Differentiation of Staphylococcus sciuri strains isolated from free-living rodents and insectivores

Twenty-nine Staphylococcus sciuri strains isolated from free-living insectivores and rodents were comparatively analysed for their biochemical capacities and their SmaI macrorestriction patterns. The 29 S. sciuri isolates represented 21 different biotypes and 22 different SmaI macrorestriction types. This observation confirmed that S. sciuri isolates obtained from insectivores and rodents living in natural environments constituted a heterogeneous population. Cluster analysis revealed that the macrorestriction patterns and the biochemical profiles matched in some cases. However, S. sciuri isolates that exhibited the same or closely related biochemical profiles were also found to be associated with different macrorestriction patterns. Analysis of the 29 S. sciuri isolates for their plasmid content and their sensitivity to antimicrobial agents showed that most of the isolates were plasmid-free and susceptible to all antimicrobial agents tested.     

Staphylococcus sciuri as a part of skin, nasal and oral flora in healthy dogs.

The coagulase-negative species Staphylococcus sciuri is widespread in nature and is associated with a variety of domestic and wild animals. However, the occurrence of S. sciuri in dogs has received little attention so far. In the present study, we established the prevalence of S. sciuri in a large population of healthy dogs, and characterized isolated strains. Samples from two mucous membrane sites (anterior nares and mouth), and two hair-coated sites (head and withers) were taken from 122 dogs and inoculated into STS agar, a novel selective medium that was introduced and tested in the study. In total, 116 isolates of S. sciuri were obtained from 488 specimens. S. sciuri was isolated from 56 out of 122 (46%) dogs. The occurrence of S. sciuri in the anterior nares and mouth were significantly higher than those in withers and head. No significant association of S. sciuri occurrence in dogs and factors such as sex, age, and living environment (indoor/outdoor) was found. Out of 56 dogs, which tested positive for S. sciuri, 30 (54%) would have it as a resident flora. Thus, we showed that S. sciuri was frequently present as a part of skin, nasal and oral flora in healthy dogs both as a resident and transient carriage.     

Toxinotypes of Clostridium perfringens isolated from sick and healthy avian species.

Currently, the factors/toxins responsible for Clostridium perfringens-associated avian enteritis are not well understood. To assess whether specific C. perfringens' toxinotypes are associated with avian enteritis, the isolates of C. perfringens from 31 cases of avian necrotic or ulcerative enteritis submitted between 1997 and 2005 were selected for retrospective analysis using multiplex PCR. C. perfringens was isolated from chickens, turkeys, quail, and psittacines. The toxinotypes of isolates from diseased birds were compared against the toxinotype of 19 C. perfringens isolates from avian cases with no evidence of clostridial enteritis. All C. perfringens isolates were classified as type A regardless of species or disease history. Although many isolates (from all avian groups) had the gene encoding the C. perfirngens beta2 toxin, only 54% produced the toxin in vitro when measured using Western blot analysis. Surprisingly, a large number of healthy birds (90%) carried CPB2-producing isolates, whereas over half of the cpb2-positive isolates from diseased birds failed to produce CPB2. These data from this investigation do not suggest a causal relationship between beta2 toxin and necrotic enteritis in birds.     

猪源缓慢葡萄球菌的分离鉴定及致病性研究

某猪场发生一种高发病率、高死亡率的传染病。本研究从该场濒死猪只的心脏血液及肺脏中分离出1株细菌,经镜检、菌落形态观察、培养特性观察及法国生物梅里埃API鉴定系统鉴定,确定该分离菌株为缓慢葡萄球菌。药敏试验表明该菌对多种药物具耐药性,但对氟哌酸、红霉素和强力霉素等敏感;致病性试验表明,分离菌可以引起实验小鼠的慢性死亡,猪只腹腔注射可复制病例,表明缓慢葡萄球菌具有致病性并很可能会对公共卫生安全具有潜在威胁。     

