Microplastic intrusion into the zooplankton, the base of the marine food chain: Evidence from the Arabian Sea, Indian Ocean.

Microplastics (MPs) are ubiquitous in the marine environment, yet information regarding their occurrence in the food web is limited. We investigated the concentration and composition of MPs in water and diverse zooplankton groups from the Arabian Sea basin. Forty-one zooplankton tows were collected with a bongo net (330 µm mesh) from the Arabian Sea in January 2019. MPs in the surface water varied between 0 and 0.055 particles/m3, with a relatively higher concentration (0.013 ± 0.002 particles/m3) in the central Arabian Sea. Though fibrous MPs were most abundant in the seawater (77.14 %), zooplankton prefers small fragments (55.3 %). The size of MPs was distinctly smaller (277.1 ± 46.74 µm) in zooplankton than that in seawater (864.32 ± 73.72 µm), and MPs bioaccumulation was observed in almost all the zooplankton functional groups. Polymer composition revealed polyamide, polyethylene, polypropylene, and PVC were abundant in water and zooplankton, suggesting that the textile, fishing, shipping, and packaging industries are significant sources. The prevailing northeasterly winds, strong West India Coastal Current, and conducive westward radiated Rossby wave during January 2019 have carried the microplastic contaminated water mass away from the coast, posing a threat to the open ocean ecosystems. These results demand further attention to investigate the state of plastic pollution in the Arabian Sea basin.

Fabrication and characterization of bio-nanocomposite films using κ-Carrageenan and Kappaphycus alvarezii seaweed for multiple industrial applications.

In the present study, the whole seaweed from Kappaphycus alvarezii (containing carrageenan) was used for preparation of bio-nanocomposite films by blending with metal oxide nanoparticles such as zinc oxide (ZnONPs), cupric oxide (CuONPs) and silicon dioxide (SiO2NPs) for multiple applications, and their properties were compared with standard refined κ-Carrageenan (commercial grade). Simultaneously, the antibacterial activity and biodegradation profile of the prepared bio-nanocomposite film were also studied. The incorporation of nanoparticles into the bioplastic film matrices altered the surface morphology, increased the roughness and significantly (p < 0.05) reduced the UV transmittance, water uptake ratio (WUR), moisture content and solubility in both standard carrageenan-based bio-nanocomposite films (CBF) and Kappaphycus- based bio-nanocomposite films (KBF) compared to control. The average roughness (Ra) of KBF increased compared to CBF; however, CBF showed better tensile strength compared to KBF. Both KBF and CBF loaded with nanoparticles exhibited strong antibacterial activity against Staphylococcus aureus and Escherichia coli. However, KBF performed better compared to CBF. Antimicrobial effect of nanoparticles delayed the degradation of the bio-nanocomposite films. The present study proposes that the whole seaweed (Kappaphycus alvarezii) can be used directly for multiple industrial applications.

Unraveling the plastic degradation potentials of the plastisphere-associated marine bacterial consortium as a key player for the low-density polyethylene degradation.

The omnipresent accumulation and non-degradable nature of plastics in the environment are posing an ever-increasing ecological threat. In this study, a total of 97 bacteria were isolated from macroplastic debris collected from the coastal environments of Andaman Island. The isolates were screened for LDPE degradation potential and were identified based on phenotypic, biochemical, and molecular characterization. 16S rDNA-based identification revealed that three-three isolates of each belong to the genus Oceanimonas and Vibrio, two were closely related to the genus Paenibacillus whereas, one-one was associated with the genus Shewanella, Rheinheimera, and Bacillus, respectively. A bacterial consortium was formulated using the top four isolates based on their individual LDPE degradation potentials. A significant increase (p < 0.05) in the mean LDPE degradation (47.07 ± 6.67% weight-loss) and change in thickness was observed after 120 days of incubation. FTIR spectrum, 13C NMR, and TG-DSC analyses demonstrated changes in the LDPE sheets' functional groups, crystallinity, and in thermal properties after 120 days of incubation. The SEM and AFM images confirmed bacterial attachments, an increase in surface roughness and deformities on LDPE sheets. This study reports a bacterial consortium that can efficiently degrade the plastics and can be used in providing eco-friendly mitigation of plastic waste.

