Bojanić Krunoslav, Midwinter Anne C, Marshall Jonathan C, Biggs Patrick J, Acke Els
mEpiLab, School of Veterinary Science, Massey University, Palmerston North, New Zealand (Bojanić, Midwinter, Marshall, Biggs).
IDEXX VetMedLabor, Ludwigsburg, Germany (Acke).
J Vet Diagn Invest. 2019 Jan;31(1):23-32. doi: 10.1177/1040638718820082. Epub 2018 Dec 21.
We applied 7 culture methods to 50 working farm dog fecal samples and 6 methods to 50 frozen home-killed raw meat diet samples to optimize recovery of a wide range of Campylobacter spp. Culture methods combined filtration, enrichment broths, and agars at 37°C and 42°C in conventional and hydrogen-enriched microaerobic atmospheres. Overall, a prevalence of 62% (31 of 50) and 6% (3 of 50) was detected in dog and meat samples, respectively, based on Campylobacter genus PCR. A total of 356 Campylobacter spp. isolates were recovered from dogs, with successful isolation by individual methods ranging from 2 to 25 dogs. The species detected most commonly were C. upsaliensis and C. jejuni, and less commonly C. coli and C. lari. Species isolated that are rarely reported from dogs included C. rectus, C. lari subsp. concheus, C. volucris, and Helicobacter winghamensis. Six isolates from dogs positive by Campylobacter genus PCR were confirmed, using 16S rRNA sequencing, as Arcobacter cryaerophilus (1) and Arcobacter butzleri (5). C. jejuni multi-locus sequence typing results revealed a diversity of sequence types in working dogs, with several uncommonly reported from other C. jejuni sources in New Zealand. Overall, 20 isolates from 3 meat samples were positive by Campylobacter genus PCR; 1 meat sample was positive for C. jejuni, 1 for C. rectus, and 1 isolate was subsequently identified as A. butzleri. The method using Campylobacter enrichment broth in a hydrogen-enriched environment on nonselective agar resulted in significantly reduced recovery of Campylobacter spp. from both sample types.
我们将7种培养方法应用于50份工作农场犬粪便样本,并将6种方法应用于50份冷冻的家庭宰杀生肉饮食样本,以优化多种弯曲杆菌属菌株的回收率。培养方法包括在常规和富氢微需氧环境中于37°C和42°C下进行过滤、增菌肉汤培养和琼脂培养。总体而言,基于弯曲杆菌属PCR检测,犬类样本和肉类样本中的检出率分别为62%(50份中的31份)和6%(50份中的3份)。从犬类样本中共分离出356株弯曲杆菌属菌株,个别方法成功分离的犬只数量从2只到25只不等。最常检测到的菌种是乌普萨拉弯曲杆菌和空肠弯曲杆菌,较少见的是结肠弯曲杆菌和拉氏弯曲杆菌。从犬类中分离出的罕见菌种包括直肠弯曲杆菌、拉氏弯曲杆菌亚种康氏弯曲杆菌、飞禽弯曲杆菌和温厄姆螺杆菌。通过16S rRNA测序,将6株弯曲杆菌属PCR检测呈阳性的犬类分离株确认为嗜低温嗜水气单胞菌(1株)和布氏嗜水气单胞菌(5株)。空肠弯曲杆菌多位点序列分型结果显示,工作犬中空肠弯曲杆菌的序列类型具有多样性,其中一些在新西兰其他空肠弯曲杆菌来源中很少见。总体而言,3份肉类样本中的20株分离株经弯曲杆菌属PCR检测呈阳性;1份肉类样本为空肠弯曲杆菌阳性,1份为直肠弯曲杆菌阳性,1株分离株随后被鉴定为布氏嗜水气单胞菌。在富氢环境中使用弯曲杆菌增菌肉汤在非选择性琼脂上培养的方法,导致两种样本类型中弯曲杆菌属菌株的回收率显著降低。