仓鼠源缓慢葡萄球菌的分离鉴定及生物学特性研究

从死亡仓鼠的血液及内脏器官中分离出一株细菌,经镜检、菌落形态观察、培养特性观察、生化试验、PCR鉴定及16 S rRNA基因测序和同源性分析,确定该分离菌株为缓慢葡萄球菌。动物回归试验表明,分离菌可以引起实验小鼠的慢性死亡,这表明缓慢葡萄球菌具有致病性并很可能会对公共卫生安全具有潜在威胁。药敏试验显示,该分离菌株对大多数抗生素均十分敏感,但对氟喹诺酮类药物及复方新诺明有很强的耐药性。     

1株黄颡鱼源松鼠葡萄球菌的分离鉴定及其耐药性分析

为确定导致江苏省某水产养殖户黄颡鱼(Pelteobagrus fulvidraco)细菌性感染死亡的病原,从患病黄颡鱼肝脏中分离到1株致病菌(命名为GM1508),通过培养特性观察、革兰染色、生化试验、16S rRNA基因序列分析,初步鉴定为松鼠葡萄球菌(Staphylococcus sciuri,S.sciuri)。扩增的16S rRNA基因序列长度为1 454 bp(Gen Bank登录号KX781275),用与该菌株核苷酸同源性较高的S.sciuri、表皮葡萄球菌(Staphylococcus epidermidis)、鸡葡萄球菌(Staphylococcus gallinarum)、猪葡萄球菌(Staphylococcus hyicus)和金黄色葡萄球菌(Staphylococcus aureus)代表菌株序列,进行同源比对和系统发育分析。结果发现,本分离株与13株S.sciuri的同源性为99.1%~99.7%,在系统发育树上聚为同一枝,表明该分离株为S.sciuri。以0.2 mL/只(1×108cfu/mL)菌液的剂量接种黄颡鱼,死亡率为100%(10/10),表明该分离株对黄颡鱼具有较强致病性。药敏试验结果显示分离株对头孢唑啉、青霉素、米诺环素、呋喃妥因、氯霉素等20种药物敏感。本研究为黄颡鱼S.sciuri感染的防控提供了理论依据。     

Antimicrobial susceptibility of Staphylococcus intermedius isolated from healthy and diseased dogs

A total of 90 strains of Staphylococcus intermedius isolated from dogs were examined for antimicrobial susceptibility. There were no significant differences in the distribution patterns of MICs between strains from 1982 to 1985 and those from 1999, and between strains from healthy dogs and those from diseased dogs. All of the strains were susceptible to ABPC, DMPPC, CEX, TDM, ERFX, BFLX, and FF at concentrations of 0.05 to 6.25 microg/ml. The MICs of OTC, KM, EM, AIV-TS, and LCM were distributed in a broad range of 0.1 to >100 microg/ml, indicating the existence of resistant as well as susceptible populations of S. intermedius. Thirty-three strains (36.7%) were resistant to one or more anitmicrobial agents such as OTC (n=32), KM (n=9), EM (n=7), AIV-TS (n=7), and LCM (n=7).     

耗儿鱼松鼠葡萄球菌的分离鉴定及耐药性分析

为了探讨松鼠葡萄球菌(Staphylococcus sciuri)的病原学特性,本试验从耗儿鱼(Corydoras)分离出菌株YDE0916,并对其进行生理生化鉴定、16S rDNA基因PCR扩增、序列分析及耐药性分析。结果显示,该菌株为革兰氏阳性球菌,呈串形,无芽孢,无荚膜。该菌不运动,β-半乳糖苷酶、VP反应为阴性,蔗糖、麦芽糖为阳性。扩增的16S rDNA基因序列长度为1 476 bp (GenBank登录号:KP031644),与GenBank中的松鼠葡萄球菌16S rDNA基因序列的相似性达99%。系统进化树显示,菌株YDE0916和松鼠葡萄球菌聚为一簇,由此判定菌株YDE0916为松鼠葡萄球菌。药敏试验结果显示,该菌株对诺氟沙星、杆菌肽耐药,对美满霉素、庆大霉素、氨苄西林等22种药物高度敏感。本研究为松鼠葡萄球菌的分离鉴定及预防积累参考依据。     

