RESUMO
Streptococcus suis (S. suis) is one of the important pathogens that cause bacterial meningitis in pigs and humans. Evading host immune defences and penetrating the blood-brain barrier (BBB) are the preconditions for S. suis to cause meningitis, while the underlying mechanisms during these pathogenic processes are not fully understood. By detecting the red blood and white blood cells counts, IL-8 expression, and the pathological injury of brain in a mouse infection model, a serine-rich repeat (SRR) glycoprotein, designated as SssP1, was identified as a critical facilitator in the process of causing meningitis in this study. SssP1 was exported to assemble a fimbria-like component, thus contributed to the bacterial adhesion to and invasion into human brain microvascular endothelial cells (HBMECs), and activates the host inflammatory response during meningitis but is not involved in the actin cytoskeleton rearrangement and the disruption of tight junctions. Furthermore, the deletion of sssP1 significantly attenuates the ability of S. suis to traverse the BBB in vivo and in vitro. A pull-down analysis identified vimentin as the potential receptors of SssP1 during meningitis and following Far-Western blot results confirmed this ligand-receptor binding mediated by the NR2 (the second nonrepeat region) region of SssP1. The co-localisation of vimentin and S. suis observed by laser scanning confocal microscopy with multiplex fluorescence indicated that vimentin significantly enhances the interaction between SssP1 and BBB. Further study identified that the NR216-781 and NR1711-2214 fragments of SssP1 play critical roles to bind to the BBB depending on the sialylation of vimentin, and this binding is significantly attenuated when the antiserum of NR216-781 or NR1711-2214 blocked the bacterial cells, or the vimentin antibody blocked the BBB. Similar binding attenuations are observed when the bacterial cells were preincubated with the vimentin, or the BBB was preincubated with the recombinant protein NR216-781, NR1711-2214 or sialidase. In conclusion, these results reveal a novel receptor-ligand interaction that enhances adhesion to and penetration of the BBB to cause bacterial meningitis in the S. suis infection and highlight the importance of vimentin in host-pathogen interactions.
Assuntos
Meningites Bacterianas , Infecções Estreptocócicas , Streptococcus suis , Animais , Modelos Animais de Doenças , Células Endoteliais/patologia , Humanos , Ligantes , Meningites Bacterianas/microbiologia , Camundongos , Infecções Estreptocócicas/microbiologia , Suínos , VimentinaRESUMO
Staphylococcus aureus RNAIII is one of the main intracellular effectors of the quorum-sensing system. It is a multifunctional RNA that encodes a small peptide, and its noncoding parts act as antisense RNAs to regulate the translation and/or the stability of mRNAs encoding transcriptional regulators, major virulence factors, and cell wall metabolism enzymes. In this review, we explain how regulatory proteins and RNAIII are embedded in complex regulatory circuits to express virulence factors in a dynamic and timely manner in response to stress and environmental and metabolic changes.
Assuntos
Percepção de Quorum , RNA Bacteriano/metabolismo , Regulon , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/enzimologia , Staphylococcus aureus/patogenicidade , Fatores de Virulência/genética , Animais , Proteínas de Bactérias , Regulação Bacteriana da Expressão Gênica , Humanos , RNA Bacteriano/genética , Staphylococcus aureus/genética , Staphylococcus aureus/fisiologia , Virulência , Fatores de Virulência/metabolismoRESUMO
The human opportunistic pathogen Staphylococcus aureus produces numerous small regulatory RNAs (sRNAs) for which functions are still poorly understood. Here, we focused on an atypical and large sRNA called RsaC. Its length varies between different isolates due to the presence of repeated sequences at the 5' end while its 3' part is structurally independent and highly conserved. Using MS2-affinity purification coupled with RNA sequencing (MAPS) and quantitative differential proteomics, sodA mRNA was identified as a primary target of RsaC sRNA. SodA is a Mn-dependent superoxide dismutase involved in oxidative stress response. Remarkably, rsaC gene is co-transcribed with the major manganese ABC transporter MntABC and, consequently, RsaC is mainly produced in response to Mn starvation. This 3'UTR-derived sRNA is released from mntABC-RsaC precursor after cleavage by RNase III. The mature and stable form of RsaC inhibits the synthesis of the Mn-containing enzyme SodA synthesis and favors the oxidative stress response mediated by SodM, an alternative SOD enzyme using either Mn or Fe as co-factor. In addition, other putative targets of RsaC are involved in oxidative stress (ROS and NOS) and metal homeostasis (Fe and Zn). Consequently, RsaC may balance two interconnected defensive responses, i.e. oxidative stress and metal-dependent nutritional immunity.
