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1.
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   

2.
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   

3.
Streptococcus suis capsular type 2 is still an important cause of economic losses in the swine industry. At the present time, vaccination of pigs against this infection is generally carried out with autogenous bacterins and results are equivocal. In this study, the protective effect of a live avirulent S. suis type 2 strain (#1330) which had induced a good protection in mice, was evaluated in swine. The experiment was performed in triplicate using 4 week-old piglets. A total of 15 piglets were vaccinated 3 times, 15 others were vaccinated 2 times, and 15 piglets were injected 3 times with sterile Todd-Hewitt broth. Using an indirect ELISA, an increase in the IgG response to S. suis antigens was noted in 27 of the 30 vaccinated piglets. On day 21 post-vaccination, all animals were challenged intravenously with a virulent S. suis type 2 strain (#999). In the 2 vaccinated groups, 26 animals were fully protected. Only 1 out of the 15 piglets vaccinated 3 times developed mild clinical signs. In the group vaccinated twice, 3 piglets showed clinical signs and 1 of them died after the challenge. In the control group, 7 animals died out of the 11 with clinical signs of infection. In conclusion, a protective immunity was observed in swine when using strain 1330. However, more studies are needed to assess the use of a live S. suis strain in a vaccine for pigs.  相似文献   

4.
Virulence of Streptococccus suis capsular type 2 strain 89-1591 has been controversial in literature. A standardized experimental model with specific-pathogen free piglets was used for a new evaluation of this strain. Twenty-nine piglets were allotted in 4 separated groups. Group 1 consisted of negative control animals which received broth medium. Groups 2, 3, and 4 were intravenously challenged with 2 mL of S. suis, strains 1330, 89-1591, and 166', respectively. The strain 1330 is a recognized avirulent Canadian strain. The strain 166' is a reference French virulent isolate. Pigs inoculated with strain 1330 did not present clinical signs of a S. suis infection. Contamination in organs and bacterial blood circulation were rare and lesions were almost non-existent. Infection of pigs with S. suis strain 89-1591 (group 3) and 166' (group 4) caused severe clinical problems, animals infected with S. suis 166' were the most affected. Pigs presented with clinical signs such as high body temperature, lameness, nervous symptoms, and even mortality. Lesions associated with S. suis were numerous for both strains, but more evident in animals of group 4. It can be concluded that S. suis strain 89-1591 is virulent, although its virulence seems to be lower than that of the French strain. Results of an experimental infection with strain 89-1591 may depend on different factors such as the route of inoculation and the immunological status of the animals used. Using conventional animals, with an unknown status regarding previous S. suis infections, equivocal results may be obtained, and this may explain differences reported by some authors with the same strain.  相似文献   

5.
为建立2型猪链球菌(S.suis 2)胞外蛋白组双向电泳样品的制备方法,本研究利用双向电泳(2-DE):分离S.suis 2强毒株与无致病株的胞外蛋白,分别得到它们的胞外蛋白图谱.在pH4~pH7范围内采用考马斯亮蓝R350染色法在2菌株的胞外蛋白质图谱中都分别检测出180±10个蛋白点.并且它们的蛋白质分子质量分布基本相似;在所检测到差异蛋白点中,其中有50个蛋白点只存在于无致病菌株中而强毒株中不存在,有52个蛋白点只存在于强毒株中而无致病菌株中不存在,有7个蛋白点在2菌株中的表达量相差5倍以上.我们在强毒株凝胶中挑取了10个差异蛋白点进行质谱分析,通过数据库检索,鉴定了其中的9个蛋白,另外1种与鞭毛蛋白有同源序列.这些结果为研究S.suis2的致病机理提供了蛋白质组学方面的信息.  相似文献   

