Comparative genomic and methylome analysis of non-virulent D74 and virulent Nagasaki Haemophilus parasuis isolates

Nicholson, Tracy L.; Brunelle, Brian W.; Bayles, Darrell O.; Alt, David P.; Shore, Sarah M.

doi:10.1371/journal.pone.0205700

Cited by 8 publications

(6 citation statements)

References 78 publications

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“…Previously identified virulence factors were evaluated for differential expression between growth conditions [9, 17–19] (S3 Table). Ten genes with a suspected role in virulence met the threshold of differential expression between broth and agar grown G .…”

Section: Resultsmentioning

confidence: 99%

Transcriptomic differences noted in Glaesserella parasuis between growth in broth and on agar

et al. 2019

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Glaesserella parasuis is the cause of Glӓsser’s disease in pigs and is a significant contributor to post-weaning mortality in the swine industry. Prevention of G . parasuis disease relies primarily on bacterin vaccines, which have shown good homologous protection and variable heterologous protection. Bacterin production involves large scale growth of the bacteria and proteins produced during the proliferation phase of production become important antigens that stimulate the immune response. In order to evaluate genes activated during G . parasuis growth on different media substrates, the transcriptome of broth and agar grown G . parasuis strain 29755 were sequenced and compared. The transcription of most purported virulence genes were comparable between broth and agar grown G . parasuis ; however, four virulence-associated genes, including ompA and vapD , had elevated expression under agar growth, while six virulence-associate genes had elevated expression during broth growth, including several protease genes. Additionally, there were metabolic shifts toward increased protein and lipid production and increased cellular division in broth grown G . parasuis . The results contribute to the understanding of how growth substrate alters gene transcription and protein expression, which may impact vaccine efficacy if immunogens important to the protective immune response are not produced under specific in vitro conditions. While the results of this work are unable to fully elucidate which growth medium presents a transcriptome more representative of in vivo samples or best suited for bacterin production, it forms a foundation that can be used for future comparisons and provides a better understanding of the metabolic differences in broth and agar grown bacteria.

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Section: Resultsmentioning

confidence: 99%

Transcriptomic differences noted in Glaesserella parasuis between growth in broth and on agar

et al. 2019

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“…Recently, wholegenome sequencing (WGS) has emerged as a powerful tool for predicting antibiotic resistance and pathogenic potential in G. parasuis. For instance, Li et al (2013) reported two G. parasuis strains with potential resistance towards the antibiotics ciprofloxacin, trimethoprim, and penicillin, based on the presence of associated resistance genes; Nicholson et al (2018) reported genomic differences in the toxin-antitoxin systems between phenotypically distinct G. parasuis strains from Japan and Sweden; and Bello-Orti et al (2014) noted the role of mobile genetic elements and strain-specific accessory genes in fostering high genomic diversity between pathogenic strains of the same serovar from diseased pigs in Japan, China, and the USA.…”

Section: Introductionmentioning

confidence: 99%

Whole-genome sequence analyses of Glaesserella parasuis isolates reveals extensive genomic variation and diverse antibiotic resistance determinants

