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Streptococcus suis is an emerging zoonotic pathogen that causes septicemia, meningitis, arthritis, and pneumonia in swine and humans. The present study aimed to characterize the genetic diversity of S. suis serotype 2 isolated from pigs showing signs of illness in Brazil using pulsed-field gel electrophoresis (PFGE), single-enzyme amplified fragment length polymorphism (SE-AFLP), and profiling of virulence-associated markers. A total of 110 isolates were studied, 62.7% of which were isolated from the central nervous system and 19.1% from the respiratory tract. Eight genotypes were obtained from the combination of virulence genes, with 43.6% and 5.5% frequencies for the mrp (+) /epf (+) /sly (+) and mrp (-) /epf (-) /sly (-) genotypes, respectively. The presence of isolates with epf gene variation with higher molecular weight also appears to be a characteristic of Brazilian S. suis serotype 2. The PFGE and SE-AFLP were able to type all isolates and, although they presented a slight tendency to cluster according to state and year of isolation, it was also evident the grouping of different herds in the same PFGE subtype and the existence of isolates originated from the same herd classified into distinct subtypes. No further correlation between the isolation sites and mrp/epf/sly genotypes was observed.

Streptococcus suis is an important swine pathogen and a zoonotic agent causing meningitis and septicemia. Although serotype 2 is the most virulent type, serotype 14 is emerging, and understanding of its pathogenesis is limited. To study the role of the capsular polysaccharide (CPS) of serotype 14 as a virulence factor, we constructed knockout mutants devoid of either cps14B, a highly conserved regulatory gene, or neu14C, a gene coding for uridine diphospho-N-acetylglucosamine 2-epimerase, which is involved in sialic acid synthesis. The mutants showed total loss of the CPS with coagglutination assays and electron microscopy. Phagocytosis assays showed high susceptibility of mutant Δcps14B. An in vivo murine model was used to demonstrate attenuated virulence of this non-encapsulated mutant. Despite the difference in the CPS composition of different serotypes, this study has demonstrated for the first time that the CPS of a serotype other than 2 is also an important antiphagocytic factor and a critical virulence factor.

The Streptococcus suis 103 gene product is an immunogenic and protective lipoprotein that is a component of an ATP-binding cassette transporter implicated in zinc uptake. Belonging to the same transcriptional unit and downstream of the 103 gene is a gene that encodes a homologue of the pneumococcal histidine triad (Pht) protein Pht309. In an intraperitoneal mouse model the virulence of a mutant lacking the 103 gene was more than 50 times lower than that of the wild-type (WT) parent strain, S. suis serotype 2 strain P1/7. In addition, the immunogenicity of this mutant was dramatically decreased. In striking contrast, the virulence and immunogenicity of a P1/7 mutant lacking the Pht309 gene were similar to those of the parent strain. These results demonstrate that the 103 lipoprotein is strongly involved in S. suis virulence and support the hypothesis that this lipoprotein might be an excellent candidate for vaccines aiming to achieve broad protection against streptococci.

The objective of this study was to determine the capsular serotypes and potential virulence factors of Streptococcus suis isolated from pigs with polyserositis. Among the 24 isolates evaluated, serotype 3 [7 (29%) of the isolates] and serotype 4 [5 (21%)] were the most common. The isolates were also studied for the presence of the genes mrp, epf, and sly, which encode muramidase-released protein (MRP), extracellular factor (EF), and suilysin (SLY), respectively. Of the 24 isolates, 8 carried mrp: 4 of serotype 3, 2 of serotype 2, and 2 of serotype 4. One mrp(+) isolate (serotype 2) also carried the epf gene. All 24 isolates carried the sly gene. The serotype and genotype distribution greatly differed from that reported for isolates from pigs with other clinical manifestations of S. suis infection in other countries.

Streptococcus suis serotype 2 is an important swine pathogen associated mainly with meningitis. In a previous study, we demonstrated the ability of S. suis serotype 2 to adhere to and invade immortalized porcine brain microvascular endothelial cells (PBMECs) forming the blood-brain barrier. The aim of the current work was to further characterize the mechanism(s) by which S. suis invades porcine endothelial cells. The ability of several S. suis strains to interact with PBMECs was not found to correlate with their geographic origin, virulence, host of origin, or suilysin production. Characterization studies demonstrated that proteinaceous adhesins/invasins, cell wall components, lipoteichoic acid, and serum components (including fibronectin) were involved in interactions between S. suis and PBMECs. In addition to PBMECs, S. suis was able to adhere to and invade 2 porcine aortic endothelial cell lines and primary PBMECs.

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.

