Immunisation of pigs with killed cultures of Streptococcus suis type 2

Ten intravenous inoculations of 10(10) formalin-killed pathogenic Streptococcus suis type 2 stimulated detectable levels of opsonic antibody and a strong protective response against intravenous challenge with 10(9) viable S suis type 2 of the same strain. Heat-killed organisms did not stimulate a protective response, even though the type-specific antigen was intact and opsonic antibodies were induced. When the number of formalin-killed organisms in the inoculum was increased 10-fold, one inoculation was insufficient to stimulate a protective response, but two inoculations protected pigs against intravenous challenge with live, pathogenic organisms. Four-hour and overnight cultures were equally immunogenic, and the response was not enhanced by either Freund's incomplete adjuvant or aluminium hydroxide gel adjuvant.     

Studies of Streptococcus suis type 2 infection in pigs

Investigations into the immunology, pathogenesis and epidemiology of Streptococcus suis type 2 infections were carried out in experimental pigs and in naturally-occurring field outbreaks of disease. The capsular polysaccharide from Str. suis type 2 was shown to induce opsonic antibodies in pigs when injected with Freund's incomplete adjuvant, but difficulties encountered in experimental production of the disease prevented a study of their protective effects. Problems with the bactericidal tests led to an investigation of other assays for antibodies against Str. suis type 2, namely, a phagocytic test with pig neutrophils, a mixed reverse passive antiglobulin haemagglutination test and an indirect haemagglutination test. There was evidence that with modifications both the latter tests would be useful. Transmission studies in 39 conventionally-reared and 7 hysterectomy-derived, colostrum-deprived pigs yielded interesting results with regard to the distribution of the organism in relation to the disease process. Tonsil carriage in clinically-healthy pigs was demonstrated after experimental and natural infection. Detectable carrier rates varied between 0 and 59%. The organism was shown to persist in the presence of circulating opsonic antibodies and in pigs on penicillin-medicated feed. Attempts to isolate the organism from the genital tract were unsuccessful. Medicated early weaning and classical SPF techniques applied to infected herds appeared to be effective in producing pigs free from Str. suis type 2 infection.     

The detection of pigs carrying Streptococcus suis type 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.     

Experimental infections of mice and pigs with Streptococcus suis type 2.

Five inbred strains of mice were tested for their susceptibility to Streptococcus suis type 2 including the type strain, two isolates from meningitis in pigs and two isolates from tonsils of clinically healthy pigs. C57BL/6, ICR and ddY strain mice showed lower susceptibility to all strains of S. suis type 2 than BALB/c and SS strain mice. The type strain and the isolates from diseased pigs produced septicaemia and meningitis in BALB/c and SS mice inoculated with 10(8) colony forming unit of the bacteria and 60 to 100% of these infected mice died. On the other hand, mice inoculated with the isolates from healthy pigs showed mild clinical signs but none of them died. In BALB/c mice which died or developed nervous signs, the purulent meningo-encephalitis, myocarditis, ophthalmitis, labyrinthitis and otitis media were observed. S. suis type 2 antigen was demonstrated in these lesions by immunoperoxidase staining using rabbit S. suis type 2 antiserum. These results were similar to those in the experimentally infected pigs with these virulent and avirulent strains against mice. These results indicate that BALB/c and SS strains of mice are useful as an experimental model of S. suis type 2 infections in pigs, and that there are virulent and avirulent strains against mice and pigs among the strains of S. suis type 2.     

Intestinal translocation of Streptococcus suis type 2 EF+ in pigs

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+.     

Experimental airborne transmission of Streptococcus suis capsular type 2 in pigs.

