Multidrug-resistant epi-endophytic bacterial community in Posidonia oceanica of Mahdia coast as biomonitoring factor for antibiotic contamination.

Faced with the significant disturbances, mainly of anthropogenic origin, which affect the Mediterranean coastal ecosystem, Posidonia oceanica (L.) Delile has often been used to assess the state of health of this environment. The present study aims to determine the multidrug resistance patterns among isolated and identified epi-endophytic bacterial strains in P. oceanica seagrass collected from Mahdia coastal seawater (Tunisia). To investigate the bacterial community structure and diversity from coastal seawater samples from Mahdia, total DNA extraction and 16S rRNA gene amplification were performed and analyzed by denaturing gradient gel electrophoresis (DGGE). The DGGE profiles showed that some bands were specific to a given site, while other bands were found to be common to more than one sample. In the other hand, bacterial strains were isolated from 1 mL of leaves and epiphytes suspension of P. oceanica seagrass in marine agar. Forty-three isolates were obtained, seven of them were selected and identified on the basis of 16S rRNA gene sequence analysis. These isolates belonged to the genus Bacillus, exhibiting 98-100% of identity with known sequences. Susceptibility patterns of these strains were studied toward commonly used antibiotics in Tunisia. All identified isolates were resistant to Aztreonam (72.1%), Ceftazidime (60.5%), Amoxicillin (56%) and Rifampicin (51.2%). S5-L13 strain had presented the highest multidrug resistance with a MAR index of 0.67.

Terrestrial-type nitrogen-fixing symbiosis between seagrass and a marine bacterium.

Symbiotic N2-fixing microorganisms have a crucial role in the assimilation of nitrogen by eukaryotes in nitrogen-limited environments1-3. Particularly among land plants, N2-fixing symbionts occur in a variety of distantly related plant lineages and often involve an intimate association between host and symbiont2,4. Descriptions of such intimate symbioses are lacking for seagrasses, which evolved around 100 million years ago from terrestrial flowering plants that migrated back to the sea5. Here we describe an N2-fixing symbiont, 'Candidatus Celerinatantimonas neptuna', that lives inside seagrass root tissue, where it provides ammonia and amino acids to its host in exchange for sugars. As such, this symbiosis is reminiscent of terrestrial N2-fixing plant symbioses. The symbiosis between Ca. C. neptuna and its host Posidonia oceanica enables highly productive seagrass meadows to thrive in the nitrogen-limited Mediterranean Sea. Relatives of Ca. C. neptuna occur worldwide in coastal ecosystems, in which they may form similar symbioses with other seagrasses and saltmarsh plants. Just like N2-fixing microorganisms might have aided the colonization of nitrogen-poor soils by early land plants6, the ancestors of Ca. C. neptuna and its relatives probably enabled flowering plants to invade nitrogen-poor marine habitats, where they formed extremely efficient blue carbon ecosystems7.

Production and identification of two antifungal terpenoids from the Posidonia oceanica epiphytic Ascomycota Mariannaea humicola IG100.

BACKGROUND: Marine fungi are an important repository of bioactive molecules with great potential in different technological fields, the annual number of new compounds isolated from marine fungi is impressive and the general trend indicates that it is still on the rise. In this context, the antifungal and antimicrobial activity of the marine strain Mariannaea humicola IG100 was evaluated and two active terpenoids were isolated and characterized. METHODS: Preliminary screening of activity of marine strain IG100 was carried out by agar plug diffusion methods against fungal (Penicillium griseofulvum TSF04) and bacterial (Bacillus pumilus KB66 and Escherichia coli JM109) strains. Subsequently, inhibition tests were done by using the cultural broth and the organic extract (ethyl acetate, EtOAc) by the agar well diffusion methods. The main active fractions were identified and tested for their antifungal activity against P. griseofulvum TSF04 in a 24 wells microplate at different concentrations (1000, 100, 10 and 1.0 µg/mL). Two active compounds were characterized and their relative MIC measured by the broth micro-dilution methods in a 96-well microplate against Aspergillus flavus IG133, P. griseofulvum TSF04, and Trichoderma pleuroticola IG137. RESULTS: Marine strain IG100 presented significant antifungal activity associated with two active compounds, the terpenoids terperstacin 1 and 19-acetyl-4-hydroxydictyodiol 2. Their MIC values were measured for A. flavus (MIC of 7.9 µg/mL and 31.3 µg/mL for 1 and 2, respectively), P. griseofulvum (MIC of 25 µg/mL and 100 µg/mL for 1 and 2, respectively) and T. pleuroticola (MIC > 500 µg/mL and 125 µg/mL for 1 and 2, respectively). They showed a rather good fungistatic effect. CONCLUSIONS: In this study, the first marine strain of M. humicola (IG100) was investigated for the production of bioactive molecules. Strain IG100 produced significant amounts of two bioactive terpenoids, terperstacin 1 and 19-acetyl-4-hydroxydictyodiol 2. The two compounds showed significant antifungal activities against A. flavus IG133, T. pleuroticola IG137 and P. griseofulvum TSF04. Compound 2 was identified for the first time in fungi.

