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Purpose Streptococcus suis serotype 14 is the second most prevalent serotype being highly prevalent in Southeast Asia. This study aimed to characterize genetic background, population structure, virulent genes, antimicrobial-resistant genes, and virulence of human S. suis serotype 14. Methods Genomes of 11 S. suis serotype 14 were sequenced by short- and long-read sequencing platforms. The genomes were analyzed for genetic relationship, virulence-associated genes, and antimicrobial-resistant genes. Antimicrobial susceptibility was conducted and the virulence was tested based on cell assay. Results All isolates belonged to clonal complex (CC) 1, with nine sequence type (ST) 105 isolates and each isolate of ST1 and ST237. They were susceptible to penicillin, whereas tetracycline and macrolide were resistance due to tetO and ermB . Genomic analysis revealed that the serotype 14-ST105 isolates were closely related to zoonotic serotype 14-ST105 isolates from Vietnam and the serotype 1-ST105 Thai strain. The serotype 14-ST1 isolate was closely related to pig-diseased serotype 1-ST1 isolates from UK and USA, whereas the serotype 14-ST237 isolate was related to serotype 1-ST237 strains recovered from healthy pig from Thailand. Of 150 virulence-associated genes, 13 were absent from the serotype 14 isolates, including atl1 , atlAss , hhly3 , nisK , nisR , pnuC , salK , salR , sp1 , srtG , virB4 , virD4 , and zmp . The virulence of strain 32481, a representative S. suis serotype 14-ST105 isolate showed reduced adhesion and invasion of two epithelial cell lines (A549 and HeLa) when compared to the serotype 2-ST1 strain P1/7, whereas apoptosis was similar. Conclusion This study highlighted the pathogenic potential of virulent serotype 14-ST105 strains and the need for increased monitoring of S. suis serotypes other than for serotype 2.

Background Streptococcus suis is divided into 29 serotypes based on a serological reaction against the capsular polysaccharide (CPS). Multiplex PCR tests targeting the cps locus are also used to determine S. suis serotypes, but they cannot differentiate between serotypes 1 and 14, and between serotypes 2 and 1/2. Here, we developed a pipeline permitting in silico serotype determination from whole-genome sequencing (WGS) short-read data that can readily identify all 29  S. suis serotypes. Results We sequenced the genomes of 121 strains representing all 29 known S. suis serotypes. We next combined available software into an automated pipeline permitting in silico serotyping of strains by differential alignment of short-read sequencing data to a custom S. suis cps loci database. Strains of serotype pairs 1 and 14, and 2 and 1/2 could be differentiated by a missense mutation in the cpsK gene . We report a 99 % match between coagglutination- and pipeline-determined serotypes for strains in our collection. We used 375 additional S. suis genomes downloaded from the NCBI’s Sequence Read Archive (SRA) to validate the pipeline. Validation with SRA WGS data resulted in a 92 % match. Included pipeline subroutines permitted us to assess strain virulence marker content and obtain multilocus sequence typing directly from WGS data. Conclusions Our pipeline permits rapid and accurate determination of S. suis serotype, and other lineage information, directly from WGS data. By discriminating between serotypes 1 and 14, and between serotypes 2 and 1/2, our approach solves a three-decade longstanding S. suis typing issue.

The Streptococcus suis (S. suis) is an important zoonotic pathogen that causes streptococcal disease in pigs and poses a threat to humans. This study provides an understanding of the prevalence of   S.suis in eastern China and provides guidance for clinical prophylaxis. From 2021 to 2023, a total of 143 strains of S. suis were isolated from 1642 lung tissue and nasal swabs from healthy and suspected infected pigs in Shandong Province, China, using the Phenotypic tests and PCR technique. The isolates were then tested for serotype, virulence-related genes, and resistance genes. Among the 143 isolates, type 2 was the predominant serotype with 98 isolates (98/143, 68.5%), followed by type 5 with 22 isolates (22/143, 15.3%), type 4 with 6 isolates (6/143, 4.2%), type 19 with 4 isolates (4/143, 2.8%) and type 21 with 5 isolates (5/143, 3.5%), respectively. A minimum of 78.3% of the strains exhibited the presence of virulence-related genes including pgda , dlta , mann , fbps , orf2 , and sspa , whereas the virulence-associated genes Sum , Sly , and Salkr are not widely prevalent. For the detection of resistance genes, it was found that the tetO gene had a high detection rate of 70.1% (101/143), whereas neither the pbp2b gene nor the cat1 and cat2 genes were detected. Antimicrobial susceptibility testing revealed that 96.5% (138/143) of the isolates exhibited multidrug resistance (MDR). And polypeptide B was found to be tolerated by 125 of the 143 strains (87.4%). Although we did not detect the β-lactam resistance gene in any of the 143 strains, an average of 39.2% of the strains were resistant to β-lactam antibiotics. The results of the current study is thought it may be help to understand the prevalence of S. suis and provide important insights into treatment and prevention.

