Environmental and ecological risk of microplastics in the surface waters and gastrointestinal tract of skipjack tuna (Katsuwonus pelamis) around the Lakshadweep Islands, India.

The presence of microplastics (MPs) in marine ecosystems is widespread and extensive. They have even reached the deepest parts of the ocean and polar regions. The number of articles on plastic pollution has increased in recent years, but few have investigated the MPs from oceanic islands which are biodiversity hotspots. We investigated the possible microplastic contamination their source and characteristics in surface waters off Kavaratti Island and in the gastrointestinal tract (GT) of skipjack tuna, Katsuwonus pelamis collected from Kavaratti Island of the Lakshadweep archipelago. A total of 424 MP particles were isolated from the surface water samples collected from off Kavaratti Island with an average abundance of 5 ± 1nos./L. A total of 117 MPs were recovered from the GT of skipjack tuna from 30 individual fishes. This points to a potential threat of MP contamination in seafood around the world since this species has a high value in local and international markets. Fiber and blue color were the most common microplastic morphotypes and colors encountered, respectively, both from surface water and GT of fish. Smaller MPs (0.01-1 mm) made up a greater portion of the recovered materials, and most of them were secondary MPs. Polyethylene and polypropylene were the most abundant polymers found in this study. The Pollution Load Index (1.3 ± 0.21) of the surface water and skipjack tuna (1 ± 0.7) indicates a minor ecological risk for the coral islands, while the Polymer Hazard Index highlights the ecological risk of polymers, even at low MP concentrations. This pioneer study sheds preliminary light on the abundance, properties, and environmental risks of MPs to this highly biodiverse ecosystem.

Distribution and antibiotic resistance of vibrio population in an urbanized tropical lake-the Vembanad-in the southwest coast of India.

Among the diverse Vibrio spp. autochthonous to coastal ecosystems, V. cholerae, V. fluvialis, V. vulnificus and V. parahaemolyticus are pathogenic to humans. Increasing sea-surface temperature, sea-level rise and water-related disasters associated with climate change have been shown to influence the proliferation of these bacteria and change their geographic distribution. We investigated the spatio-temporal distribution of Vibrio spp. in a tropical lake for 1 year at a 20-day interval. The abundance of Vibrio spp. was much higher during the south-west monsoon in 2018, when the lake experienced a once-in-a-century flood. The distribution of Vibrio spp. was influenced by salinity (r = 0.3, p < 0.001), phosphate (r = 0.18, p < 0.01) and nitrite (r = 0.16, p < 0.02) in the water. We isolated 470 colonies of Vibrio-like organisms and 341 could be revived further and identified using 16S rRNA gene sequencing. Functional annotations showed that all the 16 Vibrio spp. found in the lake could grow in association with animals. More than 60% of the isolates had multiple antibiotic resistance (MAR) index greater than 0.5. All isolates were resistant to erythromycin and cefepime. The proliferation of multiple antibiotic-resistant Vibrio spp. is a threat to human health. Our observations suggest that the presence of a diverse range of Vibrio spp. is favoured by the low-saline conditions brought about by heavy precipitation. Furthermore, infections caused by contact with Vibrio-contaminated waters may be difficult to cure due to their multiple antibiotic resistances. Therefore, continuous monitoring of bacterial pollution in the lakes is essential, as is the generation of risk maps of vibrio-infested waters to avoid public contact with contaminated waters and associated disease outbreaks.

First report of brittle stars (Echinodermata: Ophiuroidea) from a seamount in the eastern Arabian Sea.

This paper reports two species-Ophiozonella molesta (Koehler, 1904) and Ophiothamnus venustus Matsumoto, 1915, from a seamount in the Eastern Arabian Sea. This forms the first ever report of Ophiothamnus venustus from the Indian Ocean, and the first verified report of Ophiozonella molesta from Indian waters. This is the first ever work to document benthic invertebrates from seamounts in the Arabian Sea.

Characterization of pyomelanin secreted by Shewanella sp. and their application in metal recovery.