Distribution of Toxigenic Halomicronema spp. in Adjacent Environments on the Island of Ischia: Comparison of Strains from Thermal Waters and Free Living in Posidonia Oceanica Meadows.

Organisms adaptable to extreme conditions share the ability to establish protective biofilms or secrete defence toxins. The extracellular substances that are secreted may contain monosaccharides and other toxic compounds, but environmental conditions influence biofilm characteristics. Microorganisms that are present in the same environment achieve similar compositions, regardless of their phylogenetic relationships. Alternatively, cyanobacteria phylogenetically related may live in different environments, but we ignore if their physiological answers may be similar. To test this hypothesis, two strains of cyanobacteria that were both ascribed to the genus Halomicronema were isolated. H. metazoicum was isolated in marine waters off the island of Ischia (Bay of Naples, Italy), free living on leaves of Posidonia oceanica. Halomicronema sp. was isolated in adjacent thermal waters. Thus, two congeneric species adapted to different environments but diffused in the same area were polyphasically characterized by microscopy, molecular, and toxicity analyses. A variable pattern of toxicity was exhibited, in accordance with the constraints imposed by the host environments. Cyanobacteria adapted to extreme environments of thermal waters face a few competitors and exhibit a low toxicity; in contrast, congeneric strains that have adapted to stable and complex environments as seagrass meadows compete with several organisms for space and resources, and they produce toxic compounds that are constitutively secreted in the surrounding waters.

Combination of heterotrophic nitrifying bacterium and duckweed (Lemna gibba L.) enhances ammonium nitrogen removal efficiency in aquaculture water via mutual growth promotion.

We created a combined system using duckweed and bacteria to enhance the efficiency of ammonium nitrogen (NH4+-N) and total nitrogen (TN) removal from aquaculture wastewater. Heterotrophic nitrifying bacterium was isolated from a sediment sample at an intensive land-based aquaculture farm. It was identified as Acinetobacter sp. strain A6 based on 16S rRNA gene sequence (accession number MF767879). The NH4+-N removal efficiency of the strain and duckweed in culture media and sampled aquaculture wastewater at 15°C was over 99% without any accumulation of nitrite or nitrate. This was significantly higher than strain A6 or duckweed alone. Interestingly, the presence of NO3- increased NH4+-N removal rate by 35.17%. Strain A6 and duckweed had mutual growth promoting-effects despite the presence of heavy metals and antibiotics stresses. In addition, strain A6 colonized abundantly and possibly formed biofilms in the inner leaves of duckweed, and possessed indoleacetic acid (IAA)- and siderophore-producing characteristics. The mutual growth promotion between strain A6 and duckweed may be the reason for their synergistic action of N removal.

First evidence of Halomicronema metazoicum (Cyanobacteria) free-living on Posidonia oceanica leaves.

Cyanobacteria contribute to the ecology of various marine environments, also for their symbioses, since some of them are common hosts of sponges and ascidians. They are also emerging as an important source of novel bioactive secondary metabolites in pharmacological (as anticancer drugs) and biotechnological applications. In the present work we isolated a cyanobacteria in a free-living state from leaves of the seagrass Posidonia oceanica leaves. This newly collected strain was then cultivated under two laboratory conditions, and then characterized by combining morphological observation and molecular studies based on 16S rRNA gene sequences analysis. The strain showed 99% pairwise sequence identity with Halomicronema metazoicum ITAC101, never isolated before as a free-living organisms, but firstly described as an endosymbiont of the Mediterranean marine spongae Petrosia ficiformis, under the form of a filamentous strain. Further studies will investigate the actual role of this cyanobacterium in the leaf stratum of P. oceanica leaves, given its demonstrated ability to influence the vitality and the life cycle of other organisms. In fact, its newly demonstrated free-living stage, described in this study, indicate that Phormidium-like cyanobacteria could play important roles in the ecology of benthic and planktonic communities.

