prepared, stable R antigen in the freeze-dried form offers a convenient, simple, and reliable diagnostic method for the serological detection of canine brucellosis by the gel-diffusion test.     

Brucella melitensis B115-based complement fixation test to detect antibodies induced by Brucella rough strains

Aims:  To assess the efficiency of a Brucella melitensis B115 rough strain, naturally devoid of anticomplementary activity, used as antigen in a complement fixation test (CFT) to detect antibodies induced by Brucella strains with rough phenotype, such as Brucella abortus RB51, Brucella ovis and Brucella canis .
Methods and Results:  Complement fixation testing was performed on sera from RB51-vaccinated cattle and buffaloes, B. ovis -infected sheep and B. canis -infected dogs using B115, RB51 and the hot saline extract (HSE) as antigens. The B115-based CFT proved highly sensitive and specific in detecting rough antibodies and its efficiency was comparable with that of RB51 and HSE-based CFT.
Conclusions:  Brucella melitensis B115 can be successfully used as an antigen in CFT to detect antibodies induced by Brucella rough strains.
Significance and Impact of the Study:  Brucella melitensis B115 antigen may represent an improvement over Brucella rough strains for Brucella antibody detection by CFT, thus enhancing the efficiency of brucellosis surveillance systems. Owing to the absence of anticomplementary activity, it does not require particular growth conditions or modifications and can be accurately standardized. The B115-based CFT may constitute a suitable supplementary test for the diagnosis of human infections owing to rough Brucellae .     

用MLVA技术和多重PCR对犬种布氏菌基因分型

目的：对犬种布氏菌的遗传关系进行不同分子分型方法的对比研究,为犬布病分子流行病溯源提供有效方法。方法：采用多重PCR和多位点可变数量串联重复序列分析（MLVA）方法对24株犬种布氏菌的遗传关系进行比较研究。结果：多重PCR只鉴定出1株犬种布氏菌,其余23株均鉴定为猪种鲁氏菌,但不能鉴定型别;MLVA方法对已鉴定为猪种的布氏菌仍可再细分为型,87%（20/23）为猪3型,13%（3/20）为猪1型。结论：MLVA可以对布氏菌种（生物型）进行基因分型鉴定,可以作为传统表型鉴定方法的补充。     

Phage Typing Reactions on Brucella Species

The nature of the phage typing reactions on Brucella species was determined by rates of adsorption and infection, one-step growth experiments, and susceptibility to lysis from without. The highest rates of adsorption and infection were obtained on smooth B. abortus cultures, and large clear plaques were produced. One or a few phage particles per B. neotomae cell killed about one-half of the cells, but some went through an infective cycle and released mature phage that resulted in production of small clear plaques. With B. suis, more phage particles per cell were required to kill, replication did not occur, and plaques were not observed. Still greater numbers of phage particles were required to cause some inhibition of growth of B. melitensis lawns. Rough Brucella cultures and species, such as B. ovis and B. canis, were not affected by the highest concentrations of phage. B. abortus cultures of intermediate colonial morphology adsorbed phage, but only a few infected cells (after a delayed latent period) released mature phage. An infected culture or colony appeared normal until spontaneous phage mutants appeared which could penetrate the cell wall more effectively than the parent phage. The mutant phage multiplied more rapidly, and the colony changed to a sticky white form.