Application of protein profiling of virulent Haemophilus parasuis by MALDI-TOF mass spectrometry

Moreno, Luisa Zanolli; Silva, Givago F. R.; Gomes, Vasco T.M.; Matajira, Carlos Emílio Cabrera; Silva, Ana Paula S.; Mesquita, Renan E.; Lotto, Nicholas P.; Ferreira, Tiago Osório; Christ, Ana Paula Guarnieri; Sato, Maria Inês Z.; Gherpelli, Y.; Dottori, M.; Bonilauri, Paolo; Luppi, Andrea; Moreno, Andréa Micke

doi:10.3855/jidc.7787

Cited by 5 publications

(7 citation statements)

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“…A well-characterized representative strain of H. parasuis LM15 (serotype 5) was used for ex vivo timekill experiments and PK/PD modeling because serotype 5 is the primary serotype found in China [3,28,29]. Species identification was performed by using MALDI-TOF mass systems (Axima-Assurance-Shimadzu) as previously described [30]. H. parasuis strains were cultured with Haemophilus test medium (HTM; Becton Dickinson, Sparks, MD) broth and agar containing 15 μg/mL β-NAD and 5% porcine hematin.…”

Section: Antibiotics and Bacterial Strainsmentioning

confidence: 99%

Epidemiological and PK/PD cutoff values determination and PK/PD-based dose assessment of gamithromycin against Haemophilus parasuis in piglets

Zhou

Liu

et al. 2020

BMC Vet Res

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Background: Gamithromycin is a macrolide approved for the treatment of bovine and swine respiratory diseases. Our study aims to establish the clinical breakpoint and optimum dose regimen for gamithromycin against Haemophilus parasuis in piglets. Results: Gamithromycin was well absorbed and fully bioavailable (87.2-101%) after intramuscular and subcutaneous administrations. The MICs of gamithromycin for 192 clinical H. parasuis isolates ranged from 0.008 to 128 mg/L and the epidemiological cutoff (ECOFF) was calculated as 1.0 mg/L. A large potentiation effect of serum on in vitro susceptibility of gamithromycin was observed for H. parasuis, with broth/serum ratios of 8.93 for MICs and 4.46 for MBCs, respectively. The postantibiotic effects were 1.5 h (1 × MIC) and 2.4 h (4 × MIC), and the postantibiotic sub-MIC effects ranged from 2.7 to 4.3 h. Gamithromycin had rapid and concentration-dependent killing against H. parasuis, and the AUC 24h /MIC ratio correlated well with ex vivo efficacy (R 2 = 0.97). The AUC 24h /MIC targets in serum associated with bacteriostatic, bactericidal and eradication activities were 15.8, 30.3 and 41.2, respectively. The PK/PD-based population dose prediction indicated a probability of target attainment (PTA) for the current marketed dose (6 mg/kg) of 88.9% against H. parasuis. The calculated gamithromycin dose for a PTA ≥ 90% was 6.55 mg/kg. Based on Monte Carlo simulations, the PK/PD cutoff (CO PD) was determined to be 0.25 mg/L. Conclusion: The determined cutoffs and PK/PD-based dose prediction will be of great importance in gamithromycin resistance surveillance and serve as an important step in the establishment of optimum dose regimen and clinical breakpoints.

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Section: Antibiotics and Bacterial Strainsmentioning

confidence: 99%

Epidemiological and PK/PD cutoff values determination and PK/PD-based dose assessment of gamithromycin against Haemophilus parasuis in piglets

Zhou

Liu

et al. 2020

BMC Vet Res

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“…Phylogeny and population structure of H. parasuis were widely studied, and H. parasuis strains could be divided into two groups in a variety of ways (Oliveira & Pijoan, 2004; Olvera, Cerda-Cuellar & Aragon, 2006b; Turni & Blackall, 2010; Howell et al, 2014; Moreno et al, 2016; Wang et al, 2016). Results of current study on correlation between population structure and virulence were contradictory.…”

Section: Discussionmentioning

confidence: 99%

“…Strain classification of H. parasuis has been widely studied serotypically, and genotypically, since differentiation of non-virulent strains from virulent strains is essential for diagnosis and control of the disease (Kielstein & Rapp-Gabrielson, 1992; Turni & Blackall, 2005; Olvera, Calsamiglia & Aragon, 2006a; Moreno et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

