Phylogenetic Diversity of Bacterial Community Associated with the Marine Sponge Halichondira nigrocutis Collected off Southwest Indian Coast

This study aimed to evaluate the bacterial communities associated with the marine sponge Halichondria nigrocutis present in Indian waters by both cultivation and cultivation-independent techniques. Results using cultivation method showed that bacteria belonging to Bacillus, Acinetobacter and Vibrio spp. to be the predominant groups. Metagenomic study of sponge-associated bacteria by cultivation independent approach, involving cloning and sequencing of the 16S rDNA gene generated sequences that were subjected to phylogenetic analysis. Results demonstrated the community structure to be represented by the phyla Proteobacteria (alpha-, gamma- and delta-classes), Cyanobacteria, Actinobacteria and Firmicutes, with isolates belonging to alpha-proteobacterial group to be predominating. To our knowledge this study appears to be the first to record cultivable and uncultivable bacterial groups associated with H.nigrocutis from Indian waters

Effect of Ammonia Concentration on the Nitrification Potential of Ammonia Oxidizing Bacterial Isolates from Fish Processing Waste Effluents

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Future climate change conditions may compromise metabolic performance in juveniles of the mud crab Scylla serrata

This work was supported by a PhD scholarship granted by the University of Plymouth to EA (PI LMT).Research characterising the effects of future climate change on the marine environment remains heavily focussed on that of temperate regions and organisms. Furthermore, little is known of these effects on the early life stages of many marine species. Tropical regions are already experiencing an increase in sea surface temperature and decrease in sea surface salinity, conditions favoured by pathogenic bacteria such as Vibrio spp. The early life stages of crabs are known to be particularly vulnerable to both the direct physiological effects of climate change and exposure to harmful microorganisms, yet there are limited data on these effects on juveniles of many tropical crustacean species. This study assessed the metabolic responses of mud crab (Scylla serrata) juveniles to warming and/or freshening in the presence or absence of pathogenic bacteria in southwest India. Juvenile crabs were exposed to either ambient (28 °C/30 PSU) or one of three projected climate change regimes (28 °C/20 PSU (freshening), 32 °C/30 PSU (warming), 32 °C/20 PSU (warming + freshening)) for 10 days, in either the presence or absence of the pathogenic bacteria Vibrio parahaemolyticus. Results show that simulated climate change conditions, especially freshening, caused a significant increase in oxygen consumption rates (MO2), and that these were further increased when juveniles were exposed to V. parahaemolyticus. These results suggest that the effects of future climate change conditions could have significant implications for the conservation of wild stocks and commercial farming of this species in South Asia.Publisher PDFPeer reviewe

Differential expression of akirin gene in black tiger shrimp Penaeus monodon in response to immunostimulant administration and infections with Vibrio harveyi and white spot syndrome virus

The akirin gene, which is strictly localized in the nucleus, plays a critical role in regulating antimicrobial peptide transcription, and has parallel functions to NF-kappa B signaling pathway in both vertebrates and invertebrates. In shrimp, the akirin gene is expressed as innate immunity in response to microbial infection. In the present study, expression of akirin gene in Penaeus monodon with respect to Vibrio harveyi and white spot syndrome virus (WSSV) infections and immunostimulant (beta-glucan) administration were investigated by quantitative polymerase chain reaction. The gene was expressed in various tissue samples of healthy shrimp. Maximum level of expression was immediately after V. harveyi infection, suggesting that it may be an early response gene. Gene expression was remarkably upregulated in the lymphoid organ, gill, and hepatopancreas, whereas downregulation was observed in hemocytes compared with the control. In the case of WSSV-infected samples, the akirin gene was significantly downregulated in the lymphoid organ but there was no significant difference in expression pattern in hemocytes compared to the control. In gill tissue, maximum expression was observed after 2 hr of infection, the same in hepatopancreas. Experimental challenge of beta-glucan fed shrimp infected with V. harveyi and WSSV resulted in significant upregulation of akirin gene expression in lymphoid and gill tissue

Regulation of virulence factors by quorum sensing in Vibrio harveyi.

