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1.
The efficacy of a muramidase-released protein (MRP) and extracellular factor (EF) vaccine in preventing infection and disease in pigs challenged either with a homologous or a heterologous Streptococcus suis serotype 2 strain (MRP+EF+) was compared with the efficacy of a vaccine containing formalin-killed bacterin of S. suis serotype 2 (MRP+EF+). The enhancement of the immune response by different adjuvants (a water-in-oil emulsion [WO] and an aluminium hydroxide-based adjuvant [AH]) and their side effects were also studied. The MRP and EF were purified by affinity chromatography. Pigs were vaccinated twice at three weeks and six weeks of age and challenged intravenously with virulent S. suis serotype 2 strains (MRP+EF+) at eight weeks of age. At challenge, the pigs vaccinated with MRP+EF/WO had high anti-MRP and anti-EF titres and were protected as effectively as pigs vaccinated with WO-formulated vaccines with bacterin. Eight of the nine pigs survived the challenge and almost no clinical signs of disease were observed. The titres obtained with the MRP+EF/AH vaccine were low and only two of the five pigs were protected. Pigs vaccinated with either MRP or EF were less well protected; three of the four pigs died after challenge but the clinical signs of disease were significantly less severe than those observed in the placebo-vaccinated pigs. The protective capacity of the bacterin/AH vaccine was very low, and the mortality among these pigs was as high as in the placebo-vaccinated pigs (80 per cent). Postmortem histological examination revealed meningitis, polyserositis and arthritis in the clinically affected pigs. The results demonstrate that a subunit vaccine containing both MRP and EF, formulated with the WO adjuvant, protected pigs against challenge with virulent S. suis type 2 strains.  相似文献   

2.
根据猪链球菌２型的mrp基因自设计和合成了一对可扩增长度为８８５bp目的片段的引物，成功地建立了检测溶菌酶释放蛋白（ＭＰＲ）的ＰＣＲ方法，用ＸbaⅠ内切酶进行酶切，获得了与预期一致的５７8bp和３０７bp２的２个片段，并进行了ＰＣＲ的特异性试验和敏感性试验。对马链球菌兽疫亚种、猪丹毒杆菌、猪肺疫巴氏杆菌和猪肺炎支原体的ＰＣＲ检测结果均呈阴性；检测的敏感度可达１００个细菌。另外，对９株从病猪体内分离的猪链球菌２型菌株进行检测了８个呈阳性；对１５份正常屠宰猪扁桃体分离物的检测结果是１份为阳性。结果表明此法特异性敏感性均很高，可作为ＭＲＰ快速诊和流行病学调查的手段。  相似文献   

3.
The production of muramidase-released protein (MRP), extracellular protein factor (EF) and hemolysin (suilysin) by 101 Canadian field strains of Streptococcus suis capsular type 2 is described. Most strains (72%) isolated from diseased pigs were MRP-EF- and only 1 strain was MRP+EF+. This strain was also the only 1 to produce the hemolysin. Thirteen strains (15%) were MRP+ EF- and only 3 strains were MRP* EF-. All the strains isolated from clinically healthy pigs as well as a bovine and 2 human isolates had a MRP-EF- phenotype. In addition, 7 strains (8%) had a MRPS phenotype, which had so far been described for S. suis capsular type 1. In conclusion, most Canadian field isolates of S. suis capsular type 2 tested in this study do not produce the virulence-related proteins described so far for this bacterial pathogen.  相似文献   

4.
The effect of vaccination with a killed whole-cell vaccine of extracellular factor-positive Streptococcus suis serotype 2 (S suis 2 EF(+)) combined with medication with amoxicillin on the presence of virulent S suis 2 EF(+) strains on the tonsils of sows and their offspring was evaluated. In two herds, 14 pregnant sows that carried these virulent strains, as detected by PCR in three consecutive tonsillar brush samples, were selected and randomly assigned to be treated or left untreated as controls. The treated sows were vaccinated at six and three weeks before the expected farrowing date and medicated from one week before expected farrowing until the end of the experiment. Two weeks before parturition, the sows were housed in individual isolation farrowing rooms, and the sow and its litter were sampled by using tonsil brushes and tonsil swabs, respectively. Approximately 27 days postpartum, the sows and their piglets were euthanased and their tonsils were collected and analysed by pcr. No S suis 2 EF(+) could be detected in the tonsils of the seven treated sows, but the tonsils of the seven untreated sows remained positive. Only one of the litters of the untreated sows became infected, five days after birth, and none of the litters of the treated sows became infected.  相似文献   

