Studies on antagonistic marine actinomycetes from the Bay of Bengal

Screening of 26 marine sediment samples near 9 islands of the Andaman Coast of the Bay of Bengal resulted in the isolation of 88 isolates of actinomycetes. On the basis of sporophore morphology and structure of the spore chain, 64 isolates were assigned to the genus Streptomyces, 8 isolates to the genus Micromonospora, 5 to the genus Nocardia, 7 to the genus Streptoverticilium and 4 to the genus Saccharopolyspora. Among 64 Streptomyces spp., 44 isolates showed antibacterial activity and 17 isolates showed antifungal activity. Three isolates showed very promising antagonistic activities against multi-drug resistant pathogens.     

Diversity and exploration of bioactive marine actinomycetes in the Bay of Bengal of the Puducherry coast of India

The present study was designed to investigate the Puducherry coast of the Bay of Bengal, India for the diversity of bioactive actinomycetes. A total of 50 actinomycete strains were isolated from the marine sediments and most of the strains were belongs to Streptomyces. These strains were identified by means of morphological physiological, biochemical and cultural characteristics. The isolates were subjected to shake flask fermentation and the secondary metabolites were extracted with ethyl acetate and screened for cytotoxicity, hemolytic activity and antimicrobial activity against selected bacterial and fungal pathogens. The cytotoxic activity was evaluated using HeLa cell lines by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, a tetrazole (MTT) assay, hemolytic activity on mouse erythrocytes and the antifungal activity was evaluated by MTT cytotoxic assay against Aspergillus niger, Aspergillus fumigatus and Candida albicans. The antibacterial activity was studied against Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Klebsiella pneumoniae. The cytotoxicity and antimicrobial activity of secondary metabolite was found to be concentration dependent and nearly 24% of isolates showed significant antimicrobial, hemolytic and cytotoxic activity. The results of our study indicate the diversity and bioactive potential of marine actinomycetes isolated in the Puducherry coast.     

海洋抑菌放线菌Y15的筛选、鉴定及其代谢产物理化性质

【目的】筛选产广谱、高效抑菌活性物质的海洋放线菌,为新型抗生素的开发奠定基础。【方法】采用琼脂块法初筛,打孔扩散法复筛,以金黄色葡萄球菌、大肠埃希氏菌、枯草芽孢杆菌和单核细胞增生李斯特菌为指示菌从海泥样品中筛选目标菌株;利用20种指示菌株考查Y10、Y11、Y15、Y16和Y21的抑菌谱;通过形态观察和16S r RNA基因序列分析确定菌株分类地位;以单核细胞增生李斯特菌的OD600值为指标探讨Y15抑菌活性物质对该菌的作用方式;以抑菌活性为指标研究Y15抑菌活性物质的理化性质。【结果】共分离12株具抑菌活性的海洋放线菌,其中Y15抑菌谱最广,对20种指示菌株中的18种具有抑菌活性,并且在4种培养基中均能产生抑菌活性物质。16S r RNA基因序列分析表明Y15属于小单孢菌属,并与Micromonospora endolithica亲缘关系最近。Y15抑菌活性物质在144 h达到最高值为480 AU/m L,在168-216 h保持平衡为320 AU/m L。Y15抑菌活性物质对单核细胞李斯特菌作用方式为杀菌。Y15抑菌活性物质在-20-60°C抑菌活性保持稳定,在80-120°C活性逐渐下降,但在120°C处理30 min仍保留37.5%的活性;在p H 7.0-10.0抑菌活性稳定,在p H 2.0-6.0和p H 11.0-12.0抑菌活性均有所损失;Y15抑菌活性物质对紫外和4种酶(蛋白酶K、胰蛋白酶、木瓜蛋白酶和α-淀粉酶)处理均保持稳定,表明活性物质为非蛋白和非多肽类的抑菌物质。【结论】Y15产生的活性物质具有良好的抑菌谱和抑菌活性,且较为稳定,具有较高的应用价值。     

Tetrodotoxin production of actinomycetes isolated from marine sediment

Ten actinomycetes isolated from various marine sediments were investigated for the production of tetrodotoxin (TTX). Tissue culture assay, high performance liquid chromatography and gas chromatography-mass spectrometry confirmed that the nine strains produced TTX. Actinomycetes may be responsible for TTX accumulation in marine environments.     

