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58. Additional file 4 of CrfP, a fratricide protein, contributes to natural transformation in Streptococcus suis

Author

Yinchu Zhu, Jiale Ma, Zhang, Yue, Xiaojun Zhong, Qiankun Bai, Wenyang Dong, Zihao Pan, Guangjin Liu, Zhang, Cun, and Yao, Huochun

Abstract

Additional file 4. Structure and Western blot analysis of His tagged CrfP protein. (A) Domain organization of the full-length CrfP. SP, signal peptide; CHAP, the catalytic module, SH3b, bacterial SH3b module. Schematic representation of CrfP overall structure. The CHAP and SH3b are colored in blue and yellow, respectively. (B) The protein was purified from pET-28a. The protein in SDS-PAGE gel that stained with Coomassie blue R250. Lane 1 was purified CrfP with HisTrap column and line 2 was crude extract from cells without purified. Line 3, M, mean protein ladder marker. (C) The protein in figure A was analyzed with Western blot. The bands indicate the 36 kD CrfP protein.

Published
2021
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59. Additional file 5 of CrfP, a fratricide protein, contributes to natural transformation in Streptococcus suis

Author

Yinchu Zhu, Jiale Ma, Zhang, Yue, Xiaojun Zhong, Qiankun Bai, Wenyang Dong, Zihao Pan, Guangjin Liu, Zhang, Cun, and Yao, Huochun

Abstract

Additional file 5. Phylogenetic analysis of CrfP homologous proteins from Streptococcus species. (A) Evolutionary relationship between murein hydrolases from different species. Three branches were formed based on amino acid differences, and the diverse serotype SS belonged to the same group and was far from the other species. A neighbour-joining tree (bootstrap n = 1000; Poisson correction) was constructed based on a ClustalW alignment of the amino acid sequences using MEGA software version 5.0. (B) CrfP protein from different S. suis strains showed high amino acid sequence similarity. The green background indicates similarity >75%, and the blue background indicates similarity >100%.

Published
2021
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60. Additional file 6 of CrfP, a fratricide protein, contributes to natural transformation in Streptococcus suis

Author

Yinchu Zhu, Jiale Ma, Zhang, Yue, Xiaojun Zhong, Qiankun Bai, Wenyang Dong, Zihao Pan, Guangjin Liu, Zhang, Cun, and Yao, Huochun

Abstract

Additional file 6. Biological characteristics analysis between WT and ΔcrfP. (A) PCR results for deletion of crfP gene. (B) Microscopy observation of wild type and mutant. The SS was stain with crystal. (C) Growth curve analysis between wild type and mutant. (D) The biofilms were quantified using a multifunctional microplate reader at OD595. We investigated the potential roles of CrfP in S. suis biological ability. Initially, the growth properties in THB culture were compared between the wild type and mutant, and the growth curve was the same in each phase. Gram staining for microscopy showed that ZY05719 and ΔcrfP had similar chain lengths, suggesting that there was no correlation between CrfP activity and modulation of S. suis chain length. In addition, bacterial biofilms are composed of extracellular DNA and glycoproteins, which contribute to host immune defence or antimicrobial resistance. These components of biofilms are products of lytic processes. To evaluate whether CrfP is involved in S. suis biofilm formation, a crystal violet staining assay was used. However, the OD595 data showed no difference between ΔcrfP and the parental strain, which suggested that CrfP did not mediate biofilm formation in S. suis.

Published
2021
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63. Pathogenic investigations of Streptococcus pasteurianus, an underreported zoonotic pathogen, isolated from a diseased piglet with meningitis.

Author

Wang, Shuoyue, Ma, Miaohang, Liang, Zijing, Zhu, Xinchi, Yao, Huochun, Wang, Liping, and Wu, Zongfu

Subjects
PIGLETS, STREPTOCOCCUS, STREPTOCOCCUS suis, MENINGITIS, LABORATORY mice, DRUG resistance in microorganisms, STREPTOCOCCUS thermophilus, ACTINOBACILLUS
Abstract

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. [ABSTRACT FROM AUTHOR]

Published
2022
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67. Identification of the RNA-binding domain-containing protein RbpA that acts as a global regulator of the pathogenicity of Streptococcus suisserotype 2