Gold nanoparticle biodissolution by a freshwater macrophyte and its associated microbiome.

Predicting nanoparticle fate in aquatic environments requires mimicking of ecosystem complexity to observe the geochemical processes affecting their behaviour. Here, 12 nm Au nanoparticles were added weekly to large-scale freshwater wetland mesocosms. After six months, ~70% of Au was associated with the macrophyte Egeria densa, where, despite the thermodynamic stability of Au0 in water, the pristine Au0 nanoparticles were fully oxidized and complexed to cyanide, hydroxyls or thiol ligands. Extracted biofilms growing on E. densa leaves were shown to dissolve Au nanoparticles within days. The Au biodissolution rate was highest for the biofilm with the lowest prevalence of metal-resistant taxa but the highest ability to release cyanide, known to promote Au0 oxidation and complexation. Macrophytes and the associated microbiome thus form a biologically active system that can be a major sink for nanoparticle accumulation and transformations. Nanoparticle biotransformation in these compartments should not be ignored, even for nanoparticles commonly considered to be stable in the environment.

Ontogenetic transition from specialized root hairs to specific root-fungus symbiosis in the dominant Mediterranean seagrass Posidonia oceanica.

Terrestrial plants typically take up nutrients through roots or mycorrhizae while freshwater plants additionally utilize leaves. Their nutrient uptake may be enhanced by root hairs whose occurrence is often negatively correlated with mycorrhizal colonization. Seagrasses utilize both leaves and roots and often form root hairs, but seem to be devoid of mycorrhizae. The Mediterranean seagrass Posidonia oceanica is an exception: its adults commonly lack root hairs and regularly form a specific association with a single pleosporalean fungus. Here we show that at two sites in the southern Adriatic, all its seedlings possessed abundant root hairs with peculiar morphology (swollen terminal parts) and anatomy (spirally formed cell walls) as apparent adaptations for better attachment to the substrate and increase of breaking strain. Later on, their roots became colonized by dark septate mycelium while root hairs were reduced. In adults, most of terminal fine roots possessed the specific fungal association while root hairs were absent. These observations indicate for the first time that processes regulating transition from root hairs to root fungal colonization exist also in some seagrasses. This ontogenetic shift in root traits may suggests an involvement of the specific root symbiosis in the nutrient uptake by the dominant Mediterranean seagrass.

Microorganisms facilitate uptake of dissolved organic nitrogen by seagrass leaves.

Microorganisms play a critical role in nitrogen cycling by mineralising dissolved organic nitrogen (DON) compounds into bioavailable inorganic forms (DIN). Although DIN is crucial for seagrass growth, the hypothesis that seagrass leaf associated-microorganisms could convert DON to forms available for plant uptake has never been tested. We conducted a laboratory-based experiment in which seagrass (Posidonia sinuosa) leaves were incubated with 15N-amino acids (aa), with and without associated microorganisms. Samples were collected after 0.5, 2, 6 and 12 h. Both bulk stable isotope and nanoscale secondary ion mass spectrometry (NanoSIMS) analysis showed high accumulation of 15N within seagrass leaf tissues with an associated microbiota, but not in plants devoid of microorganisms. These results significantly change our understanding of the mechanisms of seagrass nitrogen use and provide evidence that seagrass microbiota increase nitrogen availability for uptake by seagrass leaves by mineralising aa, thus enhancing growth and productivity of these important coastal ecosystems.

Different Approaches to Discover Mycovirus Associated to Marine Organisms.

Here we describe the protocols to characterize the virome associated to fungi isolated from marine organisms assessed on the seagrass Posidonia oceanica and on the marine animal Holothuria poli. We provide detailed protocols for fungal isolation, fungal growth, and total RNA extraction. Ribosomal RNA depletion, cDNA library synthesis and normalization, and sequencing runs on different platforms are part of the protocols that are generally outsourced and therefore are not described in this chapter. We describe, instead, how raw reads are assembled into contigs and how to search for putative viral sequences. Furthermore, we detail qualitative checks to infer the existence of the virus as a replicative biological entity.

Sediment anoxia limits microbial-driven seagrass carbon remineralization under warming conditions.

