Fluorene-induced stress in the benthic oligochaete Tubifex tubifex: A multi-biomarker assessment of toxicological pathways and mechanisms under acute and subchronic exposures.

This study examined the multifaceted impacts of fluorene exposure on Tubifex tubifex, encompassing acute (survival analysis and behavioral responses) and subchronic exposure regimens (antioxidant enzyme response and histopathology), molecular docking studies, and generalized read-across analysis. Survival analysis revealed concentration-dependent increases in toxicity over varying time intervals, with LC50 values decreasing from 30.072 mg/L at 24 h to 12.365 mg/L at 96 h, emphasizing the time-sensitive and concentration-responsive nature of the stressor. Behavioral responses were both concentration- and duration-dependent. While Erratic Movement and Clumping Tendency exhibited earlier responses (within 24 h) at lower concentrations, the wrinkling effect and mucus secretion) exhibited delayed onset, suggesting intricate regulatory mechanisms underlying adaptability to environmental challenges; moreover, the wrinkling effect was consistently induced at higher concentrations, indicating greater sensitivity to the toxic effects of fluorene. With sublethal environmentally relevant concentrations-1.24 mg/l and 2.47 mg/L i.e., 10% and 20% 96 h, respectively-the antioxidant enzyme response (i.e., upregulation of SOD, CAT, and GST) with increasing fluorene concentration, revealing a nonlinear, hormetic response, suggested adaptive protection at lower doses but inhibition at higher concentrations. Histopathological examination indicated that higher fluorene concentrations caused cellular proliferation, inflammation, and severe tissue damage in the digestive tract and body wall. Molecular docking studies demonstrated robust interactions between fluorene and major stress biomarker enzymes, disrupting their functions and inducing oxidative stress. Interactions with cytochrome c oxidase suggested interference with cellular energy production. Generalized Read-Across (GenRA) analysis unveiled shared toxicity mechanisms among fluorene and its analogs, involving the formation of reactive epoxides and the influence of cytochrome P450 enzymes. The diverse functional groups of these analogs, particularly chlorine-containing compounds, were implicated in toxicity through lipid peroxidation and membrane damage. Adverse outcome pathways and broader consequences for aquatic ecosystem health are discussed.

Mitigating the Public Health Issues Caused by the Filarial Vector, Culex quinquefasciatus (Diptera: Culicidae) Through Phytocontrol and Larval Source Marker Management.

Failure of conventional mosquito control strategies to curb the population of vectors have made the humans vulnerable to serious medical problems transmitted by them. This effect has been compounded by global climate change enabling the mosquitoes to cross geographical boundaries and cause trouble in regions where they were initially not found. As such, the scientific community has been compelled to devise alternative and innovative strategies of mosquito control that can be integrated with the conventional practices to implement multi-phasic approach of vector management. Culex quinquefasciatus is one such mosquito species that is reported to be one of the primary vectors of lymphatic filariasis and many other diseases of global health concern. However, not much is known about its breeding habitat ecology and microbial properties that have enabled the species to achieve reproductive success in urbanized habitats. The current investigation was carried out at Digha, West Bengal, India. The region, despite being endemic for lymphatic filariasis, has rarely been explored for its mosquito diversity and/or their breeding habitat characteristics. Therefore, these were attempted. For survey and sampling, seven villages were chosen, namely, Duttapur, Jatimati, Champabani, Padima, Gobindabasan, Bhagibaharampur and Palsandapur. The study showed that Cx. quinquefasciatus is the dominant mosquito species at the sampling sites with the highest density of their larvae being recorded from man-made structures like drains and pools close to human habitations and livestock. The study was, therefore, restricted to Cx. quinquefasciatus. Seasonal abundance showed that they were most prevalent in the monsoon followed by summer. The physicochemical characterization showed their larvae to prefer almost neutral pH (6.9 to 7.3), low chloride concentration (98 to 258 ppm) and turbidity. As far as other parameters are concerned, they were tolerant towards a wide range allowing them to adapt varied habitats in the study areas. The bacterial profiling of their natural habitat waters revealed the presence of Paenibacillus nanensis DGX1(OQ690670), Bacillus cereus DGX2(OQ690675), Bacillus sp. DGX3(OQ690700) and Escherichia coli DGX4(OQ690701). Bacillus cereus was found to have high oviposition attractant properties in oviposition assays. Bacillus cereus was also obtained from the midgut of third instar larvae indicating that they had entered from the surrounding medium and colonized the larval gut. Subsequent tests exhibited the roles of B. cereus in larval development. Numerous plant products have been reported either as insecticides for killing larvae or adult mosquitoes or as repellents for mosquito biting and the best alternatives for mosquito control. Larvicidal potential of emulsified neem oil formulation against the field collected 3rd instar larvae of Culex quinquefasciatus mosquito under laboratory conditions was also evaluated. The information thus obtained can be pooled to generate larval source markers and larval source management practices by altering their habitats that cannot be removed. Furthermore, the time of implementation of these strategies can also be planned.