Author

Zhong, Xiaojun, Ma, Jiale, Bai, Qiankun, Zhu, Yinchu, Zhang, Yue, Gu, Qibing, Pan, Zihao, Liu, Guangjin, Wu, Zongfu, and Yao, Huochun

Abstract

ABSTRACTStreptococcus suisserotype 2 (SS2), an emerging zoonotic pathogen, causes swine diseases and human cases of streptococcal toxic shock syndrome. RNA-binding proteins (RBPs) can modulate gene expression through post-transcriptional regulation. In this study, we identified an RBP harbouring an S1 domain, named RbpA, which facilitated SS2 adhesion to host epithelial cells and contributed to bacterial pathogenicity. Comparative proteomic analysis identified 145 proteins that were expressed differentially between ΔrbpAstrain and wild-type strain, including several virulence-associated factors, such as the extracellular protein factor (EF), SrtF pilus, IgA1 protease, SBP2 pilus, and peptidoglycan-binding LysM’ proteins. The mechanisms underlying the regulatory effects of RbpA on their encoding genes were explored, and it was found that RbpA regulates gene expression through diverse mechanisms, including post-transcriptional regulation, and thus acts as a global regulator. These results partly reveal the pathogenic mechanism mediated by RbpA, improving our understanding of the regulatory systems of S. suisand providing new insights into bacterial pathogenicity.

Published
2022
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68. The antimicrobial systems of Streptococcus suispromote niche competition in pig tonsils

Author

Liang, Zijing, Wu, Huizhen, Bian, Chen, Chen, Hao, Shen, Yanling, Gao, Xueping, Ma, Jiale, Yao, Huochun, Wang, Liping, and Wu, Zongfu

Abstract

ABSTRACTStreptococcus suiscan cause severe infections in pigs and humans. The tonsils of pigs are major niches for S. suis, and different serotypes of S. suiscan be found in the same tonsil. Pig tonsil colonization by S. suisis believed to be an important source of infection for humans and pigs. However, how S. suiscompetes for a stable tonsil niche is unknown. Here, we found that S. suisstrain WUSS351, isolated from a healthy pig tonsil, is virulent and multidrug-resistant. The ABC transporter system SstFEG, conferring resistance to bacitracin, was reported to confer a competitive survival advantage in vivo. In addition, strain WUSS351 has several antimicrobial systems, including a novel type VII secretion system (T7SS), lantibiotic bacteriocin, and lactococcin972-like bacteriocin Lcn351. Bacterial competition experiments demonstrated T7SS-mediated cell contact-dependent antagonism of S. suis. Antibacterial activity analysis and 16S rRNA gene sequencing of the culture-independent and culture-dependent pig tonsillar microbiome revealed that Lcn351 mainly targets S. suis, one of the core microbiomes in pig tonsils. Taken together, our results revealed the mechanism of the stable persistence of S. suisin the tonsil niche, which might have important implications for S. suisepidemiology, potentially influencing strain prevalence and disease progression.

Published
2022
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75. The Novel Streptococcal Transcriptional Regulator XtgS Negatively Regulates Bacterial Virulence and Directly Represses PseP Transcription

Author

Liu, Guangjin, primary, Gao, Tingting, additional, Zhong, Xiaojun, additional, Ma, Jiale, additional, Zhang, Yumin, additional, Zhang, Shiyu, additional, Wu, Zongfu, additional, Pan, Zihao, additional, Zhu, Yinchu, additional, Yao, Huochun, additional, Liu, Yongjie, additional, and Lu, Chengping, additional

Published
2020
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78. Screening virulence factors of porcine extraintestinal pathogenic Escherichia coli (an emerging pathotype) required for optimal growth in swine blood.

