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標題: 中部地區豬胸膜肺炎放線桿菌抗藥性及血清型研究
Investigation of antimicrobial resistance and serotyping of Actinobacillus pleuropneumoniae in central Taiwan
作者: 謝一美
Hsieh, Yi-Mei
關鍵字: actinobacillus pleuropneumoniae;豬胸膜肺炎放線桿菌;antimicrobial resistance;serotyping;抗藥性;血清型
出版社: 獸醫公共衛生學研究所
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Actinobacillus pleuropneumoniae(APP) 是豬隻重要的地方流行性呼吸道疾病,造成養豬產業重大經濟損失。本研究於2007年7月至2008年7月於彰化地區養豬場採集疑似APP症狀之豬隻肺臟,共計分離56株APP,以快速平板凝集試驗法及PCR進行血清型分型結果為︰1型44株(79 %)、2型3株(5 %)、5型3株(5 %)、7型2株(4 %)、8型4株(7 %),目前台灣流行的APP血清型仍以第1型為主。以紙錠擴散法進行21種抗菌劑感受性試驗,結果顯示96%之分離株對5種以上之抗生素具有抗藥性,各藥對APP抗藥性比率以streptomycin (98 %)、oxytetracycline(93 %)為最高;敏感性方面,則以ceftiofur (98 %)、colistin(68 %)、florocol (61 %)、chloramphenicol (61 %)有效比例較高。另外,比較同一養豬場中,分離到的APP與腸內菌科細菌(沙門氏菌及大腸桿菌)之抗藥性數量,並無顯著差異(p>0.05)。本研究中,設計四對引子對以PCR法檢測APP之gyrA、gyrB、parC、parE基因之QRDRs(Quinolone Resistance Determining Regions)中的點突變,探討QRDRs突變與APP對quinolone抗菌劑產生抗藥性的關聯,結果顯示對quinolone具抵抗性的APP菌株,在gyrA基因的第83個氨基酸(Ser-83-to- Phe or Val or Tyr)或第87個氨基酸(Asp-87-to- His or Asn or Tyr)具有單一點突變;而對fluoroquinolone具抵抗性之菌株則在第83個氨基酸和第87個氨基酸具有雙重點突變(Ser-83-to-Phe and Asp-87-to-Gly, Ser-83-to-Phe and Asp-87-to-Asn, Ser-83-to-Phe and Asp-87-to-Tyr)。實驗結果顯示目前在台灣的養豬業,APP與腸內菌科已產生多重抗藥性,且APP菌株與已知腸內菌科細菌具有相似的QRDRs突變模式,因抗藥性基因可能在不同菌種間傳播,抗藥性的形成不僅會影響畜牧業的經濟效益,當抗藥性基因傳遞到人畜共通的細菌時,將嚴重影響人體健康,形成公共衛生學的威脅,因此,必需在養豬業建立一套監測系統以監控多重抗藥性菌株的出現,畜禽業抗菌劑的使用也應受到嚴格的限制與管控。

Actinobacillus pleuropneumoniae (APP) causes porcine pleuropneumoniae, a worldwide disease of swine that results in severe economic losses. A total of 56 strains of APP were isolated from the lungs of pigs that died of pleuropneumoniae from farms in Changhua county during July 2007 and July 2008. Of the 56 APP serotyped by a rapid slide agglutination test and PCR , 44 isolates (79 %) were of serotype1; 3 each (5 %) were of serotype 2, 5; 2 (4 %) were of serotype 7; and 4 (7%) were of serotype 8. Serotype 1 was still the most prevalent in Taiwan. The isolates were tested for their susceptibility to 21 antimicrobials using an agar disk diffusion method. Ninety-six percent of the isolates were resistant to at least five antimicrobials. A high rate of resistance was observed to streptomycin (98 %) and oxytetracycline (93 %). Higher rate of susceptibility was observed to ceftiofur (98 %), colistin (68 %), florocol (61 %) and chloramphenicol (61 %). When comparing the resistant numbers of antimicrobials, there was no significantly difference (p>0.05) between APP and genus of Enterobacteraceae (Salmonella and E. coli) recovered from the same farm. In order to identify QRDRs (Quinolone Resistance Determining Regions) mutations in quinolone-resistant isolates of APP, we designed four pairs of primers to amplify the QRDRs genes of gyrA, gyrB, parC, parE and sequenced. It appears that that one mutation in the QRDRs of gyrA, leading to amino acid changes in either codon 83 (Ser-83-to- Phe or Val or Tyr) or codon 87 (Asp-87-to- His or Asn or Tyr), contribute to the resistance of quinolone. Instead, strains carried double mutations in both codon 83 and 87 (Ser-83-to-Phe and Asp-87-to-Gly, Ser-83-to-Phe and Asp-87-to-Asn, Ser-83-to-Phe and Asp-87-to-Tyr) become FQ-resistant. This study demonstrates that APP and genus of Enterobacteraceae were resistant to multidrugs in Taiwan. Similar mutation patterns of QRDRs were also observed in APP as genus of Enterobacteraceae in previous works. The widespread use of antibiotics for treatment of pleuropneumonia in swine may lead to selective survival of strains with resistant genes that can be transformed to other zoonotic bacteria which may be hazards for animal health and public health. Thus, use of antibiotics must be cautiously approached and a surveillance system is indispensable for the swine industry to monitor the multidrug resistant bacterial pathogens in Taiwan.
其他識別: U0005-2201200912235500
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