Seagrass ecosystems are significant carbon sinks, and their resident microbial communities ultimately determine the quantity and quality of carbon sequestered. However, environmental perturbations have been predicted to affect microbial-driven seagrass decomposition and subsequent carbon sequestration. Utilizing techniques including 16S-rDNA sequencing, solid-state NMR and microsensor profiling, we tested the hypothesis that elevated seawater temperatures and eutrophication enhance the microbial decomposition of seagrass leaf detritus and rhizome/root tissues. Nutrient additions had a negligible effect on seagrass decomposition, indicating an absence of nutrient limitation. Elevated temperatures caused a 19% higher biomass loss for aerobically decaying leaf detritus, coinciding with changes in bacterial community structure and enhanced lignocellulose degradation. Although, community shifts and lignocellulose degradation were also observed for rhizome/root decomposition, anaerobic decay was unaffected by temperature. These observations suggest that oxygen availability constrains the stimulatory effects of temperature increases on bacterial carbon remineralization, possibly through differential temperature effects on bacterial functional groups, including putative aerobic heterotrophs (e.g. Erythrobacteraceae, Hyphomicrobiaceae) and sulfate reducers (e.g. Desulfobacteraceae). Consequently, under elevated seawater temperatures, carbon accumulation rates may diminish due to higher remineralization rates at the sediment surface. Nonetheless, the anoxic conditions ubiquitous to seagrass sediments can provide a degree of carbon protection under warming seawater temperatures.

High rates of microbial dinitrogen fixation and sulfate reduction associated with the Mediterranean seagrass Posidonia oceanica.

Seagrass meadows of Posidonia oceanica represent hotspots of productivity in the oligotrophic Mediterranean Sea. The lack of dissolved inorganic nitrogen (DIN) in the seawater suggests that the N-demand of these meadows might be in part supported by microbial dinitrogen (N2) fixation. However, currently there are no direct N2 fixation measurements available for this productive marine macrophyte. Here we investigated N2 fixation activity associated with P. oceanica leaf, rhizome and root pieces. In 15N2 incubations, the roots exhibited highest rates of N2 fixation. The rates varied considerably between replicates, presumably due to a patchy microbial colonization of the roots. Additions of organic carbon compounds (acetate, glucose, sucrose or algal lysate) did not enhance the N2 fixation rates. Sulfate reduction rates measured alongside were also highest in root incubations. Correspondingly, sequences of the nifH gene (a marker gene for the iron protein of the N2-fixing enzyme nitrogenase) related to known sulfate-reducing bacteria were retrieved from P. oceanica roots. Other nifH sequences clustered with known heterotrophic diazotrophs previously identified in other marine macrophytes. In particular, many sequences obtained from P. oceanica roots were similar (>94%) to a saltmarsh rhizosphere-associated heterotrophic diazotroph, indicating that heterotrophic lifestyle might be common among marine macrophyte-associated diazotrophs.

Communities of Cultivable Root Mycobionts of the Seagrass Posidonia oceanica in the Northwest Mediterranean Sea Are Dominated by a Hitherto Undescribed Pleosporalean Dark Septate Endophyte.

Seagrasses, a small group of submerged marine macrophytes, were reported to lack mycorrhizae, i.e., the root-fungus symbioses most terrestrial plants use for nutrient uptake. On the other hand, several authors detected fungal endophytes in seagrass leaves, shoots, rhizomes, and roots, and an anatomically and morphologically unique dark septate endophytic (DSE) association has been recently described in the roots of the Mediterranean seagrass Posidonia oceanica. Nevertheless, the global diversity of seagrass mycobionts is not well understood, and it remains unclear what fungus forms the DSE association in P. oceanica roots. We isolated and determined P. oceanica root mycobionts from 11 localities in the northwest Mediterranean Sea with documented presence of the DSE association and compared our results with recent literature. The mycobiont communities were low in diversity (only three species), were dominated by a single yet unreported marine fungal species (ca. 90 % of the total 177 isolates), and lacked common terrestrial and freshwater root mycobionts. Our phylogenetic analysis suggests that the dominating species represents a new monotypic lineage within the recently described Aigialaceae family (Pleosporales, Ascomycota), probably representing a new genus. Most of its examined colonies developed from intracellular microsclerotia occupying host hypodermis and resembling microsclerotia of terrestrial DSE fungi. Biological significance of this hitherto overlooked seagrass root mycobiont remains obscure, but its presence across the NW Mediterranean Sea and apparent root intracellular lifestyle indicate an intriguing symbiotic relationship with the dominant Mediterranean seagrass. Our microscopic observations suggest that it may form the DSE association recently described in P. oceanica roots.

Cultivation-dependent analysis of the microbial diversity associated with the seagrass meadows in Xincun Bay, South China Sea.

