Houf K, Devriese L A, De Zutter L, Van Hoof J, Vandamme P
Department of Veterinary Food Inspection, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.
Int J Food Microbiol. 2001 Dec 30;71(2-3):189-96. doi: 10.1016/s0168-1605(01)00605-5.
None of the presently available selective supplements for the specific isolation of Arcobacter species allows the growth of Arcobacter butzleri, A. cryaerophilus and A. skirrowii and at the same time fully suppresses the accompanying flora present in poultry and poultry products. Furthermore, little is known about the contamination levels of poultry with Arcobacter species. In this study, a new selective supplement comprising amphotericin B (10 mg/l), cefoperazone (16 mg/l), 5-fluorouracil (100 mg/l), novobiocin (32 mg/l) and trimethoprim (64 mg/l) was developed. With a new isolation procedure, including enrichment in Arcobacter broth with the selective supplement, incubated for 24 to 48 h at 28 degrees C under microaerobic conditions, arcobacters were isolated from 100% (n = 34) of neck skin of laying hens and from 90% (n = 71) of similar samples from broilers. Of the broiler breast meat samples examined (n = 52), 65% were found to be contaminated with these bacteria. In 64% of the samples, A. butzleri was the only Arcobacter species isolated. In 9% of the samples, A. cryaerophilus was the only species present, while 11% of the samples were positive for both species simultaneously. Using direct isolation on the selective agar medium developed in this study, incubated for 24 to 48 h under microaerobic conditions at 28 degrees C. 32 out of 45 broiler carcasses and 6 out of 25 broiler breast meat samples carried a bacterial load of arcobacters of 10(2) to 10(3) cfu/g. The prevalence of Arcobacter in Belgian poultry was found higher than the prevalence of thermophilic Campylobacter species in each of the poultry categories examined. The enrichment procedure and the direct plating method were validated for the isolation of A. skirrowii. For this species, growth performance was less than the other two Arcobacter species and it was not isolated nor detected by m-PCR from the naturally contaminated poultry samples examined. This new protocol provides a fast and reliable method for the isolation of Arcobacter species from poultry and can contribute to more comprehensive epidemiological investigations.
目前尚无用于特异性分离弓形杆菌属菌种的选择性补充剂能使布氏弓形杆菌、嗜低温弓形杆菌和斯氏弓形杆菌生长，同时又能完全抑制家禽及家禽产品中伴生菌群的生长。此外，对于家禽被弓形杆菌属菌种污染的程度了解甚少。在本研究中，开发了一种新的选择性补充剂，其包含两性霉素B（10毫克/升）、头孢哌酮（16毫克/升）、5-氟尿嘧啶（100毫克/升）、新生霉素（32毫克/升）和甲氧苄啶（64毫克/升）。采用一种新的分离程序，包括在含有该选择性补充剂的弓形杆菌肉汤中进行富集，在微需氧条件下于28℃孵育24至48小时，从100%（n = 34）的蛋鸡颈部皮肤以及90%（n = 71）的肉鸡类似样本中分离出了弓形杆菌。在所检测的肉鸡胸肉样本（n = 52）中，65%被发现被这些细菌污染。在64%的样本中，布氏弓形杆菌是唯一分离出的弓形杆菌属菌种。在9%的样本中，嗜低温弓形杆菌是唯一存在的菌种，而11%的样本同时对这两种菌种呈阳性。使用在本研究中开发的选择性琼脂培养基进行直接分离，在微需氧条件下于28℃孵育24至48小时，45个肉鸡 carcasses 中的32个以及25个肉鸡胸肉样本中的6个携带的弓形杆菌菌量为10²至10³ 菌落形成单位/克。发现比利时家禽中弓形杆菌的流行率高于所检测的各家禽类别中嗜热弯曲杆菌属菌种的流行率。富集程序和直接平板接种法经验证可用于斯氏弓形杆菌的分离。对于该菌种，生长性能低于其他两种弓形杆菌属菌种，在所检测的自然污染家禽样本中未分离到该菌种，也未通过多重聚合酶链反应检测到。这种新方案为从家禽中分离弓形杆菌属菌种提供了一种快速可靠的方法，有助于进行更全面的流行病学调查。