Backgrounds Streptococcus suis is an emerging zoonotic bacterial disease with increasing prevalence in the human population and is one of the most important bacterial infections in pig husbandry. There is still a lack of a thorough understanding of S. suis metabolism and the connection between metabolism and virulence. Results A genome-scale metabolic model i NX525, which included 525 genes, 708 metabolites, and 818 reactions, was manually constructed with a 74% overall MEMOTE score. The flux balance analysis results of the model exhibited good agreement with growth phenotypes under different nutrient conditions and genetic disturbances. The model predictions aligned with 71.6%, 76.3%, and 79.6% of the gene essentiality predictions from three mutant screens. The model was then used to analyze virulence factors and related synthetic pathways. One hundred and thirty-one virulence-linked genes were found by comparing to virulence factor databases, and among them, seventy-nine virulence-linked genes were in 167 metabolic reactions in model i NX525. One hundred and one of the metabolic genes were predicted to affect the formation of nine virulence-linked small molecules. Complex interrelationships between growth- and virulence-associated pathways were evaluated, and 26 genes were found to be essential for both cell growth and virulence factor production. Among these, eight enzymes and metabolites were identified as antibacterial drug targets, focusing on the biosynthesis of capsular polysaccharides and peptidoglycans. Conclusion Overall, the metabolic model i NX525 provides a high-quality platform for systematic elucidation of the metabolism of S. suis .

Mobile genetic elements (MGEs) enable bacteria to acquire novel genes and traits. However, the functions of cargo genes within MGEs remain poorly understood. The cadmium resistance operon cadDX is present in many gram-positive bacteria. Although cadDX has been reported to be involved in metal detoxification, its regulatory mechanisms and functions in bacterial pathogenesis are poorly understood. This study revealed that cadDX contributes to cadmium resistance, oxidative stress resistance, and virulence in Streptococcus suis , an important zoonotic pathogen in pigs and humans. CadX represses cadD expression by binding to the cadDX promoter. Notably, cadX responds to H 2 O 2 stress through an additional promoter within the cadDX operon , mitigating the harmful effect of excessive cadD expression during oxidative stress. cadDX resides within an 11 K integrative and mobilizable element that can autonomously form circular structures. Moreover, cadDX is found in diverse MGEs, accounting for its widespread distribution across various bacteria, especially among pathogenic streptococci. Transferring cadDX into another zoonotic pathogen, Streptococcus agalactiae , results in similar phenotypes, including resistance to cadmium and oxidative stresses and increased virulence of S. agalactiae in mice. The new functions and regulatory mechanisms of cadDX shed light on the importance of the cadDX system in driving evolutionary adaptations and survival strategies across diverse gram-positive bacteria.

is a major bacterial pathogen in pigs and an emerging zoonotic pathogen. Different serotypes exhibit diverse characteristics in population structure and pathogenicity. Surveillance data highlight the significance of serotype 4 (SS4) in swine streptococcusis, a pathotype causing human infections. However, except for a few epidemiologic studies, the information on SS4 remains limited. In this study, we investigated the population structure, pathogenicity, and antimicrobial characteristics of SS4 based on 126 isolates, including one from a patient with septicemia. We discovered significant diversities within this population, clustering into six minimum core genome (MCG) groups (1, 2, 3, 4, 7-2, and 7-3) and five lineages. Two main clonal complexes (CCs), CC17 and CC94, belong to MCG groups 1 and 3, respectively. Numerous important putative virulence-associated genes are present in these two MCG groups, and 35.00% (7/20) of pig isolates from CC17, CC94, and CC839 (also belonging to MCG group 3) were highly virulent (mortality rate ≥ 80%) in zebrafish and mice, similar to the human isolate ID36054. Cytotoxicity assays showed that the human and pig isolates of SS4 strains exhibit significant cytotoxicity to human cells. Antimicrobial susceptibility testing showed that 95.83% of strains isolated from our labs were classified as multidrug-resistant. Prophages were identified as the primary vehicle for antibiotic resistance genes. Our study demonstrates the public health threat posed by SS4, expanding the understanding of SS4 population structure and pathogenicity characteristics and providing valuable information for its surveillance and prevention.

