OIE Reference Laboratory for Swine Streptococcosis

Nanjing, China

OIE Reference Laboratory for Swine Streptococcosis

Nanjing, China
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Xiao G.,Nanjing Agricultural University | Xiao G.,Key Laboratory of Animal Bacteriology | Xiao G.,OIE Reference Laboratory for Swine Streptococcosis | Wu Z.,Nanjing Agricultural University | And 14 more authors.
Current Microbiology | Year: 2017

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. © 2016, Springer Science+Business Media New York.


Lai L.,Nanjing Agricultural University | Lai L.,Key Laboratory of Animal Bacteriology | Lai L.,OIE Reference Laboratory for Swine Streptococcosis | Dai J.,Nanjing Agricultural University | And 23 more authors.
Veterinary Microbiology | Year: 2017

Streptococcus suis (SS), an important pathogen for pigs, is not only considered as a zoonotic agent for humans, but is also recognized as a major reservoir of antimicrobial resistance contributing to the spread of resistance genes to other pathogenic Streptococcus species. In addition to serotype 2 (SS2), serotype 9 (SS9) is another prevalent serotype isolated from diseased pigs. Although many SS strains have been sequenced, the complete genome of a non-SS2 virulent strain has been unavailable to date. Here, we report the complete genome of GZ0565, a virulent strain of SS9, isolated from a pig with meningitis. Comparative genomic analysis revealed five new putative virulence or antimicrobial resistance-associated genes in strain GZ0565 but not in SS2 virulent strains. These five genes encode a putative triacylglycerol lipase, a TipAS antibiotic-recognition domain protein, a putative TetR family transcriptional repressor, a protein containing a LPXTG domain and a G5 domain, and a type VII secretion system (T7SS) putative substrate (EsxA), respectively. Western blot analysis showed that strain GZ0565 can secrete EsxA. We generated an esxA deletion mutant and showed that EsxA contributes to SS virulence in a mouse infection model. Additionally, the antibiotic resistance gene vanZSS was identified and expression of vanZSS conferred resistance to teicoplanin and dalbavancin in Streptococcus agalactiae. We believe this is the first experimental demonstration of the existence of the T7SS putative substrate EsxA and its contribution to bacterial virulence in SS. Together, our results contribute to further understanding of the virulence and antimicrobial resistance characteristics of SS. © 2017 Elsevier B.V.


Shao J.,Key Laboratory of Animal Bacteriology | Shao J.,OIE Reference Laboratory for Swine Streptococcosis | Zhang W.,Key Laboratory of Animal Bacteriology | Zhang W.,OIE Reference Laboratory for Swine Streptococcosis | And 4 more authors.
Current Microbiology | Year: 2014

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′, Sbp2″), has not been explored. In this study, isogenic mutants (Δsbp2′, Δsbp2″) 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 Δsbp2″. Our data suggest that Sbp2′ could be involved in SS2 pathogenesis despite absence of its pilus shaft. © 2014 Springer Science+Business Media New York.


Wu Z.,Nanjing Agricultural University | Wu Z.,Key Laboratory of Animal Bacteriology | Wu Z.,OIE Reference Laboratory for Swine Streptococcosis | Wang W.,Nanjing Agricultural University | And 25 more authors.
Gene | Year: 2014

Streptococcus suis (SS) is an important swine pathogen worldwide that occasionally causes serious infections in humans. SS infection may result in meningitis in pigs and humans. The pathogenic mechanisms of SS are poorly understood. Here, we provide the complete genome sequence of S. suis serotype 2 (SS2) strain SC070731 isolated from a pig with meningitis. The chromosome is 2,138,568. bp in length. There are 1933 predicted protein coding sequences and 96.7% (57/59) of the known virulence-associated genes are present in the genome. Strain SC070731 showed similar virulence with SS2 virulent strains HA9801 and ZY05719, but was more virulent than SS2 virulent strain P1/7 in the zebrafish infection model. Comparative genomic analysis revealed a unique 105. K genomic island in strain SC070731 that is absent in seven other sequenced SS2 strains. Further analysis of the 105. K genomic island indicated that it contained a complete nisin locus similar to the nisin U locus in S. uberis strain 42, a prophage similar to S. oralis phage PH10 and several antibiotic resistance genes. Several proteins in the 105. K genomic island, including nisin and RelBE toxin-antitoxin system, contribute to the bacterial fitness and virulence in other pathogenic bacteria. Further investigation of newly identified gene products, including four putative new virulence-associated surface proteins, will improve our understanding of SS pathogenesis. © 2013 Elsevier B.V.