In vitro biochemical characterization and identification of hemolytic bacteria associated with life history of Culicoides peregrinus (Diptera: Ceratopogonidae), a vector of bluetongue virus.

Gut bacterial communities in insects provide several beneficial roles like nutrition, digestion, fecundity, and survival of the host. The microbial communities of Culicoides spp. (Diptera: Ceratopogonidae) vary with parity, developmental stages, and environmental factors. Previous studies have revealed the presence of hemolytic bacteria in adult Culicoides peregrinus Kieffer (Diptera: Ceratopogonidae), an important vector of bluetongue virus (BTV). Our objectives were (i) to identify bacterial communities with hemolytic activities associated with all life stages and (ii) to compare between reared and field-collected adults including age graded females. Bacterial identification followed Sanger sequencing of 16S rRNA. In vitro biochemical characterizations including antibiotic sensitivity tests were also done. The majority of bacterial species were beta hemolytic with one, Alcaligenes faecalis, showing alpha hemolysis. Most bacterial species were observed in field-collected adults except Proteus spp. Throughout the life history of the vector, Bacillus cereus (CU6A, CU1E) and Paenibacillus sp. (CU9G) were detected indicating their possible role in blood digestion within the gut of this vector species. In vivo hemolytic activities of these culturable bacterial communities within this vector may be addressed in future. These hemolytic bacterial communities may be targeted to develop novel and effective strategies for vector control.

Combined effect of physico-chemical and microbial quality of breeding habitat water on oviposition of malarial vector Anopheles subpictus.

Mosquitoes prefer diverse water bodies for egg laying and larval survival. Present study was performed with an objective to characterize physico-chemical properties and microbial profiling of breeding habitat water bodies of Anopheles subpictus mosquitoes. A field survey was accomplished to check the presence of An. subpictus larvae to record per dip larval density in various breeding habitats throughout the year. Physico-chemical and bacteriological properties in relation to mosquito oviposition were assessed. Dissolved oxygen content, pH and alkalinity were found to have major impacts and ponderosity on the prevalence of An. subpictus larvae. Larval density showed significant positive correlation with dissolved oxygen content of water and significant negative correlation with pH and alkalinity of habitat water. Comparatively higher population (cfu/mL) of Bacillus spp. competent with starch hydrolyzing and nitrate reducing properties were recorded all the breeding habitat water bodies of An. subpictus. Higher amplitude of anopheline larvae was portrayed during monsoon and post-monsoon season in clear water with an inclining trend to high dissolved oxygen content and neutral pH. B. cereus, B. megaterium, B. subtilis and B. tequilensis prevalent in all habitat water bodies were marked as oviposition attractants of gravid An. subpictus mosquitoes. Microbial population played key roles in the modulation of physico-chemical parameters of habitat water with a view to enhance its acceptability by gravid mosquitoes in relation to their oviposition. Better understanding of the interactions along with the control of oviposition attractant bacterial strains from mosquito breeding habitats might contribute to the vector management programme.

Behavioral toxicity, histopathological alterations and oxidative stress in Tubifex tubifex exposed to aromatic carboxylic acids- acetic acid and benzoic acid: A comparative time-dependent toxicity assessment.

