Heavy metal accumulation analysis using bivalve, sponge, sea urchin, and gastropod species as bioindicators.

A comparative assessment of heavy metal accumulation potential in four distinct marine benthic bioindicators: the bivalve Perna perna, the sponge Callyspongia fibrosa, the sea urchin Tripneustes gratilla, and the gastropod Purpura bufo were conducted. These organisms were collected from the same location, and the concentration of ten heavy metals was analyzed in water, sediment and various body parts of the organisms. The bioaccumulation potential was evaluated using the bio-water accumulation factor and bio-sediment accumulation factor. There was significant variation in the bioaccumulation potential of each organism with respect to different metals. The sponge proved to be a reliable indicator of Cd with a highest concentration of 2.60 µg/g. Sea urchin accumulated high concentrations of Cr (16.98 µg/g) and Pb (4.80 µg/g), whereas Cu was predominant (21.05 µg/g) in gastropod, followed by bivalve (17.67 µg/g). The concentration of metals in hard parts was found to be lower than in the tissues.

Comparative analysis of temporal variation of heavy metal accumulation by two sea urchin species from a harbour region, including pre and post COVID 19 lock down period.

Sea urchins are marine invertebrates belonging to phylum Echinodermata, recognized as relevant biological tool for assessing environmental pollution. In the present study, we assessed the bioaccumulation potential of different heavy metals by two sea urchin species, Stomopneustes variolaris Lamarck, 1816 and Echinothrix diadema Linnaeus, 1758, collected from a harbour region, along the south west coast of India, during four different sampling periods for 2 years, from the same sea urchin bed. Heavy metals like Pb, Cr, As, Cd, Co, Se, Cu, Zn, Mn and Ni were analysed from water, sediment and different body parts of sea urchins, such as shell, spine, tooth, gut and gonad. The sampling periods also included the pre and post COVID 19 lockdown period during which the harbour activities were closed. The bio-water accumulation factor (BWAF), bio-sediment accumulation factor (BSAF) and the metal content/test weight index (MTWI) were calculated, in order to compare the bioaccumulation of metals by both the species. The results showed that S. variolaris had higher bioaccumulation potential than E. diadema, for metals like Pb, As, Cr, Co and Cd especially in the soft body parts like gut and gonad. The hard parts of S. variolaris like shell, spine, and tooth also accumulated more Pb, Cu, Ni and Mn than E. diadema. Following the lockdown period, there was a decline in the concentration of all heavy metals in water, whereas in sediment, Pb, Cr, and Cu levels were reduced. The gut and gonad tissues of both the urchins showed a decrease in the concentration of most of the heavy metals following the lockdown phase and no significant reduction was observed in the hard parts. This study reveals the use of S. variolaris as an excellent bioindicator of heavy metal contamination in the marine environment which can be employed for coastal monitoring programs.

Functional characterization of a histone H2A derived antimicrobial peptide HARRIOTTIN-1 from sicklefin chimaera, Neoharriotta pinnata.

Antimicrobial peptides (AMPs) are gene encoded short peptides which play an important role in the innate immunity of almost all living organisms ranging from bacteria to mammals. Histones play a very important role in defense as precursors to bioactive peptides. The present study is an attempt to decipher the antimicrobial activity of a histone H2A derived peptide, Harriottin-1 from sicklefin chimaera, Neoharriotta pinnata. Analysis in silico predicted the molecule with potent antibacterial and anticancer property. The Harriottin-1 was recombinantly produced and the recombinant peptide rHar-1 demonstrated potent antibacterial activity at 25 µM besides anticancer activity. The study strongly suggests the importance of histone H2A derived peptides as a model for the design and synthesis of potent peptide drugs.

A collection of crabs (Crustacea, Brachyura) from the southwestern coast of India, with a discussion of the systematic position of Nectopanope Wood-Mason in Wood-Mason & Alcock, 1891 (Euryplacidae).

A report on the brachyuran crabs collected from the southwestern coast of India by the Indian research vessel FORV Sagar Sampada is presented. The material consists of 13 species from three genera and five families, of which four are new records for India: Heteroplaxmaldivensis (Rathbun, 1902) (Euryplacidae), Cryptopodiacollifer Flipse, 1930 (Parthenopidae), Thalamitamacrodonta Borradaile, 1903 (Portunidae), and Paraxanthodescumatodes (MacGilchrist, 1905) (Xanthidae). The cruise also obtained the first known male of Nectopanoperhodobaphes Wood-Mason in Wood-Mason & Alcock, 1891 (type species of Nectopanope Wood-Mason in Wood-Mason & Alcock, 1891), and its characters show that it is in fact a member of the Euryplacidae Stimpson, 1871. The genus had previously been incorrectly classified in the Xanthidae MacLeay, 1838.

