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评估用于从活体肉鸡直接培养弯曲杆菌属的不同平板培养基。

Evaluation of different plate media for direct cultivation of Campylobacter species from live broilers.

作者信息

Potturi-Venkata L-P, Backert S, Lastovica A J, Vieira S L, Norton R A, Miller R S, Pierce S, Oyarzabal O A

机构信息

Department of Poultry Science, Auburn University, Alabama 36849, USA.

出版信息

Poult Sci. 2007 Jul;86(7):1304-11. doi: 10.1093/ps/86.7.1304.

Abstract

Accurate identification and optimal culturing procedures for Campylobacter spp. from live broilers are needed for epidemiological studies. Because there is no standardized protocol, we designed and conducted studies to evaluate different selective media for the culturing and isolation of Campylobacter spp. from cecal and fecal samples obtained from battery-reared and commercial broilers. Five media selective for Campylobacter were evaluated: Campylobacter agar base, Campylobacter, Campy-Line, modified Campy-Cefex, and modified charcoal cefoperazone deoxycholate agar. With contaminated broilers reared in battery cages, Campylobacter agar base, Campylobacter, modified Campy-Cefex, and modified charcoal cefoperazone deoxycholate agar revealed similar isolation rates (P > 0.05), whereas Campy-Line showed a lower efficacy (P < 0.05). With commercial live broilers, modified Campy-Cefex agar was more consistent for the isolation of Campylobacter from feces, whereas modified Campy-Cefex and modified charcoal cefoperazone deoxycholate agar showed similar isolation rates from cecal samples. Campy-Line agar showed a lower identification rate (P < 0.05) for both fecal and cecal samples. A multiplex PCR assay used for identification showed that Campylobacter jejuni and Campylobacter coli DNA was present in the samples. Pulsed field gel electrophoresis restriction profiles differed among samples collected from different commercial farms but were similar for isolates from the same farm, suggesting clonal differences. No variation was seen in pulsed field gel electrophoresis patterns among isolates cultured on different media. Our data suggest that the choice of plate medium may influence the efficiency of isolating Campylobacter spp. from broiler chickens by direct plating from fecal or cecal samples.

摘要

进行流行病学研究需要准确鉴定来自活肉鸡的弯曲杆菌属,并采用最佳培养程序。由于没有标准化方案,我们设计并开展了研究,以评估用于从笼养和商业肉鸡的盲肠及粪便样本中培养和分离弯曲杆菌属的不同选择性培养基。评估了五种对弯曲杆菌有选择性的培养基:弯曲杆菌琼脂基础培养基、弯曲杆菌培养基、Campy-Line培养基、改良弯曲杆菌-头孢克肟培养基和改良木炭头孢哌酮脱氧胆酸盐琼脂培养基。对于笼养的受污染肉鸡,弯曲杆菌琼脂基础培养基、弯曲杆菌培养基、改良弯曲杆菌-头孢克肟培养基和改良木炭头孢哌酮脱氧胆酸盐琼脂培养基显示出相似的分离率(P>0.05),而Campy-Line培养基的效果较差(P<0.05)。对于商业活肉鸡,改良弯曲杆菌-头孢克肟琼脂培养基从粪便中分离弯曲杆菌的一致性更高,而改良弯曲杆菌-头孢克肟培养基和改良木炭头孢哌酮脱氧胆酸盐琼脂培养基从盲肠样本中显示出相似的分离率。Campy-Line琼脂培养基对粪便和盲肠样本的鉴定率均较低(P<0.05)。用于鉴定的多重PCR检测表明,样本中存在空肠弯曲杆菌和结肠弯曲杆菌的DNA。脉冲场凝胶电泳限制性图谱在不同商业养殖场采集的样本之间存在差异,但同一养殖场的分离株图谱相似,表明存在克隆差异。在不同培养基上培养的分离株之间,脉冲场凝胶电泳图谱未见差异。我们的数据表明,平板培养基的选择可能会影响通过直接从粪便或盲肠样本中接种来分离肉鸡弯曲杆菌属的效率。

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