Metagenomic analysis of pathogenic bacteria and virulence factor genes in coastal sediments from highly urbanized cities of India.

A metagenomic approach was employed to investigate the diversity and distribution of Virulence Factors Genes (VFGs) and Pathogenic Bacteria (PB) in sediment samples collected from highly urbanized cities along the Indian coastline. Among the study locations, Mumbai, Veraval and Paradeep showed a higher abundance of PB, with Vibrio and Pseudomonas as dominant at the genus level, and Escherichia coli and Pseudomonas aeruginosa at the species level. In total, 295 VFGs were detected across all sediment samples, of which 40 VFGs showed a similarity of ≥90 % with the Virulence Database (VFDB) and were focused in this study. Among the virulent proteins, twitching motility protein and flagellar P-ring were found to be prevalent and significantly associated with Vibrio spp., and Pseudomonas spp., indicating potential bacterial pathogenicity. This investigation serves as the basis for future studies and provides insights into the comprehensive taxonomic profiles of PB, VFGs and their associated PB in the coastal sediments of India.

Microplastics in the benthic invertebrates from the coastal waters of Kochi, Southeastern Arabian Sea.

This study examined microplastic particles present in the benthic invertebrates Sternaspis scutata, Magelona cinta (deposit feeders) and Tellina sp. (suspension feeder) from the surface sediments of off-Kochi, southwest coast of India. The microplastic particles and thread-like fibres detected in these organisms were identified to be polystyrene by using DXR Raman microscope. Examination of the microplastic particle in Sternaspis scutata by epifluorescent microscopy showed fragmentation marks on the surface suggesting that the microplastic particle was degraded/weathered in nature. The study provides preliminary evidence of the presence of microplastics in benthic fauna from the coastal waters of India. However, further studies are required to understand the sources, distribution, fate and toxicity of the different types of microplastics in benthic invertebrates in order to identify any potential threats to higher trophic level organisms.

"How can our children learn from us about our way of life or understand who they are?": Residential schools and their impact on the wellbeing of Indigenous youth in Attapadi, South India.

Residential schools are commonly used in India to provide education for Indigenous youth, which requires young people to stay for long periods at distance from their families and communities. Internationally, there is clear evidence for the deleterious effects of residential schools on the mental health and social and community outcomes of Indigenous children, however little is known about the Indian Indigenous experience. This study examined the impact of residential schooling on Indigenous children's wellbeing and that of their communities, using data from an ethnographic research project in Attapadi, Kerala, including interviews, focus group discussions and participant observation with Indigenous communities. Key outcomes from residential schooling reported by the participants include the fear of losing Indigenous identity, shame of being Indigenous, change in the attitude of young people when they returned from schools, and feelings of confusion and stress that young Indigenous participants felt trying to fit into their communities on their return. Findings suggest that these Indigenous youth felt disconnected from several factors that are known to promote resilience for Indigenous communities including a strong cultural identity, connection to the land and ancestors, thereby making them more vulnerable to poor mental health and negative impacts on their overall wellbeing. Addressing these concerns requires a detailed understanding of the specific factors influencing outcomes for Indigenous youth within the Indian residential schooling system, and designing and implementing data-informed conceptual, structural and policy change including the provision of culturally safe mental health services.

First report on the occurrence of Gonyaulax polygramma bloom during the onset of Noctiluca scintillans bloom along the Tuticorin coast, southeast coast of India.

Dense and green-coloured patches were encountered on the sea surface waters of the Tuticorin coast on 22nd October 2022. Microscopic investigation revealed that the discoloration is caused by plankton, green Noctiluca scintillans. In order to find out the causes that trigger the bloom of N. scintillans, plankton samples were collected for 5 days in fourteen days duration from 22nd October to 4th November. During the peak bloom period, the abundance and biovolume of N. scintillans reached 1.56 × 104 cells/L and 21.8 × 1010µm3/L, respectively. The highest concentration (73.65 mg/m3) of chlorophyll-a was recorded during blooming period that was caused by Gonyaulax polygramma and endosymbiont, Pedinomonas noctilucae in N. scintillans. Formation of G. polygramma bloom is being reported for the first time in Tuticorin, southeast coast of India, with a species abundance of 36.9 × 104 cells/L. Present study concluded that besides the optimum hydrological conditions and eutrophic nature of the system, abundant prey (G. polygramma) facilitated the N. scintillans bloom.

