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1.
Alber J El-Sayed A Estoepangestie S Lämmler C Zschöck M 《Veterinary microbiology》2005,109(1-2):135-141
Bacterial superantigens are one of the major virulence factors produced by Streptococcus pyogenes and Staphylococcus aureus. The two novel superantigen encoding genes seeM and seeL were described for S. equi subsp. equi which is known as the causative agent of strangles in equids. In the present study previously characterized S. equi subsp. equi strains and strains of various other animal pathogenic streptococcal species and subspecies were investigated for the presence of the superantigen encoding genes seeM and seeL by polymerase chain reaction. According to these studies seeL and seeM appeared to be a constant characteristic of all investigated S. equi subsp. equi strains. Surprisingly, one S. equi subsp. zooepidemicus strain (S.z. 122) was also positive for both genes. The species identity of this S. equi subsp. zooepidemicus strain could additionally be confirmed by sequencing the 16S rRNA gene and the 16S-23S rDNA intergenic spacer region. The superantigen encoding genes could not be found among additionally investigated S. equi subsp. zooepidemicus strains or among strains of seven other streptococcal species. The seeL and seeM genes of the S. equi subsp. equi strain S.e. CF32 and the genes szeL and szeM of the S. equi subsp. zooepidemicus strain S.z. 122 were cloned and sequenced. A sequence comparison revealed a high degree of sequence homology between seeL, szeL, speL and seeM, szeM and speM, respectively. The superantigenic toxins L and M seemed to be widely distributed virulence factors of S. equi subsp. equi, rare among S. equi subsp. zooepidemicus but did not occur among a number of other animal pathogenic streptococcal species. 相似文献
2.
The 16S rRNA gene of 39 S. equi subsp. zooepidemicus strains and two S. equi subsp. equi strains was amplified by polymerase chain reaction and subsequently digested with the restriction enzyme Hinc II. A restriction profile with two fragments with sizes of 1250 bp and 200 bp could be observed for both S. equi subsp. equi strains and for 30 of the 39 S. equi subsp. zooepidemicus strains indicating a sequence variation within the V2 region of the 16S rRNA gene of the remaining nine S. equi subsp. zooepidemicus isolates. A segment of the 16S rRNA gene including the hypervariable V2 region of 11 S. equi subsp. zooepidemicus and two S. equi subsp. equi could be amplified by PCR and sequenced. The sequence of the V2 region of eight S. equi subsp. zooepidemicus strains appeared to be identical or almost identical to the sequence of the two S. equi subsp. equi strains. The sequence of the remaining three S equi subsp. zooepidemicus strains differed significantly from the sequence of S. equi subsp. equi. These differences allowed a division of S. equi subsp. zooepidemicus strains into two 16S rRNA types and might possibly have consequences for the taxonomic position of these phenotypically indistinguishable strains of one subspecies. A molecular typing could additionally be performed by amplification of the gene encoding the 16S-23S rRNA spacer region. A single amplicon of the spacer gene of 1100 bp could be observed for one S. equi subsp. zooepidemicus, an amplicon of 950 bp for two S. equi subsp. equi strains and 10 S. equi subsp. zooepidemicus strains, a amplicon of 780 bp for 27 S. equi subsp. zooepidemicus strains and a single amplicon of 600 bp for one S. equi subsp. zooepidemicus strain. The variations of the V2 region of the 16S rRNA gene and the size variations of the 16S-23S rRNA spacer gene were not related to each other. Both variations could be used for molecular typing of this species, possibly useful in epidemiological aspects. 相似文献
3.
根据现有细菌分类鉴定方法,采用血清学、Biolog、16S rRNA序列分析和全基因组序列分析等方法对羊败血性链球菌病疫苗生产检验用菌种CVCC 553、55001和55002进行系统鉴定。结果显示,菌种CVCC 553、55001和55002血清群为兰氏C群;Biolog鉴定为马链球菌反刍亚种;16S rRNA序列分析与马链球菌反刍亚种模式菌株CECT 5772的同源性最高(均为99.72%),且在系统发育树中位于同一分支;与马链球菌反刍亚种模式菌株CCUG 47520的DNA-DNA的杂交值大于70%,分别为80.5%、80.2%、80.5%,与马链球菌反刍亚种模式菌株CECT 5772的平均核苷酸同源性分别为97.73%、97.64%、97.77%。因此本研究认为羊败血性链球菌病疫苗生产用菌种CVCC 553、55001和55002应为马链球菌反刍亚种。 相似文献
4.