Streptococcus suis serotype 2 is a significant zoonotic pathogen that poses a serious threat to the health of humans and various animals. Ribosomal protein S1 (RPSA), acting as a key virulence factor, mediates SS2 adhesion to host cells and facilitates its penetration of the blood–brain barrier. In this study, we expressed the SS2 RPSA recombinant protein and used it to immunize BALB/c mice. Through screening, we obtained a high-affinity hybridoma clone and produced a specific anti-RPSA monoclonal antibody (designated mAb-RPSA-5E2). We found that mAb-RPSA-5E2 exhibited potent antibacterial activity, significantly inhibiting the proliferation of SS2 in vitro. In SS2-challenged mice, treatment with mAb-RPSA-5E2 significantly reduced mortality and alleviated pathological damage in the lung and brain tissues. Furthermore, evaluating the immunoprotective effect of the SS2-RPSA protein as a subunit vaccine revealed that this vaccine provided good protection against SS2 infection. These findings indicate that RPSA is a promising candidate target for developing SS2 subunit vaccines and targeted therapeutic biologics. Key points • The SS2 RPSA monoclonal antibody can inhibit the proliferation of SS2 in vitro. • The SS2 RPSA monoclonal antibody can reduce the mortality rates of mice. • The SS2-RPSA subunit vaccine provides significant immunoprotective effects in mice.

Streptococcus suis, a zoonotic bacterial pathogen circulated through swine, can cause severe infections in humans. Because human S. suis infections are not notifiable in most countries, incidence is underestimated. We aimed to increase insight into the molecular epidemiology of human S. suis infections in Europe. To procure data, we surveyed 7 reference laboratories and performed a systematic review of the scientific literature. We identified 236 cases of human S. suis infection from those sources and an additional 87 by scanning gray literature. We performed whole-genome sequencing to type 46 zoonotic S. suis isolates and combined them with 28 publicly available genomes in a core-genome phylogeny. Clonal complex (CC) 1 isolates accounted for 87% of typed human infections; CC20, CC25, CC87, and CC94 also caused infections. Emergence of diverse zoonotic clades and notable severity of illness in humans support classifying S. suis infection as a notifiable condition.

Streptococcus suis is an economically important pathogen of pigs as well as a zoonotic cause of human disease. Serotyping is used for further characterization of isolates; some serotypes seem to be more virulent and more widely spread than others. This study characterizes a collection of German field isolates of Streptococcus suis from pigs dating from 1996 to 2016 with respect to capsular genes (cps) specific for individual serotypes and pathotype by multiplex PCR and relates results to the clinical background of these isolates. The most prominent finding was the reduction in prevalence of serotype-2/serotype-1/2 among invasive isolates during this sampling period, which might be attributed to widely implemented autogenous vaccination programs in swine against serotype 2 in Germany. In diseased pigs (systemically ill; respiratory disease) isolates of serotype-1/serotype-14, serotype-2/serotype-1/2, serotype 3 to 5 and 7 to 9 were most frequent while in carrier isolates a greater variety of cps types was found. Serotype-1/serotype-14 seemed to be preferentially located in joints, serotype 4 and serotype 3 in the central nervous system, respectively. The virulence associated extracellular protein factor was almost exclusively associated with invasive serotype-1/serotype-14 and serotype-2/serotype-1/2 isolates. In contrast, lung isolates of serotype-2/serotype-1/2 mainly harbored the gene for muramidase-released protein. Serotype 4 and serotype 9 isolates from clinically diseased pigs most frequently carried the muramidase-released protein gene and the suilysin gene. When examined by transmission electron microscopy all but one of the isolates which were non-typable by molecular and serological methods showed various amounts of capsular material indicating potentially new serotypes among these isolates. Given the variety of cps types/serotypes detected in pigs, not only veterinarians but also medical doctors should consider other serotypes than just serotype 2 when investigating potential human cases of Streptococcus suis infection.

Background Streptococcus suis ( S. suis ) is an important swine and human pathogen. A recent study reported the first isolate of S. suis capable of infecting fish, designated as S. suis strain 3112. The bacterium was isolated from snakeskin gourami ( Trichopodus pectoralis ), an economically important fish species native to Southeast Asia, and it was previously shown that it can infect and cause lethal streptococcosis in the fish. Results In this study, we present the complete genome of S. suis 3112. Molecular sequence analysis revealed that it belongs to serotype 6, sequence type 2340. Phylogenetic analysis showed that the bacterium clustered with healthy-pig S. suis isolates, suggestive of an ultimate swine (as opposed to human) origin of the bacterium. Two fluoroquinolone resistance genes are present in the bacterial genome, namely patA and patB. Our results showed that both genes are expressed in our bacterium, and the bacterium is resistant to norfloxacin, but is still sensitive to other fluoroquinolones, including ciprofloxacin, enrofloxacin, and sparfloxacin. Additionally, the bacterium is sensitive to β-lactams, tetracyclines, sulphonamides, and an aminoglycoside. Conclusions This study reports and describes the complete genome of S. suis 3112, the first isolate of S. suis known to infect fish, and provides further insights into the bacterial isolate, particularly regarding its drug resistance profile. These results will facilitate further investigations of the comparative genomics and pathogenic characteristics of S. suis , as well as the development of control strategies against this newly-identified fish pathogen.