植物叶片内生放线菌的分离、分类与拮抗活性

以四川采集的芍药和北京采集的三叶草的叶片为材料,经表面消毒程序后,分离出内生放线菌15株。通过表观特征比较和BOX-PCR基因指纹图谱分析的方法,将分离出的放线菌归并于12个不同的基因型,其中6个来自芍药,6个来自三叶草。结合16S rRNA基因序列分析可知,分离菌株除C4、C5属于假诺卡氏菌外,其余的13株都是链霉菌;菌株C12与最近模式种的16S rRNA基因序列相似性较低,为96.6%。对分离菌株的抗菌活性实验表明,有11株在一个或一个以上的测试中呈阳性;其中6株对植物致病菌立枯丝核菌有明显抗性,占阳性菌株的55%。     

灯台树内生放线菌多样性及抗菌活性评价

从云南西双版纳采集7份灯台树样品,经过表面消毒,用4种分离培养基分离得到105株内生放线菌。经16S rRNA基因序列分析鉴定,分布于7个科9个属。利用5种植物病原真菌指示菌对所有菌株的发酵液进行抗菌活性检测。结果显示,有12.4%、14.3%、11.4%、12.4%、8.6%的菌株分别对镰刀霉、疫霉、赤星霉、苹果炭疽、白色念珠菌有抗性。对3株具有广谱抗菌活性的菌株进行再次发酵和抗菌活性复筛,结果显示这3株的抗菌活性稳定,并可能含有生物碱类化合物。     

Length weight relationships of five marine fishes from the Bay of Bengal

Length weight relationships (LWR) of five marine fishes were estimated from the Bay of Bengal, Bangladesh. Fish samples were collected occasionally from fisher's catch with gill net (mesh size: 3 cm) during November 2019 to October 2020. Fishing was done overnight. Total length (0.1 cm) and body weight (0.01 g) of each individual were measured. We recorded maximum total length for P. maculatus (55.0 cm) and P. parmatus (39.0 cm).     

Filamentous fungal population and species diversity from the continental slope of Bay of Bengal,India

Filamentous fungal diversity from the sediments of the continental slope of Bay of Bengal was studied. Sediment samples were collected during two voyages in 2004 and 2005. Filamentous fungal population from both the cruises showed a range of 5.17–59.51 CFU/g and 3.47–29.68 CFU/g, respectively. Totally 16 fungal genera were recorded from both the cruises. Aspergillus was found to be the dominant genus and the overall percentage occurrence was as follows: Deuteromycotina 74%, Ascomycotina 17%, Basidiomycotina 4% and non-sporulating 5%. Diversity indices were calculated and during both the cruises species richness (d) varied from 0.912 to 3.622 and 1.443 to 4.588; evenness (J′) varied from 0.9183 to 1.000 and 0.8322 to 1.000 and Shannon–Wiener index (H′ log 2) varied from 0.9183 to 1.000 and 1.000 to 3.690. The higher diversity was found in Divipoint transect (northern Bay of Bengal). 95% confidence interval and ellipse showed that the stations were well lying within the funnel. Cluster analysis and MDS grouped the northern transects which showed higher diversity. BVSTEP resulted in isolation of 23 species which were most influential in the marine filamentous fungal diversity of the continental slope of Bay of Bengal. Thus, a lower population range and higher diversity of marine filamentous marine fungi in the sediments of the continental slope of Bay of Bengal was recorded.     