眼镜蛇缓慢葡萄球菌的分离鉴定与药敏试验

对广西南宁市某眼镜蛇养殖场送检的患病眼镜蛇进行病理剖检及病原菌分离鉴定,根据临床发病症状、病理变化、病理组织学检查、病原分离鉴定及致病力试验结果,确诊为缓慢葡萄球菌感染.该分离菌对小鼠有一定的致病性;药敏试验结果显示,分离菌对阿莫西林钠舒巴坦钠、头孢哌酮钠舒巴坦钠、氨苄西林钠、阿莫西林钠、苯唑西林钠、哌拉西林钠、阿米卡星敏感,对头孢曲松钠、氧氟沙星、利福平中介,对链霉素、林可霉素、磺胺间甲氧嘧啶、氟苯尼考、土霉素、诺氟沙星耐药.选择敏感抗生素阿莫西林钠舒巴坦钠添加在水和料中预防发病,患病眼镜蛇肌肉注射头孢哌酮钠舒巴坦钠,46条病蛇经一个疗程治疗,仅5条死亡,效果显著,蛇群无新病例发生,1周后逐渐恢复采食.     

Isolation,Identification and Drug Resistance Analysis of Staphylococcus sciuri of Corydoras

To explore etiology characteristics of Staphylococcus sciuri, strain YDE0916 was isolated and identified from Corydoras by cultivation, biochemical identification, 16S rDNA gene PCR amplification and sequence analysis, and resistance analysis.The result showed this strain was gram-positive, serial, spore, without capsule.The physicochemical properties showed immobility, β-galactosidase-negative, VP reaction-negative, sugar and maltose-positive.The nucleotide sequence of 16S rDNA was 1 476 bp and its GenBank accession number was KP031644.This 16S rDNA showed 99% homology with Staphylococcus sciuri.Phylogenetic tree showed that strain YDE0916 and several Staphylococcus sciuri strains gathered into one cluster.So the strain was identified as Staphylococcus sciuri.The antibiotic sensitive test showed that strain YDE0916 was resistant to norfloxacin and bacitracin, and was highly sensitive to 22 kinds of drugs, such as minocycline, gentamicin and ampicillin.This study could provide a reference for identification and prevention of Staphylococcus sciuri.     

Lysogeny and other characteristics of Staphylococcus hyicus isolated from chickens

Lysogeny was readily demonstrated among strains of Staphylococcus hyicus that were isolated from chickens. Susceptibility to phage lysis was affected by prophage immunity, but lipase activity and erythromycin resistance were not affected by the presence of temperate phage. In contrast to previously published results, lipase-negative strains of S. hyicus were relatively common and the use of selective media based on lipase activity would have been unsuitable for detection of the S. hyicus strains examined.     

Comparative assessment of genotypic and phenotypic correlates of Staphylococcus pseudintermedius strains isolated from dogs with otitis externa and healthy dogs

Staphylococcus pseudintermedius is considered a primary pathogen of canine skin and soft tissue infections, and the rapid emergence of methicillin-resistant S. pseudintermedius worldwide is a major issue. In the current study, genotypic and phenotypic correlates associated with S. pseudintermedius causing canine otitis externa were evaluated using 41 S. pseudintermedius strains isolated from dogs with otitis externa (n = 26) and healthy dogs (n = 15). The S. pseudintermedius strains were subjected to a comparative analysis of (i) genotypes (multilocus sequence typing, agr, and spa types), (ii) methicillin resistance and SCCmec types, (iii) multidrug resistance (MDR), (iv) biofilm formation, and (v) susceptibility to canine cathelicidin (K9CATH). A high degree of genetic diversity was observed in both groups of S. pseudintermedius strains, regardless of methicillin resistance. Almost all methicillin-resistant strains (>95%) harbored SCCmec V and displayed MDR. Although there was no difference in biofilm formation, S. pseudintermedius strains derived from otitis externa exhibited enhanced resistance to cationic antimicrobial peptide (K9CATH) compared with strains from healthy dogs. The high degree of heterogeneity in MLST, agr, and spa types prevented the identification of correlations between any specific genotype and virulence phenotype in otitis externa caused by S. pseudintermedius, These findings provide an important basis for monitoring and treating canine skin and soft tissue infections in Korea.     

