Oueriaghli Nahid, Castro David J, Llamas Inmaculada, Béjar Victoria, Martínez-Checa Fernando
Microbial Exopolysacharide Research Group, Department of Microbiology, Faculty of Pharmacy, University of Granada, Granada, Spain.
Institute of Biotechnology, University of Granada, Granada, Spain.
Front Microbiol. 2018 Jun 21;9:1377. doi: 10.3389/fmicb.2018.01377. eCollection 2018.
We studied the bacterial community in Rambla Salada in three different sampling sites and in three different seasons and the effect of salinity, oxygen, and pH. All sites samples had high diversity and richness (Rr > 30). The diversity indexes and the analysis of dendrograms obtained by DGGE fingerprint after applying Pearson's and Dice's coefficient showed a strong influence of sampling season. The Pareto-Lorenz (PL) curves and analysis indicated that the microbial communities were balanced and despite the changing environmental conditions, they can preserve their functionality. The main phyla detected by DGGE were (39.73%), (28.43%), (8.23%), and (5.14%). The majority of the sequences corresponding to uncultured bacteria belonged to phylum. Within , the main genera detected were and . The environmental factors which influenced the community in a higher degree were the salinity and oxygen. The bacteria belonging to and were positively influenced by salinity. Nevertheless, bacteria related to and classes and phylum showed a positive correlation with oxygen and pH but negative with salinity. The phylum were less influenced by the environmental variables. The bacterial community composition of Rambla Salada was also studied by dilution-to-extinction technique. Using this method, 354 microorganisms were isolated. The 16S sequences of 61 isolates showed that the diversity was very different to those obtained by DGGE and with those obtained previously by using classic culture techniques. The taxa identified by dilution-to-extinction were (81.92%), (11.30%), (4.52%), and (2.26%) phyla with as predominant class (65.7%). The main genera were: (38.85%), (20.2%), and (11.2%). Nine of the 61 identified bacteria showed less than 97% sequence identity with validly described species and may well represent new taxa. The number of bacteria in different samples, locations, and seasons were calculated by CARD-FISH, ranging from 54.3 to 78.9% of the total prokaryotic population. In conclusion, the dilution-to-extinction technique could be a complementary method to classical culture based method, but neither gets to cultivate the major taxa detected by DGGE. The bacterial community was influenced significantly by the physico-chemical parameters (specially the salinity and oxygen), the location and the season of sampling.
我们研究了拉布拉·萨拉达河三个不同采样点和三个不同季节的细菌群落,以及盐度、氧气和pH值的影响。所有采样点的样本都具有高度的多样性和丰富度(丰富度指数Rr>30)。应用皮尔逊系数和迪西系数后,通过DGGE指纹图谱获得的多样性指数和聚类图分析表明,采样季节有很大影响。帕累托-洛伦兹(PL)曲线及分析表明,微生物群落是平衡的,尽管环境条件不断变化,但它们仍能保持其功能。通过DGGE检测到的主要门类为(39.73%)、(28.43%)、(8.23%)和(5.14%)。大多数与未培养细菌对应的序列属于门。在门内,检测到的主要属为和。对群落影响程度较高的环境因素是盐度和氧气。属于和的细菌受到盐度的正向影响。然而,与纲和门相关的细菌与氧气和pH值呈正相关,但与盐度呈负相关。门受环境变量的影响较小。还通过稀释至灭绝技术研究了拉布拉·萨拉达河的细菌群落组成。使用这种方法,分离出354种微生物。61株分离株的16S序列显示,其多样性与通过DGGE获得的多样性以及先前使用经典培养技术获得的多样性有很大不同。通过稀释至灭绝鉴定出的分类单元为(81.92%)、(11.30%)、(4.52%)和(2.26%)门,以纲为主导类群(65.7%)。主要属为:(38.85%)、(20.2%)和(11.2%)。61株已鉴定细菌中有9株与有效描述的物种的序列同一性低于97%,很可能代表新的分类单元。通过CARD-FISH计算不同样本、地点和季节中的细菌数量,占原核生物总数的54.3%至78.9%。总之,稀释至灭绝技术可以作为基于经典培养方法的补充方法,但都无法培养出DGGE检测到的主要分类单元。细菌群落受到物理化学参数(特别是盐度和氧气)、采样地点和季节的显著影响。