Author

Ma, Jiale, Cheng, Zhixin, Bai, Qiankun, Zhao, Kejie, Pan, Zihao, and Yao, Huochun

Subjects
SWINE growth, ESCHERICHIA coli, DRUG target, CARBON metabolism, ANAEROBIC metabolism
Abstract

Porcine extraintestinal pathogenic Escherichia coli (ExPEC) is occurring with increasing frequency in China, which causes acute septicemia and sudden death in pigs leading to significant economic losses. Bacterial survival and even proliferation within host bloodstream are a common manifestation of a number of bacterial septicemias, including porcine ExPEC diseases. However, the underlying pathogenesis for this novel pathotype of ExPEC has not been explored deeply. Here, we used a conjunction with transposon mutagenesis to identify the mechanisms of bacterial fitness involved in optimal growth of porcine ExPEC in swine serum ex vivo under static culture. Our work identified 28 genes involved in nucleotide biosynthesis, extracellular polysaccharide biosynthesis, regulators Fur and FNR, acid/zinc resistance, and Deley‐Douderoff carbon metabolism that are required for the serum fitness. Subsequent functional analyses revealed that either interruption of de novo nucleotide biosynthesis or blocking of several extracellular polysaccharide biosynthesis including O2‐antigen, Lipid A‐core, and ECA significantly affect porcine ExPEC's growth in swine serum and proliferation in host bloodstream. Furthermore, the reasonable regulations of iron and anaerobic metabolisms in response to host stimuli by global regulators Fur and FNR also play key roles during systemic infection of porcine ExPEC. These findings provide compelling evidences that de novo nucleotide biosynthesis may enable porcine ExPEC to adapt to swine blood‐specific nutrient availability, and the effective assembly of O‐antigen, lipid A‐core, and ECA is required to resist the bactericidal activity of swine serum. These studies contribute to better understand the underlying mechanisms employed by porcine ExPEC to survive, grow in the swine bloodstream, and cause disease. These related factors may serve as therapeutic targets for countering or preventing ExPEC serum resistance in the clinic. [ABSTRACT FROM AUTHOR]

Published
2021
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83. Identification of six novel capsular polysaccharide loci ( NCL ) from Streptococcus suis multidrug resistant non‐typeable strains and the pathogenic characteristic of strains carrying new NCL s

Author

Huang, Jinhu, primary, Liu, Xi, additional, Chen, Hao, additional, Chen, Li, additional, Gao, Xueping, additional, Pan, Zihao, additional, Wang, Jian, additional, Lu, Chengping, additional, Yao, Huochun, additional, Wang, Liping, additional, and Wu, Zongfu, additional

Published
2019
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86. Immunoproteomic analysis of bacterial proteins of Actinobacillus pleuropneumoniae serotype 1

Author

Wu Zongfu, Wang Yang, Zhai Zhipeng, Lu Yan, Tang Fang, Liu Guangjin, Shao Jing, Zhang Wei, Yao Huochun, and Lu Chengping

Subjects
Cytology, QH573-671
Abstract

Abstract Background Actinobacillus pleuropneumoniae (APP) is one of the most important swine pathogens worldwide. Identification and characterization of novel antigenic APP vaccine candidates are underway. In the present study, we use an immunoproteomic approach to identify APP protein antigens that may elicit an immune response in serotype 1 naturally infected swine and serotype 1 virulent strain S259-immunized rabbits. Results Proteins from total cell lysates of serotype 1 APP were separated by two-dimensional electrophoresis (2DE). Western blot analysis revealed 21 immunoreactive protein spots separated in the pH 4-7 range and 4 spots in the pH 7-11 range with the convalescent sera from swine; we found 5 immunoreactive protein spots that separated in the pH 4-7 range and 2 in the pH 7-11 range with hyperimmune sera from S259-immunized rabbits. The proteins included the known antigens ApxIIA, protective surface antigen D15, outer membrane proteins P5, subunit NqrA. The remaining antigens are being reported as immunoreactive proteins in APP for the first time, to our knowledge. Conclusions We identified a total of 42 immunoreactive proteins of the APP serotype 1 virulent strain S259 which represented 32 different proteins, including some novel immunoreactive factors which could be researched as vaccine candidates.

Published
2011
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100. Streptococcus suis serotype 9 strain GZ0565 contains a type VII secretion system putative substrate EsxA that contributes to bacterial virulence and a vanZ- like gene that confers resistance to teicoplanin and dalbavancin in Streptococcus agalactiae

Author

Lai, Liying, primary, Dai, Jiao, additional, Tang, Huanyu, additional, Zhang, Shouming, additional, Wu, Chunyan, additional, Qiu, Wancen, additional, Lu, Chengping, additional, Yao, Huochun, additional, Fan, Hongjie, additional, and Wu, Zongfu, additional

Published
2017
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