Assuntos
Proteínas de Bactérias/genética , Estresse Oxidativo/genética , Infecções Estafilocócicas/genética , Staphylococcus aureus/genética , Proteínas de Bactérias/química , Regulação Bacteriana da Expressão Gênica/genética , Homeostase/genética , Humanos , Manganês/química , Oxirredução , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/patogenicidade , Inanição , Superóxido Dismutase/química , Superóxido Dismutase/genéticaRESUMO
Streptococcus agalactiae (group B streptococcus [GBS]) has received continuous attention for its involvement in invasive infections and its broad host range. Transcriptional regulators have an important impact on bacterial adaptation to various environments. Research on transcriptional regulators will shed new light on GBS pathogenesis. In this study, we identified a novel XRE-family transcriptional regulator encoded on the GBS genome, designated XtgS. Our data demonstrate that XtgS inactivation significantly increases bacterial survival in host blood and animal challenge test, suggesting that it is a negative regulator of GBS pathogenicity. Further transcriptomic analysis and quantitative reverse transcription-PCR (qRT-PCR) mainly indicated that XtgS significantly repressed transcription of its upstream gene pseP Based on electrophoretic mobility shift and lacZ fusion assays, we found that an XtgS homodimer directly binds a palindromic sequence in the pseP promoter region. Meanwhile, the PseP and XtgS combination naturally coexists in diverse Streptococcus genomes and has a strong association with sequence type, serotype diversification and host adaptation of GBS. Therefore, this study reveals that XtgS functions as a transcriptional regulator that negatively affects GBS virulence and directly represses PseP expression, and it provides new insights into the relationships between transcriptional regulator and genome evolution.
Assuntos
Proteínas de Bactérias/metabolismo , Infecções Estreptocócicas/microbiologia , Streptococcus agalactiae/patogenicidade , Fatores de Transcrição/metabolismo , Animais , Proteínas de Bactérias/genética , Regulação Bacteriana da Expressão Gênica , Humanos , Filogenia , Regiões Promotoras Genéticas , Ligação Proteica , Infecções Estreptocócicas/metabolismo , Streptococcus/classificação , Streptococcus/genética , Streptococcus agalactiae/genética , Fatores de Transcrição/genética , Transcrição Gênica , Virulência/genética , Peixe-ZebraRESUMO
Streptococcus suis is a major pathogen of pigs and also an important zoonotic agent for humans. A S. suis protein containing Mac-1 domain (designated Mac) is a protective antigen, exclusively cleaves porcine IgM, and contributes to complement evasion with the presence of high titers of specific porcine anti-S. suis IgM, but its role in S. suis virulence has not been investigated in natural healthy host without specific IgM. In this study, a mac deletion mutant was constructed by homologous recombination in S. suis serotype 2 virulent reference strain P1/7. Deletion of mac did not significantly influence phagocytosis or intracellular survival within murine macrophages RAW264.7, or the oxidative-burst induction of RAW264.7 and murine neutrophils. Furthermore, the mutant is as virulent as the wild-type strain in pig, mouse, and zebrafish infection models. Our data suggest that Mac is not essential for S. suis virulence in strain P1/7 in natural healthy host without specific IgM, and the immunogenicity of Mac does not appear to correlate with its significance for virulence.