Thraustochytrids of Mangrove Habitats from Andaman Islands: Species Diversity, PUFA Profiles and Biotechnological Potential.

Thraustochytrids are the most promising microbial source for the commercial production of docosahexaenoic acid (DHA) for its application in the human health, aquaculture, and nutraceutical sectors. The present study isolated 127 thraustochytrid strains from mangrove habitats of the south Andaman Islands, India to study their diversity, polyunsaturated fatty acids (PUFAs), and biotechnological potential. The predominant strains were identified as belonging to two major genera (Thraustochytrium, Aurantiochytrium) based on morphological and molecular characteristics. The strain ANVKK-06 produced the maximum biomass of 5.42 g·L-1, while ANVKK-03 exhibited the maximum total lipid (71.03%). Omega-3 PUFAs such as eicosapentaenoic acid (EPA) accumulated up to 11.03% in ANVKK-04, docosapentaenoic acid (DPA) up to 8.65% in ANVKK-07, and DHA up to 47.19% in ANVKK-06. ANVKK-06 showed the maximum scavenging activity (84.79 ± 2.30%) while ANVKK-03 and ANVKK-10 displayed the highest antibacterial activity against human and fish pathogens, S. aureus (18.69 ± 1.2 mm) and V. parahaemolyticus (18.31 ± 1.0 mm), respectively. All strains were non-toxic as evident by negative blood agar hemolysis, thus, the thraustochytrids are suggested to be a potential source of DHA for application in the health care of human and fish.

Dietary supplementation of curcumin-loaded chitosan nanoparticles stimulates immune response in the white leg shrimp Litopenaeus vannamei challenged with Vibrio harveyi.

Pathogen infections in shrimps trigger the release of reactive oxygen species (ROS) as a part of immune response. The excessive accumulation of ROS causes the production of oxidative stress, which leads to oxidative damage of the biomolecules in the host cells. The inclusion of dietary antioxidants is known to mitigate oxidative stress and stimulate immunity. Curcumin, a potential antioxidant was encapsulated in chitosan nanoparticles to surge its bioavailability and was administered orally to Vibrio harveyi challenged and non-challenged Litopenaeus vannamei. The non-challenged shrimps fed with curcumin-loaded chitosan nanoparticles (Cur-CSNPs) showed a significant increase (p ≤ 0.05) in the specific growth rate, daily growth coefficient and survival rate. A significant increase (p ≤ 0.05) in the phenoloxidase activity, total hemocyte count and superoxide dismutase activity was observed in both the challenged and non-challenged shrimps fed with Cur-CSNPs. Additionally, a significant increase (p ≤ 0.05) in the relative mRNA expression of lysozyme, cMnSOD and lectin was observed in the Cur-CSNPs fed shrimps. The findings of this research suggest that Cur-CSNPs reinforce the immune system of L. vannamei against V. harveyi infection. Moreover, the non-challenged shrimps showed improvement in the growth parameters in addition to immunostimulation. Thereby a routine inclusion of dietary Cur-CSNPs could mitigate the oxidative damage caused by the incidence of environmental or pathogen-mediated oxidative stress.

Deep-sea mercury resistant bacteria from the Central Indian Ocean: A potential candidate for mercury bioremediation.

Deep-sea bacteria when grown in normal environmental conditions get morphologically and genetically adapted to resist the provided culture conditions for their survival, making them a possible aspirant in mercury bioremediation. In this study, seawater samples were collected from different depths of the Central Indian Ocean and seven mercury resistant bacteria (resistant to 100 mg L-1 concentration of inorganic Hg as HgCl2) were isolated. Based on 16S rRNA gene sequencing, the identified isolates belong to the genera Pseudomonas, Bacillus and Pseudoalteromonas. The presence of the merA gene in the isolates contributes to the effective volatilization of mercury. The Inductively Coupled Plasma Mass-Spectroscopy analysis revealed that the isolates can reduce up to >80% of inorganic mercury. Moreover, Fourier Transform Infrared spectrum analysis indicates that functional groups play a key role in the mechanism of adaptation towards Hg2+ reduction. Thus, the deep-sea bacteria expressed significant tolerance and reduction potential towards ionic mercury.