药用昆虫喙尾琵琶甲肠道抗菌活性放线菌的筛选及鉴定

【目的】探究药用昆虫喙尾琵琶甲Blaps rynchopetera肠道具有抗菌活性的放线菌,为抗菌药物开发提供新的放线菌资源。【方法】结合稀释涂布法和选择培养法从喙尾琵琶甲成虫肠道分离放线菌。以耐甲氧西林金黄色葡萄球菌(methieillin-resistant Staphylococus aureus)、金黄色葡萄球菌、大肠杆菌Escherichia coli、粪肠球菌Enterococcus faecalis、鼠伤寒沙门氏菌Salmonella typhimurium 和铜绿假单胞菌Pseudomonas aeruginosa 6株致病细菌以及黑曲霉Aspergillus niger、青霉菌Penicillium expansum和白色念珠菌Canidia albicans 3株致病真菌为指示菌,通过牛津杯琼脂扩散法测试放线菌次生代谢产物的抗菌活性。随后,采用分子生物学方法进行16S rRNA序列分析鉴定活性显著的18株放线菌并构建系统发育树。【结果】从喙尾琵琶甲成虫肠道中分离到176株共生放线菌,初步活性筛选结果显示其中46株放线菌表现出不同程度的抗菌活性。多株放线菌对致病菌具有广谱的抑菌作用且抑菌圈直径超过阳性对照药。选择抑菌圈直径大于15 mm的18株放线菌进行分子鉴定,结果显示均为链霉属Streptomyces。【结论】喙尾琵琶甲肠道含有丰富的抗菌活性放线菌资源。     

Diversity and antimicrobial activity of endophytic fungi isolated from Nyctanthes arbor-tristis, a well-known medicinal plant of India

Endophytic fungi from Nyctanthes arbor-tristis were isolated and evaluated for their antimicrobial activity. A total of 19 endophytic fungi were isolated from 400 segments of healthy leaf and stem tissues of N. arbor-tristis. Eighteen endophytic fungi were obtained from leaf, while only ten from stem. Alternaria alternata had the highest colonization frequency (15.0%) in leaf, whereas Cladosporium cladosporioides ranked first in stem with a colonization frequency of 12%. The diversity and species richness were found higher in leaf tissues than in stem. The similarity indices between leaf and stem were 0.473 for Jaccard’s and 0.642 for the Sorenson index, respectively. Of 16, 12 (75%) endophytic fungal extracts showed antibacterial activity against either one or more pathogenic bacteria. The endophytic Nigrospora oryzae showed maximum inhibition against Shigella sp. and Pseudomonas aeruginosa. The leaf endophytes Colletotrichum dematium and Chaetomium globosum exhibited a broad range of anibacterial activity and were active against Shigella flexnii, Shigella boydii, Salmonella enteritidis, Salmonella paratyphi, and P. aeruginosa. Nine out of 16 (56.25%) endophytic fungi exhibited antifungal activity to one or more fungal pathogens. Colletotrichum dematium inhibited 55.87% of the radial growth of the phytopathogen Curvularia lunata. The antimicrobial activity of these endophytic microorganisms could be exploited in the biotechnological, medicinal, and agricultural industries.     

Photobacterium marinum sp. nov., a marine bacterium isolated from a sediment sample from Palk Bay,India

The novel, cream colored, Gram-staining-negative, rod-shaped, motile bacteria, designated strains AK15^T and AK18, were isolated from sediment samples collected from Palk Bay, India. Both strains were positive for arginine dihydrolase, lysine decarboxylase, oxidase, nitrate reduction and methyl red test. The major fatty acids were C_16:0, C_{18:1 ω7c}, C_{16:1 ω7c} and/or C_{16:1 ω6c} and/or iso-C_15:0 2-OH (summed feature 3). Polar lipids content of strains AK15^T and AK18 were found to bephosphatidylethanolamine (PE), two unidentified phospholipids (PL1 and PL2) and three unidentified lipids (L1-L3). The 16S rRNA gene sequence analysis indicated strains AK15^T and AK18 as the members of the genus Photobacterium and closely related to the type strain Photobacterium jeanii with pair-wise sequence similarity of 96.7%. DNA–DNA hybridization between strain AK15^T and AK18 showed a relatedness of 87%. Based on data from the current polyphasic study, strains AK15^T and AK18 are proposed as novel species of the genus Photobacterium, for which the name Photobacterium marinum sp. nov. is proposed. The type strain of Photobacterium marinum is AK15^T (=MTCC 11066^T = DSM 25368^T).     