Impact of untreated sewage and thermal effluent discharges on the air-sea CO2 fluxes in a highly urbanized tropical coastal region.

Various biotic and abiotic factors regulate carbon dioxide (CO2) absorption and emission in coastal waters. Factors controlling the regional air-sea CO2 fluxes were studied in Tuticorin Bay, a highly urbanized region along the southeast coast of India. Significant spatial heterogeneity in the distribution of inorganic carbon components in the bay was observed based on the exposure to different anthropogenic pressures. Thermal effluent discharges made the south zone of the bay a strong CO2 source by enhancing heterotrophy. Untreated sewage discharges in the middle zone mediated eutrophic conditions leading to strong autotrophy and restricting the zone as a weak source of CO2. Irrespective of the anthropogenic stressors, biological processes dominated the air-sea CO2 fluxes in the Tuticorin Bay. The results indicated that micro-level studies are needed in understanding the carbon cycle in environments with multiple anthropogenic stressors.

Assessing the in situ bacterial diversity and composition at anthropogenically active sites using the environmental DNA (eDNA).

In this study, we identified the in situ bacterial groups and their community structure in coastal waters influenced by anthropogenic inputs. The use of environmental DNA (eDNA) and high throughput sequencing (HTS) were employed to derive accurate and reliable information on bacterial abundance. The V3 and V4 hypervariable regions of the 16S rRNA gene were amplified and the sequences were clustered into operational taxonomic units to analyze the site-specific variations in community composition. The percentage composition within the bacterial orders varied significantly among nearshore anthropogenic hotspots and offshore (5 km) samples. The microbial network constructed taking the bacterial abundance as nodes displayed strong positive and negative correlations within the bacterial families. Overall, the use of eDNA coupled with HTS is an incredible means for monitoring and assessing the abundance of bacterial communities and also serves as a biomonitoring tool to understand the degree of anthropogenic contamination in coastal waters.

Blooming of Gonyaulax polygramma along the southeastern Arabian Sea: Influence of upwelling dynamics and anthropogenic activities.

The influence of upwelling on the phytoplankton community was examined during the upwelling-relaxation period in the southeastern Arabian Sea. Elevated upwelling intensity during the summer monsoon season of 2016 resulted in the re-suspension of harmful dinoflagellates into the surface water. Further, the surplus of phosphorus (P) inputs into the coastal waters from estuarine runoff during the upwelling-relaxation period induced blooming of Gonyaulax polygramma (4.9 × 106 cells L-1). Results from canonical correspondence analysis revealed that elevated upwelling intensity, P and salinity during the year 2016 likely triggered the bloom of G. polygramma in the study region. HABs like G. polygramma threaten fish stocks such as sardines which have a vital role in the ecosystem. Studies on phytoplankton communities and nutrient dynamics in upwelling systems would be useful in predicting the incidence/toxic effects of harmful algal blooms as these regions have a high potential for fisheries.

Loricate ciliates as an indicator of eutrophication status in the estuarine and coastal waters.

Owing to the environmental sensitivity of ciliate species, the monitoring and assessment of these communities can indicate the health of the aquatic environment. Sampling for loricate ciliates was carried out during different seasons at three different locations in the estuarine and coastal waters of Kochi, Southwest coast of India. The loricate ciliate community comprising of 27 species belonging to 10 genera revealed significant differences among the three sampling locations. The maximum numerical abundance and less species diversity of loricate ciliates were recorded at the estuarine locations whereas the minimum abundance and more species diversity were encountered in the coastal waters. Multivariate statistical analyses revealed that Tintinnopsis beroidea and Tintinnopsis uruguayensis were correlated positively with nutrients in the estuarine environment, which receives large amount of anthropogenic inputs. Thus, we propose that these two loricate ciliates may be considered as a potential bioindicator of eutrophication status in marine ecosystems.

Synthesis, characterization, and antimalarial activity of the glucuronides of the hydroxylated metabolites of arteether.