6.
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   

7.
A murine model of Streptococcus suis type 2 meningitis in the pig   总被引:7,自引:0,他引:7  
When young or adult mice were infected experimentally with isolates of Streptococcus suis type 2 of known pathogenicity for pigs, the organisms produced disease, young mice being more susceptible than adults. An isolate of S suis type 2 less pathogenic for pigs also appeared less pathogenic in mice. The organism could be reisolated from infected mice for up to 42 days. Inoculum size was one factor influencing the likelihood of an individual mouse developing meningitis following intravenous inoculation. Transmission of the organism from inoculated to in-contact mice occurred in young mice after intravenous or oral administration. These studies indicate that the behaviour of S suis type 2 in mice resembles its behaviour in pigs.  相似文献   

8.
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   

9.
Pigs (9 [+/- 1] weeks old) were inoculated with Streptococcus suis type 2, pseudorabies virus (PRV), or both. For each pig of groups A, B, and C the inoculum of S suis was 10(9) colony-forming units. For each pig of groups A, B, and D the inoculum of PRV was 5 x 10(3) TCID50 of either PRV strain 4892 (group A, n = 9) or PRV isolate B (group B, n = 9). The PRV strain 4892 is a highly virulent strain; isolate B causes mild clinical signs of infection in inoculated pigs. Group-C pigs (n = 9) were given S suis alone, and group-D pigs (n = 3) were inoculated only with PRV isolate B. Clinical signs of infection and development of lesions were readily seen in pigs of groups A, B, and C. Duration and severity of clinical signs of disease and lesions were reduced in pigs of group C, compared with those of the other 2 groups. Lesions, such as polyarthritis and fibrinous pericarditis, were more abundant and acute in the groups of pigs given mixed challenge exposure, compared with pigs inoculated exclusively with S suis type 2 (group C). The group of pigs inoculated with PRV isolate B alone did not manifest clinical signs of disease or lesions. Average daily gain for group-C pigs was higher, compared with that of other groups; the difference was statistically significant at P less than 0.02 and P less than 0.05 for groups B and D, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

10.
In this study, the IgG response of mice injected with two virulent strains and one avirulent Streptococcus suis capsular type 2 strain was compared by Western blotting. The serum from mice immunized against the avirulent strain could recognize most proteins of the various strains tested and similar results were obtained with serum from mice injected with virulent strains. The live avirulent strain was injected twice (days 0 and 10) to groups of five mice, and four virulent strains from different geographical origins were used to challenge the animals. All mice, except one in one group, survived the challenge. These results suggest that a live avirulent strain could be used for immunization of swine, the natural host.  相似文献   

11.
Three swine commercial farms with high mortality rates in nursery pigs due to Streptococcus suis serotype 2 were studied. Brain samples from diseased animals were collected for a period of 6 to 10 mo and used to isolate the strain that was responsible for the mortality (virulent strain) in each farm. Tonsil swabs from piglets at 5, 10 and 15 d were taken to assess both total colonization and colonization by the virulent strain. The effect of sow vaccination against S. suis on colonization was evaluated in 1 of the farms. All suspect tonsil isolates were identified biochemically and then tested against serotype 2. The genomic patterns of serotype 2 isolates were compared to that of the virulent strain using Rep-PCR. Results showed that total colonization by S. suis occurred very early in the pigs' life, with most animals being colonized by weaning age. Prevalence of colonization by serotype 2 strains was much lower than total colonization. After comparing serotype 2 isolates with the virulent strains, only 1 tonsillar isolate had the same genomic pattern as the virulent strain and it belonged to a 4-week-old weaned pig. The genomic pattern of the virulent strain was not found in any tonsillar isolate from 15-day-old or younger pigs. Although limited by sample size, sow vaccination against S. suis increased total colonization at the same time significantly decreasing colonization by serotype 2 strains. Even though most pigs are colonized early in age by S. suis, colonization by the virulent strain is of low prevalence and delayed in time. This could constitute a risk factor for developing the disease later in time, because animals would be colonized when maternal immunity is no longer present, allowing the organism to become systemic.  相似文献   