Existence in cellulose shelters: industrial and pharmaceutical leads of symbiotic actinobacteria from ascidian Phallusia nigra, Andaman Islands.

The diversity of actinobacteria associated with marine ascidian Phallusia nigra from Andaman Islands was investigated. A total of 10 actinobacteria were isolated and based on the biochemical and molecular characterization, the isolates were assigned to 7 different actinobacterial genera. Eight putatively novel species belonging to genera Rhodococcus, Kineococcus, Kocuria, Janibacter, Salinispora and Arthrobacter were identified based on 16S rDNA sequence similarity with the NCBI database. The organic extracts of ten isolates displayed considerable bioactivity against test pathogens, which were Gram-positive and Gram-negative in nature. PCR-based screening for type I and type II polyketide synthases (PKS-I, PKS-II) and nonribosomal peptide synthetases (NRPS) revealed that, 10 actinobacterial isolates encoded at least one type of polyketide synthases biosynthesis gene. Majority of the isolates found to produce industrially important enzymes; amylase, protease, gelatinase, lipase, DNase, cellulase, urease, phosphatase and L-asparaginase. The present study emphasized that, ascidians are a prolific resource for novel bioactive actinobacteria with potential for novel drug discovery. This result expands the scope to functionally characterize the novel ascidian associated marine actinobacteria and their metabolites could be a source for the novel molecules of commercial interest.

Taxonomic Composition and Biological Activity of Bacterial Communities Associated with Marine Ascidians from Andaman Islands, India.

Marine invertebrates, particularly ascidians, constitute an important source of potential active and biofunctional natural products. The microbial diversity associated with ascidians is little recognized, although these microorganisms play a vital role in marine ecosystems. The objective of this study was to investigate bacterial population diversity in four ascidian samples: Phallusia nigra, Phallusia fumigata, Eudistoma viride, and Rhopalaea macrothorax, collected from the North Bay, Andaman and Nicobar Islands. Microbial strains identified up to the species level revealed 236 distinct species/ribotypes out of 298 bacterial strains. Of 298 ascidian-associated bacteria, 72 isolates belong to the class Gammaproteobacteria and the genus Endozoicomonas. The results from this investigation will contribute a broaden knowledge of microbial diversity associated to marine ascidians, and as a promising source for the discovery of new natural products.

Microplastics particles in seafloor sediments along the Arabian Sea and the Andaman Sea continental shelves: First insight on the occurrence, identification, and characterization.

Microplastics (MPs) are widely-recognized contaminants and marine sediments act as a sink of MPs and therefore may cause a potential threat to benthic communities. We aim to analyze the MPs abundances and characteristics in the seafloor sediments from the continental shelves of the Arabian and Andaman seas. Twenty-two seafloor sediments were collected from 8 and 14 locations of the Arabian and Andaman seas, respectively. MPs concentrations varied from not detected (ND) to 267 particles kg-1 with mean values of 128.02 ± 33.92 and 15.36 ± 2.61 particles kg-1, respectively for the Arabian and Andaman seas. Among different shapes, fiber had the highest distribution over fragments and pellet. FT-IR analysis revealed acrylic was most dominant polymer, followed by polyethylene, and nylon. Mean MP concentration at the Arabian Sea was significantly higher (p < 0.001) than in the Andaman Sea. The present study revealed the wide-spread occurrence of MPs throughout the Indian seas.

Molecular characterization, phylogenetic and in silico sequence analysis data of trehalose biosynthesis genes; otsA and otsB from the deep sea halophilic actinobacteria, Streptomyces qinglanensis NIOT-DSA03.