5.
猪链球菌2型的PCR快速检测   总被引：37，自引：4，他引：33  
根据猪链球菌 2型的荚膜多糖抗原基因 cps2 J,合成 1对可扩增长度为 6 75 bp目的片段的引物 ,建立了检测猪链球菌 2型的 PCR法。应用 PCR对 9株经玻片凝集试验检测为猪链球菌 2型的菌株进行了检测 ,均呈阳性 ;而对马链球菌兽疫亚种 (C群 )、猪葡萄球菌、猪丹毒杆菌、猪肺疫巴氏杆菌、猪肺炎霉形体等检测结果均呈阴性 ,表明了本方法的特异性。用此法对 88份正常猪的扁桃体样品的细菌分离物进行了检测 ,36份呈阳性 ,同时用玻片凝集试验进行对照检测 ,也全部呈阳性。而此法不需进行细菌的纯分离培养 ,即可用于猪链球菌 2型的快速诊断以及流行病学调查。  相似文献   

6.
Two cases of meningitis due to Streptococcus suis in humans are reported here. A butcher and an abattoir worker were referred to a health centre in Castellón (Spain) with fever and symptoms of meningitis. After adequate treatment, a slight hipoacusia persisted as sequelae in both cases. Colonies of S. suis group R, serotype 2 and phenotype MRP+EF+ were isolated from cerebroespinal fluid. Epidemiological studies showed that both workers had in common the handling of pork meat of slaughtered healthy pigs from three closed farms. A study of the tonsils from apparently healthy, slaughtered pigs was carried out. A total of 234 tonsillar samples were obtained and 81 strains of S. suis were isolated from them. Serotype 2 appeared to be the most frequent (50.6%), and the analysis for phenotype showed a high percentage of tonsillar strains with the phenotype MRP+EF+ (35.9%). The humans and 28 tonsillar swine strains showed a similar profile (S. suis group R, serotype 2 and phenotype MRP+EF+). A total of 26 of the swine isolates were analysed by ribotyping using EcoRI. The human strains showed the same six-band hybridization pattern that shared five bands with the pattern most frequently shown by most of the tonsillar N. suis group R, serotype 2 and phenotype MRP+EF+ strains, differing only in the lightest, faintest band which was slightly less anodical in human (> or = 1.8 kb) than in swine (approximately 1.8 kb). From these results, both groups of strains, humans and porcine, showed differences; how can these differences in the pattern of ribotyping be explained if they should have the same origin? Is it possible that they have undergone an adaptation to the new host or perhaps the modification is due to other unknown causes? Further studies in this area are required in order to answer these questions.  相似文献   

7.
A total of 142 strains from different serotypes of Streptococcus suis isolated in Spain from diseased pigs (88 strains) and healthy carrier pigs (54 strains) were studied for the presence of a muramidase released protein (MRP) and an extracellular factor (EF). The following five phenotypes: MRP+EF+, MRP+EF-, MRP-EF+, MRP+EF* and MRP*EF- were detected. A high percentage of S. suis serotype 2 strains isolated from diseased pigs (84 per cent) belonged to phenotype MRP+EF+, but this phenotype has also been noticed in other serotypes (serotypes 1, 1/2 and 14). Both proteins were detected in S. suis serotype 2 strains (26%) isolated from healthy carrier pigs and one of both proteins in serotypes 1 and 14 (phenotype MRP+EF*). The isolation of S. suis strains from healthy pigs which have shown both proteins may support the epidemiological significance of these carriers in the maintenance, transmission and distribution of virulent strains within and between swine farms.  相似文献   

8.
A standardized model of Streptococcus suis type 2 infection in specific-pathogen-free piglets, housed in high-security barns, was used to compare the virulence of 3 French field strains of S. suis serotype 2 isolated from tonsils of a healthy pig (strain 65) or from diseased pigs (meningitis, strain 166', or septicemia, strain 24). In one of the 2 trials, 7-week-old pigs, in 3 groups of 8, were inoculated intravenously with 2 x 10(8) colony-forming units of S. suis type 2. In each group, 1 uninfected animal was a sentinel. Eight animals were also used as negative control group. The experiment was repeated under similar conditions with strains 65 and 166'. Virulence differed markedly among these S. suis strains when clinical signs, zootechnical performances, lesions, and bacteriological data were analyzed. Strain 65 did not induce clinical signs in inoculated pigs. In contrast, pigs infected with the other 2 strains exhibited clinical signs and typical lesions of S. suis type 2 infections. Differences in virulence were also observed between the 2 virulent strains. Sentinel animals exhibited the same manifestations as those recorded in inoculated piglets. Results were similar in the second trial, indicating that under the present experimental conditions, results were reproducible. The standardized conditions described in this study could be a useful tool to further study about the S. suis infection.  相似文献   