Pan Z.,Nanjing Agricultural University | Pan Z.,OIE Reference Laboratory for Swine Streptococcosis | Ma J.,Nanjing Agricultural University | Ma J.,OIE Reference Laboratory for Swine Streptococcosis | And 7 more authors.
Applied and Environmental Microbiology | Year: 2015

Streptococcus suis is an emerging zoonotic pathogen causing severe infections in pigs and humans. In previous studies, 33 serotypes of S. suis have been identified using serum agglutination. Here, we describe a novel S. suis strain, CZ130302, isolated from an outbreak of acute piglet meningitis in eastern China. Strong pathogenicity of meningitis caused by strain CZ130302 was reproduced in the BALB/c mouse model. The strain showed a high fatality rate (8/10), higher than those for known virulent serotype 2 strains P1/7 (1/10) and 9801 (2/10). Cell adhesion assay results with bEnd.3 and HEp2 cells showed that CZ130302 was significantly close to P1/7 and 9801. Both the agglutination test and its complementary test showed that strain CZ130302 had no strong cross-reaction with the other 33 S. suis serotypes. The multiplex PCR assays revealed no specified bands for all four sets used to detect the other 33 serotypes. In addition, genetic analysis of the whole cps gene clusters of all serotypes was performed in this study. The results of comparative genomics showed that the cps gene cluster of CZ130302, which was not previously reported, showed no homology to the gene sequences of the other strains. Especially, the wzy, wzx, and acetyltransferase genes of strain CZ130302 are phylogenetically distinct from strains of the other 33 serotypes. Therefore, this study suggested that strain CZ130302 represents a novel variant serotype of S. suis (designated serotype Chz) which has a high potential to be virulent and associated with meningitis in animals. © 2015, American Society for Microbiology.


Wu Z.,Nanjing Agricultural University | Wu Z.,Key Laboratory of Animal Bacteriology | Wu Z.,OIE Reference Laboratory for Swine Streptococcosis | Wu C.,BGI Shenzhen | And 24 more authors.
RNA | Year: 2014

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. © 2014 Wu et al.


PubMed | OIE Reference Laboratory for Swine Streptococcosis, Nanjing Agricultural University and China Animal Health and Epidemiology Center
Type: Journal Article | Journal: Applied and environmental microbiology | Year: 2015

Streptococcus suis is an emerging zoonotic pathogen causing severe infections in pigs and humans. In previous studies, 33 serotypes of S. suis have been identified using serum agglutination. Here, we describe a novel S. suis strain, CZ130302, isolated from an outbreak of acute piglet meningitis in eastern China. Strong pathogenicity of meningitis caused by strain CZ130302 was reproduced in the BALB/c mouse model. The strain showed a high fatality rate (8/10), higher than those for known virulent serotype 2 strains P1/7 (1/10) and 9801 (2/10). Cell adhesion assay results with bEnd.3 and HEp2 cells showed that CZ130302 was significantly close to P1/7 and 9801. Both the agglutination test and its complementary test showed that strain CZ130302 had no strong cross-reaction with the other 33 S. suis serotypes. The multiplex PCR assays revealed no specified bands for all four sets used to detect the other 33 serotypes. In addition, genetic analysis of the whole cps gene clusters of all serotypes was performed in this study. The results of comparative genomics showed that the cps gene cluster of CZ130302, which was not previously reported, showed no homology to the gene sequences of the other strains. Especially, the wzy, wzx, and acetyltransferase genes of strain CZ130302 are phylogenetically distinct from strains of the other 33 serotypes. Therefore, this study suggested that strain CZ130302 represents a novel variant serotype of S. suis (designated serotype Chz) which has a high potential to be virulent and associated with meningitis in animals.