The hydroxylated metabolites (log P 2.6-2.7) of beta-arteether (1) in rat liver microsomes that retain their endoperoxide moiety showed comparable in vitro antimalarial activity to that of the parent drug arteether (log P = 3.89). The search for analogs of artemisinin (7) more suitable for intravenous use led to the synthesis of the glucuronide conjugates of the phase I hydroxylated metabolites of arteether which were found to have good water solubility, yet retained moderate lipophilicity (log P = 0.6-1.8). While a strong correlation was observed between the log P value of the glucuronides, the phase I metabolites, and the parent compound, it was found that 9 beta-hydroxyarteetherglucuronide (26) was the most active and the most polar (log P = 0.61) of the glucuronides. While the in vitro antimalarial activity of 26 (IC50 = 89.3 ng/mL) was found to be much less than that for the parent compound, the activity of 26 was within a range that would have potential therapeutic use.

Epidemiology of acute filarial episodes caused by Wuchereria bancrofti infection in two rural villages in Tamil, Nadu, south India.

This year-long study investigated the epidemiology of acute filarial episodes due to Wuchereria bancrofti in 2 rural villages in south India. The annual incidence of 96.5 episodes/1000 population was significantly higher in males (108.5) than females (84.1) an strongly age dependent. First occurrence of acute disease was observed in 0.86% of the population, and the average duration of each episode was 3.6 +/- 2.0 d. Although more than half (63.5%) of the affected individuals suffered only 1 episode, a few experienced as many as 8 over the one-year period. Individuals with chronic disease were more prone to acute attacks, with 82.9% of the total episodes occurring in this group. No seasonal pattern was observed in the frequency of episodes. Probit analysis showed that the number of episodes per affected person was dependent on sex and chronic condition. Swelling of lymph nodes in the inguinal region and fever were the most common symptoms of acute disease. The high incidence and resulting debility observed in this study suggest that acute episodes are a significant health problem associated with lymphatic filariasis. There is clearly a need for more studies on this acute form of filarial disease to aid the understanding of the aetiology and epidemiology of acute episodes, in planning appropriate control interventions, and in evaluating the resulting health burden.

Identification of the glucuronides of the hydroxylated metabolites of the antimalarial arteether in rat plasma and urine by thermospray high-performance liquid chromatography/mass spectrometry.

A method was developed to quantitate the glucuronide conjugates (phase II metabolites, log P = 0.6-1.8) of the phase I hydroxylated metabolites (log P = 2.6-2.7) of beta-arteether (2) (log P = approximately 4.0) in rat plasma and urine. The biological fluids were extracted with a C-18 solid-phase cartridge, then subjected to analysis by thermospray high-performance liquid chromatography/mass spectroscopy with a C-18 reversed-phase analytical column in the selected ion monitoring mode. The detection limits for these glucuronide conjugates in plasma and urine ranged from 25 to 60 ng/mL, and the detection limits were appropriate for the biological activity of these glucuronide conjugates in plasma. Following intravenous administration of arteether (11.6 mg/kg), the formation of these glucuronide conjugates in rat urine was undetected (below detection limit) over 24 h. However, in rat plasma, formation of 9alpha-hydroxyarteether glucuronide (13; 255 ng/mL, IC50 = 6560 ng/mL, log P = 0.93) was noted at 15 min post-injection. Although several of the glucuronide metabolites of arteether retained sufficient in vitro antimalarial activity to be potentially useful antimalarial agents if administered as prodrugs, the levels of the glucuronides formed from arteether were too low to contribute to the antimalarial activity observed for the parent compound.

Cost of mass annual single dose diethylcarbamazine distribution for the large scale control of lymphatic filariasis.