Thirteen strains of Streptococcus equi subsp. ruminatorum from free-ranging spotted hyenas (Crocuta crocuta) and plains zebras (Equus burchelli) in Tanzania were characterized by biochemical and molecular-biological methods. Although the colony appearance of the S.e. ruminatorum wildlife strains differed from that of the S.e. ruminatorum type strain CECT 5772(T), all biochemical reactions of the wildlife strains were similar to those of the type strain. In addition, all wildlife strains produced hyaluronidase and were capable of hydrolysing arginine, three strains (23%) synthesized acetoin, but only eight strains (62%) produced acid from ribose. rep-PCR indicated that different clones of S.e. ruminatorum were distributed among the hyena and zebra populations in the study area. Identical rep-PCR patterns in hyena and zebra strains suggest that a direct transmission of S.e. ruminatorum between these species may occur. The presence of a M-like protein (SrM) gene was demonstrated in all S.e. ruminatorum strains including the type strain. Sequencing of the M-like protein gene revealed a hypervariable region within the deduced amino acid sequence. Most of the strains clustered with previously described strains based on the hypervariable region of the S.e. zooepidemicus SzP protein. Sequencing also demonstrated that identical SrM protein sequences were shared among S.e. ruminatorum strains from different host species. 相似文献
5.
Alber J El-Sayed A Lämmler C Hassan AA Weiss R Zschöck M 《Journal of veterinary medicine. B, Infectious diseases and veterinary public health》2004,51(10):455-458
The closely related streptococcal species Streptococcus equi subsp. zooepidemicus and S. equi subsp. equi were identified by polymerase chain reaction using oligonucleotide primers designed according to species-specific parts of the superoxide dismutase A encoding gene sodA. A further differentiation of both subspecies could be performed by amplification of the genes seeH and seeI encoding the exotoxins SeeH and SeeI, respectively, which could be detected for S. equi subsp. equi but not for S. equi subsp. zooepidemicus. A further simplification of the identification and differentiation of both subspecies was conducted by sodA-seeI multiplex polymerase chain reaction. 相似文献
6.
Younan M Estoepangestie AT Cengiz M Alber J El-Sayed A Lämmler C 《Journal of veterinary medicine. B, Infectious diseases and veterinary public health》2005,52(3):142-146
Seventeen Streptococcus equi subsp. zooepidemicus strains isolated from camels and camel milk in Kenya and Somalia were identified by their cultural characteristics, by biochemical and serological reactions with the help of commercial identification systems and by molecular studies using a multiplex PCR. The isolates were further characterized by a PCR-mediated detection of size polymorphisms in the 16S-23S rDNA intergenic spacer region and the virulence gene szp and by amplification of the virulence gene cne. These molecular analysis are potentially useful in identifying and characterizing S. equi subsp. zooepidemicus strains of this origin and could possibly be valuable in epidemiological investigations. 相似文献
7.
Evasion of phagocytosis is an important virulence determinant of Streptococcus equi (S. equi subsp. equi), the cause of equine strangles and distinguishes it from the closely related but much less virulent S. zooepidemicus (S. equi subsp. zooepidemicus). We describe Se18.9, a novel H factor binding protein secreted by S. equi but not by S. zooepidemicus that reduces deposition of C3 on the bacterial surface and significantly reduces the bactericidal activity of equine neutrophils suspended in normal serum for both S. equi and S. zooepidemicus. Se18.9 is secreted abundantly by actively dividing cells and is also bound to the bacterial surface. Strong serum and mucosal antibody responses are elicited in S. equi infected horses. Although a gene identical to se18.9 was not detected in S. zooepidemicus, sequences encoding proteins of similar size with similar signal peptide sequences were found in 3 of 12 randomly selected strains. Since Se18.9 is unique to S. equi, and immunoreactive with convalescent sera and mucosal IgA, it has potential for immunodiagnosis and for study of mucosal antibody response to S. equi. 相似文献
8.