This study evaluated Acetic acid (AA) and Benzoic acid's (BA) acute and sublethal toxicity by observing mortality, behavioral responses, and changes in the levels of oxidative stress enzymes in Tubifex tubifex. Exposure-induced changes in antioxidant activity (Catalase, Superoxide dismutase), oxidative stress (Malondialdehyde concentrations), and histopathological alterations in the tubificid worms were also noted across exposure intervals. The 96 h LC50 values of AA and BA to T. tubifex were 74.99 and 37.15 mg/l, respectively. Severity in behavioral alterations (including increased mucus production, wrinkling, and reduction in clumping) and autotomy showed concentration-dependent trends for both toxicants. Although histopathological effects also showed marked degeneration in the alimentary and integumentary systems in highest exposure groups (worms exposed to 14.99 mg/l for AA and 7.42 mg/l for BA) for both toxicants. Antioxidant enzymes (catalase and superoxide dismutase) also showed a marked increase of up to 8-fold and 10-fold for the highest exposure group of AA and BA respectively. While species sensitivity distribution analysis revealed T. tubifex as most sensitive to AA and BA compared to other freshwater vertebrates and invertebrates, General Unified Threshold model of Survival (GUTS) predicted individual tolerance effects (GUTS-IT), with slower potential for toxicodynamic recovery, as a more likely pathway for population mortality. Study findings demonstrate BA with greater potential for ecological effects compared to AA within 24 h of exposure. Furthermore, ecological risks to critical detritus feeders like T. tubifex may have severe implications for ecosystem services and nutrient availability within freshwater habitats.

Neem-based products as potential eco-friendly mosquito control agents over conventional eco-toxic chemical pesticides-A review.

Mosquitoes cause serious health hazards for millions of people across the globe by acting as vectors of deadly communicable diseases like malaria, filariasis, dengue and yellow fever. Use of conventional chemical insecticides to control mosquito vectors has led to the development of biological resistance in them along with adverse environmental consequences. In this light, the recent years have witnessed enormous efforts of researchers to develop eco-friendly and cost-effective alternatives with special emphasis on plant-derived mosquitocidal compounds. Neem oil, derived from neem seeds (Azadirachta indica A. Juss, Meliaceae), has been proved to be an excellent candidate against a wide range of vectors of medical and veterinary importance including mosquitoes. It is environment-friendly, and target-specific at the same time. The active ingredients of neem oil include limonoids like azadirachtin A, nimbin, salannin and numerous other substances that are still waiting to be discovered. Of these, azadirachtin has been shown to be very effective and is mainly responsible for its toxic effects. The quality of the neem oil depends on its azadirachtin content which, in turn, depends on its manufacturing process. Neem oil can be used directly or as nanoemulsions or nanoparticles or even in the form of effervescent tablets. When added to natural breeding habitat waters they exert their mosquitocidal effects by acting as ovicides, larvicides, pupicides and/or oviposition repellents. The effects are generated by impairing the physiological pathways of the immature stages of mosquitoes or directly by causing physical deformities that impede their development. Neem oil when used directly has certain disadvantages mainly related to its disintegration under atmospheric conditions rendering it ineffective. However, many of its formulations have been reported to remain stable under environmental conditions retaining its efficiency for a long time. Similarly, neem seed cake has also been found to be effective against the mosquito vectors. The greatest advantage is that the target species do not develop resistance against neem-based products mainly because of the innumerable number of chemicals present in neem and their combinations. This makes neem-based products highly potential yet unexplored candidates of mosquito control agents. The current review helps to elucidate the roles of neem oil and its various derivatives on mosquito vectors of public health concern.

Multifaceted entrancing role of glucose and its analogue, 2-deoxy-D-glucose in cancer cell proliferation, inflammation, and virus infection.