Remote sensing of bacterial response to degrading phytoplankton in the Arabian Sea.

A remote sensing technique has been developed to detect physiological condition of phytoplankton using in situ and moderate imaging spectroradiometer (MODIS)-Aqua data. The recurring massive mixed algal bloom of diatom and Noctiluca scintillans in the Northern Arabian Sea during winter-spring was used as test bed to study formation, growth and degradation of phytoplankton. The ratio of chlorophyll (chl) to particulate organic carbon (POC) was considered as an indicator of phytoplankton physiological condition and used for the approach development. Algal blooms represent the areas of new production, and therefore, knowledge of their degradation is important to the study microbial loop and export carbon flux. Relation of chl/POC ratio with bacterial abundance revealed Gaussian distribution. Bacteria were strongly correlated with POC, and hence, the latter which is available from satellite data could be used as a proxy for remote assessment of bacteria. Thresholds for active and degrading phytoplankton were determined using the ratio computed from the satellite data. The criteria were implemented on MODIS data to generate an image representing distribution of degrading algal bloom. Bacteria abundance data from two validation cruises during dinoflagellate and cyanobacteria bloom confirmed well match up of phytoplankton degradation information from the satellite. Comparison of environmental parameters during decay phase of dinoflagellate (N. scintillans bloom (winter) and Trichodesmium bloom (summer) revealed that degradation after active Trichodesmium bloom was more severe as compared to the N. scintillans. The present study also highlights the prediction capability of phytoplankton degradation using a time series of satellite retrieved chlorophyll/POC images.

Pyocyanin induced in vitro oxidative damage and its toxicity level in human, fish and insect cell lines for its selective biological applications.

Pyocyanin is a redox active phenazine pigment produced by Pseudomonas aeruginosa, with broad antibiotic activity having pharmacological, aquaculture, agriculture and industrial applications. In the present work cytotoxicity induced by pyocyanin is demonstrated in a human embryonic lung epithelial cell line (L-132), a rainbow trout gonad cell line (RTG-2) and a Spodoptera frugiperda pupal ovarian cell line (Sf9). For toxicity evaluation, cellular morphology, mitochondrial function (XTT), membrane leakage of lactate dehydrogenase, neutral red uptake, affinity of electrostatic binding of protein with sulforhodamine B dyes, glucose metabolism, and reactive oxygen species, were assessed. Results showed that higher pyocyanin concentration is required for eliciting cytotoxicity in L-132, RTG-2 and Sf9. The microscopic studies demonstrated that the cell lines exposed to pyocyanin at higher concentrations alone showed morphological changes such as clumping and necrosis. Among the three cell lines L-132 showed the highest response to pyocyanin than the others. In short, pyocyanin application at concentrations ranging from 5 to 10 mg l(-1) were not having any pathological effect in eukaryotic systems and can be used as drug of choice in aquaculture against vibrios in lieu of conventional antibiotics and as biocontrol agent against fungal and bacterial pathogens in agriculture. This is besides its industrial and pharmacological applications.

Impaired telomerase activity hinders proliferation and in vitro transformation of Penaeus monodon lymphoid cells.

Retaining terminal transferase activity of telomerase, the ribonucleoprotein enzyme which add telomeric repeats on chromosome end is thought to be required to prevent cellular ageing. Additionally, telomerase considered as a marker for cell proliferation and immortalization in eukaryotes. We examined telomerase activity in tissues and lymphoid cell culture of Penaeus monodon. Along with telomerase activity, telomere repeats and an attempt on identification of telomerase reverse transcriptase (PmTERT) were made. Telomeric repeat amplification protocol revealed that telomerase-dependent telomeric lengthening has been taking place in P. monodon and the adult tissues were retaining this capacity throughout their lifespan with the highest activity in ovary, testis and lymphoid organ. However, telomerase activity could not be detected in lymphoid cells in culture. The canonical telomeric repeats added by telomerase of lymphoid tissue extract were identified as TTAGG, but pentameric repeats GGTTA and AGGTT were also added by the telomerase. PmTERT protein sequence (partial) shared 100 % identity with the TERT sequence of Daphnia pulex, 27 % sequence identity with Purple sea urchin and 24-25 % with Zebra fish. Undetectable telomerase activity in lymphoid cell culture supports the hypothesis that the inadequate telomerase activity or gene expression may be a reason that prevents neoplastic transformation and spontaneous immortalization of the cells in vitro. Thus, it is envisaged that telomerase activation in lymphoid cells may surmount cellular ageing for in vitro transformation and cell line establishment.