12.
对来自广西边境地区部分发病猪场及屠宰场的370份猪组织进行细菌分离鉴定,共分离到16株链球菌,对这16株链球菌进行了形态学观察、PCR鉴定及小鼠致病性试验。结果表明,其形态、染色、生化特性均符合猪链球菌的特点,通过GDH鉴定与测序,证实这16株菌株均为猪链球菌,分别命名为GX1~GX16。通过特异性引物PCR分型(35个血清型),其中有1株为1型,1株为2型,1株为5型,1株为29型,5株为8型,7株未分出型。用这16株猪链球菌对昆明小鼠和BALB/c小鼠进行动物致病性试验,结果发现,菌株GX10对昆明小鼠和BALB/c小鼠致死率均为100%;菌株GX11对BALB/c小鼠致死率为80%;其他菌株对昆明小鼠和BALB/c小鼠均无致病性。菌株GX10、GX11对BALB/c小鼠的LD50分别为4.5×105和3.6×109 CFU。本试验结果表明,广西边境地区存在致病性猪链球菌8型。BALB/c小鼠对猪链球菌的敏感性高于昆明小鼠,更适合作为猪链球菌致病性研究的动物模型。  相似文献   

13.
16 strains of Streptococcus were isolated and identified from 370 pig tissues of clinically morbidity farms and slaughter houses in Guangxi border area.Morphological observation, PCR identification and pathogenicity test were carried out. The results showed that the morphology,dyeing,biochemical characteristics were coincidence with the characteristics of Streptococcus suis,confirmed that 16 strains were Streptococcus suis,and named as GX1 to GX16. Further by divided type identification (35 serotypes) by GDH,1 strains was type 1,1 strains was type 2,1 strains was type 5,1 strain was type 29,5 strains were type 8 and 7 strains could not differentiate types. The pathogenicity of 16 strains Streptococcus suis were detected by Kunming mice and BALB/c mice. The results showed that the letal rate of GX10 for Kunming mice and BALB/c mice was 100%;GX11 for BALB/c mice was 80%;Other strains were apathogenicity to the Kunming mice and BALB/c mice. The LD50 of GX10 and GX11 was 4.5×105 and 3.6×109 CFU for BALB/c mice,respectively. The results showed that pathogenic Streptococcus suis type 8 occured in Guangxi border area. The sensitivity of BALB/c mice to Streptococcus suis was higher than Kunming mice, BALB/c mice was more suitable for the pathogenic study of Streptococcus suis.  相似文献   

14.
Streptococcus suis serotypes associated with disease in weaned pigs   总被引:5,自引:0,他引:5  
Streptococcus suis was recovered from 9 outbreaks of septicaemia and meningitis in weaned pigs between 1979 and 1983. Fifteen isolates from 7 outbreaks were identified as S. suis type 9, and 3 isolates from 2 outbreaks as S. suis type 2. Three further isolates of S. suis type 2 and an isolate of S. suis type 3 were recovered from cases of bronchopneumonia in weaned pigs from 4 other piggeries.  相似文献   

15.
Streptococcus suis is an important pathogen of swine, causing meningitis, arthritis, polyserositis, septicemia, and sudden death in weaning piglets as well as fattening pigs. Recently, 3 molecular tests have been developed in our laboratory: a multiplex polymerase chain reaction (m-PCR) assay for the detection of S. suis species and serotypes 2 and 1/2, and 2 molecular typing methods, pulsed-field gel electrophoresis and an approach based on PCR amplification of a fragment of rRNA genes, including a part of the 16S and 23S genes and the 16S-23S rDNA intergenic spacer region (ISR), followed by restriction fragment length polymorphism (RFLP) analysis (ISR-RFLP). In the present study, we used these tests to analyze tonsil samples from clinically healthy pigs and to identify individual isolates of S. suis during epidemiologic investigations of 8 related herds with a history of septicemia caused by S. suis serotype 2. Capsular typing showed that 58% of the strains were nontypable. Of the 17 serotypes present, serotype 22 was the most prevalent. In the 7 farms without clinical signs on the day of sampling, we detected S. suis serotype 2 or 1/2, or both, in less than 5% of the pigs by m-PCR or by bacteriologic culture. In the 8th farm, on which 2 pigs had clinical signs of septicemia on the day of sampling, we detected S. suis serotype 2 or 1/2, or both, by m-PCR in the tonsils of 40% of fattening pigs (21 wk old) that lacked symptoms. Molecular typing of the serotype 2 strains showed a common origin of contamination in these herds, given that 1 pattern (C1) was detected in the isolates from 6 of the 8 herds. However, up to 4 patterns were associated with septicemia and sudden death. Several patterns of S. suis serotype 2 can be responsible for disease in the same herd. These molecular tools may be useful for confident studies of the transmission of S. suis, thereby contributing to the control of S. suis infection.  相似文献   