Screening of marine actinobacteria for amylase enzymes inhibitors

Amylase inhibitor producing actinobacteria were isolated and characterized from terrestrial environment and there is no much report found from marine environment, hence in the present study, 17 strains isolated from the rhizosphere sediments of mangroves were tested for their amylase inhibition ability. Seawater requirement test for the growth of actinobacteria found that the strains SSR-3, SSR-12 and SSR-16 requires at least 50% and SSR-6 requires at least 25% seawater for their growth. The inhibition activity of both prokaryotic and eukaryotic amylase was tested by using Bacillus subtilis and Aspergillus niger. The maximum amylase activity (40mm) produced by the A. niger was taken as positive control, when the test actinobacteria strains grown in the medium they inhibited amylase activity and was evidenced by the reduction in inhibition zone (14–37 mm) similarly the amylase produced by the Bacillus subtilis was also recorded maximum (35 mm) amylase activity and was taken as positive control, and the test atinobacterial strains reduced enzyme action(12–33 mm) it varied levals. This indicates that the actinobacteria strains were controlled amylase enzyme activity in both the cases. The strain SSR-10 was highly effective and SSR-8 was less effective in inhibiting eukaryotic amylase produced by A. niger. The strain SSR-2 was effective and SSR-6 showed very less effect in inhibiting the prokaryotic amylase produced by the B subtilis.     

番茄灰霉病拮抗内生放线菌的筛选、鉴定及其活性评价

对安徽省淮北市番茄植株根、茎、叶中内生放线菌进行了分离、筛选,并测定了其抑菌活性。结果表明:番茄根、茎和叶中的内生放线菌的数量分别为5.66×104、0.67×104和0.39×104CFU.g-1鲜重。根据分离部位和表型特征,从健康番茄植株体内分离到93株内生放线菌,通过对峙实验,筛选到7株对番茄灰霉病菌有拮抗作用的菌株,占所分离内生放线菌总数的7.5%。来自根组织中的菌株HNU-EA27的抑菌效果最佳,抑菌圈直径达28.3mm。根据形态特征、培养特征、生理生化特性、细胞壁组分和16SrDNA序列分析,将菌株HNU-EA27鉴定为毒三素链霉菌(Streptomyces toxytricini)。室内测定菌株HNU-EA27发酵滤液对灰霉病菌菌丝生长及分生孢子萌发的抑制作用,结果表明:菌株HNU-EA27发酵滤液可以抑制灰霉病菌菌丝生长和分生孢子萌发,且浓度越高,抑制能力越强;当发酵滤液浓度为30%时则完全抑制灰霉病菌菌丝生长和分生孢子萌发。盆栽防效试验结果表明:30%菌株HNU-EA27发酵滤液对番茄灰霉病的预防与治疗效果分别为80.6%和73.8%,均高于50%多菌灵可湿性粉剂600倍液。本研究表明,菌株HNU-EA27是防治番茄灰霉病潜在的优良生防菌株,具有良好的开发应用价值。     