Economic analysis of the revised strategy to control lymphatic filariasis with mass annual single dose diethylcarbamazine (DEC) at 6 mg/kg body weight launched in one of the districts of Tamil Nadu in 1996 was carried out. This exploratory study, proposed for five years in 13 districts under 7 states on a pilot scale through the Department of Public Health is an additional input of the existing National Filaria Control Programme in India. A retrospective costing exercise was undertaken systematically from the provider's perspective following the completion of the first round of drug distribution. The major activities and cost components were identified and itemized cost menu was prepared to estimate the direct (financial) and indirect (opportunity) cost related to the implementation of the Programme. The total financial cost of this Programme to cover 22.7 lakh population in the district was Rs. 22.05 lakhs. The opportunity cost of labour and capital investment was calculated to be Rs. 7.98 lakhs. The total per capita cost was Rs. 1.32, with Rs. 0.97 and Rs. 0.35 as financial and opportunity cost respectively. Based on these estimates, the implementation cost of the Programme at Primary Health Centre (PHC) level was calculated and projected for five years. The additional financial cost for the existing health care system is estimated to be Rs. 27,800 per PHC every year. DEC tablets (50 mg) was the major cost component and sensitivity analysis showed that the cost of the Programme could be minimized by 20 per cent by switching over to 100 mg tablets. The analysis indicates that this Programme is a low-cost option and the results are discussed in view of its operational feasibility and epidemiological impact.

Spatial distribution of hexabromocyclododecanes (HBCDs), polybrominated diphenyl ethers (PBDEs) and organochlorines in bivalves from Japanese coastal waters.

In order to elucidate the spatial distribution of brominated flame retardants (BFRs) in the Japanese coastal environment, hexabromocyclododecanes (HBCDs), polybrominated diphenyl ethers (PBDEs) and organochlorines (OCs: PCBs, DDTs, CHLs, HCHs, HCB) were determined in bivalves (oysters and mussels) collected from Japanese coastal waters. HBCDs and PBDEs were detected in all samples analyzed. Concentration ranges of HBCDs were 12-5200 ng g(-1) lipid wt., followed by PCBs (20-3100 ng g(-1))>PBDEs (3.1-86 ng g(-1) lipid wt.). The highest concentration of HBCDs was found in the Osaka region. This result indicates that HBCDs are ubiquitous and predominant compounds in bivalves from the Japanese coastal waters. Since no species differences between oysters and mussels were observed for the bioaccumulation properties of HBCDs and PBDEs, oysters could be utilized for BFR contamination monitoring worldwide as an alternative to mussels. Global comparisons between oysters and mussels showed that HBCD concentrations in Japan are among the highest levels reported from Asia and Europe. Estimated dietary exposures of HBCDs and PBDEs through seafood were 0.45-34 ng kg body weight(-1)d(-1), and 0.054-6.8 ng kg body weight(-1)d(-1), respectively. These exposure levels were more than 1000 times lower than the lowest observable effects or no observable adverse effects levels for HBCDs and PBDEs, respectively.

Organochlorine pesticides, polychlorinated biphenyls, and polybrominated diphenyl ethers in Irrawaddy dolphins from India.

The Irrawaddy dolphin (Orcaella brevirostris) is at risk of extinction throughout its range as a result of incidental catches, habitat degradation, and pollution. Populations of Irrawaddy dolphins are constrained by the species' narrow habitat requirement-lagoons, estuaries, rivers, and lakes-and are therefore particularly vulnerable to the effects of human activities. In this study, for the first time, concentrations of organochlorine (OC) pesticides, polychlorinated biphenyls (PCBs), and polybrominated diphenyl ethers (PBDEs) were determined in tissues of Irrawaddy dolphins collected from Chilika Lake, India, to understand the status of contamination. Dichlorodiphenyltrichloroethane and its metabolites (DDTs) were the predominant contaminants found in Irrawaddy dolphins; the highest concentration found was 10,000 ng/g lipid weight in blubber. Hexachlorocyclohexanes (HCHs) were the second most prevalent contaminants in dolphin tissues. Concentrations of PCBs, chlordanes, hexachlorobenzene, tris(4-chlorophenyl)methane, and tris(4-chlorophenyl)methanol were in the ranges of few ng/g to few hundreds of ng/g on a lipid-weight basis. In general, concentrations of OC pesticides and PCBs in Irrawaddy dolphins were lower than the concentrations reported for coastal and riverine dolphins collected in Asia. PBDEs were detected in the blubber of Irrawaddy dolphins at concentrations ranging from 0.98 to 18 ng/g lipid weight. BDE congener 47 accounted for 60% to 75% of the total PBDE concentrations. Although these results establish the baseline levels of persistent organic pollutants in Irrawaddy dolphins, efforts should be made to decrease the sources of contamination by DDTs and HCHs in Chilika Lake.