16.
Streptococcus suis isolated from pigs in Finland   总被引:6,自引:0,他引:6  
A total of 58 Streptococcus suis strains were isolated from deceased pigs submitted to the National Veterinary Institute, Regional Laboratory in Kuopio, Finland, over a 3 1/2 year period, most frequently from cases of pneumonia. The bacteria were isolated from cases of meningitis, sepsis, rhinitis, endocarditis and abortion. S. suis was also isolated from nasal cavity, lung and brain of some sick piglets without signs of inflammation. Further S. suis was detected in 12 out of 107 tonsils of healthy fatteners tested. S. suis strains were identified by biochemical methods followed by typing. The most common capsular types were 7, 3 and 2, respectively. Only one type 1 strain and no types 6 and 9 strains were found. All S. suis strains tested were sensitive to penicillin and ampicillin.S. suis is not uncommon in Finnish pig herds. S. suis may be regarded as a potentially pathogenic organism which under certain predisposing conditions may cause serious disease.  相似文献   

17.
A total of 323 isolates of Streptococcus suis recovered from diseased or healthy pigs in France were serotyped. The presence of virulence-related proteins, Muraminidase-Released Protein (MRP), Extracellular Factor (EF) and Suilysin was also studied in 122 isolates of capsular types 2, 1/2, 9, 7 and 3 to evaluate their implication in virulence of S. suis. Capsular types 2, 1/2, 9, 7 and 3 were the most frequently detected (93%), with 69% for the capsular type 2 alone. Capsular types 2, 1/2, 9, 7, 3, 1, 4, 8, 18, 10 and 12 were isolated from diseased pigs, whereas types 2, 7, 9, 1/2, and 3 originated from the nasal cavities or tonsils of healthy animals. Most of the S. suis type 2 isolates recovered from diseased pigs carried MRP+ EF- Suilysin- (46%) or MRP+ EF+ Suilysin+ (28%) phenotypes. The MRP+ EF- Suilysin- phenotype was also detected in 67% of S. suis type 2 strains isolated from healthy pigs. The production of the virulence-related proteins was less frequently found in S. suis types 1/2, 9, 7 and 3 recovered either from diseased or healthy pigs. In this study, all the capsular type 1/2 strains were MRP+ EF- Suilysin- and all the S. suis type 7 harboured an MRP- EF- Suilysin- phenotype. The MRP- EF- Suilysin- phenotype was found in S. suis types 2, 3, 7 and 9 isolated from septicaemia, meningitis, pneumonia, and pleurisy. These results suggest that the presence of these proteins should not be used as a single condition for classifying the virulence of a field isolate in France.  相似文献   

18.
We developed a PCR assay for the rapid and sensitive detection of virulent Streptococcus suis type 2 and highly virulent S. suis type 1 in tonsillar specimens from pigs. The PCR primers were based on the sequence of the gene encoding the EF-protein of virulent S. suis type 2 strains (MRP+EF+) and highly virulent S. suis type 1 strains (MRP(s)EF+) and of the EF protein of weakly virulent S. suis type 2 strains (MRP+EF). The latter strains give rise to larger PCR products than the virulent strains of S. suis type 1 and 2. A positive control template was included in the assay to identify false negative results. The PCR was evaluated using tonsillar specimens from herds known (or suspected) to be infected and herds without an S. suis history. The results obtained with the PCR assay were compared with the results obtained with a newly developed bacteriological examination. In this bacteriological examination we were able to identify the EF-positive strains directly in the tonsillar specimens. From the 99 tonsils examined, 48 were positive in the PCR and 51 negative. All specimens from which EF-positive S. suis strains were isolated were also positive in the PCR assay. Three samples were positive in the PCR, but negative by bacteriological examination. The results demonstrated that the PCR is a highly specific and sensitive diagnostic tool for the detection of pigs carrying virulent strains of S. suis type 2 and highly virulent strains of type 1. Application of the assay may contribute to the control of S. suis infections.  相似文献   