胶州湾沉积物可培养细菌的多样性及其抑菌活性

【目的】海洋微生物在活性物质开发方面具有巨大的应用前景。为了研究胶州湾微生物的多样性和抑菌活性,选取胶州湾9个观测站点的沉积物进行了细菌多样性及抑菌活性分析。【方法】采用YPD和Z2216E培养基分离细菌,通过16S r RNA基因测序对分离菌株进行分类鉴定,继而采用牛津杯法考察分离菌株对7株指示菌的抑菌活性,最后用PCR方法筛选16株代表菌的PKSI、NRPS、CYP、Phz E和d TGD基因。【结果】从胶州湾沉积物中共分离出76株细菌,通过对16S r RNA序列分析,将其归为8个科11个属:节杆菌属、考克氏菌属、微球菌属、微杆菌属、假交替单胞菌属、Oceanisphaera、海单胞菌属、葡萄球菌属、芽孢杆菌属、硫胺素芽孢杆菌属和短芽孢杆菌属,其中分离细菌中有34株至少对1种指示菌有抑菌活性。所选取的16株菌均能检测到一种功能基因,且其中5株菌能同时检测到3种以上的功能基因。【结论】以上研究表明胶州湾海域有丰富的微生物资源,在生物活性次级代谢产物合成上有较大潜力。     

Reproductive biology of the sawtooth barracuda,Sphyraena putnamae,in the Bay of Bengal,India

Environmental Biology of Fishes - With the lack of information available globally on the reproductive biology of Sphyraena putnamae, the present study was performed using 908 individuals landed...     

Patterns of antimicrobial activities from soil actinomycetes isolated under different conditions of pH and salinity

AIMS: To evaluate the patterns of the production of antimicrobial compounds by diverse collection of actinomycetes isolated from different geographies under alternative conditions of pH and salinity in the media. METHODS AND RESULTS: Actinomycetes were grouped based on their method of isolation and their phenotype diversity was determined by total fatty acid analysis. A total of 335 representative isolates, including 235 Streptomyces species and 100 actinomycetes from other taxa, were screened for the production of antimicrobial activities against a panel of bacteria, filamentous fungi and yeasts, including some of clinical relevance. Production of antimicrobial activities was detected in 230 strains. In the case of the genus Streptomyces, 181 antimicrobial activities (77% of the tested isolates) were recorded. The activities observed among the other actinomycetes taxa were lower (49% of the tested isolates). CONCLUSIONS: The results of this study support the idea that species of actinomycetes isolated in alternative selective conditions of pH and salinity present a significant capacity to produce compounds with antibacterial or antifungal activity. The best group of isolates in terms of production of active secondary metabolites was the one isolated in saline conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: The results demonstrate that these actinomycetes strains isolated in alternative selective conditions of pH and salinity and collected from diverse geographical locations present a significant capacity to produce compounds with antibacterial or antifungal activity.     

Screening of selected West Bengal plants for antifungal activity

A systematic screening of 170 different West Bengal plants was carried out to find antifungal antibiotics, usingAspergillus niger andTrichophyton rubrum as test organisms. From them, four plant species, viz.Curcuma zedoaria, C. aromatica, C. amada, and aBrassica sp. have been found to contain active antibiotic principles with strong inhibitory effects against both test organisms.     

A method to type the potential angucycline producers in actinomycetes isolated from marine sponges

Angucyclines are aromatic polyketides with antimicrobial, antitumor, antiviral and enzyme inhibition activities. In this study, a new pair of degenerate primers targeting the cyclase genes that are involved in the aromatization of the first and/or second ring of angucycline, were designed and evaluated in a PCR protocol targeting the jadomycin cyclase gene of Streptomyces venezuelae ISP5230. The identity of the target amplicon was confirmed by sequencing. After validation, the primers were used to screen 49 actinomycete isolates from three different marine sponges to identify putative angucycline producers. Seven isolates were positively identified using this method. Sequence analysis of the positive amplicons confirmed their identity as putative angucycline cyclases with sequence highly similar to known angucycline cyclases. Phylogenetic analysis clustered these positives into the angucycline group of cyclases. Furthermore, amplifications of the seven isolates using ketosynthase-specific primers were positive, backing the results using the cyclase primers. Together these results provided strong support for the presence of angucycline biosynthetic genes in these isolates. The specific primer set targeting the cyclase can be used to identify putative angucycline producers among marine actinobacteria, and aid in the discovery of novel angucyclines.