Molecular characterization of Pasteurella multocida pfhaB1 gene fragment from buffalo and cattle isolates from Nusa Tenggara Timur Indonesia

Ine Karni, D. Handijatno, L. T. Suwanti, K. Kusnoto, J. Rahmahani, W. Tyasningsih
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Abstract

Almost all regions in Nusa Tenggara Timur East Nusa Tenggara (NTT) Province Indonesia are endemic areas of Haemorragic Septicaemia (HS), which is caused by Pasteurella multocida  (P. multocida ) Serotypes B: 2. The fragment  pfhaB1gene is one of the virulence factors of P. multocida.The objective of this study was to determaine the phylogenetic, homology of P. multocidapfhaB1gene fragment of isolatedfrom Buffalo and Cattle in NTT. The P. multocida isolateswere re-cultured and further microscopic examined the biochemical tests, PCR, sequencing, homology, and phylogenetic relatedness test. P. multocida was observed as gram negative, coccobacilus, no growth on MacConkey Agar, does not produce H2S and gas, nonmotile and indole positive, does not produce urease enzymes, does not use citrate as a carbon source, does not ferment maltose and lactose but it does ferment glucose, sucrose and mannitol. ThepfhaB1gene fragmentfrom buffalo and cattle NTT isolates and also Katha strain vaccine showed DNA band 506 bp. P.multocida isolates from buffalo and cattle in NTT have 91% - 99% score homology with the comparative isolate. The isolate P. multocida from buffalo and cattle in NTT are in one cluster and their phylogenetic relatedness is close to isolates from Iran and India. It is concluded that the  pfhaB1gene fragmentof P. multocida from buffalo and cattle isolates have phylogenetic relatedness close and homolog with the other comparative isolates.    Keywords: Haemorrhagic Septicaemia; Nusa Tenggara Timur; Pasteurella multocida;  pfhaB1gene
印尼努沙登加拉水牛和牛分离株多杀性巴氏杆菌pfhaB1基因片段的分子特征
印度尼西亚东努沙登加拉省几乎所有地区都是由多杀性巴氏杆菌血清型B: 2引起的出血性败血症(HS)流行区。pfhab1基因片段是多杀假单胞菌的毒力因子之一。本研究的目的是确定NTT水牛和牛分离的P. multicidapfhab1基因片段的系统发育和同源性。对多杀假单胞菌分离株进行再培养,进一步进行生化试验、PCR、测序、同源性和系统发育相关性试验。多杀假单胞菌为革兰氏阴性,球芽孢杆菌在麦康基琼脂上不生长,不产生H2S和气体,无运动和吲哚阳性,不产生脲酶,不以柠檬酸盐为碳源,不发酵麦芽糖和乳糖,但发酵葡萄糖、蔗糖和甘露醇。从水牛和牛NTT分离株和Katha株疫苗中分离出的pfhab1基因片段显示出506 bp的DNA带。从NTT水牛和牛中分离出的多杀假单胞菌与比较分离物的同源性评分为91% ~ 99%。来自NTT水牛和牛的多杀假单胞菌分离株属于一个聚类,其系统发育亲缘性与来自伊朗和印度的分离株接近。结果表明,从水牛和牛分离的多杀假单胞菌pfhab1基因片段与其他比较分离株具有密切的系统亲缘关系和同源性。关键词:出血性败血症;努沙登加拉帖木儿;巴斯德菌multocida;pfhaB1gene
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