Laboratory of Foodborne Parasites, Department of Translational Physiology, Infectiology and Public Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium.
Department of Veterinary and Biosciences, Faculty of Veterinary Medicine, Ghent University, Heidestraat 19, 9820 Merelbeke, Belgium; Laboratory of Microbiology, Department of Biochemistry and Microbiology, Faculty of Sciences, Ghent University, Karel Lodewijk Ledeganckstraat 35, 9000 Ghent, Belgium.
J Microbiol Methods. 2022 Dec;203:106599. doi: 10.1016/j.mimet.2022.106599. Epub 2022 Oct 13.
Consumers demand more fresh, safe, and high-quality food. As this is partiallycorrelated to the microbial profile, several microbiological examination tools are available. Incontrast to meat, no microbiological normalized methods to assess the microbiological quality of fresh marine fish have been agreed on. As a result, studies on the detection and diversity of spoilage associated organisms (SAOs) in fish often apply various detection, isolation, and identification techniques. This complicates the comparison and interpretation of data reported, and often results in different or inconclusive results. Therefore, the present review aimed to present a critical overview of the isolation/cultivation and detection techniques currently applied in fish microbiology. After a comprehensive search in the PubMed, Web of Science and Scopus databases, a total of 111 studies fulfilled the review selection criteria. Results revealed that when relying on culture media for the isolation of SAOs in fish, it is essential to include a salt-containing medium next to plate count agar that is currently used as the reference medium for the enumeration of bacteria on fish. In terms of identification, MALDI-TOF MS and 16S rRNA gene sequencing are currently the most promising tools, though other housekeeping genes should be targeted as well, and, the biggest challenge at this point is still the lack of comprehensive proteomic and sequence databases for SAOs. A full replacement of cultivation by next generation sequencing is difficult to recommend due to the absence of a standardized experimental methodology, especially for fish, and the relatively high sequencing costs. Additionally, a discrepancy between culture-dependent and independent methods in revealing the bacterial diversity, and abundancy, from marine fish was demonstrated by several authors. It is therefore recommended to consider both approaches as complements of one another, rather than substitutes, and to include them simultaneously to yield more complete results regarding the SAOs in fresh marine fish. As such, a thorough understanding of the biology of spoilage organisms and process will be obtained to prolong the shelf-life and deliver a high-quality product.
消费者需要更多新鲜、安全和高质量的食品。由于这部分与微生物特征相关，因此有多种微生物检测工具可供使用。与肉类不同，目前尚未就评估新鲜海鱼微生物质量达成标准化的微生物方法。因此，关于鱼类中腐败相关生物体（SAO）的检测和多样性的研究通常采用各种检测、分离和鉴定技术。这使得报告的数据的比较和解释变得复杂，并经常导致不同的或不确定的结果。因此，本综述旨在对目前应用于鱼类微生物学的分离/培养和检测技术进行批判性概述。在对 PubMed、Web of Science 和 Scopus 数据库进行全面搜索后，共有 111 项研究符合综述选择标准。结果表明，当依靠培养基分离鱼类中的 SAO 时，除了目前用作鱼类细菌计数的参考培养基的平板计数琼脂外，还必须包含含盐培养基。在鉴定方面，MALDI-TOF MS 和 16S rRNA 基因测序目前是最有前途的工具，尽管也应该针对其他管家基因，目前最大的挑战仍然是缺乏针对 SAO 的全面蛋白质组学和序列数据库。由于缺乏标准化的实验方法，特别是对于鱼类，以及相对较高的测序成本，因此，很难推荐通过下一代测序完全替代培养。此外，几位作者证明了依赖培养和独立方法在揭示海洋鱼类细菌多样性和丰度方面存在差异。因此，建议将这两种方法视为彼此的补充，而不是替代品，并同时使用它们，以获得关于新鲜海鱼中 SAO 的更完整的结果。通过这种方式，可以更深入地了解腐败生物和过程的生物学特性，从而延长保质期并提供高质量的产品。