Chronic exposure to high glucose inside the human body helps in the progression of cancer by activating various signaling pathways including PI3K, Akt, mTOR, Ras, Raf, MAPK, and PKC. Hyperglycemia induces ROS and AGE production and decreases the functional activities of the cellular antioxidant system. By downregulating the prolyl hydroxylase, it stabilizes HIF-α leading to EMT-induced cancer progression and inhibition of apoptosis. High glucose level increases inflammation by creating a pro-inflammatory environment through the production of certain pro-inflammatory mediators (cytokines, chemokines, leukotrienes), and by influencing the recruitment of immune cells, leukocytes in the inflamed region. High glucose impairs the immune response and dysregulates ROS formation through the alteration in ETC and glutaminolysis which makes hyperglycemic patients more susceptible to viral infection. 2-DG is a modified form of D-glucose, that shows anticancer, anti-inflammatory, and anti-viral effects. It enters the cells through GLUT transporters and is converted into 2-deoxy-D-glucose-6-phosphate with the help of hexokinase. It inhibits the glycolysis, the TCA cycle, and the pentose phosphate pathway leading to ATP depletion. By downregulating glucose uptake and energy (ATP) production it halts various pathways responsible for cancer progression. It promotes the formation of anti-inflammatory mediators, and macrophage polarization, and also modulates immune function, which decreases inflammation. 2-DG inhibits PI3K/Akt/mTOR and upregulates the AMPK pathway, causing activation of the SIRT-4 gene that reduces lipogenesis, glucose uptake, nucleotide formation, and alters viral replication thus reducing the chances of infection.

Antibacterial potency of cytocompatible chitosan-decorated biogenic silver nanoparticles and molecular insights towards cell-particle interaction.

In the study, leaf extract of Carica papaya was utilized for the biogenic fabrication process of chitosan functionalized silver nanoparticles (Ag-Chito NPs). HRTEM analysis revealed that the fabricated Ag-Chito NPs was spherical in shape, with an average particle size of 13.31 (±0.07) nm. FTIR, UV-Vis, DLS, and other characterizations were also performed to analyze the diverse physicochemical properties of the particles. The antibacterial potency of the synthesized Ag-Chito NPs was tested against the two clinically isolated multidrug resistant uropathogenic bacterial strains, i.e. MLD 2 (Escherichia coli) and MLD 4 (Staphylococcus aureus) through MIC, MBC, time and concentration dependent killing kinetic assay, inhibition of biofilm formation assay, fluorescence and SEM imaging. Significantly, Ag-Chito NPs showed the highest sensitivity against the MLD 2 (MIC value of 12.5 µg/mL) strain, as compared to the MLD 4 (MIC value of 15 µg/mL) strain. From the hemolysis assay, it was revealed that Ag-Chito NPs exerted no significant toxicity up to 50 µg/mL against healthy human blood cells. Additionally, in silico analysis of chitosan (functionalized on the surface of AgNPs) and bacterial cell membrane protein also evidently suggested a strong interaction between Ag-Chito NPs and bacterial cells, which might be responsible for bacterial cell death.

Comparative Study on Bacterial Population Dynamics of Foregut, Midgut, and Hindgut Content of Perionyx excavatus (Perrier) Isolated from Eco-friendly, Non-hazardous Vermicompost.

The ideal condition of earthworm gut promotes growth and multiplication of beneficial soil microorganisms eliminating pathogens and converts organic wastes into nutrients rich compost. The present study has been carried out to determine the population dynamics of earthworm gut bacteria and to find out relative abundance of different functional bacterial groups in the foregut, midgut, and hindgut of earthworm Perionyx excavatus. To assess bacterial diversity, a viable plate count method was adopted. In the different gut region of earthworm, aerobic heterotrophic, amylolytic, Bacillus, Gram-negative, proteolytic, fat hydrolyzing, nitrate-reducing, nitrifying, asymbiotic nitrogen-fixing, Azotobacter, and phosphate solubilizing bacterial populations ranged from 22.2 to 241.6 × 106, 8.0 to 171.60 × 106, 1.83 to 2.79 × 106, 10.68 to 23.04 × 104, 3.70 to 5.52 × 104, 59.60 to 208.40 × 104, 1.86 to 7.34 × 104, 10.94 to 19.78 × 104, 0.80 to 3.42 × 104, 7.83 to 13.70 × 104, 1.31 to 2.67 × 104 cfu/ml gut suspension, respectively. The results of the one-way ANOVA revealed that the bacterial load of most of the bacterial groups was significantly higher (p < 0.05) in the hindgut region, followed by midgut and foregut. Only the density of the proteolytic group was significantly higher (p < 0.05) in the midgut region followed by foregut and hindgut. Starch hydrolyzing bacteria constitute the largest group of bacteria in the gut content. From principal component analysis, two components were extracted with the eigenvalues of 8.485 and 1.132. Agglomerative hierarchical cluster analysis revealed that the bacterial populations were clustered into four different groups. Quantitative variation among bacterial groups in earthworm's gut seems to determine the soil health and composting efficiency; from this point of view, the present study will provide a better understanding about different functional bacterial groups of earthworm's guts and might be helpful in sustainable agriculture and waste management.

