Institute of Veterinary Food Science, Justus-Liebig-University Gießen, Frankfurter Street 92, D-35392 Gießen, Germany; Department of Veterinary Public Health (DVPH), College of Veterinary Medicine, Mosul University, Mosul, Iraq.
Agriculture and Agri-Food Canada, Ottawa Research and Development Centre, Ottawa, ON K1A 0C6, Canada.
J Microbiol Methods. 2020 Aug;175:105968. doi: 10.1016/j.mimet.2020.105968. Epub 2020 May 29.
The standard procedure for the improved cultural recovery of viable Mycobacterium spp. from diverse samples mainly depends on reducing the viability of background microbiota using different chemical compounds. This study was designed to i) evaluate the efficacy and comparison between N-Acetyl-l-Cysteine-Sodium hydroxide (NALC-2% NaOH) and hexadecylpyridinium chloride (0.75% HPC) treatment and exposure time on reducing the viability of undesirable microorganisms with minimal impact on colostrum consistency; and ii) assess the impact of NALC-2% NaOH on improved and enhanced recovery of Mycobacterium avium subsp. paratuberculosis (MAP) in spiked postpartum colostrum samples and consistency of colostrum. A total of 40 samples, each treated with NALC-2% NaOH for 15 min or 0.75% HPC for 5 h, were investigated for total mesophilic aerobic bacteria (MAB) and enterobacteria (EB) (CFU mL). The results showed that treatment of colostrum samples with NALC-2% NaOH completely eliminated EB and significantly reduced MAB (3.6 log CFU mL). Conversely, samples treated with 0.75% HPC produced a complex mixture following interaction with the colostrum protein and showed non-significant and variable results. In addition, the spiked colostrum treated with NALC-2% NaOH for 15 min revealed recovery of viable MAP cells with a minimum limit of detection of 1.36 log CFU 10 mL where no change in the consistency of colostrum was observed. In conclusion, 15-min NALC-2% NaOH treatment of colostrum may significantly reduce the viability of undesirable microorganisms and help to enhance the efficient recovery of MAP without impacting the consistency of high quality postpartum colostrum. This rapid procedure is suitable for efficient recovery and early detection of MAP as well as preventing its transmission to neonates and young calves in MAP infected herds.
从不同样本中提高 viable Mycobacterium spp. 的文化回收率的标准程序主要依赖于使用不同的化合物降低背景微生物群落的活力。本研究旨在:i)评估 N-乙酰-L-半胱氨酸-氢氧化钠(NALC-2% NaOH)和十六烷基氯化吡啶(0.75% HPC)处理的效果和比较,并在对初乳稠度影响最小的情况下,评估其减少不需要的微生物活力的效果;ii)评估 NALC-2% NaOH 对添加产后初乳样本中 Mycobacterium avium subsp. paratuberculosis(MAP)的改进和增强回收以及初乳稠度的影响。总共研究了 40 个样本,每个样本用 NALC-2% NaOH 处理 15 分钟或 0.75% HPC 处理 5 小时,以检测总嗜温需氧细菌(MAB)和肠杆菌(EB)(CFU mL)。结果表明,NALC-2% NaOH 处理初乳样本可完全消除 EB,并显著减少 MAB(3.6 log CFU mL)。相反,用 0.75% HPC 处理的样本与初乳蛋白相互作用后产生了复杂的混合物,且结果不显著且变化较大。此外,用 NALC-2% NaOH 处理 15 分钟的添加初乳显示出有活力的 MAP 细胞的回收,最低检测限为 1.36 log CFU 10 mL,且初乳稠度没有变化。总之,NALC-2% NaOH 处理初乳 15 分钟可能会显著降低不需要的微生物的活力,并有助于增强 MAP 的有效回收,而不会影响高质量产后初乳的稠度。该快速程序适用于 MAP 的高效回收和早期检测,以及防止其在 MAP 感染牛群中传播给新生牛和小牛。