Microbial communities have largely existed in the seagrass meadows. A total of 496 strains of the bacteria in the seagrass meadows, which belonged to 50 genera, were obtained by the plate cultivation method from three sites of Xincun Bay, South China Sea. The results showed that Bacillales and Vibrionales accounted for the highest proportions of organisms in all communities. The diversity of the bacteria in the sediment was higher than that associated with seagrass. Thalassia hemperichii possessed the highest abundance of bacteria, followed by Enhalus acoroides and Cymodocea rotundata. Robust seasonal dynamics in microbial community composition were also observed. It was found that microbial activities were closely tied to the growth stage of the seagrass. The microbial distribution was the lowest in site 3. The abundance of the bacteria was linked to the interactions between bacteria and plants, the condition of plant and even the coastal water quality and the nutrition level in the sediment.

Anatomically and morphologically unique dark septate endophytic association in the roots of the Mediterranean endemic seagrass Posidonia oceanica.

Roots of terrestrial plants host a wide spectrum of soil fungi that form various parasitic, neutral and mutualistic associations. A similar trend is evident in freshwater aquatic plants and plants inhabiting salt marshes or mangroves. Marine vascular plants (seagrasses), by contrast, seem to lack specific root-fungus symbioses. We examined roots of two Mediterranean seagrasses, Posidonia oceanica and Cymodocea nodosa, in the northwestern Mediterranean Sea for fungal colonization using light and scanning and transmission electron microscopy. We found that P. oceanica, but not C. nodosa, is regularly associated with melanized septate hyphae in a manner resembling colonization by the ubiquitous dark septate endophytes (DSE) in roots of most terrestrial plants. P. oceanica roots were found to be colonized by sparse dematiaceous running hyphae as well as dense parenchymatous nets/hyphal sheaths on the root surface, intracellular melanized microsclerotia and occasionally also intra- and intercellular hyphae. The colonization was most prominent in the thick-walled hypodermis of the thinnest healthy looking roots, and the mycobiont seemed to colonize both living and dead host cells. Dark septate hyphae infrequently occurred also inside rhizodermal cells, but never colonized vascular tissues. The biological significance of this overlooked marine symbiosis remains unknown, but its morphology, extent, distribution across the NW Mediterranean Sea and absence in C. nodosa indicate an intriguing relationship between the dominant Mediterranean seagrass and its dark septate root mycobionts.

Lulwoana sp., a dark septate endophyte in roots of Posidonia oceanica (L.) Delile seagrass.

Posidonia oceanica is the most common, widespread and important monocotyledon seagrass in the Mediterranean Basin, and hosts a large biodiversity of species, including microorganisms with key roles in the marine environment. In this study, we ascertain the presence of a fungal endophyte in the roots of P. oceanica growing on different substrata (rock, sand and matte) in two Sicilian marine meadows. Staining techniques on root fragments and sections, in combination with microscope observations, were used to visualise the fungal presence and determine the percentage of fungal colonisation (FC) in this tissue. In root fragments, statistical analysis of the FC showed a higher mean in roots anchored on rock than on matte and sand. In root sections, an inter- and intracellular septate mycelium, producing intracellular microsclerotia, was detected from the rhizodermis to the vascular cylinder. Using isolation techniques, we obtained, from both sampling sites, sterile, slow-growing fungal colonies, dark in colour, with septate mycelium, belonging to the dark septate endophytes (DSEs). DNA sequencing of the internal transcribed spacer (ITS) region identified these colonies as Lulwoana sp. To our knowledge, this is the first report of Lulwoana sp. as DSE in roots of P. oceanica. Moreover, the highest fungal colonisation, detected in P. oceanica roots growing on rock, suggests that the presence of the DSE may help the host in several ways, particularly in capturing mineral nutrients through lytic activity.

Bathymetric variation of epiphytic assemblages on Posidonia oceanica (L.) Delile leaves in relation to anthropogenic disturbance in the southeastern Mediterranean.

A survey of the epiphytic leaves of Posidonia oceanica was conducted along a depth transect at both the control station Attaya in the Kerkennah Islands and the disturbed Mahres station on the Sfax coast (Tunisia). Samples were collected by scuba divers at depths of 5, 10, 15, and 20 m in July 2008. We evaluated whether the pattern of spatial variability of the macroepiphyte assemblages of leaves of Posidonia oceanica differed in relation to anthropogenic interference. The results indicate that the decrease in shoot density and leaf length according to depth was low at Mahres. The biomass of epiphytic leaves and the percentage cover of epiphytic assemblages decreased with depth for both stations and heavily at Mahres, this decline being related to anthropogenic disturbance. This study shows that the highest values of epifauna and epiflora were detected at the disturbed station Mahres. Macroalgae assemblages decreased with depth at both stations and were dominated by Rhodophyta, whereas the percentage cover of the epifauna leaf that decreases according to depth was dominated by Hydrozoa and Bryozoa. Changes in epiphyte assemblages, epiphytic biomass, percentage cover, and species richness in proportion to Heterokontophyta, Rhodophyta, Cyanobacteria, Hydrozoa, Porifera, and Tunicata between the two stations constitute promising tools for detecting environmental disturbance.