Experimental airborne transmission of Streptococcus suis type 2 was studied in specific pathogen free piglets. Forty piglets were allotted to five groups of eight 7-week-old animals and housed in three separated units. Negative control pigs (group 1) were housed in unit A and infected batches were housed in units B (group 2) and C (groups 4). In units B and C, non-inoculated groups (groups 3 and 5, respectively), 40 cm distant from the respective inoculated group and without any physical contact between them, also took place. Six animals of groups 2 and 4 were inoculated intravenously with 2 x 10(8) colony forming units (cfu) of a mild and a high virulent S. suis strains, respectively. The remaining animals in these groups and pigs from groups 1, 3, 5 received broth medium in the same way. Differences among virulence of S. suis capsular type 2 were observed in inoculated pigs of groups 2 and 4. Pigs from group 2 became carriers, showing only mild symptoms. By contrast, animals from group 4 presented an acute form of the disease. All the indirect contact pigs in groups 3 and 5 had S. suis in palatine tonsils from day 6 after the infection and they presented clinical manifestations similar to those observed in experimentally infected pigs. Two direct contact animals were also contaminated in the upper respiratory tract but surprisingly they did not show any symptoms. Airborne transmission of S. suis in experimentally pigs was demonstrated in the present study. Indirect infections, as described in this study, are a more realistic way to infect pigs than other experimental procedures and may be used to further study the pathogenesis of the infection caused by this important pathogen.     

Immunization of pigs against Streptococcus suis serotype 2 infection using a live avirulent strain.

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.     

Inoculation of pigs with Streptococcus suis type 2 alone or in combination with pseudorabies virus.

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)     

Detection and molecular typing of Streptococcus suis in tonsils from live pigs in France

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.     

Detection of virulent strains of Streptococcus suis type 2 and highly virulent strains of Streptococcus suis type 1 in tonsillar specimens of pigs by PCR.

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.     

Streptococcus suis type 2 infections in pigs in the Netherlands (Part two)

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.     

Streptococcus suis type 2 infections in pigs in the Netherlands (part two)

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.     

Passive immunization of pigs against experimental infection with Streptococcus suis serotype 2

The safety and protective efficacy of a horse antiserum raised against inactivated whole cell preparations of Streptococcus suis serotype 2 was investigated in pigs by experimental challenge. The antiserum was evaluated in two similar experiments each comprising 12 4-week-old pigs treated with 6 ml of antiserum the day before challenge and four pigs used as challenge controls. Pigs were infected by subcutaneous injection with approximately 10(11) colony forming units of S. suis serotype 2. Clinical disease in the pigs that could be attributed to infection with S. suis was reduced from 88 to 35% (P = 0.015). The percentage of pigs with lesions that could be associated with S. suis was reduced from 88 to 22% (P = 0.002) and isolation of S. suis serotype 2 was reduced from five (63%) out of eight pigs in the combined challenge control groups to 3 (13%) out of 23 pigs in the combined treatment groups. These results indicate that passive immunization of pigs may be a way to reduce or control S. suis serotype 2 infections in pigs.     

Rapid detection of Streptococcus suis serotype 2 in weaned pigs

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)     

四川猪2型链球菌病的PCR检测

2005年7月中下旬,四川贵阳等地人、猪陆续发生一种发病惠、死亡快、高度散发的疾病,从病死猪组织中分离出19株链球菌,设计合成3对引物,分别对分离菌株进行了猪链球菌16SrRNA、CPS2J、MRP基因片段扩增,有15株菌3个基因的扩增结果均为阳性,表明此次疲情的病原为猪2型链球菌。     

猪链球菌2型毒力相关蛋白

猪链球菌 (Streptococcus suis)是重要的猪病原菌 ,并能感染人 ,可致人和猪死亡。猪链球菌病多发于 3～ 12周龄断奶后的仔猪 ,几乎世界各国都有发生 ,对养猪业危害严重 ,同时事关公共卫生。目前已鉴定了 35种血清型的猪链球菌 ,其中最重要、最常见的是 2型 [1 ] 。我国于 1991年在广东省首次证实此菌的存在 ,1998年在江苏的人群和猪群中暴发由 2型所致的链球菌病 [2 ] ,其毒力因子的研究 ,尤其值得关注。1　毒力相关蛋白的发现　　基于传统观念 ,最初有人提出荚膜多糖决定猪链球菌 2型的侵袭力 [3 ,4] ,但后来的研究不能证实荚膜多糖是该菌…     

人感染猪链球菌2型的研究进展

猪链球菌2型是一种重要的人畜共患病原菌,可引起人的脑膜炎、败血症等疾病.2005年6月发生的四川疫情是由猪链球菌2型引起的人猪共同感染,流行广,发展快,人猪患病数量多.本文从病原、流行特点、感染途径、临床表现、实验室鉴定(包括培养特性、形态特征、生化特性、血清凝集、毒力基因)等方面做了详细的综述,初步探讨了2型猪链球菌病的防治方法.