Homology modeling and virtual characterization of cytochrome c nitrite reductase (NrfA) in three model bacteria responsible for short-circuit pathway, DNRA in the terrestrial nitrogen cycle.

NrfA is the molecular marker for dissimilatory nitrate reduction to ammonium (DNRA) activity, catalysing cytochrome c nitrite reductase enzyme. However, the limited study has been made so far to understand the structural homology modeling of NrfA protein in DNRA bacteria. Therefore, three model DNRA bacteria (Escherechia coli, Wolinella succinogenes and Shewanella oneidensis) were chosen in this study for in-silico protein modeling of NrfA which roughly consists of similar length of amino acids and molecular weight and they belong to two contrasting taxonomic families (γ-proteobacteria with nrfABCDEFG and ε-proteobacteria with nrfHAIJ operon). Multiple bioinformatic tools were used to examine the primary, secondary, and tertiary structure of NrfA protein using three distinct homology modeling pipelines viz., Phyre2, Swiss model and Modeller. The results indicated that NrfA protein in E. coli, W. succinogenes and S. oneidensis was mostly periplasmic and hydrophilic. Four conserved Cys-X1-X2-Cys-His motifs, one Cys-X1-X2-Cys-Lys haem-binding motif and Ca ligand were also identified in NrfA protein irrespective of three model bacteria. Moreover, 11 identical conserved amino acids sequence was observed for the first time between serine and proline in NrfA protein. Secondary structure of NrfA revealed that α-helices were observed in 77.9%, 73.4%, and 77.4% in E. coli, W. succinogenes and S. oneidensis, respectively. Ramachandran plot showed that number of residue in favored region in E. coli, W. succinogenes and S. oneidensis was 97.03%, 97.01% and 97.25%, respectively. Our findings also revealed that among three pipelines, Modeller was considered the best in-silico tool for prediction of NrfA protein. Overall, significant findings of this study may aid in the identification of future unexplored DNRA bacteria containing cytochrome c nitrite reductase. The NrfA system, which is linked to respiratory nitrite ammonification, provides an analogous target for monitoring less studied N-retention processes, particularly in agricultural ecosystems. Furthermore, one of the challenging research tasks for the future is to determine how the NrfA protein responds to redox status in the microbial cells.

Characterizations of Larval Gut Bacteria of Anopheles subpictus Grassi (1899) and their Role in Mosquito Development in Hooghly, West Bengal, India.