Melanin is a biopolymer with versatile structural and functional properties and diverse applications in recovering toxic chemicals from water and wastewater, biomedical imaging, and as theragnostic agent. We report the structural characterization and biosynthetic pathway of an extracellular pyomelanin secreted by a sponge-associated bacterium, Shewanella sp. (Shewanella-melanin), and their potential application in metal recovery from liquid. Pyomelanin particles of > 50 µm size were found in the culture medium within 48 h of growth, which were formed through the self-polymerization of benzoquinone molecule produced through homogentisic acid pathway. The aspC and hppD genes involved in the biosynthetic pathway of pyomelanin were detected in the whole genome sequence of Shewanella sp. The FT-IR spectra of Shewanella-melanin, at ~ 3300-3420 cm-1 corresponding to the stretching vibration of -NH and -OH, was in good agreement with that of Sepia melanin, while its elemental composition (C/N/H/S of 29.2:8.23:6.41:1.58) was unique. Shewanella-melanin showed ~ 300 and ~ 950 times increased chelation of manganese and iron from a liquid medium supplemented with 2 mM of MnSO4 and FeSO4, respectively, compared to a control. The FT-IR spectrum showed the binding of metal ions to the carboxylic acid, hydroxyl, and amine groups of Shewanella-melanin. The Shewanella-melanin, with its excellent metal biosorption, could be a potential candidate for removing toxic compounds from water, in turn contributing to the fulfillment of sustainable development goal (SDG) 6.

Analysis of non-pharmaceutical interventions and their impacts on COVID-19 in Kerala.

In the absence of an effective vaccine or drug therapy, non-pharmaceutical interventions are the only option for control of the outbreak of the coronavirus disease 2019, a pandemic with global implications. Each of the over 200 countries affected has followed its own path in dealing with the crisis, making it difficult to evaluate the effectiveness of measures implemented, either individually, or collectively. In this paper we analyse the case of the south Indian state of Kerala, which received much attention in the international media for its actions in containing the spread of the disease in the early months of the pandemic, but later succumbed to a second wave. We use a model to study the trajectory of the disease in the state during the first four months of the outbreak. We then use the model for a retrospective analysis of measures taken to combat the spread of the disease, to evaluate their impact. Because of the differences in the trajectory of the outbreak in Kerala, we argue that it is a model worthy of a place in the discussion on how the world might best handle this and other, future, pandemics.

The Influence of Photodynamic Antimicrobial Chemotherapy on the Microbiome, Neuroendocrine and Immune System of Crustacean Post Larvae.

Photodynamic antimicrobial chemotherapy (PACT), employing a combination of light and natural photosensitizer molecules such as curcumin, has been accepted as a safe modality for removing aquatic pathogens which cause diseases such as cholera in humans and vibriosis in aquatic animals. Curcumin and its photodegradation products are generally considered as safe to animals, but the impact of reactive oxygen species (ROS) generated by these products on the growth and survival of organisms at a cellular level has not been studied in detail. The ROS generated by curcumin on photoexcitation using blue light (λmax 405 nm, 10 mW cm-2) disinfects more than 80% of free-living Vibrio spp. in the rearing water of Penaeus monodon. However, it is less effective against Vibrio spp. colonized inside P. monodon because the carapace of the animal prevents the transmission of more than 70% of light at the 400-450 nm range and thus reduces the formation of ROS. The influence of curcumin and photoexcited curcumin on the microbiome of P. monodon were revealed by nanopore sequencing. The photoexcited curcumin induced irregular expression of genes coding the moult-inhibiting hormone (MIH), Crustacean hyperglycaemic hormone (CHH)), prophenoloxidase (ProPO), and crustin, which indicates toxic effects of ROS generated by photoexcited curcumin on the neuroendocrine and immune systems of crustaceans, which could alter their growth and survival in aquaculture settings. The study proposed the cautious use of photodynamic therapy in aquaculture systems, and care must be taken to avoid photoexcitation when animals are experiencing moulting or environmental stress.