Trehalose, a non-reducing disaccharide (α-D-glucopyranosyl-(1→1)-α-D-glucopyranoside) is a natural compound, which serves as a protective substance in halophilic bacterial cells. Trehalose biosynthesis genes (otsA and otsB) were PCR amplified from the genomic DNA of deep sea actinobacteria, Streptomyces qinglanensis NIOT-DSA03. The amplified genes were cloned and nucleotide sequences were determined. In silico sequence and phylogenetic analysis of nucleotides and amino acids of otsA and otsB sequences of S. qinglanensis were also determined. The experimental data described in this study will be helpful to develop a recombinant expression system to produce trehalose for biotechnological applications.

Studies on the development and characterization of bioplastic film from the red seaweed (Kappaphycus alvarezii).

Seaweeds rich in polysaccharides are considered commercially important because of their wide range of industrial applications. In this study, the red seaweed Kappaphycus alvarezii was used for bioplastic film production. Plasticizer polyethylene glycol (PEG) to seaweed biomass ratio was optimized and a thin bioplastic film with higher tensile strength was produced. The films obtained were characterized by its thickness, tensile strength, colour (L, a, b), elongation at break (EAB), water vapour transmission rate (WVTR) and oxygen transmission rate (OTR). TG-DSC, AFM, SEM and FTIR spectroscopy analysis were performed to assess the composition, phase transitions and chemical reaction capabilities of the film. The bioplastic film obtained from 4% K. alvarezii (whole seaweed) showed better physical and mechanical properties, whereas TG-DSC, FTIR and AFM showed similar kind of bioplastic properties in all the concentrations. Decrease in OTR was observed against decreasing wall thickness of the film. The present study suggests that the seaweed would be a potential alternate source for bioplastic production which may reduce the usage of non-degradable plastics.

A multiplex PCR kit for the detection of three major virulent genes in Enterococcus faecalis.

A multiplex PCR kit that detects three major virulence genes, gelE, hyl and asaI, in Enterococcus faecalis was developed. Analyses of the available sequences of three major virulence genes and designed primers allowed us to develop the three-gene, multiplex PCR protocol that maintained the specificity of each primer pair. The resulting three amplicon bands for gelE, hyl and asaI were even and distinct with product sizes of 213, 273 and 713 bp, respectively. The multiplex PCR procedure was validated with a total of 243 E. faecalis strains that included 02 ATCC strains, 109 isolates from marine samples (sediment, water and sea foods), 22 isolates from cattle fodder, 79 isolates fresh water samples and 31 isolates from nosocomial samples. Specificity of the kit was indicated by amplification of only three major virulent genes gelE, hyl and asaI without any nonspecific bands. Tests for the limit of detection revealed that amplified genes from the sample with a minimum of 104 CFU/g or CFU/mL (10 cells/reaction) of E. faecalis and lower cell load samples, after a 3 h enrichment in NIOT-E. faecalis enrichment medium at 37 °C, a sensitivity level of 10 CFU/g or CFU/mL was achieved.

Characteristics and dynamics of Salmonella diversity and prevalence of biomarker genes in Port Blair Bays, South Andaman, India.

Salmonella is a major cause of morbidity and mortality in humans worldwide, and the infection with multidrug-resistant strains can cause severe diseases. Many coastal cities around the world discharge their wastewaters into the marine environment. These wastewaters contain a variety of pathogenic microorganisms that may have a role in the contamination of this ecosystem and have potential risks for public health. Using an environmental approach, the present study investigated the presence of Salmonella in sediment and water samples collected from Port Blair Bays. In this environmental approach, the provided information about the diversity of the Salmonella serovars, antibiotic resistance and the prevalence of virulence factors in Salmonella, especially from the coastal waters of Port Blair Bays. The occurrence of Salmonellae was significantly higher in water column samples (2.9%) than in those taken from the marine sediments (0.7%). Of the 133 positive Salmonella strains, 22 different serovars were identified. Salmonella enterica serovar Senftenberg was the predominant serovar, being represented by 54 isolates (42.5%), followed by serovar Typhimurium (19 isolates [15%]) and serovar Agona (12 isolates [9.4%]). The presence of virulence genes (filC, sitC, hilA, invA, sipC, hilD, hilC, invF, invE, invH, sipF, aadA, pare, gyrA, spaP and parC) and susceptibility studies with 10 selected antibiotics were also performed. The results of this study revealed that all Salmonella isolates were positive for targeted virulence genes and were resistant to at least one antibiotic. Antibiotic susceptibility studies revealed the presence of multidrug resistant Salmonella strains in coastal water, which usually from land base sources end up in the marine environment and may pose a significant risk on public health.