Knowledge and beliefs about transmission, prevention and control of lymphatic filariasis in rural areas of south India.

To identify the gaps in people's knowledge of lymphatic filariasis, we collected descriptive and quantitative information in rural areas of Tamil Nadu, South India. People are well aware of filariasis in their communities and recognize its different clinical manifestations. Only 9% of those with and 20% of those without the disease knew that filariasis is caused through mosquito bites; the rest attributed it to many other causes. People's knowledge about transmission and prevention of filariasis is also very poor. The study identifies the rationale for people's misconceptions about the disease. Misconceptions and ignorance could adversely affect personal protection measures against mosquito bites and the use of appropriate treatment. Health education campaigns aimed at highlighting the role of mosquitoes in transmission and the importance of early diagnosis should help people in taking personal protection measures and seeking appropriate treatment.

Development of a high-performance liquid chromatographic-tandem mass spectrometric method for the determination of pharmacokinetics of Co 102862 in mouse, rat, monkey and dog plasma.

A method for determining concentration levels of Co 102862 in mouse, rat, monkey and dog plasma was validated in the range of 5 to 2000 ng/ml using 200 microl plasma sample volume. This validation report describes the linearity, specificity, sensitivity, reproducibility, accuracy, recovery and stability of the analytical method. The inter-day RSD ranged from 3.5 to 10.1%, intra-day RSD from 0.6 to 5.7% and intra-day accuracy (mean absolute percent difference) ranged from 2.2 to 14.9% for rat, monkey and dog plasma. A mini-validation (5-2000 ng/ml) of Co 102862 was performed in mouse plasma using the same methods. Additionally, the assay range at the low end was successfully extended to 0.5 ng/ml for monkey plasma. The method was used for the routine analysis of Co 102862 in mouse, rat, monkey and dog plasma and summary of the pharmacokinetic data are presented.

In vivo metabolism and mass balance of 4-[4-fluorophenoxy]benzaldehyde semicarbazone in rats.

The pharmacokinetics, mass balance, tissue distribution, and metabolism of Co 102862 was investigated in rats after a single oral dose. [(14)C]Co 102862 showed multiexponential pharmacokinetics in rat plasma with an extensive distribution phase. After p.o. administration (approximately 10 mg/kg), the half-lives were long for total radioactivity compared with unchanged Co 102862. Profiles of rat urine and bile suggest that Co 102862 is extensively metabolized in vivo. [(14)C]Co 102862 was extensively distributed into all tissues, with the fatty tissues and secretory glands tissues containing the highest radioactivity. Elimination of radioactivity from the tissues had an estimated half-life of 14 days. A total of 91% of the administered radioactivity was recovered in both intact and bile-duct cannulated rats over 120 and 48 h, respectively, with the majority ( approximately 74%) of the radioactivity being excreted in the urine. Approximately 10% of the total radioactivity remained in the tissues on day 5 and decreased with time to approximately 3% on day 28. Bile-duct cannulated experiments show the enterohepatic circulation is an important route of elimination and reabsorption. Six metabolites were identified in the urine and bile of which the carboxylic acid was the major metabolite. The carboxylic acid was the only metabolite found in plasma and was probably responsible for the radioactivity in the tissues.

Acrolein mercapturates: synthesis, characterization, and assessment of their role in the bladder toxicity of cyclophosphamide.