Wan

Osmundson

et al. 2020

PeerJ

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Background Glaesserella parasuis (G. parasuis) is a respiratory pathogen of swine and the etiological agent of Glässer’s disease. The structural organization of genetic information, antibiotic resistance genes, potential pathogenicity, and evolutionary relationships among global G. parasuis strains remain unclear. The aim of this study was to better understand patterns of genetic variation, antibiotic resistance factors, and virulence mechanisms of this pathogen. Methods The whole-genome sequence of a ST328 isolate from diseased swine in China was determined using Pacbio RS II and Illumina MiSeq platforms and compared with 54 isolates from China sequenced in this study and 39 strains from China and eigtht other countries sequenced by previously. Patterns of genetic variation, antibiotic resistance, and virulence mechanisms were investigated in relation to the phylogeny of the isolates. Electrotransformation experiments were performed to confirm the ability of pYL1—a plasmid observed in ST328—to confer antibiotic resistance. Results The ST328 genome contained a novel Tn6678 transposon harbouring a unique resistance determinant. It also contained a small broad-host-range plasmid pYL1 carrying aac(6’)-Ie-aph(2”)-Ia and blaROB-1; when transferred to Staphylococcus aureus RN4220 by electroporation, this plasmid was highly stable under kanamycin selection. Most (85.13–91.74%) of the genetic variation between G. parasuis isolates was observed in the accessory genomes. Phylogenetic analysis revealed two major subgroups distinguished by country of origin, serotype, and multilocus sequence type (MLST). Novel virulence factors (gigP, malQ, and gmhA) and drug resistance genes (norA, bacA, ksgA, and bcr) in G. parasuis were identified. Resistance determinants (sul2, aph(3”)-Ib, norA, bacA, ksgA, and bcr) were widespread across isolates, regardless of serovar, isolation source, or geographical location. Conclusions Our comparative genomic analysis of worldwide G. parasuis isolates provides valuable insight into the emergence and transmission of G. parasuis in the swine industry. The result suggests the importance of transposon-related and/or plasmid-related gene variations in the evolution of G. parasuis.

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“…Previous G. parasuis genome analyses have identified similarities and divergence in the genomes of different G. parasuis strains [ 13 , 14 , 15 ]. Whether HPS_06257 is present in all strains G. parasuis remains unknown.…”

Section: Introductionmentioning

confidence: 99%

Expression Analysis of Outer Membrane Protein HPS_06257 in Different Strains of Glaesserella parasuis and Its Potential Role in Protective Immune Response against HPS_06257-Expressing Strains via Antibody-Dependent Phagocytosis

Chen

Shi

Cheng

et al. 2022

Veterinary Sciences

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HPS_06257 has been identified as an important protective antigen against Glaesserella parasuis infection. However, little is known about the role of HPS_06257 in the protective immune response. A whole-genome data analysis showed that among 18 isolates of Glaesserella parasuis, 11 were positive for the HPS_06257 gene, suggesting that not every strain contains this gene. We used PCR to investigate the presence of the HPS_06257 gene among 13 reference strains and demonstrated that 5 strains contained the gene. A polyclonal antibody against HPS_06257 was generated with a recombinant protein to study the expression of HPS_06257 in those 13 strains. Consistent with the PCR data, five strains expressed HPS_06257, whereas eight strains were HPS_06257 null. We also compared the protective effects of HPS_06257 against an HPS_06257-expressing strain (HPS5) and an HPS_06257-null strain (HPS11). Immunization with HPS_06257 only protected against HPS5 and not HPS11. Moreover, phagocytosis of antibody-opsonized bacteria demonstrates that the antibody against HPS_06257 increased the phagocytosis of the HPS5 strain by macrophages but not the phagocytosis of the HPS11 strain, suggesting that antibody-dependent phagocytosis is responsible for the protective role exerted by HPS_06257 in the immune response to HPS5. Our data also show that the antibody against HPS_06257 increased the phagocytosis of the other HPS_06257-expressing strains by macrophages but not that of HPS_06257-null strains. In summary, our findings demonstrate that antibody-dependent phagocytosis contributes to the protective immune response induced by immunization with HPS_06257 against HPS_06257-expressing strains.

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Comparative genomic and methylome analysis of non-virulent D74 and virulent Nagasaki Haemophilus parasuis isolates

Cited by 8 publications

References 78 publications

Transcriptomic differences noted in Glaesserella parasuis between growth in broth and on agar

Transcriptomic differences noted in Glaesserella parasuis between growth in broth and on agar

Whole-genome sequence analyses of Glaesserella parasuis isolates reveals extensive genomic variation and diverse antibiotic resistance determinants

Expression Analysis of Outer Membrane Protein HPS_06257 in Different Strains of Glaesserella parasuis and Its Potential Role in Protective Immune Response against HPS_06257-Expressing Strains via Antibody-Dependent Phagocytosis

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