9.
Streptococcus equi (S. equi subsp. equi) is widely believed to have evolved from an ancestral strain of S. zooepidemicus (S. equi subsp. zooepidemicus) based on high sequence homology. A striking difference is the absence of phage sequences from S. zooepidemicus. In this study we show that the receptor for SeP9, a temperate bacteriophage of S. equi, is the Lancefield group C carbohydrate. However, although SeP9 binds to group C carbohydrate from S. zooepidemicus, it appears not to replicate and produce plaques. 相似文献
10.
Hobo S Niwa H Oku K 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2012,74(3):329-333
Loop-mediated isothermal amplification (LAMP) constitutes a potentially valuable diagnostic tool for rapid diagnosis of contagious diseases. In this study, we developed a novel LAMP method (seM-LAMP) to detect the seM gene of Streptococcus equi subsp. equi (S. equi), the causative agent of strangles in equids. The seM-LAMP successfully amplified the target sequence of the seM gene at 63°C within 60 min. The sensitivity of the seM-LAMP was slightly lower than the 2nd reaction of the seM semi-nested PCR. To evaluate the species specificity of the seM-LAMP, we tested 100 S. equi and 189 non-S. equi strains. Significant amplification of the DNA originating from S. equi was observed within 60 min incubation, but no amplification of non-S. equi DNA occurred. The results were identical to those of seM semi-nested PCR. To investigate the clinical usefulness of the methods, the seM-LAMP and the seM semi-nested PCR were used to screen 590 nasal swabs obtained during an outbreak of strangles. Both methods showed that 79 and 511 swabs were S. equi positive and negative, respectively, and the results were identical to those of the culture examination. These results indicate that the seM-LAMP is potentially useful for the reliable routine diagnosis of Streptococcus equi subsp. equi infections. 相似文献
11.
Akineden O Alber J Lämmler C Weiss R Siebert U Foster G Tougaard S Brasseur SM Reijnders PJ 《Veterinary microbiology》2007,121(1-2):158-162
The present study was designed to identify 15 beta-hemolytic streptococci isolated during a period between 1988 and 2005 from nine harbour seals and six grey seals from various origins of the North Sea. All isolates were identified as Streptococcus equi subsp. zooepidemicus. The bacteria were additionally investigated for relatedness by restriction fragment length polymorphism analysis of PCR amplified 16S-23S rDNA intergenic spacer region and gene szp and by macrorestriction analysis of chromosomal DNA of the strains by pulsed field gel electrophoresis. The molecular analysis yielded identical or closely related patterns within the strains of the present study and with the S. equi subsp. zooepidemicus strains isolated from harbour seals of German North Sea which were investigated previously [Akineden, O., Hassan, A.A., Alber, J., El-Sayed, A., Estoepangestie, A.T.S., L?mmler, C., Weiss, R., Siebert, U., 2005. Phenotypic and genotypic properties of S. equi subsp. zooepidemicus isolated from harbor seals (Phoca vitulina) from the German North Sea during the phocine distemper outbreak in 2002. Vet. Microbiol. 110, 147-152]. This indicates that this single or closely related bacterial clone existed during both phocine distemper virus epidemics in 1988 and 2002 and that a direct transmission of the strains has occurred between two seal species and between seal populations of far distant regions possibly with grey seals as a vector. 相似文献
12.
