Streptococcus suis type 2 (SS2), a major emerging/re-emerging zoonotic pathogen found in humans and pigs, can cause severe clinical infections, and pose public health issues. Our previous studies recognized peptidyl-prolyl isomerase (PrsA) as a critical virulence factor promoting SS2 pathogenicity. PrsA contributed to cell death and operated as a pro-inflammatory effector. However, the molecular pathways through which PrsA contributes to cell death are poorly understood. Here in this study, we prepared the recombinant PrsA protein and found that pyroptosis and necroptosis were involved in cell death stimulated by PrsA. Specific pyroptosis and necroptosis signalling inhibitors could significantly alleviate the fatal effect. Cleaved caspase-1 and IL-1β in pyroptosis with phosphorylated MLKL proteins in necroptosis pathways, respectively, were activated after PrsA stimulation. Truncated protein fragments of enzymatic PPIase domain (PPI), N-terminal (NP), and C-terminal (PC) domains fused with PPIase, were expressed and purified. PrsA flanking N- or C-terminal but not enzymatic PPIase domain was found to be critical for PrsA function in inducing cell death and inflammation. Additionally, PrsA protein could be anchored on the cell surface to interact with host cells. However, Toll-like receptor 2 (TLR2) was not implicated in cell death and recognition of PrsA. PAMPs of PrsA could not promote TLR2 activation, and no rescued phenotypes of death were shown in cells blocking of TLR2 receptor or signal-transducing adaptor of MyD88. Overall, these data, for the first time, advanced our perspective on PrsA function and elucidated that PrsA-induced cell death requires its flanking N- or C-terminal domain but is dispensable for recognizing TLR2. Further efforts are still needed to explore the precise molecular mechanisms of PrsA-inducing cell death and, therefore, contribution to SS2 pathogenicity.

Nontyphoidal Salmonella (NTS) causes global outbreaks of foodborne disease. The main source of Salmonella for humans is animal-borne foods; however, the monitoring of Salmonella in the food chain via genomic platforms was limited in China. This study evaluated the prevalence, resistome, and virulome diversity of Salmonella strains identified from pork, retail environment, aquatic products, and poultry eggs of retail markets in Yichun city, Jiangxi province. The overall incidence of Salmonella was 9.4 %, with a higher contamination rate observed in pork at 13.5 %, followed by the retail environment at 7.69 %. The genomic analysis of the isolates revealed a total of fifteen distinct serovars, with serovar Enteritidis being the most prevalent (64.3 %). The phenotypic resistance analysis conducted by the broth microdilution method, revealed that 81.12 % of the isolates exhibited multidrug resistance (MDR), with high resistance to trimethoprim/sulphonamides (100 %), followed by tetracycline (99.3 %) and streptomycin (99.3 %). Genotypic analysis of antimicrobial resistance identified 80 antimicrobial-resistant genes (ARGs), with mdf(A), aph(3')-Ib, tet(A), dfrA12, floR, bla _TEM-1B , qnrS3, and sul2, conferring resistance to different antimicrobial classes, being the predominant ARGs. Additionally, forty ESBL genes, particularly critical genes such as bla _CTX-M and bla _NDM-1, were also identified in Salmonella isolates. The IncR, IncFIB (K), and IncX1 plasmid replicons were widely prevalent and served as significant reservoirs of horizontally acquired foreign genes. Moreover, key virulence genes such as cdtB, lpf and sef were also detected, in addition to Salmonella pathogenicity islands SPI-1 and SPI-2. This study reveals the prevalence of multidrug-resistant and virulent strains of Salmonella serovars in the markets of Yichuan city, posing a risk of human infections. The gained knowledge provided essential baseline information that may be utilized for regular tracking of MDR Salmonella transmission in the food chain to minimize potential future outbreaks.

