Madar M, Slizova M, Czerwinski J, Hrckova G, Mudronova D, Gancarcikova S, Popper M, Pistl J, Soltys J, Nemcova R
1 Department of Microbiology and Immunology, University of Veterinary Medicine and Pharmacy in Kosice, Komenskeho 73, 04181 Kosice, Slovakia.
2 The Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences, Instytucka 3, 05-110 Jablonna, Poland.
Benef Microbes. 2015;6(6):899-907. doi: 10.3920/BM2015.0016. Epub 2015 Jun 30.
The study of biofilm function in vivo in various niches of the gastrointestinal tract (GIT) is rather limited. It is more frequently used in in vitro approaches, as an alternative to the studies focused on formation mechanisms and function of biofilms, which do not represent the actual in vivo complexity of microbial structures. Additionally, in vitro tests can sometimes lead to unreliable results. The goal of this study was to develop a simple approach to detect bacterial populations, particularly Lactobacillus and Bifidobacterium in biofilms, in vivo by the fluorescent in situ hybridisation (FISH) method. We standardised a new Histo-FISH method based on specific fluorochrome labelling probes which are able to detect Lactobacillus spp. and Bifidobacterium spp. within biofilms on the mucosal surface of the GIT embedded in paraffin in histological slices. This method is also suitable for visualisation of bacterial populations in the GIT internal content. Depending on the labelling probes, the Histo-FISH method has the potential to detect other probiotic strains or pathogenic bacteria. This original approach permits us to analyse bacterial colonisation processes as well as biofilm formation in stomach and caecum of BALB/c and germ-free mice.
对胃肠道（GIT）不同微环境中生物膜体内功能的研究相当有限。它在体外研究方法中使用得更为频繁，作为专注于生物膜形成机制和功能研究的替代方法，而这些研究并不能代表微生物结构实际的体内复杂性。此外，体外测试有时会导致不可靠的结果。本研究的目的是开发一种简单的方法，通过荧光原位杂交（FISH）技术在体内检测生物膜中的细菌群体，特别是乳酸杆菌和双歧杆菌。我们基于特异性荧光染料标记探针标准化了一种新的组织FISH方法，该探针能够在组织学切片中石蜡包埋的GIT黏膜表面生物膜内检测乳酸杆菌属和双歧杆菌属。该方法也适用于可视化GIT内部内容物中的细菌群体。根据标记探针的不同，组织FISH方法有潜力检测其他益生菌菌株或病原菌。这种原创方法使我们能够分析BALB/c小鼠和无菌小鼠胃和盲肠中的细菌定植过程以及生物膜形成。