MODIS-Aqua detects Noctiluca scintillans and hotspots in the central Arabian Sea.

Northern Arabian Sea is considered as an ecologically sensitive area as it experiences a massive upwelling and long-lasting algal bloom, Noctiluca scintillans (green tide) during summer and spring-winter, respectively. Diatom bloom is also found to be co-located with N. scintillans and both have an impact on ecology of the basin. In-house technique of detecting species of these blooms from Moderate Resolution Imaging Spectroradiometer (MODIS)-Aqua data was used to generate a time-series of images revealing their spatial distribution. A study of spatial-temporal variability of these blooms using satellite data expressed a cyclic pattern of their spread over a period of 13 years. An average distribution of the blooms for January-March period revealed a peak in 2015 and minimum in 2013. Subsequently, a time-series of phytoplankton species images were generated for these 2 years to study their inter-annual variability and the associated factors. Species images during active phase of the bloom (February) in 2015 indicated development of N. scintillans and diatom in the central Arabian Sea also, up to 12° N. This observation was substantiated with relevant oceanic parameters measured from the ship as well as satellite data and the same is highlight of the paper. While oxygen depletion and release of ammonia associated with N. scintillans are detrimental for waters on the western side; it is relatively less extreme and supports the entire food chain on the eastern side. In view of these contrasting eco-sensitive events, it is a matter of concern to identify biologically active persistent areas, hot spots, in order to study their ecology in detail. An ecological index, persistence of the bloom, was derived from the time-series of species images and it is another highlight of our study.

Alkaline protease from a non-toxigenic mangrove isolate of Vibrio sp. V26 with potential application in animal cell culture.

Vibrio sp. V26 isolated from mangrove sediment showed 98 % similarity to 16S rRNA gene of Vibrio cholerae, V. mimicus, V. albensis and uncultured clones of Vibrio. Phenotypically also it resembled both V. cholerae and V. mimicus. Serogrouping, virulence associated gene profiling, hydrophobicity, and adherence pattern clearly pointed towards the non-toxigenic nature of Vibrio sp. V26. Purification and characterization of the enzyme revealed that it was moderately thermoactive, nonhemagglutinating alkaline metalloprotease with a molecular mass of 32 kDa. The application of alkaline protease from Vibrio sp. V26 (APV26) in sub culturing cell lines (HEp-2, HeLa and RTG-2) and dissociation of animal tissue (chick embryo) for primary cell culture were investigated. The time required for dissociation of cells as well as the viable cell yield obtained by while administering APV26 and trypsin were compared. Investigations revealed that the alkaline protease of Vibrio sp. V26 has the potential to be used in animal cell culture for subculturing cell lines and dissociation of animal tissue for the development of primary cell cultures, which has not been reported earlier among metalloproteases of Vibrios.

Molecular characterization of a crustin-like antimicrobial peptide in the giant tiger shrimp, Penaeus monodon, and its expression profile in response to various immunostimulants and challenge with WSSV.

A crustin-like antimicrobial peptide from the haemocytes of giant tiger shrimp, Penaeus monodon was partially characterized at the molecular level and phylogenetic analysis was performed. The partial coding sequence of 299 bp and 91 deduced amino acid residues possessed conserved cysteine residues characteristic of the shrimp crustins. Phylogenetic tree and sequence comparison clearly confirmed divergence of this crustin-like AMP from other shrimp crustins. The differential expression of the crustin-like AMP in P. monodon in response to the administration of various immunostimulants viz., two marine yeasts (Candida haemulonii S27 and Candida sake S165) and two ß-glucan isolates (extracted from C. haemulonii S27 and C. sake S165) were noted during the study. Responses to the application of two gram-positive probiotic bacteria (Bacillus MCCB101 and Micrococcus MCCB104) were also observed. The immune profile was recorded pre- and post-challenge white spot syndrome virus (WSSV) by semi-quantitative RT-PCR. Expressions of seven WSSV genes were also observed for studying the intensity of viral infection in the experimental animals. The crustin-like AMP was found to be constitutively expressed in the animal and a significant down-regulation could be noted post-challenge WSSV. Remarkable down-regulation of the gene was observed in the immunostimulant fed animals pre-challenge followed by a significant up-regulation post-challenge WSSV. Tissue-wise expression of crustin-like AMP on administration of C. haemulonii and Bacillus showed maximum transcripts in gill and intestine. The marine yeast, C. haemulonii and the probiotic bacteria, Bacillus were found to enhance the production of crustin-like AMP and confer significant protection to P. monodon against WSSV infection.