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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.

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.

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.

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.

Quorum sensing (QS) is an important protective mechanism that allows bacteria to adapt to its environment. A limited number of signal molecules play the key role of transmitting information in this mechanism. Signals are transmitted between individual bacterium through QS systems, resulting in the expression of specific genes. QS plays an important role in a variety of bacterial processes, including drug resistance, biofilm formation, motility, adherence, and virulence. Most Gram-positive and Gram-negative bacteria possess QS systems, mainly the LuxS/AI-2-mediated QS system. Evidence has been brought that LuxS/AI-2 system controls major virulence determinants in Streptococcus suis and, as such, the ability of this bacterial species to cause infections in humans and pigs. Understanding the S. suis LuxS/AI-2 system may open up novel avenues for decreasing the drug resistance and infectivity of S. suis. This article focuses on the progress made to date on the S. suis LuxS/AI-2-mediated QS system.

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.

poses a significant threat to both pig farming and public health, causing severe disease such as septicemia and meningitis. The type VII secretion system (T7SS) delivers toxic effectors to play a crucial role in interbacterial competition and is vital for the zoonotic pathogen to colonize host tonsils effectively. Here, we identified a type of hypothetical T7SS effector in , which appears to be fragmented toxins lacking the N-terminal YeeF domain, redesignated as MSE-ExTs. MSE (marker for searching effectors) is a conserved sequence at the N-termini of modular effectors showing a diverse range of toxicities targeting NAD +. Cognate WXG100-like and full-length EssC proteins contribute to activate the T7SS secretion. While most MSE-ExTs (MSE-fusing exported toxins) are encoded downstream of a truncated essC and lack cognate WXG100-like genes, they are nonetheless exported and function in interbacterial antagonism, thereby conferring a competitive advantage against bacterial isolates derived from tonsil microbiota. Deletion of the truncated essC could not diminish the MSE-ExT1 delivery, while the full-length EssC1 encoded in T7SS core locus was required for the lethality of MSE-ExT1 to sensitive bacterial cells. MapC2, an upstream small helical protein, shares a nearly identical C-terminal 50-amino acid (aa) sequence with EIC-CR (C-terminal conserved region of effector-paired immunity protein). This conserved fragment harbors a \"YxxxD\" targeting signal and interacts with the D1 ATPase domain of the non-neighboring EssC, thereby activating the secretion of MSE-ExTs. This alternative strategy facilitates effectors' delivery, even for fragmented substrates, highlighting its importance in ensuring the functionality of T7SS.

Background Streptococcus suis poses a significant threat to both humans and pigs, causing severe disease such as meningitis, septicaemia, and sudden death. The type VII secretion system (T7SS) plays a crucial role in exporting effectors that mediate bacterial competition and colonization in polymicrobial environments. Although Leu-x-Gly (LXG) polymorphic toxins are among the main effectors secreted by the T7SS, the mechanism underlying their secretion remains to be fully elucidated. Results Here, we identified a complex repertoire of LXG effectors in S. suis , comprising at least five distinct LXG-domain groups and 14 C-terminal toxin types. We focused on one LXG effector, LXG1-CT3, from the virulent strain WUSS351, demonstrating its pore-forming activity and essential role in S. suis competition and virulence. Additionally, we discovered that a small protein family, DUF4176, partners with each cognate C-terminal toxin, stabilizing the corresponding LXG effector prior to secretion, thereby enhancing bacterial competitiveness. Conclusions These findings deepen our understanding of S. suis competition and pathogenesis, while offering new insights into the T7SS-mediated secretion of LXG effectors.

Background: Streptococcus suis is a zoonotic pathogen that infects pigs and can occasionally cause serious infections in humans. S. suis infections occur sporadically in human Europe and North America, but a recent major outbreak has been described in China with high levels of mortality. The mechanisms of S. suis pathogenesis in humans and pigs are poorly understood. Methodology/Principal Findings: The sequencing of whole genomes of S. suis isolates provides opportunities to investigate the genetic basis of infection. Here we describe whole genome sequences of three S. suis strains from the same lineage: one from European pigs, and two from human cases from China and Vietnam. Comparative genomic analysis was used to investigate the variability of these strains. S. suis is phylogenetically distinct from other Streptococcus species for which genome sequences are currently available. Accordingly, approximately 40% of the approximately 2 Mb genome is unique in comparison to other Streptococcus species. Finer genomic comparisons within the species showed a high level of sequence conservation; virtually all of the genome is common to the S. suis strains. The only exceptions are three approximately 90 kb regions, present in the two isolates from humans, composed of integrative conjugative elements and transposons. Carried in these regions are coding sequences associated with drug resistance. In addition, small-scale sequence variation has generated pseudogenes in putative virulence and colonization factors. Conclusions/Significance: The genomic inventories of genetically related S. suis strains, isolated from distinct hosts and diseases, exhibit high levels of conservation. However, the genomes provide evidence that horizontal gene transfer has contributed to the evolution of drug resistance.

is a significant and emerging zoonotic pathogen. ST1 and ST7 strains are the primary agents responsible for human infections in China, including the Guangxi Zhuang Autonomous Region (GX). To enhance our understanding of ST1 population characteristics, we conducted an investigation into the phylogenetic structure, genomic features, and virulence levels of 73 ST1 human strains from GX between 2005 and 2020. The ST1 GX strains were categorized into three lineages in phylogenetic analysis. Sub-lineage 3-1a exhibited a closer phylogenetic relationship with the ST7 epidemic strain SC84. The strains from lineage 3 predominantly harboured 89K-like pathogenicity islands (PAIs) which were categorized into four clades based on sequence alignment. The acquirement of 89K-like PAIs increased the antibiotic resistance and pathogenicity of corresponding transconjugants. We observed significant diversity in virulence levels among the 37 representative ST1 GX strains, that were classified as follows: epidemic (E)/highly virulent (HV) (32.4%, 12/37), virulent plus (V+) (29.7%, 11/37), virulent (V) (18.9%, 7/37), and lowly virulent (LV) (18.9%, 7/37) strains based on survival curves and mortality rates at different time points in C57BL/6 mice following infection. The E/HV strains were characterized by the overproduction of tumour necrosis factor (TNF)-α in serum and promptly established infection at the early phase of infection. Our research offers novel insights into the population structure, evolution, genomic features, and pathogenicity of ST1 strains. Our data also indicates the importance of establishing a scheme for characterizing and subtyping the virulence levels of strains.