Assuntos
Proteínas de Bactérias/metabolismo , Infecções Estreptocócicas/veterinária , Streptococcus suis/patogenicidade , Doenças dos Suínos/microbiologia , Fatores de Virulência/metabolismo , Animais , Proteínas de Bactérias/genética , Feminino , Imunoglobulina M/imunologia , Macrófagos/imunologia , Macrófagos/microbiologia , Camundongos , Camundongos Endogâmicos BALB C , Fagocitose , Células RAW 264.7 , Infecções Estreptocócicas/imunologia , Infecções Estreptocócicas/microbiologia , Streptococcus suis/genética , Streptococcus suis/metabolismo , Suínos , Doenças dos Suínos/imunologia , Virulência , Fatores de Virulência/genética , Peixe-ZebraRESUMO
Objective: In addition to Streptococcus suis serotype 2, Streptococcus suis serotype 9 (SS9) is also a currently prevalent serotype and a zoonotic pathogen. In our previous study, SS9 DNA nuclease (SsnA) was considered as a candidate virulence factor. To clarify the impact of SsnA on SS9 virulence, we constructed ssnA mutant (ΔssnA) and studied its biological functions. Methods: We evaluated the virulence of wild type strain and ΔssnA in a zebrafish infection model and compared the adherence rate to HEp-2 cells, the survival rate in pig blood, and enzymatic activity between wild type stain and ΔssnA. Results: In a zebrafish infection experiment, the 50% lethal dose value of ΔssnA was 11.2-fold higher than that of wild type strain. The adherence rate of ΔssnA to HEp-2 cells was only 60.61% of the wild strain level. The survival rate of ΔssnA in pig blood was declined to 71.88% of wild strain level. The enzymatic activity assay showed that SsnA can degrade both linear and circular DNA. Conclusion: SsnA contributes to SS9 virulence in a zebrafish infection model, the adherence to HEp-2 cells, and the survival in pig blood. SsnA is indeed an essential virulence factor for SS9.
Assuntos
Proteínas de Bactérias/metabolismo , Desoxirribonucleases/metabolismo , Infecções Estreptocócicas/veterinária , Streptococcus suis/enzimologia , Streptococcus suis/patogenicidade , Doenças dos Suínos/microbiologia , Animais , Aderência Bacteriana , Proteínas de Bactérias/genética , Desoxirribonucleases/genética , Infecções Estreptocócicas/microbiologia , Streptococcus suis/genética , Streptococcus suis/fisiologia , Suínos , Virulência , Peixe-ZebraRESUMO
Streptococcus suis (SS) is an important pathogen of pigs, and it is also recognized as a zoonotic agent for humans. SS infection may result in septicemia or meningitis in the host. However, little is known about genes that contribute to the virulence process and survival within host blood or cerebrospinal fluid (CSF). Small RNAs (sRNA) have emerged as key regulators of virulence in several bacteria, but they have not been investigated in SS. Here, using a differential RNA-sequencing approach and RNAs from SS strain P1/7 grown in rich medium, pig blood, or CSF, we present the SS genome-wide map of 793 transcriptional start sites and 370 operons. In addition to identifying 29 sRNAs, we show that five sRNA deletion mutants attenuate SS virulence in a zebrafish infection model. Homology searches revealed that 10 sRNAs were predicted to be present in other pathogenic Streptococcus species. Compared with wild-type strain P1/7, sRNAs rss03, rss05, and rss06 deletion mutants were significantly more sensitive to killing by pig blood. It is possible that rss06 contributes to SS virulence by indirectly activating expression of SSU0308, a virulence gene encoding a zinc-binding lipoprotein. In blood, genes involved in the synthesis of capsular polysaccharide (CPS) and subversion of host defenses were up-regulated. In contrast, in CSF, genes for CPS synthesis were down-regulated. Our study is the first analysis of SS sRNAs involved in virulence and has both improved our understanding of SS pathogenesis and increased the number of sRNAs known to play definitive roles in bacterial virulence.
