aroA gene PCR-RFLP diversity patterns in Haemophilus parasuis and Actinobacillus species

Rio, María-Luisa del; Gutiérrez-Martín, César B.; Navas, Jesús; Gutiérrez-Muñiz, B.; Rodrı́guez-Barbosa, José Ignacio; Ferri, Elías Fernando Rodríguez

doi:10.1016/j.rvsc.2005.03.004

Cited by 22 publications

(11 citation statements)

References 17 publications

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“…So far, 15 serovars and a considerable number of non-typeable isolates have been identified. Various molecular approaches have revealed the diversity in genotypic lineage and have shed light on the heterogeneity of pathogenic H. parasuis serovars (Olvera, Calsamiglia & Aragon, 2006; Del Río et al, 2006; Zhang et al, 2011). …”

Section: Introductionmentioning

confidence: 99%

Prevalence and seroepidemiology ofHaemophilus parasuisin Sichuan province, China

Wang

Zhao

Wei

et al. 2017

PeerJ

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Haemophilus parasuis, the causative agent of Glässer’s disease, has been reported as widespread, but little is known about its epidemiology in the Sichuan province of China. The goal of our research is to reveal the prevalence and distribution of H. parasuis in this area. Sampling and isolation were performed across Sichuan; isolates were processed using serotyping multiplex PCR (serotyping-mPCR) and agar gel diffusion (AGD) for confirmation of serovar identity. This study was carried out from January 2014 to May 2016 and 254 H. parasuis field strains were isolated from 576 clinical samples collected from pigs displaying clinical symptoms. The isolation frequency was 44.10%. Statistically very significant differences of infection incidence were found in three age groups (P < 0.01) and different seasons (P < 0.01). Serovars 5 (25.98%) and 4 (23.62%) were the most prevalent, however, non-typeable isolates accounted for nearly 7.87%. In terms of geographical distribution, serovars 5 and 4 were mostly prevalent in west and east Sichuan. The results confirmed that the combined approach was dependable and revealed the diversity and distribution of serovars in Sichuan province, which is vital for efforts aimed at developing vaccine candidates allowing for the prevention or control of H. parasuis outbreaks.

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Section: Introductionmentioning

confidence: 99%

Prevalence and seroepidemiology ofHaemophilus parasuisin Sichuan province, China

Wang

Zhao

Wei

et al. 2017

PeerJ

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“…Recently, different genotyping methods have been proposed to differentiate H. parasuis strains. The majority of them are fingerprinting methods, and, even though the reported techniques have a higher level of discrimination than serotyping, they present application problems, such as limited resolution (de la Puente-Redondo et al, del Rio et al, 2006) or difficulty in comparing results from different laboratories (Rafiee et al, 2000;Smart et al, 1988). To improve the epidemiological study of H. parasuis strains, a single-locus sequence typing method was recently used by our group (Olvera et al, 2006).…”

Section: Introductionmentioning

confidence: 99%

Study of the population structure of Haemophilus parasuis by multilocus sequence typing

2006

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Haemophilus parasuis is the aetiological agent of Glä sser's disease in swine. In addition, this bacterium causes other clinical outcomes and can also be isolated from the upper respiratory tract of healthy pigs. Isolates of H. parasuis differ in phenotypic features (e.g. protein profiles, colony morphology or capsule production) and pathogenic capacity. Differences among strains have also been demonstrated at the genetic level. Several typing methods have been used to classify H. parasuis field strains, but they had resolution or implementation problems. To overcome these limitations, a multilocus sequence typing (MLST) system, using partial sequences of the house-keeping genes mdh, 6pgd, atpD, g3pd, frdB, infB and rpoB, was developed. Eleven reference strains and 120 field strains were included in this study. The number of alleles per locus ranged from 14 to 41, 6pgd being the locus with the highest diversity. The high genetic heterogeneity of this bacterium was confirmed with MLST, since the strains were divided into 109 sequence types, and only 13 small clonal complexes were detected by the Burst algorithm. Further analysis by unweighted-pair group method with arithmetic mean (UPGMA) identified six clusters. When the clinical background of the isolates was examined, one cluster was statistically associated with nasal isolation (putative non-virulent), while another cluster showed a significant association with isolation from clinical lesions (putative virulent). The remaining clusters did not show a statistical association with the clinical background of the isolates. Finally, although recombination among H. parasuis strains was detected, two divergent branches were found when a neighbour-joining tree was constructed with the concatenated sequences. Interestingly, one branch included almost all isolates of the putative virulent UPGMA cluster. INTRODUCTIONHaemophilus parasuis is a member of the family Pasteurellaceae and the causative agent of Glässer's disease in pigs, which is pathologically characterized by fibrinous to fibrinopurulent polyserositis and polyarthritis (RappGabrielson et al., 2006). In addition to Glässer's disease, H. parasuis produces other clinical outcomes, such as pneumonia, and colonizes the upper respiratory tract of healthy animals (Rapp-Gabrielson et al., 2006). Although it is commonly accepted that one strain is responsible for each clinical outbreak, diagnosis of H. parasuis infection is complicated by the fact that it is usual to detect several strains in a farm and even within a single animal. Therefore, it is essential to determine the causative strain by its isolation from organs with the characteristic lesions of the disease.Differences among strains in phenotypic and genotypic characteristics have been reported, although no clear association with virulence could be determined (Oliveira & Pijoan, 2004;Rapp-Gabrielson et al., 2006). However, several studies have confirmed that different strains of H. parasuis have different pathogenic capacity (Kielstein & Rapp-Gabrielson, 199...

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“…However, 16S rRNA gene sequences are usually not suitable for strain differentiation due to a lack of variability below the species level. Recently, PCR-restriction fragment length polymorphism (PCR-RFLP) analyses using the sequences of tbpA (12) and aroA (13) have been proposed, but the application of these techniques does not provide sufficient information about the phylogeny between strains. Another approach to differentiating field strains is the use of enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) (41).…”

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Genotypic Diversity of Haemophilus parasuis Field Strains

Olvera

Calsamiglia

Aragón

2006

Appl Environ Microbiol

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Haemophilus parasuis is the cause of Glässer's disease and other clinical disorders in pigs. It can also be isolated from the upper respiratory tracts of healthy pigs, and isolates can have significant differences in virulence. In this work, a partial sequence from the 60-kDa heat shock protein (Hsp60) gene was assessed as an epidemiological marker. We analyzed partial sequences of hsp60 and 16S rRNA genes from 103 strains of H. parasuis and other related species to obtain a better classification of the strains and examine the correlation with virulence. The results were compared with those obtained by enterobacterial repetitive intergenic consensus PCR. Our results showed that hsp60 is a reliable marker for epidemiological studies of H. parasuis and that the analysis of its sequence is a better approach than fingerprinting methods. Furthermore, the analysis of the hsp60 and 16S rRNA gene sequences revealed the presence of a separate lineage of virulent strains and indicated the occurrence of lateral gene transfer among H. parasuis and Actinobacillus strains.

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aroA gene PCR-RFLP diversity patterns in Haemophilus parasuis and Actinobacillus species

Cited by 22 publications

References 17 publications

Prevalence and seroepidemiology ofHaemophilus parasuisin Sichuan province, China

Prevalence and seroepidemiology ofHaemophilus parasuisin Sichuan province, China

Study of the population structure of Haemophilus parasuis by multilocus sequence typing

Genotypic Diversity of Haemophilus parasuis Field Strains

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