Immortalization of shrimp lymphoid cells by hybridizing with the continuous cell line Sf9 leading to the development of 'PmLyO-Sf9 '.

Shrimp progressively gets more attention among marine invertebrates from researchers all over the world due to it being a healthy food as well as having economic importance. There were a lot of attempts to develop a continuous cell line from shrimp but none successful. In this context a novel hybrid cell line named 'PmLyO-Sf9' could be developed by fusing shrimp lymphoid organ cells with Sf9 cells after to metabolic blocking of Sf9 cells using puromycin and actinomycin D and effecting the fusion by way of PEG application. The cells are maintained and multiplied in a mixture of SCCM and TNM-FH having osmolality 550 mOsm kg-1 and pH 6.8. Transmission electron microscopy of the hybrid cells revealed the presence of two nuclei during the initial stages and a single nucleus subsequently. The cell line is with shrimp and Sf9 genomic components and shrimp specific protein and is susceptible to WSSV. Shrimp elongation factor, Sf9 beta-actin, shrimp STAT and peroxinectin could be expresses through RT-PCR in the cell line. This is the first successful report of a hybrid cell line with shrimp genomic components and envisaged to be recognized a model system for multitudes of biomedical research in vitro. The cell line is in the National Cell Line Repository of ICAR - National Bureaue of Fish Genetic Resources, Lucknow, India.

Inhibition of pathogenic Vibrio harveyi using calamenene, derived from the Indian gorgonian Subergorgia reticulata, and its synthetic analog.

We report the synthesis and antimicrobial properties of a partially reduced dihydronathphthoquinone analogue of 2-methoxy, 5-acetoxy calamenene, extracted from Subergorgia reticulata. The growth of a pathogenic Vibrio harveyi strain was effectively controlled by the calamenene derivative 1 (Cala1) and its synthetic analog 2 (Cala2). Complete mortality of V. harveyi was observed with 2.5 and 0.5 µg mL-1 concentrations of Cala1 and Cala2, respectively. The metabolic assays demonstrated that Cala1 is a bacteriostatic agent while Cala2 showed bactericidal properties. It was confirmed that translocation of Cala2 into the cytoplasm does not induce any change to the integrity of the bacterial cell wall. The Cala2 induced damage to the genetic material of 70% of cells while genetic material of 91% of cells treated with Cala1 remained intact. The Cala2 is, therefore, proposed as a potential bactericidal compound against the aquaculture pathogen V. harveyi. The fact that the Cala2 exhibited minimal cytotoxicity to Artemia nauplii indicates its potential use as an antimicrobial agent for aquaculture operations.

Neuroendocrine and immunotoxicity of polyaromatic hydrocarbon, chrysene in crustacean post larvae.

Polyaromatic hydrocarbons are a group of chemical pollutants which cause a significant threat to the living organisms in estuaries and marine ecosystems. We report the effect of chrysene, a major PAH pollutant found in Cochin Estuary along the southwest coast of India, on the neuroendocrine and immune gene expression of the post larvae (PL-25) of Penaeus monodon. The PL- 25 of P. monodon were administered with feed coated with increasing concentrations of chrysene (1, 2 and 3 µg/g) for 10 days and the gene expression was studied on 7th, 11th and 15th day. The PL exposed to chrysene showed moulting stress and changes in the levels of moult-inhibiting hormone I (MIH I) indicated by irregular moulting in the experimental tanks. At the molecular level, the higher concentration of chrysene induced two-fold upregulation of neuroendocrine (MIH I) and downregulation of immune (ProPO and crustin) gene on the 7th day of exposure. The expression of MIH I gene reduced on withdrawing the experimental feed (on 11th day), while continued downregulation of ProPO and crustin were observed on the 11th day. The results of the present study indicate that the microgram levels of PAH can impinge the neuroendocrine and immune system of the P. monodon, which may induce morbidity and mortality to the larvae in polluted coastal ecosystems. Therefore, more attention may be given to avoid PAH pollution in the estuaries to maintain a healthy ecosystem and to protect the animals from extinction.

