The field of microbiome research has evolved rapidly over the past few decades and has become a topic of great scientific and public interest. As a result of this rapid growth in interest covering different fields, we are lacking a clear commonly agreed definition of the term "microbiome." Moreover, a consensus on best practices in microbiome research is missing. Recently, a panel of international experts discussed the current gaps in the frame of the European-funded MicrobiomeSupport project. The meeting brought together about 40 leaders from diverse microbiome areas, while more than a hundred experts from all over the world took part in an online survey accompanying the workshop. This article excerpts the outcomes of the workshop and the corresponding online survey embedded in a short historical introduction and future outlook. We propose a definition of microbiome based on the compact, clear, and comprehensive description of the term provided by Whipps et al. in 1988, amended with a set of novel recommendations considering the latest technological developments and research findings. We clearly separate the terms microbiome and microbiota and provide a comprehensive discussion considering the composition of microbiota, the heterogeneity and dynamics of microbiomes in time and space, the stability and resilience of microbial networks, the definition of core microbiomes, and functionally relevant keystone species as well as co-evolutionary principles of microbe-host and inter-species interactions within the microbiome. These broad definitions together with the suggested unifying concepts will help to improve standardization of microbiome studies in the future, and could be the starting point for an integrated assessment of data resulting in a more rapid transfer of knowledge from basic science into practice. Furthermore, microbiome standards are important for solving new challenges associated with anthropogenic-driven changes in the field of planetary health, for which the understanding of microbiomes might play a key role.
As an alternative strategy to antibiotic use in aquatic disease management, probiotics have recently attracted extensive attention in aquaculture. However, the use of terrestrial bacterial species as probiotics for aquaculture has had limited success, as bacterial strain characteristics are dependent upon the environment in which they thrive. Therefore, isolating potential probiotic bacteria from the marine environment in which they grow optimally is a better approach. Bacteria that have been used successfully as probiotics belong to the genus Vibrio and Bacillus, and the species Thalassobacter utilis. Most researchers have isolated these probiotic strains from shrimp culture water, or from the intestine of different penaeid species. The use of probiotic bacteria, based on the principle of competitive exclusion, and the use of immunostimulants are two of the most promising preventive methods developed in the fight against diseases during the last few years. It also noticed that probiotic bacteria could produce some digestive enzymes, which might improve the digestion of shrimp, thus enhancing the ability of stress resistance and health of the shrimp. However, the probiotics in aquatic environment remain to be a controversial concept, as there was no authentic evidence / real environment demonstrations on the successful use of probiotics and their mechanisms of action in vivo. The present review highlights the potential sources of probiotics, mechanism of action, diversity of probiotic microbes and challenges of probiotic usage in shrimp aquaculture.
A sponge-associated marine actinomycetes Nocardiopsis alba MSA10 was screened and evaluated for the production of biosurfactant. Biosurfactant production was confirmed by conventional screening methods including hemolytic activity, drop collapsing test, oil displacement method, lipase production and emulsification index. The active compound was extracted with three solvents including ethyl acetate, diethyl ether and dichloromethane. The diethyl ether extract was fractionated by TLC and semi-preparative HPLC to isolate the pure compound. In TLC, a single discrete spot was obtained with the R (f) 0.60 and it was extrapolated as valine. Based on the chemical characterization, the active compound was partially confirmed as lipopeptide. The optimum production was attained at pH 7, temperature 30 degrees C, and 1% salinity with glucose and peptone supplementation as carbon and nitrogen sources, respectively. Considering the biosurfactant production potential of N. alba, the strain could be developed for large-scale production of lipopeptide biosurfactant.