19.
Streptococcus (S.) suis is an invasive porcine pathogen causing meningitis, septicemia, arthritis and other diseases. Studies on pathogenesis as well as vaccine trials have focused on serotype 2 strains, which are worldwide the most prevalent among invasive isolates. However, in Europe serotype 9 strains also contribute substantially to S. suis-associated invasive diseases of piglets. The objective of this study was to determine the virulence of an MRP* SLY+ serotype 9 S. suis strain in comparison to an MRP+ EF+ SLY+ serotype 2 strain. Experimental intranasal and intravenous infections of 7-8 weeks old SPF piglets were investigated with regard to clinic and pathology. In contrast to the virulent serotype 2 strain, the serotype 9 strain did not cause disease with clinical manifestations after intranasal administration. However, histological screenings of these animals revealed pathological lesions, such as mild focal suppurative meningitis. Clinical manifestations related to meningitis, arthritis and serositis could be induced by intravenous application of this serotype 9 strain. Bacteriological culture and immunohistochemistry of the brain confirmed association with the S. suis challenge strains in all cases with clinical manifestations. Interestingly, expression of MRP within meningitis lesions was demonstrated for both pathotypes via immunohistochemistry. In conclusion, this study demonstrates that MRP* SLY+ serotype 9 strains are less virulent for growers than MRP+ EF+ SLY+ serotype 2 strains. Thus, intravenous application of this serotype 9 strain is required to evaluate heterologous protection in the course of vaccine development based on serotype 2 strains in the future.  相似文献   

20.
为分析猪链球菌4型(Streptococcus suis type 4,SS4)分离株的病原生物学特性,试验对来自中国不同地区的10株猪链球菌4型进行了多位点序列分型,通过PCR方法对猪链球菌7个保守管家基因aroA、gki、dpr、mutS、recA、thrA、cpn60进行扩增,测序后将结果上传至MLST数据库查找序列型,然后制作聚类分析图来阐明菌株之间的亲缘关系;采用PCR方法对7种主要的毒力基因gdh、mrp、epf、sly、fbps、gapdh与orf2进行鉴定,通过毒力因子谱来分析猪链球菌4型毒力因子的分布;以纯化的10株细菌对BALB/c小鼠进行动物致病性试验,根据小鼠致死数量筛选出最强毒株,并进行新西兰兔致病性试验。结果显示,10株猪链球菌4型经多位点序列分型,6株为ST850型,3株为ST1006型,1株为ST94型;结合菌株分离地区分析发现,广东和江苏地区菌株有较高的同源性,江沪地区菌株出现分化现象,表现为遗传多样性;10株菌株均检测到了gdh、gapdh和orf2毒力基因,7株检测到sly基因,4株检测到fbps基因,根据毒力因子谱发现共有3个毒力基因型,gdh+sly+gapdh+orf2+型有6株,gdh+fbps+gapdh+orf2+型有3株,gdh+sly+fbps+gapdh+orf2+型仅有1株。动物致病性试验表明,10株细菌均能使BALB/c小鼠死亡,其中SH1510的半数致死量低至1×108 CFU,对小鼠的致病性最强,将纯化的SH1510菌液接种新西兰兔,可使新西兰兔出现典型的神经症状并死亡。以上结果为猪链球菌的遗传进化、毒力研究提供了新的数据,丰富了猪链球菌病的研究。  相似文献   

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