9.
Streptococcus suis type 2 infections in pigs in the Netherlands (Part two)   总被引：2，自引：0，他引：2  
Since 1983 some pig breeding and fattening farms in the Netherlands have been faced with a considerable mortality in pigs due to Streptococcus suis type 2 infections. The most predominant clinical feature of S. suis type 2 infection is meningitis, although sudden deaths often occur. It was noted that some affected farms had imported breeding stock from the United Kingdom. Tonsils of slaughter pigs were collected from herds with and without a history of S. suis type 2 infections. Bacteriological examination was done by using an elective-selective medium. No significant difference was found in carrier rates of S. suis type 2 between clinically healthy and affected herds (38% vs. 45%). A cohort study was carried out by regular bacteriological examination of tonsil biopsies on a farm with a high incidence of streptococcal meningitis. Twenty-seven percent of the pigs were carriers of S. suis type 2 at nine weeks of age. Possible methods for disease control are discussed.  相似文献   

10.
Sepsis with subsequent multisystem organ failure after translocation of bacteria from the gut is a serious risk associated with stress situations. We showed that intestinal bacterial translocation could be one of the pathways for pathogenic Streptococcus suis infections in the pig. In 24 piglets weighing 10-14 kg, free of the extracellular factor (EF+) producing phenotype of S. suis serotype 2, a silicon canula was placed in the proximal jejunum to enable intestinal inoculation and bypassing the upper alimentary tract. The pigs were individually housed. After stress induction in 18 pigs by means of a truck drive in individual cages for 1h, pigs were inoculated through the intestinal canula either with S. suis type 2 EF+ or with growth medium only, and put back in their original housing. The six not transported pigs were also inoculated with the same strain. To prevent oral self-infection, faeces were collected in a bag that was glued around the anus. Clinical and behavioral symptoms were recorded for 72 h post inoculation, and then the animals were sacrificed for pathological and bacteriological examination. In three animals, the inoculation strain was re-isolated from mesenterial lymph nodes and typically affected organs. No S. suis type 2 EF+ was detected by specific polymerase chain reaction (PCR) in any of the tonsil-swabs and -homogenates. We concluded that infection of the organs had taken place after bacterial translocation out of the gut and that the intestinal tract can be a porte d'entree for S. suis type 2 EF+.  相似文献   

11.
Tonsillar and nasal swabs were collected from weanling pigs in 50 representative Ontario swine herds and tested for the presence of 5 important bacterial upper respiratory tract pathogens. All but 1 herd (2%) tested positive for Streptococcus suis by polymerase chain reaction (PCR); 48% of herds were S. suis serovar 2, 1/2 positive. In all but 2 herds there was evidence of Haemophilus parasuis infection. In contrast, toxigenic strains of Pasteurella multocida were detected by a P. multocida--enzyme-linked immunosorbant assay (PMT-ELISA) in only one herd. Seventy-eight percent of the herds were diagnosed positive for Actinobacillus pleuropneumoniae by apxIV PCR. Sera from finishing pigs on the same farms were also collected and tested by ELISA for the presence of A. pleuropneumoniae antibodies. Seventy percent of the herds tested had evidence of antibodies to A. pleuropneumoniae including serovars 1-9-11 (2%), 2 (4%), 3-6-8-15 (15%), 5 (6%), 4-7 (26%), and 12 (17%). This likely represents a shift from previous years when infection with A. pleuropneumoniae serovars 1, 5, and 7 predominated. At least 16% and possibly as many as 94% of the herds tested were Actinobacillus suis positive; only 3 of the 50 herds were both A. pleuropneumoniae and A. suis negative as judged by the absence of a positive PCR test for apxII. Taken together, these data suggest that over the past 10 years, there has been a shift in the presence of pathogenic bacteria carried by healthy Ontario swine with the virtual elimination of toxigenic strains of P. multocida and a move to less virulent A. pleuropneumoniae serovars. As well, there appears to be an increase in prevalence of S. suis serovar 2, 1/2, but this may be a reflection of the use of a more sensitive detection method.  相似文献   

