Surface hydrophobicity of Staphylococcus intermedius and Staphylococcus hyicus.

Surface hydrophobicity of 90 Staphylococcus intermedius and 55 S hyicus isolates was evaluated using the hexadecane adherence assay and the ammonium sulphate salt aggregation test. A strongly positive hydrocarbon adherence in the hexadecane adherence assay was demonstrated in 11 per cent of the S intermedius isolates and 7 per cent of the S hyicus isolates. Bacterial aggregation in 1.6 M, or less, ammonium sulphate was observed in 28 per cent of the S intermedius isolates and 37 per cent of the S hyicus isolates. There was no statistical correlation between the two assays. The adherence of both bacterial species to hexadecane was eliminated when the cells were first treated with pronase and trypsin, while it was mildly enhanced by prior heat treatment (60 degrees C and 95 degrees C for up to three hours). In contrast, aggregation of S intermedius in ammonium sulphate was not influenced by trypsin pretreatment, and aggregation of both bacterial species was diminished, or eliminated, with pronase or prior 95 degrees C heat treatment. Surface hydrophobicity, as measured in both assays, appeared to have no relationship with growth patterns in serum soft agar or production of slime. Similarly, the presence or absence of substantial surface receptor activity to fibrinogen, fibronectin or IgG did not appear to be related to surface hydrophobicity.     

Isolation and characterization of immunoglobulin binding proteins from Staphylococcus intermedius and Staphylococcus hyicus.

125-I-IgG binding activities were observed with 15 (17%) of 90 S. intermedius isolates from dogs and 39 (95%) of 41 S. hyicus isolates from pigs. Binding activities were not detected with S. hyicus isolates from cows. The IgG binding proteins of 2 S. intermedius, 2 S. hyicus, and protein A from S. aureus Cowan I were isolated from their cell surfaces. The proteins precipitated with IgG preparations from human, rabbit, pig, dog and horse, but not with IgG from cow, mouse and chicken. This indicated that these IgG binding proteins could be classified as type I receptors. In addition, the isolated proteins from all 3 staphylococcal species precipitated with polyclonal chicken anti-protein A antiserum. SDS-PAGE, Western blotting and gel isoelectric focussing of the proteins revealed numerous bands in the 42,000 D range and acid isoelectric points. The isoelectric point of the isolated proteins from both S. intermedius cultures was slightly more acidic than those from S. hyicus and S. aureus. The present results indicate a close functional and antigenic similarity, if not identity, between IgG binding proteins of S. intermedius and S. hyicus, and protein A of S. aureus.     

一株鸡源猪葡萄球菌的分离与鉴定

为进一步研究引起蛋鸡产蛋下降的病原菌的致病机理,本研究从患有输卵管炎的蛋鸡输卵管中分离到一株革兰氏阳性球菌,通过培养形态观察和生化试验进行初步鉴定,并采用PCR扩增其16S rRNA基因序列进行测序,与GenBank中登录的已知序列进行比对分析,结果显示该菌与猪葡萄球菌(Staphylococcus hyicus)在进化关系上的距离最近,同源性达到99.5％以上,表明该分离株为S.hyicus.并通过小鼠毒力试验和蛋鸡回归试验对该分离株的致病力研究,表明该菌对小鼠无致病性,但可以引起生殖系统病变,导致产蛋能力下降.     

Antimicrobial resistance in staphylococci from animals with particular reference to bovine Staphylococcus aureus, porcine Staphylococcus hyicus, and canine Staphylococcus intermedius.

