Ongör H, Cetinkaya B, Açik M N, Atabay H I
Department of Microbiology, Faculty of Veterinary Medicine, University of Firat, Elazig, Turkey.
Lett Appl Microbiol. 2004;38(4):339-44. doi: 10.1111/j.1472-765x.2004.01494.x.
To investigate the presence of Arcobacter spp. in minced beef meat (n = 97) and rectal faecal samples (n = 200) collected from cattle immediately after slaughter at a local abattoir in Turkey.
Meat samples were examined using three different isolation procedures (CAT-supplemented media, de Boer arcobacter isolation method and membrane filtration method), but only one method (CAT-supplemented media) was employed for faecal samples. The isolated Arcobacter strains were identified by genus- and species-(multiplex) specific PCR assays. Arcobacter spp. were isolated from 5 and 9.5% of meat and faecal samples respectively. Although the only Arcobacter sp. found in meat samples was Arcobacter butzleri, all three pathogenic species--A. butzleri, A. cryaerophilus and A. skirrowii--were detected in the rectal swabs. No Arcobacter was isolated when the de Boer method was used for minced meat samples but the same five meat samples were found positive for arcobacters when CAT-supplemented media and membrane filtration method were used.
The membrane filtration method was found to be superior to the CAT-supplemented media, because it led to a reduction in competing microflora. However, the necessity for one filter and medium for each sample makes this method somewhat expensive. The multiplex-PCR (m-PCR) assay shortened significantly the time required for the identification of Arcobacter spp. and also removed the possibility of false positive results due to other campylobacteria.
This study reports the isolation of Arcobacter spp. in cattle for the first time in Turkey. The m-PCR assay enables the identification and differentiation of all arcobacters simultaneously in one-step PCR.
调查在土耳其当地一家屠宰场宰杀后立即采集的97份牛肉末样本和200份直肠粪便样本中弓形杆菌属细菌的存在情况。
肉类样本采用三种不同的分离程序(添加CAT的培养基、德布尔弓形杆菌分离法和膜过滤法)进行检测,但粪便样本仅采用一种方法(添加CAT的培养基)。分离出的弓形杆菌菌株通过属特异性和种特异性(多重)PCR检测进行鉴定。分别从5%的肉类样本和9.5%的粪便样本中分离出弓形杆菌属细菌。虽然在肉类样本中发现的唯一弓形杆菌是布氏弓形杆菌,但在直肠拭子中检测到了所有三种致病菌种——布氏弓形杆菌、嗜低温弓形杆菌和斯氏弓形杆菌。当德布尔方法用于牛肉末样本时未分离出弓形杆菌,但使用添加CAT的培养基和膜过滤法时,相同的5份肉类样本被发现弓形杆菌呈阳性。
发现膜过滤法优于添加CAT的培养基,因为它减少了竞争性微生物群落。然而,每个样本需要一个过滤器和一种培养基,这使得该方法成本较高。多重PCR(m-PCR)检测显著缩短了鉴定弓形杆菌属细菌所需的时间,也消除了因其他弯曲杆菌导致假阳性结果的可能性。
本研究首次报道了在土耳其牛中分离出弓形杆菌属细菌。m-PCR检测能够在一步PCR中同时鉴定和区分所有弓形杆菌。