Malaria is a serious vector borne disease transmitted by different species of Anopheles mosquitoes. The present study was aimed to isolate and characterize the bacterial flora from the gut of larvae of An. subpictus Grassi (1899) prevalent in Hooghly and explore their roles in host survival and development. Mosquito larvae and adults were collected from field and were maintained in laboratory. Bacterial load in the larval mid-gut was determined, and predominant strains were isolated and characterized by polyphasic approach. Role of these bacteria in larval survival and development were assayed. Bacterial load in the gut of larvae was found to vary in field-collected and lab-reared mosquitoes in different seasons. Morphological, bio-chemical, and molecular analyses explored four common bacterial isolates, namely Bacillus subtilis, Bacillus pumilus, Bacillus cereus, and Proteus vulgaris in the larval gut throughout the year. Larval survival rate was greatly reduced (0.06) and time of pupation was prolonged (17.8 ± 0.57) [days] in the absence of their gut bacteria. Total tissue protein (7.78 ± 0.56) [µg/mg], lipid (2.25 ± 0.19) [µg/mg] & carbohydrate (16.5 ± 0.79) [µg/mg] contents of larvae, and body weight & wing length of adult male (0.17 ± 0.02 & 1.74 ± 0.43) [mm] & female (0.19 ± 0.02 & 1.99 ± 0.46) [mm] mosquitoes were also found to be greatly reduced in the absence of gut bacteria. Developmental characteristics were restored with the introduction of culture suspension of all four resident gut bacterial isolates. Present study indicates that the mosquitoes largely depend on their gut bacteria for their survival and development. So, manipulation or control of this gut bacterial communities might inhibit survival and development of vector mosquitoes.

Commonly used surfactants sodium dodecyl sulphate, cetylpyridinium chloride and sodium laureth sulphate and their effects on antioxidant defence system and oxidative stress indices in Cyprinus carpio L.: an integrated in silico and in vivo approach.

The present study evaluated the homology modelling, in silico prediction and characterization of Cyprinus carpio cytochrome P450, as well as molecular docking experiments between the modelled protein and the surfactants sodium dodecyl sulphate (SDS), sodium laureth sulphate (SLES) and cetylpyridinium chloride (CPC). Homology modelling of cytochrome P450 was performed using the best fit template structure. The structure was optimized with 3D refine, and the ultimate 3D structure was checked with PROCHEK and ERRATA. ExPASy's ProtParam was likewise used to analyse the modelled protein's physiochemical and stereochemical attributes. To establish the binding pattern of each ligand to the targeted protein and its effect on the overall protein conformation, molecular docking calculations and protein-ligand interactions were performed. Our in silico analysis revealed that hydrophobic interactions with the active site amino acid residues of cytochrome p450 were more prevalent than hydrogen bonds and salt bridges. The in vivo analysis exhibited that exposure of fish to sublethal concentrations (10% and 30% of 96 h LC50) of SDS (0.34 and 1.02 mg/l), CPC (0.002 and 0.006 mg/l) and SLES (0.69 and 2.07 mg/l) at 15d, 30d and 45d adversely affected the oxidative stress and antioxidant enzymes (CAT, SOD, GST, GPx and MDA) in the liver of Cyprinus carpio. As a result, the study suggests that elicited oxidative stress, prompted by the induction of antioxidant enzymes activity, could be attributable to the stable binding of cytochrome P450 with SDS, CPC and SLES which ultimately leads to the evolution of antioxidant enzymes for its neutralization.

The influence of IL1RN VNTR polymorphism on HPV infection among some tribal communities.

Persistent infection of human Papillomavirus is the main etiological factor for cervical cancer. Austro-Asiatic tribes are early settlers in India and they have unique genetic variations compared to other people. The immunological response is crucial for the prevention of viral associated diseases. Interleukin-1 receptor antagonist (IL-1RN) is considered being an important regulator of host immune surveillance. A total of 45 Santali tribal women and 10 Kora tribal women were enrolled in the present study and demographic variables were recorded during collection. Genomic DNA was extracted from cervical/vaginal swab samples. IL1RN variable number of tandem repeats (VNTR) polymorphisms and HPV types were determined by PCR-based assay. Association between IL1RN VNTR polymorphisms with the HPV infections among the tribal communities was determined by logistic regression analysis. HPV18 prevalence was significantly higher among tribal women. We observed that the polymorphism A2*A2 (p = 0.022; odds ratio [OR] (95% confidence interval [CI]) = 0.16 (0.03-0.86)] were more resistant to oncogenic HPV infection. Use of oral contraceptives was associated with higher relative risk (p = 0.008; OR [95% CI] = 5.39 [1.47-19.8]) for oncogenic HPV18 positivity among the tribal women. The A2 allele homozygosity of IL1RN VNTR was identified to be associated with the protection from oncogenic HPV infection among various tribal communities of West Bengal and therefore may be a useful marker of host immune response among them.

