Graça Ana Patrícia, Viana Flávia, Bondoso Joana, Correia Maria Inês, Gomes Luis, Humanes Madalena, Reis Alberto, Xavier Joana R, Gaspar Helena, Lage Olga M
Department of Biology, Faculty of Sciences, University of Porto Porto, Portugal ; Centre of Marine and Environmental Research (CIIMAR) Porto, Portugal.
Department of Biology, Faculty of Sciences, University of Porto Porto, Portugal ; Centro de Química e Bioquímica e Departamento de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa Lisboa, Portugal.
Front Microbiol. 2015 May 7;6:389. doi: 10.3389/fmicb.2015.00389. eCollection 2015.
Interest in the study of marine sponges and their associated microbiome has increased both for ecological reasons and for their great biotechnological potential. In this work, heterotrophic bacteria associated with three specimens of the marine sponge Erylus deficiens, were isolated in pure culture, phylogenetically identified and screened for antimicrobial activity. The isolation of bacteria after an enrichment treatment in heterotrophic medium revealed diversity in bacterial composition with only Pseudoalteromonas being shared by two specimens. Of the 83 selected isolates, 58% belong to Proteobacteria, 23% to Actinobacteria and 19% to Firmicutes. Diffusion agar assays for bioactivity screening against four bacterial strains and one yeast, revealed that a high number of the isolated bacteria (68.7%) were active, particularly against Candida albicans and Vibrio anguillarum. Pseudoalteromonas, Microbacterium, and Proteus were the most bioactive genera. After this preliminary screening, the bioactive strains were further evaluated in liquid assays against C. albicans, Bacillus subtilis and Escherichia coli. Filtered culture medium and acetone extracts from three and 5 days-old cultures were assayed. High antifungal activity against C. albicans in both aqueous and acetone extracts as well as absence of activity against B. subtilis were confirmed. Higher levels of activity were obtained with the aqueous extracts when compared to the acetone extracts and differences were also observed between the 3 and 5 day-old extracts. Furthermore, a low number of active strains was observed against E. coli. Potential presence of type-I polyketide synthases (PKS-I) and non-ribosomal peptide synthetases (NRPSs) genes were detected in 17 and 30 isolates, respectively. The high levels of bioactivity and the likely presence of associated genes suggest that Erylus deficiens bacteria are potential sources of novel marine bioactive compounds.
由于生态原因及其巨大的生物技术潜力,对海洋海绵及其相关微生物群落的研究兴趣日益增加。在这项工作中,从海洋海绵Erylus deficiens的三个标本中分离出异养细菌,进行纯培养,进行系统发育鉴定并筛选其抗菌活性。在异养培养基中进行富集处理后分离细菌,结果显示细菌组成具有多样性,只有两个标本中存在假交替单胞菌。在83株选定的分离株中,58%属于变形菌门,23%属于放线菌门,19%属于厚壁菌门。针对四种细菌菌株和一种酵母进行生物活性筛选的扩散琼脂试验表明,大量分离出的细菌(68.7%)具有活性,特别是对白色念珠菌和鳗弧菌。假交替单胞菌属、微杆菌属和变形杆菌属是生物活性最强的属。经过初步筛选后,对生物活性菌株在液体试验中针对白色念珠菌、枯草芽孢杆菌和大肠杆菌进行了进一步评估。对3天和5天龄培养物的过滤培养基和丙酮提取物进行了检测。证实了水提取物和丙酮提取物对白色念珠菌均具有高抗真菌活性,而对枯草芽孢杆菌无活性。与丙酮提取物相比,水提取物的活性水平更高,并且在3天和5天龄提取物之间也观察到差异。此外,观察到对大肠杆菌有活性的菌株数量较少。分别在17株和30株分离株中检测到I型聚酮合酶(PKS-I)和非核糖体肽合成酶(NRPSs)基因的潜在存在。高生物活性水平以及相关基因的可能存在表明,Erylus deficiens细菌是新型海洋生物活性化合物的潜在来源。