Besides their role as commensals on the skin and mucosal surfaces, staphylococci may be involved in a wide variety of diseases in animals. Staphylococcal infections in animals are mainly treated with antimicrobial agents and as a consequence, staphylococci from animal sources have developed and/or acquired resistance to the respective antimicrobial agents. Resistance statistics obtained from national monitoring programmes on staphylococci from cattle and pigs, but also from surveillance studies on staphylococci involved in diseases in dogs are reported and reviewed with regard to their comparability. This review mainly focusses on the genetic basis of antimicrobial resistance in staphylococci of animal origin. Particular attention is paid to resistance to those antimicrobial agents which are most frequently used in veterinary medicine, but also to antimicrobial agents, such as chloramphenicol and mupirocin, which are used in specific cases for the control of staphylococcal infections in pets and companion animals. In addition, plasmids and transposons associated with the respective resistance properties and their ways of spreading between members of the same or different staphylococcal species, but also between staphylococci and other gram-positive bacteria, are described.     

Staphylococcus hyicus in cattle

Staphylococcus hyicus (subsp hyicus) was found to be a frequently occurring inhabitant of the skin of cattle. Significant numbers of this bacterium were found to be present in mange lesions of cattle between one and three years old. Experimental inoculations revealed that S hyicus had a pathogenic effect on the superficially scarified skin of young cattle.     

猪葡萄球菌437-2株的分离鉴定及生物学特性

从发生渗出性皮炎的仔猪体内分离到1株细菌,根据其形态特征和生化特性,结合16S rRNA核苷酸序列分析,确定为猪葡萄球菌,并命名为437-2株.用1×109 CFU/mL剂量的该菌株经皮下感染小鼠、仔猪,均能引起注射部位皮肤脱毛、破渍、有炎性渗出物、结痂等病变,病理学观察可见表皮局部上皮细胞消失、皮下小血管充血和炎症细...     

Cytotoxic activity of Staphylococcus hyicus

Culture supernatants from a number of Staphylococcus hyicus strains caused toxic effects to both murine fibroblast and porcine keratinocyte cells in culture. The extent of cytotoxicity was shown to differ between strains and may provide an indication of strain virulence. Purification of cytotoxic activity produced by S. hyicus (strain P119) using preparative isoelectric-focussing demonstrated it to be cytolytic, haemolytic and non-proteolytic. The cytotoxin demonstrates certain properties in common with the delta haemolysin of Staphylococcus aureus.     

Bacteriophage typing of Staphylococcus hyicus subsp hyicus isolated from pigs

Four phages were isolated and used for typing Staphylococcus hyicus subsp hyicus isolated from pigs with or without exudative epidermatitis (EE) in Japan. Sixty-four (85.3%) of the 75 isolates examined were typeable at either routine test dilution (RTD) or 100 X RTD. Two or more kinds of phage patterns were present in the isolates from each pig with EE. All isolates from healthy pigs showed a single-phage pattern. Fourteen (32.6%) of 43 isolates and 7 (87.5%) of 8 isolates from pigs with EE in Belgium and Czechoslovakia, respectively, were typeable with the 4 phages. None of 180 isolates of S aureus, 7 (6.4%) of 110 isolates of S intermedius, and 2 (2.3%) of 86 isolates of S epidermidis were typeable.     

1株猪葡萄球菌强毒株的分离鉴定及致病性试验

为确定广东省增城某猪场产房仔猪暴发渗出性皮炎的致病菌,从病猪心脏、肺脏、病变皮肤分离并纯化细菌,经形态学观察,镜检,生化特性及gap基因的序列分析,确定为猪葡萄球菌,命名为ZC-4株。药敏试验结果显示该菌株对头孢拉定、头孢唑啉、新生霉素高度敏感,对头孢噻肟、万古霉素、庆大霉素中度敏感,对青霉素、阿莫西林、磺胺类药物、链霉素、恩诺沙星等均耐药。对分离株毒素基因进行检测,结果显示,分离株携带ExhA毒素基因,测序结果表明,其编码区全长为822bp,编码274个氨基酸;氨基酸序列与丹麦分离株ExhA基因(AF515453)的同源性最高,为99.6%;遗传进化分析显示,分离株所携带的毒素基因与参考菌株猪葡萄球菌ExhA毒素基因聚类紧密,位于一个小的分支内。分离株致病性试验结果显示,分离株接种25日龄健康断奶仔猪后24h,病猪耳根、四肢内侧、背部、腹部、臀部、尾根等部位出现蜕皮并露出有液体渗出的鲜红创面,继而皮肤形成一层厚厚的结痂,说明该菌株具有较强的致病性。本试验为猪葡萄球菌疫苗的研制及致病机制研究提供了理论依据。     