Four different experimental models for Streptococcus suis-induced disease were compared to find a model that closely mimics naturally occurring disease in conventional pigs. Fourteen, 2-week old pigs free of S. suis type 2 were used in 2 experiments. In experiment 1, 3 pigs were inoculated intravenously (IV) and 3 pigs intranasally (IN) with S. suis. Two out of 3 of the IV-inoculated pigs exhibited signs of severe central nervous system disease (CNS) and were euthanized. Streptococcus suis type 2 was isolated from whole blood, joints, and serosal surfaces of both pigs. No clinical signs and no growth of S. suis were detected in the IN-inoculated pigs. In experiment 2, 4 pigs were inoculated IV and another 4 were inoculated IN with the same isolate as in experiment 1. One hour before inoculation the IN-inoculated pigs were given 5 mL of 1% acetic acid intranasally (IN-AA). All the IV-inoculated pigs showed CNS disease and lameness, and 2 of the pigs became severely affected and were euthanized. All the IN-AA inoculated pigs exhibited roughened hair coats and 2 pigs developed severe CNS disease and were euthanized. Streptococcus suis was isolated from the joints and blood of 3 pigs in the IV-inoculated group. Streptococcus suis was isolated from blood of 2 pigs, meninges of 3 pigs, and joints of 1 pig in the IN-AA inoculated group. Natural exposure to S. suis most likely occurs by the intranasal route. The IN-AA model should serve as a good model for S. suis-induced disease, because the natural route of exposure is intranasal and the IN-AA model was effective in inducing disease that mimics what is observed in the field.

Streptococcus suis is considered one of the most important bacterial swine pathogens worldwide. The distribution of the 35 described serotypes in diseased animals may vary in different regions. Data regarding S. suis isolation from pigs in South America is not available. In the present study, 51 isolates of S. suis recovered in pure culture or as the predominant species from diseased animals in Brazil, were analyzed. These isolates were classified as serotypes 2 (58.8%), 3 (21.5%), 7 (13.7%), 1 (3.9%), and 14 (2%). Serotype 2 isolates were further studied for their production of virulence-related proteins muramidase-released protein (MRP), extracellular factor (EF), and suilysin. In addition, the genetic diversity was studied by randomly amplified polymorphic DNA. All but 1 of the serotype 2 isolates showed a clonal distribution of an atypical phenotype (MRP+, EF*, suilysin+), different from the known European (MRP+, EF+, suilysin+), and North American (MRPv, EF-, suilysin-), phenotypes.

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.

Haemophilus parasuis and Streptococcus suis are both major causes of losses during the nursery period, especially in herds using the segregated early weaning system. In this system, only a few piglets may be colonized with the herd's prevalent systemic strain, which results in infection of naive penmates late in the nursery. In view of these factors, the objectives of this study were: (1) to evaluate the early colonization of piglets with the farm's prevalent systemic strain of H. parasuis and S. suis as an alternative method for disease prevention; and (2) to evaluate 2 different protocols for experimental colonization: direct colonization of piglets and colonization of piglets through nose-to-nose contact with inoculated sows. Haemophilus parasuis and S. suis isolates recovered from diseased nursery pigs were characterized by the rep-PCR technique and the herd's prevalent strains were used for colonization. Piglets in the experimentally colonized groups were inoculated at 5 days of age by the oral route using a spray pump. Sows were colonized at 2 weeks prior to farrowing using a similar protocol. Although both colonization protocols were successful in getting the piglets colonized, direct inoculation of 5-day-old piglets with the herd's systemic strains of H. parasuis and S. suis tended to be more effective in reducing the morbidity and the mortality than the colonization of piglets by nose-to-nose contact with inoculated sows.

Control of Streptococcus suis infections and associated disease have proven to be a difficult challenge under most farm conditions. The objective of this study was to experimentally expose young pigs with a pathogenic strain of S. suis serotype 2 as a means of controlling the disease in a commercial swine farm. Prior to the start of the study, the pathogenic S. suis strain responsible for mortality in the farm was identified and used to experimentally inoculate baby piglets. Over a 3-week period, groups of pigs were selected (100 pigs/wk) and divided into 2 groups: control (50 pigs/week) and experimentally exposed (50 pigs/week). Pigs in the experimentally exposed group were inoculated at 5 d old by tonsillar swabbing with the pathogenic S. suis farm isolate. The effect of exposure with this pathogenic strain was evaluated during the nursery and finishing stages and was based on: morbidity (pigs with central nervous signs (CNS) and/or lameness), mortality and number of treatments required by pigs that had either CNS or lameness. The relative risk (RR) of acquiring disease due to S. suis infection was also calculated. Results showed that morbidity in the experimentally exposed groups was lower than in the control group and these results were statistically different (P = 0.006). Experimentally exposed pigs also showed a statistically significant reduction in lameness problems (P = 0.012), but not in CNS (P = 0.20) or mortality (P = 0.59). Pigs in the control group had an increased RR of 4.76, 8.77 and 2.7 for morbidity, to have lameness or to have CNS signs, respectively. In conclusion, experimental exposure of young pigs with the farm's pathogenic S. suis strain at a young age, had a positive effect in reducing clinical signs characteristics of S. suis infection. This method constitutes a novel approach to the control of S. suis infections in swine farms.

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.

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.