First evidence of microplastics bioaccumulation by marine organisms in the Port Blair Bay, Andaman Islands.

Microplastic (MP) pollution has become a global concern. We aim to quantify the extent of MP pollution in the coastal ecosystem of the Port Blair Bay, A&N Islands. Water, sediment, zooplankton, finfish, and shellfish samples were collected from the Port Blair Bay and analyzed for the presence of MP. Average concentrations of MP in water, sediment, zooplankton, finfish, and shellfishes were found to be 0.93 ± 0.59 particles per m3, 45.17 ± 25.23 particles per kilogram, 0.12 ± 0.07 pieces per zooplankter and 10.65 ± 7.83 particles per specimen, respectively. High amount of MP retention was observed in the zooplankton community. Maximum MP ingestion was observed in adult Carangoides malabaricus. Fiber was most abundant in water, sediment, and fish samples, followed by fragment and pellet. However, fragments were predominant in zooplankton. Nylon, acrylic, and ionomer surlyn were most abundant polymer types in the bay environment. These results demand further attention to combat plastic pollution in the coastal ecosystem.

Ectoine biosynthesis genes from the deep sea halophilic eubacteria, Bacillus clausii NIOT-DSB04: Its molecular and biochemical characterization.

Ectoine, the most prominent osmolyte in nature, is a vital compatible solute present in halophilic bacterium. It protects the cellular biomolecules of the halophilic bacteria and retains their intrinsic function from extreme circumstances. In the current research, ectoine biosynthesis gene cluster (ectABC) in Bacillus clausii NIOT-DSB04 was expressed heterologically in E. coli M15 (pREP4). RP-HPLC resolved several fractions of the purified recombinant product, one of which had been confirmed as ectoine. The recombinant ectoine was further characterized by 1H and 13C Nuclear Magnetic Resonance. The purified recombinant ectoine was also authenticated by FT-IR studies with the existence of ester carbonyl and C-H group. In IPTG induced E. coli M15 transgenic cells, the enzymatic activity of the ectA, B and C genes were found to be higher than that of uninduced cells.

Assessment of trace metal contamination in the marine sediment, seawater, and bivalves of Parangipettai, southeast coast of India.

Industrial and domestic discharge into the coastal environment has driven us to propose an integrated approach to delineate stations contaminated with metals on the Parangipettai coast by collecting sediment and seawater samples monthly at 18 stations from 2015 to 2017. Descriptive statistics revealed that the concentrations of some metals in the sediment and seawater samples were beyond the permissible level. Further, factor analysis showed a sampling adequacy of 0.90 with high positive loading for Ni (0.94), Cd (0.91), Co (0.90), Pb (0.89), and Zn (0.87) in sediment samples. The degree of contamination by metals was evaluated using pollution indices. The results of the contamination index revealed that some stations in the study area were moderately polluted, and those of the ecological index showed that open sea was under low risk while other stations were in the moderate-to-high-risk category. The results obtained are essential to establish the reference condition for a comparative study in similar environments in the tropical regions.

Studies on diversity of Vibrio sp. and the prevalence of hapA, tcpI, st, rtxA&C, acfB, hlyA, ctxA, ompU and toxR genes in environmental strains of Vibrio cholerae from Port Blair bays of South Andaman, India.