Acrolein is the metabolite of cyclophosphamide (CP) believed to be involved in the bladder toxicity associated with this anticancer drug. The mechanism by which this extremely reactive intermediate is delivered to the bladder is not known. Glutathione (GSH) readily conjugates with acrolein, and the acrolein mercapturate S-(3-hydroxypropyl)-N-acetylcysteine (3-hydroxy-PrMCA) has been found in the urine of animals and man given CP. The objectives of this study were to prepare and characterize synthetic standards of the GSH acrolein adduct (3-oxopropyl)glutathione (3-oxoPrGSH), the acrolein mercapturates S-(3-oxopropyl)-N-acetylcysteine (3-oxoPrMCA) and 3-hydroxyPrMCA, and the S-oxidation product of 3-oxoPrMCA (3-oxoPrMCA S-oxide). In addition, the release of acrolein from, and the bladder toxicity of, these conjugates was determined. 3-OxoPrGSH and 3-oxoPrMCA were prepared with a 99% yield by condensing acrolein with GSH and N-acetylcysteine, respectively. 3-HydroxyPrMCA was prepared with a 63% yield by refluxing 3-chloropropanol and N-acetylcysteine in a basic medium. Oxidation of 3-oxoPrMCA with H2O2 was used to prepare 3-oxoPrMCA S-oxide. By decreasing the reaction time to 1 h, and adjusting the ratio of 3-oxoPrMCA to H2O2, the yield of 3-oxoPrMCA S-oxide was increased to 96%. The anhydrous aldehyde, 3-oxoPrMCA, afforded characteristic aldehydic proton resonances (1H NMR) in deuterated dimethyl sulfoxide. New resonances were observed in deuterated water, indicating a 75% hydration of the aldehyde to the corresponding geminal diol. This phenomenon was enhanced with 3-oxoPrMCA S-oxide where approximately 100% hydration of the aldehyde to the corresponding geminal diol was observed. When incubated at 25 degrees C in 100 mM potassium phosphate buffer containing 1 M KCl, pH 8.0, 3-oxoPrMCA released approximately 6% and 3-oxoPrMCA S-oxide released approximately 16-18% of the theoretical maximum yield of acrolein after 30 min, as indicated by an increase in absorbance at 210 nm and confirmed by trapping this aldehyde as a semicarbazone. There was less than a 2% yield of acrolein from 3-hydroxyPrMCA or 3-oxoPrGSH under similar conditions. At pH 7.4 the release of acrolein from 3-oxoPrMCA and 3-oxoPrMCA S-oxide was decreased by 50%. An assay where aldehydes are reacted with m-aminophenol in acid media produced fluorescence consistent with 72%, 46%, 23%, and 1% yields of acrolein from 3-oxoPrMCA S-oxide, 3-oxoPrMCA, 3-oxoPrGSH, and 3-hydroxyPrMCA, respectively.(ABSTRACT TRUNCATED AT 400 WORDS)

Studies on the basis for the toxicity of acrolein mercapturates.

Acrolein, 3-oxopropyl glutathione (oxoPrGSH), and S-3-hydroxypropyl N-acetylcysteine (hydroxyPrMCA) are confirmed metabolic products of cyclophosphamide. Other potential metabolites include the mercapturic acid S-3-oxopropyl N-acetylcysteine (oxoPrMCA), its sulfoxide, and their corresponding diacid forms. The reactivity of acrolein would appear to preclude its movement from the main site of formation (liver) to the sites of toxicity (lung and bladder). However, the rerelease of acrolein from various thiol conjugates via a beta-elimination reaction is possible. It is also possible that the parent conjugate is directly toxic. The current study examined the toxicity of various acrolein-thiol conjugates and related analogs to human lung adenoma A549 cells. The expected enhancement of acrolein, oxoPrMCA, and oxoPrMCA S-oxide toxicity (assessed as cell proliferation by the alamarBlue assay) following a 2-hr exposure of cells treated with diethyl maleate (DEM) to deplete GSH was observed. OxoPrGSH was toxic when present for 24 hr, and this toxicity was also enhanced by pretreatment with DEM. When treated with these conjugates alone, the depletion of intracellular GSH only occurred at doses above those needed to significantly inhibit cell proliferation. There were no changes in protein thiols as determined using the membrane impermeant fluorescent thiol probe para-sulfobenzoyloxybromobimane. The diacid conjugate was not toxic to A549 cells indicating further oxidation products of the mercapturic acids are not a factor in toxicity. ButanoneMCA, an analog of oxoPrMCA that cannot exist in the geminal diol form, inhibited A549 cell growth only slightly less effectively than oxoPrMCA, suggesting the geminal diol is not toxicologically significant. OxoBuMCA, which cannot undergo beta-elimination of acrolein, showed no toxicity to these cells, suggesting that the release of acrolein could be required. Exogenous GSH provided protection from all toxic compounds suggesting that the toxic species is electrophilic. Overall, the data suggest that toxicity, in terms of an inhibition of cell proliferation, is not due to the parent molecule but rather is the result of rereleased acrolein that then affects factors necessary for cell proliferation.