Çin’de Kanatlı Hayvanlardan İzole Edilen Pasteurella multocida Suşlarının Moleküler Karakterizasyonu ve tonB Geni Açısından Genetik Analizi

Abstract

Pasteurella multocida (P. multocida) causes fowl cholera in birds and the pathogenesis and virulence factors involved are still poorly understood. TonB protein is a periplasmic protein prevalent in a large number of Gram-negative bacteria, as in Pasteurella, which is believed to be responsible for the import of ferric iron complexes across the outer membrane and plays a role as an important virulence factor to help bacteria to obtain nutrients. In this study, a total of 23 isolates of P. multocida were obtained from 172 cases of clinical respiratory disease in duck and geese, giving an isolation rate of 13.4%. P. multocida Serogroup A was isolated from 22 cases (95.7%), whereas serogroup F was from 1 case (4.3%). All isolates were analyzed for their susceptibility to 15 antibiotics and the presence of 18 genes for virulence factors. The susceptibility profiles suggested that Tetracycline, Cephalosporin, Enrofloxacin, and Aminoglycosides were the drugs most likely to be active against P. multocida. However, 78.3% and 52.2% of poultry strains were resistant to Penicillin and Sulfisoxazole, respectively. PCR results showed that ptfA, fimA, tonB, fur, hgbA, hgbB, sodA, sodC, pmHAS, nanH, nanB, plpB and ompH genes occur in most poultry strains of P. multocida. The virulence genes such as toxA, tadD, hsf-1, pfhA and ompA were each present in <73.9% of strains. The tonB gene was detected in all 23 clinical strains of P. multocida which were recovered from different hosts and harboring different serotypes. Phylogenetic analysis indicated that these tonB genes of P. multocida from different hosts and harboring different serotypes have close genetic relationship with a high similarity. Multiple sequence alignment demonstrated that TonB protein of Gram-negative bacteria exists in multiple conserved sites, such as -xSSGx-, -YP-, -LD-and xA[A/V]Lx motif. These findings provide clinical data into the epidemiological and molecular characteristics of avian P. multocida isolates and provide a reference for the researches of drug target in P. multocida. Keywords: Antimicrobial susceptibility, Capsular antigens, Pasteurella multocida, tonB gene, Virulence gene, Cloning, Phylogenetic analysis Çin'de Kanatlı Hayvanlardan İzole Edilen Pasteurella multocida Sışlarının Moleküler Karakterizasyonu ve tonB Geni Açısından Genetik Analizi ÖzetPasteurella multocida (P. multocida) kuşlarda kanatlı kolerasına neden olur ve hastalıkta virulans faktörleri hala tam olarak anlaşılamamıştır. TonB proteini Pasteurella da dahil pek çok Gram-negatif bakteride yaygın olarak bulunan periplazmik bir protein olup membran dışına ferrik demir komplekslerini taşımakta görevlidir ve bakterilerde besin sağlamaya yardım etmek suretiyle önemli bir virulans faktörü olarak rol oynar. Bu çalışmada, 172 klinik solunum hastalıklı ördek ve kazdan toplam 23 P. multocida izolatı elde edilerek %13.4'lük izolasyon oranı sağlandı. P. multocida Serogrup A 22 vakadan izole edilirken (%95.7), serogrup F 1 vakadan izole edildi (%4.3). Tüm izolatlar 15 antibiyotiğe kar...

Molecular Studies on Pasteurella multocida in Ducks

“…These findings were similar to Many authors; Ewers et al, (2006), Shayegh et al, (2010 and Khamesipour et al, (2014) who recorded tox A genes in P. multocida isolates. Then all 16 P. multocida isolates that were applied for tox A gene detection were submitted for capsular typing by multiplex PCR and found that all isolates were positive to capsular type A with amplified fragment 1044 pb, these results were in agreement with Zhangcheng et al, (2018) who recorded that the capsular types of P. multocida were confirmed by multiplex PCR. As the results of multiplex PCR specific for capsular antigens of P. multocida, 22 strains (95.7%) were found type A with a 1044 bp amplified fragment, whereas 1 strain (4.3%) was type F with 851 bp length fragment.…”

“…As for postmortem lesions, they include: haemorrhages in the heart, liver, gizzard and intestines besides white spots and necrotic foci in the liver and spleen as described by Abd-El-Rahman et al, (2009). In the present study, out of 220 ducks which suffer from duck septicemia examined, sixteen samples were positive to P. multocida (7.3 %) as obtained with Eid et al, (2019) who isolated P. multocida from ducks with proportion of (7.3%), but it is lower than the level reported by Zhangcheng et al, (2018) who isolated (13.4%) of P. multocida from ducks in china, (Bhattacharya, 2005) who recorded 18 isolates of P. multocida with a high percentage (77.77%) of P. multocida isolates that were recovered from ducks, Walaa and Lamyaa (2016) who isolated (63%) of P. multocida in duck flocks and who isolated (34%) P. multocida from sick ducks. These results are higher than the level reported by (Mbuthia et al, 2008) who recorded an isolation rate of 6.2% and (Muhairwa et al, 2001) who recorded that The prevalence of P. multocida was 7%.…”