Vibrio species are widely distributed in the estuarine and coastal waters that possess the greatest threat to human health worldwide. In this study it is aimed to isolate and observe the abundance of Vibrio sp. and prevalence of biomarker genes and antibiotic resistance profile of V. cholerae isolated from the Port Blair bays of South Andaman. A total of 56 water samples were collected from the seven sampling stations of Port Blair bays in which maximum number of Vibrio sp. population density (1.78 × 104) was recorded in Phoenix Bay. Among the 786 isolates 57.38% of the isolates were confirmed as Vibrio sp., Vibrio cholerae and Vibrio parahaemolyticus. PCR results revealed that the prevalence of biomarker genes was recorded maximum in the isolates from Phoenix Bay and Junglighat Bay samples. Upon further analysis, it was observed that the prevalence of hlyA gene (215 bp), was found to be the most widespread biomarker determinant in 84.17% of isolates. Major virulence determinants; ctxA, ompU and toxR genes were not detected in V. cholerae isolates from Port Blair bays. Maximum antibiotic resistance pattern was observed in Phoenix Bay isolates and maximum number of V. cholerae isolates was resistance to tetracycline (60.76%). Cluster and Principal Component Analysis were employed to understand the diversity and distribution of Vibrio isolates and its biomarker genes. Upon PCA analysis seasonal influence was not much perceived in Vibrio species diversity in Port Blair bays and the lack of significant difference in the detection of species diversity in this study is due to resemblance in geographical conditions and sources of pollution.

Biodiversity and antibacterial potential of cultivable halophilic actinobacteria from the deep sea sediments of active volcanic Barren Island.

Deep Sea sediment cores were collected from the surrounding of active volcanic Barren Island, Andaman & Nicobar Islands. A total of 123 cultivable marine actinobacteria were isolated and identified based on their biochemical and 16S rDNA sequences. The isolates were categorized under 10 genera, of which Streptomyces sp., Dietzia sp. and Brevibacterium sp. are the dominant genera. Of 123 isolates, 73 isolates exhibited appreciable inhibitory activity against both Gram positive and Gram negative bacteria. High frequencies of positive PCR amplification were obtained for PKS-I (39.13%), PKS-II (56.52%) and NRPS (69.57%). The highest levels of biosynthetic systems were observed for NRPS and PKS-II. Majority of the actinobacterial isolates revealed excellent potential for bioprospecting of novel byproducts with industrial and pharmaceutical importance.

Functional characterization of a major compatible solute in Deep Sea halophilic eubacteria of active volcanic Barren Island, Andaman and Nicobar Islands, India.

Ectoine, a cyclic tetrahydropyrimidine (2-methyl-1,4,5,6-tetrahydropyrimidine-4-carboxylic acid) is a compatible solute, serves as a protective compound in many halophilic eubacterial cells under stress. In this study, the ectoine biosynthesis genes (ectA, B and C) from the genomic DNA of a deep sea eubacteria, Bacillus clausii NIOT-DSB04 was PCR amplified, cloned into the expression vector pQE30 with a 6 × histidine tag and expressed in M15 cells. The lysates of induced cells with diaminobutyric acid aminotransferase and ectoine synthase disclosed two clear expressed bands with molecular masses of 46 kDa and 15 kDa as estimated by SDS-PAGE. The recombinant ectoine synthase activity of the expressed cells was at higher level than that of uninduced cells. In silico sequence and phylogenetic analysis of nucleotides and amino acids revealed that the ectA, B and C sequences of Bacillus clausii NIOT-DSB04 were conserved in many eubacteria.

Evaluation of the environmental quality of Parangipettai, Southeast Coast of India, by using multivariate and geospatial tool.

The anthropogenic pressure in recent years has driven us to investigate the environmental quality at 22 stations in Parangipettai by collecting seawater samples monthly from 2014 to 2015. The sampling stations were grouped into three different environments, namely, Vellar Estuary (VE), Coleroon Estuary (CE), and Open Sea (OS). Factor analysis showed a total variance of 65.63% and exhibited a strong factor loading for atmospheric temperature (0.914), water temperature (0.917), ammonia (0.767), inorganic phosphate (0.897), total phosphorus (0.783), and phytoplankton (0.829). The index value showed water quality was good in OS (74.18), whereas it was moderate in VE (69.73) and CE (68.47). The visual model developed using Geographical Information System (GIS) displayed a spatial pattern of water temperature and phytoplankton dispersion in a distinct manner. The results obtained through multivariate analysis and GIS-based model are imperative to establish reference for a comparative study with other similar ecosystem for better planning and management of tropical seawaters.