Association betweeniscR-based phylogeny, serovars and potential virulence markers ofHaemophilus parasuis

et al. 2019

PeerJ

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Haemophilus parasuis is an economically important bacterial pathogen of swine. Extensive genetic and phenotypic heterogeneity among H. parasuis strains have been observed, which hinders the deciphering of the population structure and its association with clinical virulence. In this study, two highly divergent clades were defined according to iron–sulphur cluster regulator (iscR)-based phylogeny analysis of 148 isolates. Clear separation of serovars and potential virulence markers (PVMs) were observed between the two clades, which are indicative of independent evolution of the two lineages. Previously suggested virulence factors showed no correlation with clinical virulence, and were probably clade or serovar specific genes emerged during different stage of evolution. PVMs profiles varied widely among isolates in the same serovar. Higher strain diversity in respect of PVMs was found for isolates from multi-strain infected farms than those from single strain infected ones, which indicates that multi-strain infection in one farm may increase the frequency of gene transfer in H. parasuis. Systemic isolates were more frequently found in serovar 13 and serovar 12, while no correlation between clinical virulence and iscR-based phylogeny was observed. It shows that iscR is a reliable marker for studying population structure of H. parasuis, while other factors should be included to avoid the interference of gene exchange of iscR between isolates. The two lineages of H. parasuis may have undergone independent evolution, but show no difference in clinical virulence. Wide distribution of systemic isolates across the entire population poses new challenge for development of vaccine with better cross-protection. Our study provides new information for better deciphering the population structure of H. parasuis, which helps understanding the extreme diversity within this pathogenic bacterium.

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“…Phylogeny and population structure of H.parasuis were widely studied, and H.parasuis strains could be divided into two groups in a variety of ways (Oliveira and Pijoan, 2004;Olvera et al, 2006b;Turni and Blackall, 2010;Howell et al, 2014;Moreno et al, 2016;Wang et al, 2016). Results of current study on correlation between population structure and virulence were contradictory.…”

Section: Discussionmentioning

confidence: 99%

“…Strain classification of H.parasuis has been widely studied serotypically, and genotypically, since differentiation of non-virulent strains from virulent strains is essential for diagnosis and control of the disease (Kielstein and Rapp-Gabrielson, 1992;Turni and Blackall, 2005;Olvera et al, 2006a;Moreno et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

Peer Review #2 of "Association between iscR-based phylogeny, serovars and potential virulence markers of Haemophilus parasuis (v0.1)"

2019

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Haemophilus parasuis is an economically important bacterial pathogen of swine. Extensive genetic and phenotypic heterogeneity among H.parasuis strains have been observed, which hinders the deciphering of the population structure and its association with clinical virulence. In this study, two highly divergent clades were defined according to iscR-based phylogeny analysis of 148 isolates. Clear separation of serovars and potential virulence markers (PVMs) were observed between the two clades, which are indicative of independent evolution of the two lineages. Previously suggested virulence factors showed no correlation with clinical virulence, and were probably clade or serovar specific genes emerged during different stage of evolution. PVMs profiles varied widely among isolates in the same serovar. Higher strain diversity in respect of PVMs was found for isolates from multi-strain infected farms than those from single strain infected ones, which indicates that multi-strain infection in one farm may increase the frequency of gene transfer in H.parasuis. Systemic isolates were more frequently found in serovar 13 and serovar 12, while no correlation between clinical virulence and iscR-based phylogeny was observed. It shows that iscR is a reliable marker for studying population structure of H.parasuis, while other factors should be included to avoid the interference of gene exchange of iscR between isolates. The two lineages of H.parasuis may have undergone independent evolution, but show no difference in clinical virulence. Wide distribution of systemic isolates across the entire population poses new challenge for development of vaccine with better cross-protection. Our study provides new information for better deciphering the population structure of H.parasuis, which helps understanding the extreme diversity within this pathogenic bacterium.

show abstract

Application of protein profiling of virulent Haemophilus parasuis by MALDI-TOF mass spectrometry

Abstract: This item has no abstract. Follow the links below to access the full text.

Cited by 5 publications

References 15 publications

Epidemiological and PK/PD cutoff values determination and PK/PD-based dose assessment of gamithromycin against Haemophilus parasuis in piglets

Epidemiological and PK/PD cutoff values determination and PK/PD-based dose assessment of gamithromycin against Haemophilus parasuis in piglets

Association betweeniscR-based phylogeny, serovars and potential virulence markers ofHaemophilus parasuis

Peer Review #2 of "Association between iscR-based phylogeny, serovars and potential virulence markers of Haemophilus parasuis (v0.1)"

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