12.
Virulence-associated gene profiling of Streptococcus suis isolates by PCR   总被引：11，自引：0，他引：11  
Definition of virulent Streptococcus suis strains is controversial. One successful approach for identification of virulent European strains is differentiation of capsular serotypes (or the corresponding cps types) and subsequent detection of virulence-associated factors, namely the extracellular factor (EF, epf), the muramidase-released protein (MRP, mrp) and the hemolysin suilysin (SLY, sly). In this work we present a novel multiplex PCR (MP-PCR) and an mrp variant PCR for identification and characterization of virulent S. suis strains. These new methods were used to identify association of disease with particular profiles of virulence-associated genes. The MP-PCR allowed identification of S. suis through detection of the housekeeping gene gdh, differentiation of four cps types (1, 2, 7 and 9), and detection of epf, mrp, sly and arcA (arginine deiminase from S. suis). Furthermore, this study describes the first PCR assay for differentiation of at least six mrp variants. Expression of the corresponding size variants of MRP was shown for four of the six mrp variants, but was undetectable for the two larger mrp variants in the particular strains investigated. The results of this study suggest that cps7 strains are associated with pneumonia and that variation of mrp is very pronounced among these strains. Gene profiles of invasive, pneumonia and carrier S. suis isolates by combination of PCR assays allowed differentiation of 24 different genotypes among cps1, 2, 7 and 9 strains. Forty-five percent of the invasive S. suis diseases investigated in this study were caused by only two of these genotypes, namely cps2/mrp+/epf+/sly+ and cps9/mrp(*)/epf-/sly+. Thus, this study demonstrates for the first time a uniform profile of the particular virulence-associated genes for the vast majority of the investigated invasive cps9 strains.  相似文献   

13.
根据猪链球菌2型荚膜多糖和马链球菌兽疫亚种类M蛋白的保守区序列分别设计了2对简并引物,建立了一种能同时检测猪链球菌2型和马链球菌兽疫亚种的双重PCR方法。结果显示,该双重PCR能从100个细菌的混合纯培养物中扩增出2条目的片段。而且可以直接从病料组织中检测到相应的病原菌。用建立的双重PCR方法和细菌分离培养法平行检测人工感染的组织病料,PCR方法与细菌培养法的阳性检出率基本一致,但PCR方法的特异性好、敏感性高,简便易行,可以用于猪链球菌病的流行病学调查和实验室的快速鉴别诊断。  相似文献   

14.
猪链球菌2型对PK-15细胞的黏附动力学   总被引：1，自引：1，他引：0  
为研究携带不同毒力因子的猪链球菌2型对细胞侵袭作用的差异,本实验室从全国各地分离100多株猪链球菌,PCR鉴定出10株链球菌2型,其中携带毒力因子菌株为7-4［MRP+EF+］、8-3［MRP+EF-］、ZH-1［MRP-EF+］,另外选择1株LN-2 ［MRP-EF-］与PK 15细胞进行黏附动力学试验,结果表明MPR毒力因子与菌株对PK-15细胞黏附具有密切相关性。  相似文献   

15.
Summary

Since 1983 some pig breeding and fattening farms in the Netherlands have been faced with a considerable mortality in pigs due to Streptococcus suis type 2 infections. The most predominant clinical feature of S. suis type 2 infection is meningitis, although sudden deaths often occur. It was noted that some affected farms had imported breeding stock from the United Kingdom.

Tonsils of slaughter pigs were collected from herds with and without a history of S. suis type 2 infections. Bacteriological examination was done by using an elective‐selective medium. No significant difference was found in carrier rates of S. suis type 2 between clinically healthy and affected herds (38% vs. 45%).

A cohort study was carried out by regular bacteriological examination of tonsil biopsies on a farm with a high incidence of streptococcal meningitis. Twenty‐seven percent of the pigs were carriers of S. suis type 2 at nine weeks of age. Possible methods for disease control are discussed.  相似文献   

16.
猪链球菌2型及其毒力因子检测多重PCR的建立与应用   总被引：8，自引：3，他引：8  
根据相关文献设计并合成引物，建立了能同时检测猪链球菌2型（cps）及其重要毒力因子溶菌酶释放蛋白（mrp）和细胞外因子（epf）的多重PCR方法。目的片段的大小分别为885bp（mrp）、675bp（cps）和443bp（epf）。对参考菌株、人工攻毒病料和临床收集病料的检测结果显示，该多重PCR特异性强、敏感性高，可直接从临床病料中检测出猪链球菌2型，并能鉴定其毒力因子表型。  相似文献   