1株奶牛源松鼠葡萄球菌的分离鉴定及耐药性分析

为确定奶牛垫料中是否含有病原菌并深入了解垫料中优势生长菌的类型、耐药性及致病性等情况,本试验进行了细菌分离培养、革兰染色镜检、生化试验鉴定、16S rDNA序列分析及同源性比对、药敏试验、小鼠致病性试验。结果显示,分离菌在普通营养琼脂平板上形成圆形、表面光滑的白色菌落,血琼脂平板上形成黏稠、较大的白色菌落。革兰染色镜检结果显示,分离菌为革兰阳性,球型,呈葡萄串状排列,或单个散落。生化试验结果显示,分离菌对木糖、葡萄糖、麦芽糖、果糖、乳糖、硝酸盐还原等呈阳性反应,而蜜二糖、木糖醇、山梨醇、尿素、V-P反应等呈阴性反应。16S rDNA序列分析结果显示,扩增的16S rDNA序列长度为1 298 bp,与松鼠葡萄球菌的核苷酸同源性达99.85%~100%;系统进化树结果显示,分离菌与松鼠葡萄球菌处于同一分支。动物试验结果表明,分离菌株对试验小鼠有较强的致病性,以0.2 mL/只（7.9×10⁸ CFU/mL）菌液的剂量接种小鼠,在48 h内死亡率为60%（3/5）。药敏特性分析结果显示,分离菌株对复方新诺明、氨苄西林等7种药物敏感,对克林霉素、头孢噻肟和头孢呋辛中度敏感,对青霉素、红霉素和林可霉素耐药。本研究为奶牛源松鼠葡萄球菌的分离鉴定及防控提供了参考依据。     

不同动物源性维氏气单胞菌生物学特性比较研究

为深入了解维氏气单胞菌,本试验选取8株不同动物源性的维氏气单胞菌,并对其染色特性、理化特性、培养特性、耐药性、不同p H值、温度、渗透压对其生长的影响以及16S r RNA序列的相似性等生物学特性进行比较研究。结果显示,多数维氏气单胞菌在体外培养的老龄培养物中呈现多形性;理化特性大致相同;可以在TSA培养基、RS培养基、S.S.培养基、BS培养基、HE培养基上生长良好;可以在4℃、25℃~40℃、p H值6~9、氯化钠含量0%~4%中生长;对青霉素G、克林霉素、万古霉素、多粘菌素B、氨苄西林、新生霉素等表现出耐药性;经比对,不同维氏气单胞菌菌株的16S r RNA基因序列具有较高的相似性,但存在一定的差异。研究结果将为维氏气单胞菌的检验检疫、疾病预防奠定基础,同时为该菌的深入研究提供数据支持。     

Differentiation of Staphylococcus sciuri Strains Isolated from Free‐Living Rodents and Insectivores

Twenty‐nine Staphylococcus sciuri strains isolated from free‐living insectivores and rodents were comparatively analysed for their biochemical capacities and their SmaI macrorestriction patterns. The 29 S. sciuri isolates represented 21 different biotypes and 22 different SmaI macrorestriction types. This observation confirmed that S. sciuri isolates obtained from insectivores and rodents living in natural environments constituted a heterogeneous population. Cluster analysis revealed that the macrorestriction patterns and the biochemical profiles matched in some cases. However, S. sciuri isolates that exhibited the same or closely related biochemical profiles were also found to be associated with different macrorestriction patterns. Analysis of the 29 S. sciuri isolates for their plasmid content and their sensitivity to antimicrobial agents showed that most of the isolates were plasmid‐free and susceptible to all antimicrobial agents tested.     

Detection of a novel chloramphenicol resistance plasmid from "equine" Staphylococcus sciuri

A small chloramphenicol resistance (Cm) plasmid of 4.65 kB could be detected in an "equine" Staphylococcus sciuri-culture. This plasmid, designated as pSC3, was identified by interspecific protoplast transformation. On the basis of restriction endonuclease analyses a detailed restriction map of pSC3 could be constructed. This allowed structural comparisons of pSC3 with Cm-plasmids of other staphylococcal species from infections of humans and animals and identification of pSC3 as a member of the pC 221-family of staphylococcal Cm-plasmids. The pSC3-plasmid encoded an inducible chloramphenicol acetyltransferase as confirmed by enzymatic assays. This enzyme could be demonstrated in cell-free lysates of Cm-induced pSC3-transformants.     

