Martelli F, Gosling R, McLaren I, Wales A, Davies R
Department of Bacteriology, Animal Health and Veterinary Laboratories Agency, Addlestone, Surrey, UK.
Lett Appl Microbiol. 2014 Oct;59(4):443-8. doi: 10.1111/lam.12299. Epub 2014 Jul 14.
Salmonella-contaminated poultry house dust plus 10 g chicken faeces inoculated with Salmonella Enteritidis and then frozen for storage and transport were used as candidate external quality assurance test samples. Variations in faeces sample preparation, storage and culture were examined initially. This indicated that, within modest limits, the age of the inoculating culture and of the faeces did not affect detection, nor did swirling the pre-enrichment culture or extending its duration. Under optimal conditions of preparation and storage, Salmonella numbers of 70 colony-forming units (CFU) and above were reliably detected at the originating laboratory. A ring trial was performed, involving 13 external UK laboratories plus the originating laboratory. Faeces samples inoculated with Salmonella Enteritidis were frozen, transported on dry ice and tested by the ISO 6579:2002 (Annex D) method. Detection by the originating laboratory was consistent with the previously established lower limit for reliability of 70 CFU. However, the sensitivity of detection by the external laboratories was apparently poorer in several cases, with significant interlaboratory variation seen at the lowest inoculum level, using Fisher's exact test. Detection of Salmonella in poultry house dust appeared to be more sensitive and uniform among laboratories. Significance and impact of the study: Salmonella surveillance and control regimes in the European poultry industry and elsewhere require sensitive culture detection of Salmonella in environmental samples, including poultry faeces. A ring trial was conducted, and the results highlighted that some of the participating laboratories failed to identify Salmonella. This suggests that contaminated frozen faeces cubes could be beneficial to assess proficiency, according to the results of this preliminary study. The data obtained in this study can be used as an indication for the design of realistic external quality assurance for laboratories involved in official testing of Salmonella in poultry flocks.
受沙门氏菌污染的禽舍灰尘加上接种肠炎沙门氏菌后冷冻用于储存和运输的10克鸡粪,被用作外部质量保证测试候选样本。最初研究了粪便样本制备、储存和培养过程中的差异。结果表明,在一定适度范围内,接种培养物和粪便的保存时间并不影响检测结果,预增菌培养物的振荡或延长预增菌培养时间也不影响检测结果。在制备和储存的最佳条件下,源实验室能够可靠地检测到70个及以上菌落形成单位(CFU)的沙门氏菌数量。开展了一项能力验证试验,有13家英国外部实验室以及源实验室参与。接种肠炎沙门氏菌的粪便样本经冷冻后,用干冰运输,并采用ISO 6579:2002(附录D)方法进行检测。源实验室的检测结果与之前确定的70 CFU可靠性下限一致。然而,在一些情况下,外部实验室的检测灵敏度明显较差,使用Fisher精确检验发现在最低接种水平存在显著的实验室间差异。禽舍灰尘中沙门氏菌的检测在各实验室之间似乎更灵敏且更一致。研究的意义和影响:欧洲及其他地区的家禽业沙门氏菌监测和控制体系要求对包括家禽粪便在内的环境样本中的沙门氏菌进行灵敏的培养检测。开展了一项能力验证试验,结果突出表明一些参与实验室未能鉴定出沙门氏菌。根据这项初步研究的结果,这表明受污染的冷冻粪块可能有助于评估检测水平。本研究获得的数据可作为为参与家禽群沙门氏菌官方检测的实验室设计实际外部质量保证的参考依据。