Assuntos
Sangue/microbiologia , Líquido Cefalorraquidiano/microbiologia , RNA Bacteriano/genética , Infecções Estreptocócicas/genética , Streptococcus suis/genética , Transcrição Gênica/genética , Virulência/genética , Animais , Estudo de Associação Genômica Ampla/métodos , Óperon/genética , Deleção de Sequência/genética , Infecções Estreptocócicas/microbiologia , Suínos , Regulação para Cima/genética , Peixe-Zebra/genética , Peixe-Zebra/microbiologiaRESUMO
Protein phosphorylation is a major mechanism of signal transduction in bacteria. Here, we analyzed the proteome and phosphoproteome of a wild-type strain of the food-borne pathogen Listeria monocytogenes that was grown in either chemically defined medium or rich medium containing glucose. We then compared these results with those obtained from an isogenic prfA* mutant that produced a constitutively active form of PrfA, the main transcriptional activator of virulence genes. In the prfA* mutant grown in rich medium, we identified 256 peptides that were phosphorylated on serine (S), threonine (T), or tyrosine (Y) residues, with a S/T/Y ratio of 155:75:12. Strikingly, we detected five novel phosphosites on the virulence protein ActA. This protein was known to be phosphorylated by a cellular kinase in the infected host, but phosphorylation by a listerial kinase had not previously been reported. Unexpectedly, SILAC experiments with the prfA* mutant grown in chemically defined medium revealed that, in addition to previously described PrfA-regulated proteins, several other proteins were significantly overproduced, among them were several proteins involved in purine biosynthesis. This work provides new information for our understanding of the correlation among protein phosphorylation, virulence mechanisms, and carbon metabolism.
Assuntos
Proteínas de Bactérias/metabolismo , Listeria monocytogenes/metabolismo , Fatores de Terminação de Peptídeos/metabolismo , Fosfoproteínas/metabolismo , Proteoma/metabolismo , Proteômica/métodos , Proteínas de Bactérias/análise , Proteínas de Bactérias/genética , Sítios de Ligação/genética , Cromatografia Líquida , Meios de Cultura/química , Meios de Cultura/farmacologia , Glucose/farmacologia , Listeria monocytogenes/genética , Listeria monocytogenes/patogenicidade , Mutação , Fatores de Terminação de Peptídeos/análise , Fatores de Terminação de Peptídeos/genética , Peptídeos/análise , Peptídeos/genética , Peptídeos/metabolismo , Fosfoproteínas/análise , Fosfoproteínas/genética , Fosforilação/efeitos dos fármacos , Proteoma/análise , Proteoma/genética , Purinas/biossíntese , Serina/genética , Serina/metabolismo , Espectrometria de Massas em Tandem , Treonina/genética , Treonina/metabolismo , Tirosina/genética , Tirosina/metabolismo , Virulência/genéticaRESUMO
Streptococcus suis serotype 2 (SS2) is an emerging zoonotic agent responsible for a number of infections in pigs and humans. Pili have been proposed as virulence factors in Gram-positive bacteria. However, due to the abolition of pili production, the function of the srtBCD pilus cluster, especially the truncated major pilin subunit Sbp2 (Sbp2', Sbp2â³), has not been explored. In this study, isogenic mutants (Δsbp2', Δsbp2â³) were constructed by homologous replacement in SS2 strain P1/7. Deletion of sbp2' attenuated the virulence in a zebrafish model as shown by more than an eightfold increase in the LD50 of Δsbp2', compared with that of the parent strain. In addition, the adhesion of Δsbp2' to HEp-2 cell monolayers decreased significantly. Compared with the parent strain, no obvious differences in virulence and adherence efficiency were observed for Δsbp2â³. Our data suggest that Sbp2' could be involved in SS2 pathogenesis despite absence of its pilus shaft.