17.
Fifteen newborn germ-free pigs were inoculated with 2 strains, D-282 and T-15, of Streptococcus suis type II. Some pigs also were preinoculated with Bordetella bronchiseptica, which successfully predisposed them to S suis infection. The 2 streptococcal strains were differentiated by muramidase treatment, which released certain high molecular-weight proteins, termed muramidase-released proteins (MRP), from the cell wall of strain D-282, but not from the cell wall of strain T-15. Only strain D-282 (MRP-positive) induced clinical signs of disease and markedly increased neutrophil numbers in pigs. Streptococci were more frequently isolated from fecal swab specimens obtained from pigs inoculated with strain D-282 (MRP-positive) than from specimens obtained from pigs inoculated with strain T-15 (MRP-negative). Both strains were isolated from nasal swab specimens obtained from all infected pigs. Postmortem examination revealed fibrinopurulent meningitis, polyserositis, and polyarthritis in pigs inoculated with strain D-282; this strain was isolated from the CNS, serosae, visceral organs, heart, and joints. Whereas strains D-282 caused several pathologic changes, strain T-15, isolated from the lungs, caused only pneumonia. Both strains were isolated from the tonsils of all pigs. Virulence differed distinctly between the MRP-positive and the MRP-negative strains.  相似文献   

18.
The detection of pigs carrying Streptococcus suis type 2   总被引：2，自引：0，他引：2  
An indirect fluorescent antibody test (I.F.A.T.) was performed on tonsillar swabs collected at slaughter and on nasal swabs from electrically stunned pigs prior to exsanguination. The identification of carriers of Streptococcus suis type 2 using the I.F.A.T. was compared with bacteriological isolation. If it is assumed that 100% of pigs were carrying the bacterium, 76% of 89 carriers were detected when the I.F.A.T. was performed on the bacterial growth from blood agar cultures of tonsillar swabs, compared with only 15% detected using cultural techniques. Similarly, I.F.A.T. on nasal swabs increased the sensitivity of the detection of carriers. Nasal swabbing, although of lower sensitivity than tonsillar swabbing, allows for the detection of S. suis type 2 carriers in the live animal.  相似文献   

19.
In order to identify gene sequences unique to the virulent strains, suppression subtractive hybridization (SSH) was conducted using virulent Streptococcus suis type 2 (SS2) strain HA9801 and avirulent S. suis type 2 strain T15. Thirty genomic regions were absent in T15, and the DNA sequences of these regions in HA9801 were determined. These DNA fragments, containing putative virulence genes, encoded 28 proteins that were homologous to proteins involved in various aspects of cellular surface structure, molecular synthesis, energy metabolism, regulation, transport systems and others of unknown function. According to the published SS2 genomic sequence of the Chinese strain 98HAH33, PCR primers for 14 significant DNA fragments were designed and used for detection of the distribution of these fragments in S. suis strains from different sources, serotypes, regions, groups and times. The results showed that these 14 DNA fragments were widely distributed in 37 detected SS2 strains, yet were absent among the avirulent strain T15. Moreover, these fragments could be detected in other serotypes of S. suis, but each serotype had a different distribution of the fragments.  相似文献   

20.
Rapid detection of Streptococcus suis serotype 2 in weaned pigs   总被引：5，自引：0，他引：5  
A survey to detect Streptococcus suis serotype 2 in 1,716 weaned pigs was done in Quebec. Forty-nine sow herds were included in this survey: in 26 herds, S suis serotype 2 had been isolated during the preceding 12 months and in 23 herds (control), the organism had not been detected during a previous study. Swab specimens of the nasal cavity and tonsils of pigs were obtained for bacteriologic culture, and S suis serotype 2 was easily detected by the use of brain-heart infusion agar containing a Streptococcus-selective supplement and 5% goat antiserum raised against S suis serotype 2. After measurement of the diameter of the precipitation zone of 539 isolates, a slide agglutination test was performed to identify the S suis serotype 2 isolates. The mean precipitation zone diameter obtained for group S suis serotype 2 was larger (P less than 0.001) than that for the group designated as "others". With slide agglutination test results as reference and on the basis of discriminant analysis to stimulate detection of S suis serotype 2, 93.1% of all isolates were correctly classified, using the precipitation zone diameter as unique classification criterion. Relative specificity was 94.5% and relative sensitivity was 88.7%. Use of the precipitation zone diameter on a quantitative basis led to the proposal of a simple and reliable technique to screen swine herds for S suis serotype 2 in weaned pigs. Nasal and tonsillar swab specimens were obtained and analyzed concurrently for S suis serotype 2. The organism was found in both sites in only 20.4% of 103 carrier pigs.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

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