He S.,Nanjing Agricultural University | He S.,Key Laboratory of Animal Bacteriology | He S.,OIE Reference Laboratory for Swine Streptococcosis | Shi J.,Chinese Academy of Agricultural Sciences | And 6 more authors.
Microbes and Infection | Year: 2015

In early 2013, a Bengal tiger (Panthera tigris) in a zoo died of respiratory distress. All specimens from the tiger were positive for HPAI H5N1, which were detected by real-time PCR, including nose swab, throat swab, tracheal swab, heart, liver, spleen, lung, kidney, aquae pericardii and cerebrospinal fluid. One stain of virus, A/Tiger/JS/1/2013, was isolated from the lung sample. Pathogenicity experiments showed that the isolate was able to replicate and cause death in mice. Phylogenetic analysis indicated that HA and NA of A/Tiger/JS/1/2013 clustered with A/duck/Vietnam/OIE-2202/2012 (H5N1), which belongs to clade 2.3.2.1. Interestingly, the gene segment PB2 shared 98% homology with A/wild duck/Korea/CSM-28/20/2010 (H4N6), which suggested that A/Tiger/JS/1/2013 is a novel reassortant H5N1 subtype virus. Immunohistochemical analysis also confirmed that the tiger was infected by this new reassortant HPAI H5N1 virus. Overall, our results showed that this Bengal tiger was infected by a novel reassortant H5N1, suggesting that the H5N1 virus can successfully cross species barriers from avian to mammal through reassortment. © 2014 Institut Pasteur.


Tang F.,Nanjing Agricultural University | Tang F.,OIE Reference Laboratory for Swine Streptococcosis | Li D.,Nanjing Agricultural University | Li D.,OIE Reference Laboratory for Swine Streptococcosis | And 8 more authors.
Applied and Environmental Microbiology | Year: 2015

Streptococcus suis and Streptococcus equi subsp. zooepidemicus are capable of infecting humans and various animals, causing significant problems for the worldwide swine industry. As antibiotic resistance has increased, lysosomal enzymes encoded by phages have shown potential for use against pathogenic bacteria. In this study, a novel bacteriophage lysin, Ply30, encoded by the S. suis prophage phi30c, was recombinantly expressed and purified. Ply30 showed high bacteriolysis activity on S. suis and S. equi subsp. zooepidemicus in vitro. The ratio of the optical density at 600 nm (OD600) with treatment versus the OD600 with no treatment for most tested S. suis and S. equi subsp. zooepidemicus strains decreased from 1 to <0.3 and <0.5, respectively, within 1 h. The results of plate viability assays showed that treated bacteria suffered a 1- to 2-log decrease in CFU within 1 h. The optimal concentration of Ply30 was 50 μg/ml, and the optimal pH was 7. Moreover, Ply30 maintained high activity over a wide pH range (pH 6 to 10). The MICs of Ply30 against Streptococcus strains ranged from 16 to 512 μg/ml. In vivo, a 2-mg dose of Ply30 protected 90% (9/10 mice) of mice from infection with S. equi subsp. zooepidemicus and 80% (8/10 mice) of mice from infection with S. suis. Seven days after lysin Ply30 treatment, bacterial loads were significantly decreased in all tested organs and blood compared with those at 1 h postinfection without Ply30 treatment. Ply30 showed in vitro and in vivo antimicrobial efficiency and protected mice against two kinds of bacterial infections, indicating that Ply30 may be an effective therapeutic against streptococci. © 2015, American Society for Microbiology.

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