Assuntos
Proteínas de Fímbrias/metabolismo , Streptococcus suis/patogenicidade , Fatores de Virulência/metabolismo , Animais , Adesão Celular , Linhagem Celular , Modelos Animais de Doenças , Proteínas de Fímbrias/genética , Deleção de Genes , Hepatócitos/microbiologia , Humanos , Dose Letal Mediana , Família Multigênica , Subunidades Proteicas/metabolismo , Infecções Estreptocócicas/microbiologia , Infecções Estreptocócicas/patologia , Streptococcus suis/genética , Análise de Sobrevida , Peixe-ZebraRESUMO
Cell-penetrating peptides (CPPs) are promising carriers to effectively transport antisense oligonucleotides (ASOs), including peptide nucleic acids (PNAs), into bacterial cells to combat multidrug-resistant bacterial infections, demonstrating significant therapeutic potential. Streptococcus suis, a Gram-positive bacterium, is a major bacterial pathogen in pigs and an emerging zoonotic pathogen. In this study, through the combination of super-resolution structured illumination microscopy (SR-SIM), flow cytometry analysis, and toxicity analysis assays, we investigated the suitability of four CPPs for delivering PNAs into S. suis cells: HIV-1 TAT efficiently penetrated S. suis cells with low toxicity against S. suis; (RXR)4XB had high penetration efficiency with inherent toxicity against S. suis; (KFF)3K showed lower penetration efficiency than HIV-1 TAT and (RXR)4XB; K8 failed to penetrate S. suis cells. HIV-1 TAT-conjugated PNA specific for the essential gyrase A subunit gene (TAT-anti-gyrA PNA) effectively inhibited the growth of S. suis. TAT-anti-gyrA PNA exhibited a significant bactericidal effect on serotypes 2, 4, 5, 7, and 9 strains of S. suis, which are known to cause human infections. Our study demonstrates the potential of CPP-ASO conjugates as new antimicrobial compounds for combating S. suis infections. Furthermore, our findings demonstrate that applying SR-SIM and flow cytometry analysis provides a convenient, intuitive, and cost-effective approach to identifying suitable CPPs for delivering cargo molecules into bacterial cells.
RESUMO
Streptococcus suis is an important zoonotic pathogen that can cause meningitis and septicemia in swine and humans. Among numerous pathogenic serotypes, S. suis serotype 8 has distinctive characteristics such as a high detection rate and causing multi-host infection. There is no complete genome of serotype 8 strains so far. In this study, the complete genome of two S. suis serotype 8 strains, virulent strain 2018WUSS151 and non-virulent strain WUSS030, were sequenced. Comparative genomic analysis showed that the homology of the two genomes reaches 99.68%, and the main difference is the distinctive prophages. There are 83 genes unique to virulent strain 2018WUSS151, including three putative virulence-associated genes (PVGs). Two PVGs, padR and marR, are passenger genes in ISSsu2 family transposons that are able to form circular DNA intermediates during transposition, indicating the possibility of horizontal transmission among S. suis strains. The deletion mutant of PVGs marR or atpase attenuated the virulence of serotype 2 virulent SC070731 in a mouse infection model, confirming their role in S. suis virulence. These findings contribute to clarifying the genomic characterization of S. suis serotype 8 and S. suis pathogenesis.