Streptococcus suis type 2 (SS2) is an important zoonotic pathogen for swine and humans. The increasing prevalence of antimicrobial resistance in S. suis isolates poses a threat to public health. This study investigated the antimicrobial activity and therapeutic potential of cinnamaldehyde (CA), a natural compound from cinnamon, against SS2. CA showed significant antimicrobial activity with a minimal inhibition concentration of 0.25 µg/mL and prolonged post-antibiotic effect of over 7 h in SS2. Increased bacterial cell membrane permeability and blocked protein synthesis of SS2 were observed after being treated with CA. CA could effectively prevent biofilm formation. CA treatment reduced the crucial virulence factor of suilysin expression and secretion in SS2 cells through a probable interaction with the suilysin protein. CA treatment could prominently alleviate both epithelial HEp-2 and phagocytic RAW264.7 cell cytotoxicity induced by SS2. The pathogenic SS2 strain was attenuated by CA, as demonstrated by the diminished adherence in HEp-2 cells, increased clearance by RAW264.7 and mice whole blood, and improved survival rate in a mouse infection model. CA possessed therapeutic efficacy since the CA treatment exhibited a 50% improvement in mouse survival rate, which surpassed the traditional ampicillin therapy control group. Alleviated clinical symptoms and histopathological phenotypes, with reduced bacterial burden in mouse organs after CA treatment, were examined. Overall, this study identified cinnamaldehyde as a novel antibacterial compound against SS2 with potential therapeutic protective effects, offering an alternative drug for controlling SS2 prevalence and infection. IMPORTANCE: Widespread infections caused by Streptococcus suis type 2 (SS2) have garnered significant attention in the realm of public health due to their zoonotic nature. In recent years, antimicrobial resistance phenotypes in SS2 have emerged and intensified within the context of animal husbandry. Herbal compounds and medicinal plants are increasingly recognized as promising therapeutic alternatives for mitigating or addressing the challenges posed by antimicrobial resistance. The aim of this present study was to explore a novel compound of cinnamaldehyde, which obtained significant antimicrobial activity and potential therapeutic protective effect against SS2 infection. The research has made an innovative discovery that the bactericidal effect of cinnamaldehyde is associated with its antivirulence strategies, such as targeting the key virulence factors of SS2 and countering the bacterial infection process.

Streptococcus suis serotype 2 (S. suis type 2, SS2) is one of the zoonotic pathogens known to induce meningitis, septicemia, and arthritis in both pigs and humans, resulting in public health concerns. CbpD, also termed CrfP, is one of the choline-binding proteins (CBPs) that was found as a murein hydrolase in SS2 and plays crucial roles in natural genetic transformation under the control of ComRS-ComX regulatory system by a previous study. Nonetheless, the possible functions of CbpD in virulence and pathogenesis in SS2 remain unclear. In this study, a cbpD gene mutant (ΔcbpD) with its complemental strain (cΔcbpD) was constructed and further used to examine the pathogenic roles of CbpD in SS2 infection. The results showed that the CbpD deficiency leads to increased bacterial chain elongation and aggregation with little impact on the growth capability of SS2. The ΔcbpD strain represented more vulnerable to a thermo, acid, or oxidative stress. Elevated adhesion to human epithelial HEp-2 cells, decreased invasion into bEND3.0 cells, and more easily phagocytosed by murine RAW264.7 macrophages of ΔcbpD were found. The virulence of cbpD mutant was attenuated in a mouse infection model. Enhanced susceptibility within mice blood and impaired ability to colonize organs with alleviated histopathological lesions were also demonstrated as compared with wild-type SS2. It is noteworthy that the discrepant expression of multiple virulence-associated factors including serine/threonine phosphorylase Stp, anti-phagocytosis factor of transglutaminase TGase and adhesin of chaperon DnaJ, were examined resulting from the deletion of cbpD. Overall, these findings provided evidence that the CbpD factor contributes to SS2 infection and is involved in bacterial adhesion, invasion, and anti-phagocytosis processes by modulating crucial virulence-associated factors expression.

Streptococcus suis type 2 (S. suis type 2, SS2), an infectious pathogen which is zoonotic and can induce severely public health concern. Our previous research identified a newly differential secreted effector of tagatose-bisphosphate aldolase (LacD) mediated by VirD4 factor within the putative type IV secretion system of SS2, whereas the functional basis and roles in virulence of LacD remain elusive. Here in this study, the LacD was found enzymatic and can be activated to express under oxidative stress. Gene mutant and its complemental strain (ΔlacD and cΔlacD) were constructed to analyze the phenotypes, virulence and transcriptomic profiles as compared with the parental strain. The lacD gene deletion showed no effect on growth capability and cells morphology of SS2. However, reduced tolerance to oxidative and heat stress conditions, increased antimicrobial susceptibility to ciprofloxacin and kanamycin were found in ΔlacD strain. Further, the LacD deficiency led to weakened invasion and attenuated virulence since an easier phagocytosed and more prone to be cleared of SS2 in macrophages were shown in ΔlacD mutant. Distinctive transcriptional profiling in ΔlacD strain and typical down-regulated genes with significant mRNA changes including alcohol dehydrogenase, GTPase, integrative and conjugative elements, and iron ABC transporters which were mainly involved in cell division, stress response, antimicrobial susceptibility and virulence regulation, were examined and confirmed by RNA sequencing and real time qPCR. In summary, the results demonstrated for the first time that LacD was a pluripotent protein mediated the metabolic, stress and virulent effect of SS2.