A Printable Device for Measuring Clarity and Colour in Lake and Nearshore Waters.

Two expanding areas of science and technology are citizen science and three-dimensional (3D) printing. Citizen science has a proven capability to generate reliable data and contribute to unexpected scientific discovery. It can put science into the hands of the citizens, increasing understanding, promoting environmental stewardship, and leading to the production of large databases for use in environmental monitoring. 3D printing has the potential to create cheap, bespoke scientific instruments that have formerly required dedicated facilities to assemble. It can put instrument manufacturing into the hands of any citizen who has access to a 3D printer. In this paper, we present a simple hand-held device designed to measure the Secchi depth and water colour (Forel Ule scale) of lake, estuarine and nearshore regions. The device is manufactured with marine resistant materials (mostly biodegradable) using a 3D printer and basic workshop tools. It is inexpensive to manufacture, lightweight, easy to use, and accessible to a wide range of users. It builds on a long tradition in optical limnology and oceanography, but is modified for ease of operation in smaller water bodies, and from small watercraft and platforms. We provide detailed instructions on how to build the device and highlight examples of its use for scientific education, citizen science, satellite validation of ocean colour data, and low-cost monitoring of water clarity, colour and temperature.

Diversity and seasonal distribution of ammonia-oxidizing archaea in the water column of a tropical estuary along the southeast Arabian Sea.

Diversity and distribution pattern of ammonia-oxidizing archaea (AOA) were studied across a salinity gradient in the water column of Cochin Estuary (CE), a tropical monsoonal estuary along the southeast Arabian Sea. The water column of CE was found to be nutrient rich with high bacterial (3.7-6.7 × 108 cells L-1) and archaeal abundance (1.9-4.5 × 108 cells L-1). Diversity and seasonal variation in the distribution pattern of AOA were studied using clone library analysis and Denaturing gradient gel electrophoresis (DGGE). Clone library analysis of both the amoA and 16S rRNA gene sequences showed similar diversity pattern, however the diversity was more clear when the 16S rRNA gene sequences were analyzed. More than 70% of the sequences retrieved were clustered under uncultured Thaumarchaeota group 1 lineage and the major fractions of the remaining sequences were grouped into the Nitrosopumilus lineage and Nitrosopelagicus lineage. The AOA community in the CE was less adaptable to changing environmental conditions and its distribution showed seasonal variations within the DGGE banding pattern with higher diversity during the pre-monsoon period. The distribution of AOA also showed its preference to intermediate salinity for their higher diversity. Summer monsoon associated runoff and flushing played a critical role in regulating the seasonality of AOA distribution.

Role of heavy metals in structuring the microbial community associated with particulate matter in a tropical estuary.

Particulate matter (PM), which are chemically and biochemically complicated particles, accommodate a plethora of microorganisms. In the present study, we report the influence of heavy metal pollution on the abundance and community structure of archaea and bacteria associated with PM samples collected from polluted and non-polluted regions of Cochin Estuary (CE), Southwest coast of India. We observed an accumulation of heavy metals in PM collected from CE, and their concentrations were in the order Fe > Zn > Mn > Cr > Pb > Cu > Cd > Co > Ni. Zinc was a major pollutant in the water (4.36-130.50 µgL-1) and in the particulate matter (765.5-8451.28 µgg-1). Heavy metals, Cd, Co, and Pb were recorded in the particulate matter, although they were below detectable limits in the water column. Statistical analysis showed a positive influence of particulate organic carbon, nitrogen, PM-Pb, PM-Zn and PM-Fe on the abundance of PM-archaea and PM-bacteria. The abundance of archaea and bacteria were ten times less in PM compared with planktonic ones. The abundance of PM-archaea ranged between 4.27 and 9.50 × 107and 2.73 to 3.85 × 107 cellsL-1 respectively for the wet and dry season, while that of PM-bacteria was between 1.14 and 6.72 × 108 cellsL-1 for both seasons. Community structure of PM-bacteria varied between polluted and non-polluted stations, while their abundance does not show a drastic difference. This could be attributed to the selective enrichment of bacteria by heavy metals in PM. Such enrichment may only promote the growth of metal resistant archaea and bacteria, which may not participate in the processing of PM. In such cases, the PM may remain without remineralization in the system arresting the food web dynamics and biogeochemical cycles.