An integrated in silico and in vivo approach to determine the effects of three commonly used surfactants sodium dodecyl sulphate, cetylpyridinium chloride and sodium laureth sulphate on growth rate and hematology in Cyprinus carpio L.

The purpose of this work is to evaluate the homology modeling, in silico prediction, and characterization of somatotropin and erythropoietin from Cyprinus carpio as well as molecular docking and simulation experiments between the modeled proteins and surfactants sodium dodecyl sulfate (SDS), sodium laureth sulfate (SLES) and cetylpyridinium chloride (CPC). Using the best fit template structure, homology modeling of somatotropin and erythropoietin of Cyprinus carpio respectively was conducted. The model structures were improved further with 3Drefine, and the final 3D structures were verified with PROCHEK, ERRATA and ProQ. The physiochemical, as well as the stereochemical parameters of the modeled proteins, were evaluated using ExPASy's ProtParam. Molecular docking calculations, protein-ligand interactions, and protein flexibility analysis were carried out to determine the binding pattern of each ligand to the targeted proteins and their effect on the overall proteins' conformation. Our in silico analysis showed that hydrophobic interactions with the active site amino acid residues of the modeled proteins (somatotropin and erythropoietin) were more prevalent than hydrogen bonds and salt bridges that affect the flexibility and stability of the somatotropin and erythropoietin as revealed from our protein flexibility analysis. The in vivo analysis showed that sublethal concentrations of SDS, SLES, and CPC negatively affected the growth and hematological parameters of Cyprinus carpio. Hence, it may be inferred from the study that the alterations in the flexibility of somatotropin and erythropoietin of Cyprinus carpio upon addition of SDS, CPC and SLES might be attributable to the reduction in growth and hematological parameters.

λ cyhalothrin induced toxicity and potential attenuation of hematological, biochemical, enzymological and stress biomarkers in Cyprinus carpio L. at environmentally relevant concentrations: A multiple biomarker approach.

The present study was aimed to evaluate the toxic effects of a commonly used synthetic pyrethroid, λ cyhalothrin on the common carp, Cyprinus carpio L. The results depicted that 96 h LC50 value of λ cyhalothrin to the fish was 1.48 µg l-1. During 45 days of chronic exposure a significant reduction (p < 0.05) in the RBC, hemoglobin, and hematocrit value of fish was observed in λ cyhalothrin treated fish. Blood glucose, cholesterol and creatinine levels increased significantly, while total protein and albumin were significantly decreased (p < 0.05) in the exposed fish. Moreover, alanine aminotransferase and aspartate aminotransferase levels in the blood also increased significantly (p < 0.05) in the treated fish. In gills and liver, glutathione S-transferase (GST) and glutathione peroxidase (GPx) and in liver GST exhibited a significant initial augmentation followed by a subsequent reduction while catalase (CAT), superoxide dismutase (SOD), and malondialdehyde (MDA) level increased markedly with incrementing concentrations of λ cyhalothrin in both the organs. Acetylcholinesterase (AchE) activity in both gills and liver decreased in exposed fish upon addition λ cyhalothrin. However, the hazardous effects of λ cyhalothrin on C. carpio were characterized and portrayed by the development of integrated biomarker response (IBR), and biomarker response index (BRI). GUTS-SD and IT modeling were implied for a better interpretation of the toxicity. These results indicate that exposure to λ cyhalothrin alters the survivability at the acute level and the activity of hematological, plasma biochemical as well as enzymological and stress parameters (in gills and liver) at the sublethal level in C. carpio.

Characterization and mosquitocidal potency of a Bacillus thuringiensis strain of rice field soil of Burdwan, West Bengal, India.