Vibrio harveyi is an important aquatic pathogen that produces several virulence factors. In this study, the effect of quorum sensing, bacterial cell-to-cell communication, on the production of the virulence factors caseinase, gelatinase, lipase, hemolysin, and phospholipase, was investigated. The activity of virulence factors was studied through enzymatic plate assays using V. harveyi wild type and mutants with constitutively maximal or minimal quorum sensing activity. The results showed that quorum sensing negatively regulates phospholipase activity as higher activity was observed in mutants with minimal quorum sensing activity than in the mutant with maximal quorum sensing activity. Reverse transcriptase real-time PCR with specific primers revealed that the expression level of three phospholipase genes was 2-fold lower in the mutant with minimal quorum sensing activity than in the mutant with maximal quorum sensing activity. As far as we know, this is the first report of quorum sensing regulation of phospholipase. Finally, caseinase and gelatinase activity were positively regulated by quorum sensing, which is consistent with previous reports, and lipase and hemolysin activity were found to be independent of quorum sensing. Hence, the regulation is different for different virulence factors, with some being either positively or negatively regulated, and others being independent of quorum sensing. This might reflect the need to produce the different virulence factors at different stages during infection

Diagnosis, Clinical and Molecular Delineation of Human Plasmodium Species from Mangaluru, Southwest India

Malaria is a global threat and a never-ending battle without appropriate identification and differentiation of the parasite species. This work compared the diagnostic methods including the thick film microscopy technique, quantitative buffy coat, and polymerase chain reaction. The inaccuracy of species determination by microscopy and the consequent treatment regime underlines the necessity to upgrade routine diagnostic methods with molecular techniques. In the study, 436 samples were collected; venous blood was processed for the quantitative buffy coat technique followed by classical Giemsa staining of thin and thick smears and nested Polymerase Chain Reaction (nPCR) for the genus-specific region of Plasmodium targeting 18S rDNA followed by species-specific identification. Of 436 samples screened for malaria, results in PCR showed 78.7% (100/127) to be P. vivax, 4.8% (6/127) as P. falciparum and 16.5% (21/127) to be mixed infection (P. vivax + P. falciparum). The prevalence of malaria was 0.29, and there was good concordance between the methods for detecting Plasmodium (Kappa:0.77). In our investigation, nested PCR and TFM exhibited a sensitivity of 97.7% and a specificity of 100% for malaria detection compared to QBC. Clinical parameters- thrombocytopenia and anemia, were compared in this study. A positive association was observed between thrombocytopenia and malaria (p<0.05), but the association between anemia and malaria infection remains unclear. Primer cross-reactions were also observed in the primer sequence of P. ovale and P. knowlesi, but sequencing confirmed it as P. vivax and the study of phylogeny paved a new way in analyzing the relatedness of the sequences

Toxic Algae Silence Physiological Responses to Multiple Climate Drivers in a Tropical Marine Food Chain

Research on the effects of climate change in the marine environment continues to accelerate, yet we know little about the effects of multiple climate drivers in more complex, ecologically relevant settings – especially in sub-tropical and tropical systems. In marine ecosystems, climate change (warming and freshening from land run-off) will increase water column stratification which is favorable for toxin producing dinoflagellates. This can increase the prevalence of toxic microalgal species, leading to bioaccumulation of toxins by filter feeders, such as bivalves, with resultant negative impacts on physiological performance. In this study we manipulated multiple climate drivers (warming, freshening, and acidification), and the availability of toxic microalgae, to determine their impact on the physiological health, and toxin load of the tropical filter-feeding clam, Meretrix meretrix. Using a structural equation modeling (SEM) approach, we found that exposure to projected marine climates resulted in direct negative effects on metabolic and immunological function and, that these effects were often more pronounced in clams exposed to multiple, rather than single climate drivers. Furthermore, our study showed that these physiological responses were modified by indirect effects mediated through the food chain. Specifically, we found that when bivalves were fed with a toxin-producing dinoflagellate (Alexandrium minutum) the physiological responses, and toxin load changed differently and in a non-predictable way compared to clams exposed to projected marine climates only. Specifically, oxygen consumption data revealed that these clams did not respond physiologically to climate warming or the combined effects of warming, freshening and acidification. Our results highlight the importance of quantifying both direct and, indirect food chain effects of climate drivers on a key tropical food species, and have important implications for shellfish production and food safety in tropical regions.</p