The sponge-associated actinomycetes were isolated from the marine sponge Dendrilla nigra, collected from the southwest coast of India. Eleven actinomycetes were isolated depending upon the heterogeneity and stability in subculturing. Among these, Nocardiopsis dassonvillei MAD08 showed 100% activity against the multidrug resistant pathogens tested. The culture conditions of N. dassonvillei MAD08 was optimized under submerged fermentation conditions for enhanced antimicrobial production. The unique feature of MAD08 includes extracellular amylase, cellulase, lipase, and protease production. These enzymes ultimately increase the scope of optimization using broad range of raw materials which might be efficiently utilized. The extraction of the cell free supernatant with ethyl acetate yielded bioactive crude extract that displayed activity against a panel of pathogens tested. Analysis of the active thin layer chromatography fraction by Fourier transform infrared and gas chromatography-mass spectrometry evidenced 11 compounds with antimicrobial activity. The ammonium sulfate precipitation of the culture supernatant at 80% saturation yielded an anticandidal protein of molecular weight 87.12 kDa. This is the first strain that produces both organic solvent and water soluble antimicrobial compounds. The active extract was non-hemolytic and showed surface active property envisaging its probable role in inhibiting the attachment of pathogens to host tissues, thus, blocking host-pathogen interaction at an earlier stage of pathogenesis.
Sponges invariably filter a large volume of seawater and potentially accumulate heavy metals and other contaminants from the environment. Sponges, being sessile marine invertebrates and modular in body organization, can live many years in the same location and therefore have the capability to accumulate anthropogenic pollutants such as metals over a long period. Almost all marine sponges harbor large number of microorganisms within their tissues where they reside in the extra- and intra-cellular spaces. Bacteria in seawater have already been established as biological indicators of contamination. The present study was intended to find out the heavy metal resistance pattern of sponge-associated bacteria so as to develop suitable biological indicators. The bacteria associated with a marine sponge Fasciospongia cavernosa were evaluated as potential indicator organisms. The associated bacteria including Streptomyces sp. (MSI01), Salinobacter sp. (MSI06), Roseobacter sp. (MSI09), Pseudomonas sp. (MSI016), Vibrio sp. (MSI23), Micromonospora sp. (MSI28), Saccharomonospora sp. (MSI36) and Alteromonas sp. (MSI42) showed resistance against tested heavy metals. Based on the present findings, Cd and Hg emerged as the highly resistant heavy metal pollutants in the Gulf of Mannar biosphere reserve. Plasmids in varied numbers and molecular weights were found in all the isolates. Particularly the isolates MSI01 and MSI36 harbored as many as three plasmids each. The results envisaged that the plasmids might have carried the resistance factor. No correlation was observed in number of plasmids and level of resistance. The literature evidenced that the sponge-associated bacteria were seldom exploited for pollution monitoring though they have been extensively used for bioprospecting. In this background, the present findings come up with a new insight into the development of indicator models.
Biosurfactants are smart biomolecules which have wide spread application in medicines, processed foods, cosmetics as well as in bioremediation. In food industry, biosurfactants are used as emulsion stabilizing agents, antiadhesives, and antimicrobial/antibiofilm agents. Nowadays biosurfactant demands in industries has increased tremendously and therefore new bacterial strains are being explored for large scale production of biosurfactants. In this study, an actinobacterial strain MSA31 was isolated from a marine sponge Fasciospongia cavernosa which showed high activity in biosurfactant screening assays such as drop collapsing, oil displacement, lipase and emulsification. Lipopeptide produced by MSA31 was found to be thermostable which was evident in differential scanning calorimetry analysis. The spectral data obtained in the Fourier transform infrared spectroscopy showed the presence of aliphatic groups combined with peptide moiety which is a characteristic feature of lipopeptides. The stability index of lipopeptide MSA31 revealed “halo-alkali and thermal tolerant biosurfactant” which can be used in the food industry. Microtiter plate assay showed 125 μg/ml of lipopeptide was effective in reducing the biofilm formation activity of pathogenic multidrug resistant Staphylococcus aureus. The confocal laser scanning microscopic images provided further evidences that lipopeptide MSA31 was an effective antibiofilm agent. The antioxidant activity of lipopeptide MSA31 may be due to the presence of unsaturated fatty acid present in the molecule. The brine shrimp cytotoxicity assay showed lipopeptide MSA31 was non-toxic and can be used as food additives. Incorporation of lipopeptide MSA31 in muffin showed improved organoleptic qualities compared to positive and negative control. This study provides a valuable input for this lipopeptide to be used in food industry as an effective emulsifier, with good antioxidant activity and as a protective agent against S. aureus.
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