Epiphytic cyanobacteria of the seagrass Cymodocea rotundata: diversity, diel nifH expression and nitrogenase activity.

Seagrasses are photoautotrophic, ecologically important components of many globally widespread coastal ecosystems, in which combined nitrogen may limit their production. We examined the biodiversity and diazotrophic capacity of microbial epiphytes associated with the phyllosphere of the seagrass Cymodocea rotundata of the Western Indian Ocean. Light microscopy, 16S rRNA and nifH gene analysis revealed the dominance of cyanobacteria in the epiphytic microbial community. Most phylotypes were related to free-living uncultured benthic cyanobacteria, while some to cyanobacterial endosymbionts of marine diatoms. Novel and potentially diazotrophic species, some of known pantropical distribution, were also discovered. Significant diel nitrogenase activities (acetylene reduction assay) were recorded (up to 358 ± 232 nmol C2H4 g(-1) of seagrass FW h(-1)). The nifH gene expression patterns showed that heterocystous phylotypes may be the dominant diazotrophs during the day and non-heterocystous at night. These data show that C. rotundata is colonized by diverse diazotrophic cyanobacteria species and suggest that these may be beneficial partners of seagrasses in nitrogen-depleted waters.

Diversity, ecological role and potential biotechnological applications of marine fungi associated to the seagrass Posidonia oceanica.

The marine environment is characterized by high salinity and exerts a strong selective pressure on the biota, favouring the development of halo-tolerant microorganisms. Part of this microbial diversity is made up of fungi, important organisms from ecological and biotechnological points of view. In this study, for the first time, the qualitative and quantitative composition of the mycoflora associated to leaves, rhizomes, roots and matte of the seagrass Posidonia oceanica was estimated. A total of 88 fungal taxa, mainly belonging to Ascomycota, were identified by morphological and molecular methods. The most represented genera were Penicillium, Cladosporium and Acremonium. Most of the species (70) were selectively associated with one district; only two species (Penicillium chrysogenum var. chrysogenum and P. janczewskii) were isolated from all the districts. Moreover the capability to produce laccases, peroxidases and tannases by 107 fungal isolated by the different districts of P. oceanica was carried out. These results show that the mycoflora associated to P. oceanica is very rich and characterized by fungi able to produce ligninolytic enzymes and tannases useful to degrade and detoxify lignocellulose residues in presence of high salt concentrations. These fungi, hence, may play important ecological roles in marine environments but can also be very useful in different biotechnological areas.

Antagonistic properties of seagrass associated Streptomyces sp. RAUACT-1: a source for anthraquinone rich compound.

OBJECTIVE: To identify the antibacterial potential of seagrass (Syringodium isoetifolium) associate microbes against bacterial pathogens. METHODS: Eumeration of microbial associates were analyzed with leaf and root samples of Syringodium isoetifolium. MIC and MBC were calculated for bacterial pathogens with microbial associates. Phylogenetic and GC-MS analysis were calculated for Actinomycetes sp. (Act01) which was the most potent. RESULTS: Of the isolated microbial associates phosphatase producing bacterial isolates were identified as maximum [(261.78±35.09) CFU×10(4)/g] counts in root sample. Of the selected microbial isolates Actinomycete sp (Act01) showed broad spectrum of antibacterial activity against antibiotic resistant and fish bacterial pathogens. Phylogenetic analysis of Act01 showed maximum identities (99%) with the Streptomyces sp. (GU5500072). The 16s rDNA secondary structure of Act01 showed the free energy values as -366.3 kkal/mol. The GC-MS analysis Act01 showed maximum retention value with 23.742 RT and the corresponding chemical class was identified as 1, 4-dihydroxy-2-(3-hydroxybutyl)-9, 10-anthraquinone 9, 10-anthrac. CONCLUSIONS: In conclusion, Streptomyces sp. (GU045544.1) from Syringodium isoetifolium could be used as potential antibacterial agent.