Longitudinal disease studies in small-holder black tiger shrimp (Penaeus monodon) farms in Andhra Pradesh, India. I. High prevalence of WSSV infection and low incidence of disease outbreaks in BMP ponds

A longitudinal study was conducted from January to August 2005 in small-holder black tiger shrimp (Penaeus monodon) ponds in the West Godavari District of Andhra Pradesh, India (16°25′ N, 81°19′ E). The study involved 457 ponds owned by low-income farmers participating in a better management practice (BMP) programme. Disease outbreaks occurred in 16.6% of ponds. There was significant spatial clustering of disease outbreaks with 31 (40.8%) of the 76 recorded disease outbreaks occurring in a single village block. Bivariate analysis indicated a 1.6-fold higher likelihood of disease outbreaks from nursery-stocked ponds but this was not significant in multivariate analysis due to the confounding effect of pond location. There was evidence of increasing prevalence of WSSV infection during grow-out. WSSV was detected in 5.9% of 119 batches of postlarvae tested at stocking, 38.2% of 34 juvenile batches collected at the time of transfer to grow-out ponds, and 47.0% of 336 pond stock tested at normal harvest or crop failure. WSSV was detected in 43 of 59 (72.9%) disease outbreak ponds tested and 115 of 277 (41.5%) non-outbreak ponds tested. Heavy WSSV infection was detected at harvest in 116 of the 336 (34.5%) of the ponds tested, including 78 ponds for which no outbreak was recorded. Duration of crop was recorded for 431 ponds with a mean of 117.0 days and a range of 20 to 176 days. Median duration was significantly shorter for disease outbreak ponds (68.5 days) compared to nonoutbreak ponds (119.0 days). Duration of crop also varied according to WSSV detection levels at harvest, with median duration for ponds classified as heavy WSSV infection (108.5 days) significantly shorter than for ponds classified as either light WSSV infection (116.0 days) or WSSV-negative (116.5 days). The study indicated a high risk of WSSV infection during grow-out but a relatively low incidence of disease despite a high prevalence of heavy WSSV infection in non-outbreak ponds

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Ecology, Virulence and Detection of Pathogenic and Pandemic Vibrio Parahaemolyticus

Vibrio parahaemolyticus is a gram negative, halophilic bacterium that occurs in the coastal and estuarine environments worldwide and is implicated in several cases of seafood-born gastroenteritis around the globe. However, not all strains of V. parahaemolyticus are pathogenic. Clinical isolates of V. parahaemolyticus most often produce either the thermostable direct haemolysin (TDH) or TDH-related haemolysin (TRH) encoded by tdh and trh genes, respectively. A pandemic clone of O3:K6 which was first detected in Kolkata (India), has been responsible for many outbreaks in Asia and the USA. With the emergence of pandemic clone of V. parahaemolyticus, this organism has assumed significance. Although most of the V. parahaemolyticus outbreaks are invariably related to seafood consumption, pathogenic strains are rarely isolated from seafood. Virulent strains producing TDH or TRH and the pandemic clone, which is responsible for most of the outbreaks (that have occurred after 1996) have been rarely isolated from seafood and other environmental samples. This could be due to the occurrence of pathogenic strains in the estuarine environment at a lower level compared to non-pathogenic strains. Another reason can be that the pathogenic stains are more sensitive to dystropic conditions in the aquatic environment and rapidly become non-culturable. Similarity in growth kinetics between virulent and non-virulent strains also made the isolation of virulent strains from the aquatic environment difficult. Several studies were done to determine the factors responsible for an increased virulence and persistance of pandemic clone. However, none of those studies were conclusive. Several researchers have proposed various genetic markers for specific detection of pandemic clone of V. parahaemolyticus. But many of those genetic markers were found to be unreliable. Recently, seven genomic islands (VPaI-1 to VPaI-7) unique to pandemic clone were identified. This Research Topic is dedicated to improve our current understanding of ecology, pathogenesis and detection of pathogenic and pandemic clone of V. parahaemolyticus, and will also strive to identify areas of future development