为了解马源马链球菌兽疫亚种（S.zooepidemicus）新疆分离株马链球菌兽疫亚种类M蛋白（SzM）基因的分子进化与变异情况,为该菌引起的感染性疾病的防控提供依据,本试验对新疆地区某马场采集的病马淋巴结样品进行病原菌的分离培养和生化鉴定,并对分离菌株进行药物敏感性试验。根据已发表的马源SzM基因序列设计引物,对其SzM基因进行PCR扩增及序列测定。将获得的SzM序列与GenBank中不同动物源马链球菌兽疫亚种分离株序列进行同源性比对和遗传进化分析。结果显示,分离得到了一株革兰氏阳性链球菌,将其命名为马链球菌兽疫亚种ZMSY15-1。药物敏感性试验结果表明,分离菌株对青霉素、磺胺嘧啶钠耐药,对其他14种药物均敏感。序列分析结果显示,马链球菌兽疫亚种ZMSY15-1与国内外不同动物源分离株SzM基因氨基酸同源性为56.0%~70.0%。遗传进化分析结果显示,这些菌株可分为4个群。马链球菌兽疫亚种ZMSY15-1与猪源分离株SzM蛋白的氨基酸同源性为59.9%,分别属于2个不同的群,其与美国马源分离株NH55426亲缘关系最近。本试验结果可丰富国内马源马链球菌兽疫亚种SzM基因的信息数据,为马链球菌兽疫亚种的致病机制研究和预防控制提供参考依据。 相似文献
13.
14.
Akineden O Hassan A Alber J El-Sayed A Estoepangestie AT Lämmler C Weiss R Siebert U 《Veterinary microbiology》2005,110(1-2):147-152
The present study was designed to identify and compare 32 beta-hemolytic streptococci isolated from 28 different harbor seals of the German North Sea during the phocine distemper outbreak in 2002. The bacteria were identified as Streptococcus equi subsp. zooepidemicus based on cultural, biochemical, serological and molecular studies. Epidemiological investigations by PCR restriction fragment length polymorphism analysis of the 16S-23S rDNA intergenic spacer region and gene szp and by macrorestriction analysis of the chromosomal DNA of the strains by pulsed field gel electrophoresis revealed that all 32 strains appeared to be identical. These results indicate that a single bacterial clone seemed to be distributed among the harbor seal population of the German North Sea during this outbreak. 相似文献
15.
Strangles is a serious disease in horses caused by Streptococcus equi subspecies equi. In this study, genes encoding putative extracellular proteins in this subspecies have been identified using signal sequence phage display. Among these, one showed similarities to the SclB protein, a member of the collagen-like proteins of Streptococcus pyogenes. The novel gene denoted sclC encodes a protein, SclC, of 302 amino acids, containing typical features found in cell wall-anchored proteins in Gram-positive bacteria. Based on similarities to the S. pyogenes collagen-like proteins the mature SclC protein can be divided into various domains: an N-terminal non-repetitive region (A), a highly repetitive collagen-like region (CL), and a C-terminal proline-rich wall-associated region (W). Using PCR, the sclC gene was detected in all studied strains of S. equi subsp. equi and S. equi subsp. zooepidemicus. Further, antibodies against recombinant SclC were detected in a collection of sera from horses with no history of strangles as well as horses previously infected with S. equi subsp. equi. Interestingly, the sera from convalescence horses were found to have significantly increased antibody titers against the SclC protein indicating that this protein is expressed during infection of S. equi subsp. equi. 相似文献
16.
马链球菌兽疫亚种类M蛋白亚单位疫苗小鼠免疫试验 总被引：2，自引：0，他引：2
为获得控制猪链球菌病安全高效的疫苗，本试验进行了马链球菌兽疫亚种类M蛋白亚单位疫苗的研制。应用热酸法提取马链球菌兽疫亚种ATCC35246株类M蛋白，并分别采用羟基磷灰石（HAT）层析和冷酒精沉淀法进行纯化，结果得到较纯的类M蛋白。用热酸提取的粗制类M蛋白、2种纯化的类M蛋白及全菌苗免疫小鼠，结果保护率分别为92％（11／12）、100％（12／12）、100％（12／12）和83％（10／12），表明类M蛋白亚单位疫苗有进一步开发的必要。 相似文献
17.