Studies on the Virulence of Staphylococcus hyicus

Abstract— Culture supernatants from six Staphylococcus hyicus isolates were concentrated by ultrafiltration and were injected intradermally into the abdominal skin of 2-week-old piglets. Two distinct types of reaction were observed; (1) a focal erythema and (2) an exfoliative reaction with crusting. The severity of these reactions and the number of animals affected differed between isolates. The exfoliative reaction observed in these skin tests may be a good indicator of virulence. Résumé— Les surnageants de six souches de Staphylococcus hyicus ont été concentrés par ultrafiltration et furent injectés par voie intradermique dans la peau de l'abdomen de porcelets ägés de deux semaines. Deux types distincts de réactions furent observés: (1) un éryhème focal et (2) une réaction d'exfoliation avec apparition de croûtes. L'intensité de ces réactions et le nombre d'animaux atteints différaient selon les souches. La réaction d'exfoliation observée lors de ces tests pourrait être un bon indicateur de virulence. Zusammenfassung— Überstände der Kultur von sechs Staphylococcus hyicus-Isolaten wurden durch Ultra-zentrifugieren konzentriert und bei 2 Wochen alten Ferkeln intradermal in die Abdominalhaut injiziert. Es konnten zwei deutlich unterscheidbare Reaktionen beobachtet werden: (1) ein fokales Erythem und (2) eine exfoliative Reaktion mit Krustenbildung. Die Schwere dieser Reaktionen und die Zahl der betroffenen Tiere war je nach Isolat unterscheidlich. Die exfoliative Reaktion bei diesen Hauttests könnte ein guter Indikator für die Virulenz sein. Resumen Los sobrenadantes de 6 cultivos diferentes de Staphylococcus hyicus se concentraron mediante ultrafiltración y se inyectaron via intradérmica en la piel abdominal de lechones de dos semanas de edad. Se observaron dos tipos de reacción: (1) un eritem focal y (2) una reacción exfoliativa y costrosa. La gravedad de las reacciones y el nümero de animales afectados variaba en functión del cultivo utilizado. La reacción exfoliativa que se observó en estos tests intradérmicos puede ser un buen indicador de virulencia.     

Evaluation of methicillin resistance in Staphylococcus intermedius isolated from dogs

Methicillin and multi-drug resistance were investigated in 136 Staphylococcus intermedius strains of canine origin. The large majority of isolates were found to be mecA-negative by polymerase chain reaction, whereas only four strains were positive for the mecA gene. All mecA-positive strains were confirmed as methicillin-resistant by complementary tests, except for oxacillin disk diffusion, which yielded one false-negative result. A significantly higher resistance to fusidic acid, lincosamides, and cotrimoxazole was observed in methicillin-resistant S. intermedius (MRSI) compared with methicillin-susceptible strains. Although the prevalence of MRSI in dogs appeared to be low, methicillin resistance was confirmed to be associated with multi-drug resistance, suggesting the importance of antimicrobial susceptibility testing of canine S. intermedius strains.     

Comparative studies on bacteriolytic properties of Staphylococcus chromogenes and Staphylococcus hyicus

Staphylococcus chromogenes and Staphylococcus hyicus showed bacteriolytic activities towards a Micrococcus luteus reference strain. This was demonstrated on tryptone soya agar containing M. luteus cells. Both bacteriolytic enzymes could be isolated by ionic exchange chromatography and subsequent gel filtration. The isolated bacteriolysin of S. chromogenes lysed the M. luteus reference culture, was heat inactivated by 95 degrees C and precipitated specifically with antiserum produced against the bacteriolysin of S. hyicus.     