RESUMO
Zoonotic streptococci cause several invasive diseases with high mortality rates, especially meningitis. Numerous studies elucidated the meningitis pathogenesis of zoonotic streptococci, some specific to certain bacterial species. In contrast, others are shared among different bacterial species, involving colonization and invasion of mucosal barriers, survival in the bloodstream, breaching the blood-brain and/or blood-cerebrospinal fluid barrier to access the central nervous system, and triggering inflammation of the meninges. This review focuses on the recent advancements in comprehending the molecular and cellular events of five major zoonotic streptococci responsible for causing meningitis in humans or animals, including Streptococcus agalactiae, Streptococcus equi subspecies zooepidemicus, Streptococcus suis, Streptococcus dysgalactiae, and Streptococcus iniae. The underlying mechanism was summarized into four themes, including 1) bacterial survival in blood, 2) brain microvascular endothelial cell adhesion and invasion, 3) penetration of the blood-brain barrier, and 4) activation of the immune system and inflammatory reaction within the brain. This review may contribute to developing therapeutics to prevent or mitigate injury of streptococcal meningitis and improve risk stratification.
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Bacterial DeoR family transcription regulators regulate multiple physiological processes. Little is known about the function of DeoR family regulators in streptococci. Here, we identified a novel DeoR family regulator, GlpR, from Streptococcus suis, a pathogen causing severe diseases in pigs and humans. GlpR was involved in glycerol utilization and exhibited specific signature residues at positions 30-31 (KV) which are crucial for DNA binding. Deletion of glpR (ΔglpR) showed a significant increase in relative growth rate in glycerol medium compared to the wild-type (WT) and complementary strains (CΔglpR). Employing RNA-seq analysis, ß-galactosidase activity analysis, and electrophoretic mobility shift assay, we discovered that GlpR directly represses the expression of glycerol metabolism-related genes pflB2, pflA1, and fsaA, encoding pyruvate formate-lyase and its activating enzyme, and fructose-6-phosphate aldolase, respectively. Compared to WT and CΔglpR, ΔglpR showed a reduced survival rate under oxidative stress and in murine macrophages and attenuated virulence in mice. GlpR probably enhances oxidative stress resistance and virulence in S. suis by functioning as a glycerol metabolic repressor decreasing energy consumption. These findings contribute to a better understanding of S. suis pathogenesis and enrich our knowledge of the biological functions of DeoR family regulators in streptococci.
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Streptococcus suis is a major bacterial pathogen in pigs and an emerging zoonotic pathogen. Different S. suis serotypes exhibit diverse characteristics in population structure and pathogenicity. Surveillance data highlight the significance of S. suis 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.
Assuntos
Sorogrupo , Infecções Estreptocócicas , Streptococcus suis , Doenças dos Suínos , Streptococcus suis/patogenicidade , Streptococcus suis/genética , Streptococcus suis/classificação , Streptococcus suis/efeitos dos fármacos , Streptococcus suis/isolamento & purificação , Animais , Suínos , Humanos , Infecções Estreptocócicas/microbiologia , Infecções Estreptocócicas/veterinária , Doenças dos Suínos/microbiologia , Virulência , Camundongos , Genoma Bacteriano , Peixe-Zebra , Antibacterianos/farmacologia , Filogenia , Testes de Sensibilidade Microbiana , Fatores de Virulência/genéticaRESUMO
Streptococcus suis is a significant and emerging zoonotic pathogen. ST1 and ST7 strains are the primary agents responsible for S. suis human infections in China, including the Guangxi Zhuang Autonomous Region (GX). To enhance our understanding of S. suis ST1 population characteristics, we conducted an investigation into the phylogenetic structure, genomic features, and virulence levels of 73 S. suis 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 S. suis strains.
