Núñez-del Arco Alma, Salas-Téllez Enrique, de la Garza Mireya, Díaz-Aparicio Efrén, Tenorio-Gutiérrez Víctor
Cátedra de Inmunodiagnóstico de Enfermedades Bacterianas y Micóticas, Facultad de Estudios Superiores (FES) Cuautitlán, Universidad Nacional Autónoma de Mexico (UNAM), Av. Primero de mayo s/n, Cuautitlân Izcalli, Estado de Mexico 54740, Mexico.
Can J Vet Res. 2006 Jan;70(1):43-9.
Actinobacillus seminis is a gram-negative bacterium of the Pasteurellaceae family that is involved in ovine epididymitis. Looking for a protein specific to this species, we determined the protein profile of subcellular fractions of A. seminis (American Type Culture Collection number 15768): proteins from the outer membrane (OMPs), inner membrane (IMPs), and cytoplasm (CPs). These profiles provide the first data, to our knowledge, regarding subcellular fractions of A. seminis. In the OMP fraction, we identified a protein with a molecular mass of 75 kDa that proved to be immunogenic and apparently specific for A. seminis. This conclusion was based on the reaction of hyperimmune serum of rabbits inoculated with whole cells of A. seminis that was tested against sonicated complete cells of reference strains and field isolates of Brucella ovis, Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni. No protein of these bacteria cross-reacted with the 75-kDa protein of A. seminis. Furthermore, when each type of hyperimmune serum was tested against the sonicated cells and each of the subcellular fractions of A. seminis, it did not recognize the A. seminis 75-kDa protein. We also isolated and identified this protein in microvesicles released to the culture supernatant. The results suggest that the 75-kDa protein could be used to establish a diagnostic test specific for ovine epididymitis caused by A. seminis.
精液放线杆菌是巴氏杆菌科的一种革兰氏阴性细菌，可引发绵羊附睾炎。为寻找该物种特有的蛋白质，我们测定了精液放线杆菌（美国典型培养物保藏中心编号15768）亚细胞组分的蛋白质谱：外膜（OMPs）、内膜（IMPs）和细胞质（CPs）中的蛋白质。据我们所知，这些蛋白质谱提供了关于精液放线杆菌亚细胞组分的首批数据。在外膜蛋白组分中，我们鉴定出一种分子量为75 kDa的蛋白质，该蛋白质具有免疫原性，且显然是精液放线杆菌特有的。这一结论基于用精液放线杆菌全细胞接种的兔子的超免疫血清与绵羊布鲁氏菌、溶血曼氏杆菌、多杀巴斯德菌和睡眠嗜组织菌的参考菌株及田间分离株的超声破碎全细胞的反应。这些细菌的蛋白质均未与精液放线杆菌的75 kDa蛋白质发生交叉反应。此外，当每种超免疫血清与精液放线杆菌的超声破碎细胞及其各亚细胞组分进行检测时，均未识别出精液放线杆菌的75 kDa蛋白质。我们还在释放到培养上清液中的微泡中分离并鉴定了这种蛋白质。结果表明，75 kDa蛋白质可用于建立针对由精液放线杆菌引起的绵羊附睾炎的特异性诊断试验。