Streptococcus equi subsp. equi and subsp. zooepidemicus are important pathogens of the equine respiratory tract. Isolates of both subspecies were examined by pulsed-field gel electrophoresis (PFGE). With the exception of eight isolates, a unique band pattern was displayed for each of the 48 subsp. zooepidemicus isolates tested. A method to distinguish isolates of the genetically very homogeneous subsp. equi has hitherto not been available, although several methods have been tested. By the use of PFGE, 50 isolates of subsp. equi could be divided into eleven groups, each with a unique pulsotype. In addition, the recently characterised genes encoding the cell-wall proteins ZAG and FNZ of S. equi subsp. zooepidemicus strain ZV were shown by Southern blots to be present in all 98 tested isolates, including the type strains of the two subspecies. Binding assays showed that the expression of the two genes clearly differentiate between the two subspecies. 相似文献
18.
J Hewson C K Cebra 《The Canadian veterinary journal. La revue veterinaire canadienne》2001,42(6):465-467
A 7-month-old, male llama was diagnosed with peritonitis caused by Streptococcus equi subsp. zooepidemicus. Clinical findings, medical treatment, and case outcome are described. Hematogenous dissemination from suspected pneumonia is proposed as the route of infection in this case. Possible transmission of the organism through contact with horses is discussed. 相似文献
19.
I W Wibawan F H Pasaribu I H Utama A Abdulmawjood C L?mmler 《Research in veterinary science》1999,67(2):131-135
Hyaluronic acid is thought to be one of the critical virulence factors of Streptococcus equi subsp. zooepidemicus. The present study was designed to study the role of hyaluronic acid capsular material in mediating adherence and to resist the phagocytosis of the host's immune defence. The studies were performed with two encapsulated S. equi subsp. zooepidemicus and two unencapsulated phase variants. The bacteria had been previously isolated from diseased pigs and monkeys in Indonesia. The presence of capsular material was determined using the hyaluronic acid decapsulation test and by electron microscopic studies. Both encapsulated bacteria showed mucoid colonies after cultivation on blood agar, grew with diffuse colonies in soft agar media and reacted negatively in the salt aggregation test. The unencapsulated bacteria grew with small colonies on blood agar, formed compact colonies in soft agar media and reacted positively in the salt aggregation test. Adherence and phagocytosis studies revealed that the encapsulated bacteria adhered significantly more to HeLa cells and were less phagocytosed by murine macrophages compared to unencapsulated bacteria. Pretreatment of the HeLa cells using hyaluronic acid or pretreatment of the bacteria by hyaluronidase decreased the adherence value of encapsulated bacteria. Pretreatment of bacteria with pronase had no effect. The presented results strongly indicate that the hyaluronic acid capsular material contributes to adherence properties of S. equi subsp. zooepidemicus and might help the bacteria to resist phagocytosis by macrophages. 相似文献
20.
Finno C Pusterla N Aleman M Mohr FC Price T George J Holmberg T 《Journal of the American Veterinary Medical Association》2006,229(5):721-724
CASE DESCRIPTION: A 4-month-old American Paint Horse colt was evaluated because of acute onset of ataxia, left-sided head tilt, and fever and a recently noticed heart murmur. Upper respiratory tract infection caused by Streptococcus equi subsp equi had been diagnosed at 3 months of age. CLINICAL FINDINGS: Hematologic abnormalities included leukocytosis, mature neutrophilia, monocytosis, and mild anemia. Analysis of a CSF sample revealed high total protein concentration and total nucleated cell count; nucleated cells consisted mainly of degenerate neutrophils. Results of a real-time PCR assay were positive for S equi subsp equi, and a diagnosis of S equi subsp equi meningoencephalomyelitis was made. TREATMENT AND OUTCOME: Treatment included administration of potassium penicillin and fluids, but the foal developed uroperitoneum and was subsequently euthanized. Postmortem examination revealed meningoencephalomyelitis, and S equi subsp equi was cultured from a brain aspirate. Additional findings included suppurative cystitis with rupture and neutrophilic myocarditis. CLINICAL RELEVANCE: Findings suggest that S equi subsp equi meningoencephalomyelitis should be considered in the differential diagnosis for foals with neurologic signs that have a history of strangles or exposure to affected horses. 相似文献