Assuntos
Genoma Bacteriano , Ilhas Genômicas , Filogenia , Infecções Estreptocócicas , Streptococcus suis , Streptococcus suis/genética , Streptococcus suis/patogenicidade , Streptococcus suis/classificação , Streptococcus suis/isolamento & purificação , Infecções Estreptocócicas/microbiologia , Infecções Estreptocócicas/veterinária , Infecções Estreptocócicas/epidemiologia , China/epidemiologia , Humanos , Virulência , Animais , Camundongos , Feminino , Genômica , Fatores de Virulência/genéticaRESUMO
Streptococcus pasteurianus is a zoonotic pathogen causing meningitis and bacteremia in animals and humans. A lack of accurate and convenient detection methods hinders preventing and controlling diseases caused by S. pasteurianus. Additionally, there is limited knowledge about its pathogenicity and antimicrobial resistance characteristics, as there are only three complete genome sequences available. In this study, we established a multiplex PCR assay for the detection of S. pasteurianus, which was applied to six fecal samples from cattle with diarrhea and 285 samples from healthy pigs. Out of the samples tested, 24 were positive, including 5 from pig tonsils, 18 from pig hilar lymph nodes, and 1 from cattle feces. Two strains were isolated from positive samples, and their complete genomes were sequenced. The two strains were non-virulent in mice and multidrug-resistant by the antimicrobial susceptibility test. We first found the presence of genes tet(O/W/32/O) and lsa(E) in S. pasteurianus, leading to resistance to lincosamides and tetracyclines. The convenient and specific multiplex PCR assay provides essential technical support for epidemiological research, and the complete genome sequence of two non-virulent strains contributes to understanding this zoonotic bacterium's genomic characteristics and pathogenesis.
RESUMO
Streptococcus suis is a bacterium that can cause infections in pigs and humans. Although oxidative stress is common occurrence during bacterial growth and infection, the regulation networks of S. suis under oxidative stress remain poorly understood. To address this, we utilized RNA-Seq to reveal the transcriptional landscape of S. suis in response to H2O2 stress. We identified novel genes responsible for S. suis resistance to oxidative stress, including those involved in DNA repair or protection, and essential for the biosynthesis of amino acids and nucleic acids. In addition, we found that a novel aquaporin, Aagp, belonging to atypical aquaglyceroporins and widely distributed in diverse S. suis serotypes, plays a crucial role during H2O2 stress. By performing oxidative stress assays and measuring the intracellular H2O2 concentrations of the wild-type strain and Aagp mutants during H2O2 stress, we found that Aagp facilitated H2O2 efflux. Additionally, we found that Aagp might be involved in glycerol transport, as shown by the growth inhibition and H2O2 production in the presence of glycerol. Mice infection experiments indicated that Aagp contributed to S. suis virulence. This study contributes to understanding the mechanism of S. suis oxidative stress response, S. suis pathogenesis, and the function of aquaporins in prokaryotes.
Assuntos
Aquaporinas , Streptococcus suis , Humanos , Animais , Camundongos , Suínos , Peróxido de Hidrogênio/farmacologia , Streptococcus suis/genética , Glicerol , Virulência , Aquaporinas/genéticaRESUMO
Streptococcus suis, an emerging zoonotic pathogen, is important reservoirs of antibiotic resistance genes that play critical roles in the horizontal transfer of corresponding resistances. In the present study, 656 antibiotic resistance (AR) genes were detected in 154 of 155 genomes of S. suis strains isolated from the nasopharynx of slaughtered pigs and the lungs of diseased pigs in China. The AR genes were clustered into 11 categories, consisting of tetracycline, macrolides, lincosamide, streptogramin, aminoglycoside, trimethoprim, amphenicols, nucleoside, quinupristin/dalfopristin, glycopeptide, and oxazolidinones resistance genes. In order to investigate the transmission patterns of the AR genes, AR genes-associated the mobile genetic elements (MGEs) were extracted and investigated. Twenty ICEs, one defective ICE, one tandem ICE, and ten prophages were found, which mainly carried tetracycline, macrolides/lincosamides/streptogramin (MLS), and aminoglycosides resistance genes. Three types of DNA cargo with AR genes were integrated into specific sites of ICEs: integrative mobilizable elements (IMEs), cis-IMEs (CIMEs), and transposon Tn916. Obvious differences in AR gene categories were found among the three cargo types. IMEs mainly harbored tetracycline and MLS resistance genes. CIMEs mainly carried aminoglycoside resistance genes, while transposon Tn916 carried only the tet (M) gene. Nearly all AR genes in ICEs were carried by IMEs and CIMEs. IMEs were prevalent and were also detected in additional 29 S. suis genomes. The horizontal transfer of IMEs and CIMEs may play critical role in ICE evolution and AR gene transmission in the S. suis population. Our findings provide novel insights into the transmission patterns of AR genes and the evolutionary mechanisms of ICEs in S. suis.