一株猪葡萄球菌的分离鉴定

从患有渗出性皮炎的仔猪分离到1株革兰阳性球菌,生化试验、形态学观察进行初步鉴定,然后进行16 S rRNA基因片段的扩增和测序,将测序结果与GenBank上的已知序列进行同源性比较,结果与猪葡萄球菌的核酸序列同源性达到99.6%,综合可见该分离菌株为猪葡萄球菌,并将其命名为GDSH1。     

猪葡萄球菌脱落毒素研究进展

猪渗出性皮炎是一种由猪葡萄球菌引起的哺乳或断奶仔猪的急性传染病.猪葡萄球菌产生的脱落毒素被认为是引起猪渗出性皮炎的主要因子之一,不同毒株可以产生不同的毒素,分别被命名为ExhA、ExhB、ExhC和ExhD 4种.论文就近年来对猪葡萄球菌脱落毒素的结构特征、生物学功能以及在诊断和免疫学方面的研究进行综述.     

Isolation of exfoliative toxin from Staphylococcus hyicus subsp. hyicus and its exfoliative activity in the piglet

Exfoliative toxin was isolated from the sterile cell-free filtrate of 24 h culture of Staphylococcus hyicus subsp. hyicus strain P-1. The partial purification of exfoliative toxin produced by S. hyicus (shET) was performed by precipitation with 50-80% saturated ammonium sulfate, gel filtration on a Sephadex G-75 column and column chromatography on DEAE-cellulose. Partially purified shET (pp-shET) caused exfoliation in piglets at 8 to 12 h after intradermal or subcutaneous injection. However, heat-treated pp-shET did not cause exfoliation in piglets for up to 24 h after injection. On histopathological examination of the skin at 12 h after injection of pp-shET, an intraepidermal cleavage plane was shown between the stratum corneum and stratum granulosum and at the stratum granulosum.     

Development of a PCR test for the identification of Staphylococcus intermedius based on the 16S rDNA sequence

The development of a PCR assay based on the 16S ribosomal RNA gene (rDNA) sequence was carried out for the identification of Staphylococcus intermedius. Sixty-six strains of S. intermedius, 70 of Staphylococcus aureus and 2 of Staphylococcus hyicus were examined for the assay. The 16S rDNA, of which the PCR target fragment makes up 901 bp corresponding to the sequence data of the gene, was detected in all strains of S. intermedius, but it was not detected in any strains of either S. aureus or S. hyicus. These results suggest that the PCR allows a simple and precise identification of S. intermedius.     

Assay of protein A in Staphylococcus hyicus subsp. hyicus by ELISA and immunoelectron microscopy

The presence and quantity of protein A in Staphylococcus hyicus subsp. hyicus isolates were examined by an enzyme-linked immunosorbent assay (ELISA) and immunoelectron microscopy. Cell-bound protein A was demonstrated in 45 (94%) of 48 isolates from diseased pigs and in 113 (86%) of 132 isolates from healthy pigs by ELISA using peroxidase-conjugated rabbit antibody, but was not found in isolates from chickens and cows. Most of these swine isolates contained about 100 to 300 ng of cell-bound protein A/ml. Extracellular protein A was not detected in any isolates from pigs, chickens or cows. In the immunoelectron microscopy assay, swine isolates were labeled with goat anti-mouse IgG conjugated to colloidal gold particles, but chicken and cow isolates were not labeled.     

Proteolytic zymograms of Staphylococcus hyicus subsp. hyicus isolated from pigs, chickens and cows

The extracellular proteases of Staphylococcus hyicus subsp. hyicus were assayed by a zymogram showing caseinolysis and gelatinolysis. Four bands were associated with caseinolysis or with gelatinolysis. The patterns shown by strains isolated from pigs, chickens and cows were compared; isolates from pigs differed from those isolated from chickens or cows but strains isolated from diseased and healthy pigs could not be differentiated.