患病和痊愈牦牛源大肠杆菌对氨基糖苷类和磺胺类耐药性比较分析

[目的]为了探讨患病和痊愈牦牛源大肠杆菌的抗菌药物及其耐药性之间的相关性。[方法]采用微量肉汤稀释法对91株牦牛源大肠杆菌进行氨基糖苷类药物(链霉素和庆大霉素)和磺胺类药物(磺胺甲噁唑和磺胺二甲基嘧啶)敏感性试验,同时采用PCR方法检测氨基糖苷类相关耐药基因aac(6’)-Ib、aac(3’)-IV及磺胺类相关耐药基因sul2、sul3的携带情况。[结果]结果表明：39株患病牦牛源菌株对链霉素、庆大霉素、磺胺甲噁唑和磺胺二甲基嘧啶的耐药率分别为30.77%、5.13%、61.54%和87.18%,aac(6’)-Ib、aac(3’)-IV、sul2和sul3的检出率分别为43.59%、25.64%、38.46%和41.03%;52株痊愈牦牛源菌株对链霉素、庆大霉素、磺胺甲噁唑和磺胺二甲基嘧啶的耐药率分别为42.30%、21.15%、94.23%和71.15%,aac(6’)-Ib、aac(3’)-IV、sul2和sul3的检出率分别为71.15%、19.23%、69.23%和51.92%。[结论]试验结果表明,痊愈牦牛源大肠杆菌对氨基糖苷类和磺胺类药物耐药率及相关耐药基因检出率均高于患...     

奶牛乳腺炎金黄色葡萄球菌溶血素基因型和表现型分析

为调查奶牛乳腺炎金黄色葡萄球菌溶血素基因型和表现型分布,并对两者之间的关系进行研究,采用PCR方法对从奶牛乳腺炎病例中分离的33株金黄色葡萄球菌的溶血素基因进行检测,并分别用牛血琼脂平板和绵羊血琼脂平板检测其溶血性。结果表明,各溶血素hla,hlb,hld,hlg,hlg2基因的检出率分别为90.91%,100%,93.94%,78.79%,66.67%;牛血平板检测表明β溶血占84.85%,绵羊血平板检测β溶血只有57.58%,金黄色葡萄球菌在牛血和绵羊血琼脂平板上均以β血为主,但牛血检测出的β溶血比例更高,表明牛红细胞对金黄色葡萄球菌β溶血素更为敏感。综上表明,金黄色葡萄球菌溶血素基因型和表现型之间无对应关系。     

Biochemical and toxigenic characteristics of Aeromonas spp. isolated from diseased mammals, moribund and healthy fish

In this study we describe biochemical, toxigenic and surface characteristics of 33 motile Aeromonas isolated from diseased mammals, 3 from moribund marine mammals, 24 from healthy fish and 4 from moribund fish. Aeromonas hydrophila, A. caviae and A. sobria were isolated from both mammals and fish but at a different incidence. Aeromonas hydrophila was the predominant species isolated from clinical specimens; it was isolated from pneumonia, wound infections, septicemia and abortion in horses, cattle and pigs. Aeromonas sobria was isolated from one mammal and 11 healthy fish. Aeromonas caviae was isolated in 2 cases from healthy fish and in 9 cases from diseased mammals. Variations in some biochemical tests including sorbitol, amylase and citrate, were observed between isolates from different sources. However, these differences did not allow the differentiation of isolates from diseased mammals and healthy fish. The majority of A. hydrophila isolates produced different extracellular products; A. sobria isolates produced less exotoxin. With A. caviae isolates no hemolysin, protease, enterotoxin or elastase were detected. There was no quantitative difference in hemolysin, protease, enterotoxin or elastase production between isolates from mammals and fish. It is suggested that A. hydrophila could be a potential pathogen for domestic animals, and fish may represent a potential reservoir of infection.