Unicellular cyanobacteria Synechocystis accommodate heterotrophic bacteria with varied enzymatic and metal resistance properties.

The interactions between heterotrophic bacteria and primary producers have a profound impact on the functioning of marine ecosystem. We characterized the enzymatic and metal resistance properties of fourteen heterotrophic bacteria isolated from a unicellular cyanobacterium Synechocystis sp. that came from a heavy metal contaminated region of Cochin estuary, southwest coast of India. Based on 16S rRNA gene sequence similarities, the heterotrophic bacteria were grouped into three phyla: namely Actinobacteria, Firmicute, and Proteobacteria. Overall Proteobacteria showed a higher level of enzyme expression while Actinobacteria and Firmicutes showed higher tolerance to heavy metals. Among Proteobacteria, an isolate of Marinobacter hydrocarbonoclasticus (MMRF-584) showed highest activities of ß-glucosidase (1.58 ± 0.2 µMml(-1) min(-1) ) and laminarinase (1170.17 ± 95.4 µgml(-1) min(-1) ), while other two isolates of M. hydrocarbonoclasticus, MMRF-578 and 581, showed highest phosphatase (44.71 ± 0.2 µMml(-1) min(-1) ) and aminopeptidase (33.22 ± 0 µMml(-1) min(-1) ) activities respectively. Among Firmicutes, the Virgibacillus sp. MMRF-571 showed exceptional resistance against the toxic heavy metals Cd (180 mM), Pb (150 mM), and Hg (0.5 mM). Bacillus cereus, MMRF-575, showed resistance to the highest concentrations of Co (250 mM), Cd (150 mM), Pb (180 mM), Hg (0.5 mM), Ni (280 mM), and Zn (250 mM) tested. Our results show that heterotrophic bacteria with varied enzymatic and metal resistance properties are associated with Synechocystis sp. Further studies to delineate the role of these heterotrophic bacteria in protecting primary producers from toxic effects of heavy metals and their potential application in bioremediation will be appreciated.

Bacterial domination over archaea in ammonia oxidation in a monsoon-driven tropical estuary.

Autotrophic ammonia oxidizing microorganisms, which are responsible for the rate-limiting step of nitrification in most aquatic systems, have not been studied in tropical estuaries. Cochin estuary (CE) is one of the largest, productive, and monsoon-driven estuary in India opening into the southeast Arabian Sea. CE receives surplus quantities of ammonia through industrial and domestic discharges. The distribution of ammonia-oxidizing bacteria (AOB), ammonia-oxidizing archaea (AOA), and anaerobic ammonia-oxidizing bacteria (anammox) were studied using fluorescence in situ hybridization (FISH) and their relative contribution to the process as well as the governing factors were examined and reported for the first time from CE. The order of occurrence of these assemblages was ß-proteobacteria (0.79 to 2 × 10(5) cells ml(-1)) > γ-proteobacteria (0.9 to 4.6 × 10(4) cells ml(-1)) > anammox (0.49 to 1.9 × 10(4) cells ml(-1)) > AOA (0.56 to 6.3 × 10(3) cells ml(-1)). Phylogenetic analysis of DGGE bands showed major affiliation of AOB to ß-proteobacteria, while AOA was affiliated to Crenarchaeota. The abundance of AOB was mostly influenced by ammonia concentrations. The recovered ammonia oxidation rate of AOB was in the range of 45-65%, whereas for AOA, it was 15-45%, indicating that AOB were mostly responsible for the ammonia oxidation in CE during the study period. Overall, the present study provides an insight into the relevance and contribution of different groups of ammonia oxidizing bacteria in CE and emphasizes the need for further in depth studies across space and on season scale.