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Streptococcus pasteurianus, an underreported opportunistic pathogen, is considered an increasingly recognized cause of meningitis and bacteremia in many animals and humans worldwide. However, except for some epidemiological studies, there is no report about the gene-deletion mutagenesis, virulence factors, reservoir niches or animal infection models for this pathogen. In this study, we first isolated an S. pasteurianus strain from a newly weaned piglet's brain with meningitis. The genomic sequence of this swine isolate WUSP067 shared high homology with that of two human strains. The comparative genome analysis showed that strain WUSP067 contained a fucose utilization cluster absent in human strains, and it shared 91% identity with that of an integrative and conjugative element (ICE) ICEssuZJ20091101-2 from Streptococcus suis, another important swine bacterial pathogen. Strain WUSP067 was resistant to erythromycin, tulathromycin, lincomycin, clindamycin, doxycycline and gentamycin, and ICEs are vehicles for harbouring antimicrobial resistance genes. The infection model was established using the 3-week-old newly weaned ICR mice. The 50% lethal dose value of strain WUSP067 was 4.0 × 107 colony-forming units per mouse. The infected mice showed severe signs of meningitis and pathological changes in brains. Furthermore, the capsule-deficient mutant was generated using natural transformation, and we showed that capsule was an essential virulence factor for S. pasteurianus. In addition, we found that tonsils and hilar lymph nodes of healthy pigs may be reservoir niches for this bacterium. Thus, our study provided valuable information about the pathogenetic characteristics and antimicrobial resistance of S. pasteurianus and paved the way for studying its pathogenesis.
Assuntos
Meningite , Doenças dos Roedores , Infecções Estreptocócicas , Streptococcus suis , Doenças dos Suínos , Animais , Clindamicina , Doxiciclina , Eritromicina , Fucose , Gentamicinas , Humanos , Meningite/veterinária , Camundongos , Camundongos Endogâmicos ICR , Infecções Estreptocócicas/microbiologia , Infecções Estreptocócicas/veterinária , Streptococcus , Streptococcus suis/genética , Suínos , Doenças dos Suínos/microbiologia , Fatores de Virulência/genéticaRESUMO
Streptococcus suis, the leading causative agent of swine streptococcosis, is considered as a severe zoonotic and foodborne pathogen for humans. Characteristics of population structure and pathogenicity of S. suis vary significantly by serotypes. As one of the main pathogenic serotypes causing clinical disease in pigs, very little is known about the pathogenicity, population structure, and antimicrobial resistance of S. suis serotype 8 (SS8). In this study, the genome of 26 SS8 strains isolated from healthy and diseased pigs was sequenced. Together with 38 sequences from NCBI, we found that SS8 population was clustered into 12 sequence types (ST) and 4 minimum core genome (MCG) groups, linked to the geographical distribution. Noteworthily, 10 strains belonged to MCG group 1 which was defined to possess the capacity to cause global outbreaks in our previous study. We found that 75% (9/12) of representative SS8 strains were virulent in mice and zebrafish, including all ST1241 strains. No virulence indicators were identified from 67 putative virulence-associated genes mainly identified among pathogenic serotype 2 strains. Instead, we found that the genotype of some of these genes was correlated to their evolution. All 26 isolates were classified as multidrug-resistant strains by antimicrobial susceptibility testing. The high carrying rate of tetO and ermB, mainly disseminated by integrative mobilizable elements, contributed to the prevalent resistance phenotypes to macrolides, lincosamides and tetracyclines. These findings indicated that the pathogenic potential of SS8 cannot be ignored and provided valuable information for SS8 surveillance.