Bacillus thuringiensis is the most popular mosquitocidal bacteria, strains of which are effective against almost all mosquito larvae. It has host specificity and thus, has no adverse effect on non-target species of the ecosystem. Culex tritaeniorhynchus, a vector of Japanese encephalitis (JE), breeds in vast area of rice fields in Burdwan district of West Bengal, India, which has already confronted JE epidemic. Entomological investigation and ecological studies on this vector mosquito showed that JE epidemic may reoccur anytime in the area. A strain of Bt (BU55) was isolated from rice field soil, efficacy was tested against Cx. tritaeniorhynchus and mosquitocidal role was confirmed against Cx. quinquefascistus also. The LC50 of Bacillus thuringiensis BU55 against Cx. tritaeniorhynchus and Cx. quinquefascistus after 72 h was 8.59 ml (final dose 2.49 x107 CFU/ml) and 7.52 ml (final dose 2.20 x 107 CFU/ml), respectively. Insecticidal crystal protein profile of BU55 produced 136.89, 64.80, 43.45, 33.65 and 26.98 kDa bands. Among them 136.89, 64.29, 26.98 kDa proteins are comparable to actual toxins viz. Cry1Ac (138.3 kDa, Lepidoptera specific), Cry4D (68.0 kDa, Diptera specific) and Cyt (27.4 kDa, Diptera specific). The results clearly showed that the Bt strain is a potent dipteran larvicide and can be used against the JE vectors to control the disease.

Acute toxicity and sublethal effects of sodium laureth sulfate on oxidative stress enzymes in benthic oligochaete worm, Tubifex tubifex.

The present study was performed to determine the acute toxicity of sodium laureth sulfate (SLES) and its sublethal effects on oxidative stress enzymes in benthic oligochaete worm Tubifex tubifex. The results showed that 96 h median lethal concentration (LC50) value of SLES for Tubifex tubifex is 21.68 mg/l. Moreover exposed worms showed abnormal behaviours including incremented erratic movement, mucus secretion, and decreased clumping tendency at acute level. Percentage of autotomy additionally increased significantly (P < 0.05) with the increasing dose of toxicant at 96 h exposure. Sublethal concentrations of SLES (10% and 30% of 96 h LC50 value) caused paramount alterations in the oxidative stress enzymes. Superoxide dismutase (SOD), reduced glutathione (GSH), glutathione S-transferase (GST), and glutathione peroxidase (GPx) exhibited a striking initiatory increment followed by a resulting descending pattern. Moreover, during exposure times, catalase (CAT) activity and malondialdehyde (MDA) level increased markedly with incrementing concentrations of SLES. However, the effects of sodium laureth sulfate on Tubifex tubifex were characterized and portrayed by the development of a correlation matrix and an integrated biomarker response (IBR) assessment. These results indicate that exposure to this anionic surfactant alters the survivability and behavioral response at acute level and modifies changes in oxidative stress enzymes at sublethal level in Tubifex tubifex.

Oxidative stress in benthic oligochaete worm, Tubifex tubifex induced by sublethal exposure to a cationic surfactant cetylpyridinium chloride and an anionic surfactant sodium dodecyl sulfate.

The present study was assessed to determine the in vivo toxic effects of a cationic surfactant, cetylpyridinium chloride (CPC), and an anionic surfactant, sodium dodecyl sulfate (SDS) in terms of oxidative stress biomarkers in benthic oligochaete worm Tubifex tubifex for 14 days. The investigation demonstrated that sublethal concentrations of CPC (0.0213, and 0.0639 mg L-1) and SDS (1.094 and 3.092 mg L-1)induced paramount alterations in the oxidative stress enzymes in Tubifex tubifex. Superoxide dismutase (SOD), glutathione S-transferase (GST), reduced glutathione (GSH), and glutathione peroxidase (GPx) exhibited an initial notable increase in their activities in the surfactants exposed worms at 1d and 7d of exposure period followed by consequential reduction at 14d exposure period with respect to control, while catalase (CAT) and malondialdehyde (MDA) activities markedly incremented gradually throughout the exposure periods. Through the construction of the correlation matrix and integrated biomarker response (IBR), the effects of CPC and SDS on Tubifex tubifex were distinguished. These results indicate that exposure to these cationic and anionic surfactants modulates the levels of oxidative stress enzymes in Tubifex tubifex.

Acute toxicity of organophosphate pesticide profenofos, pyrethroid pesticide λ cyhalothrin and biopesticide azadirachtin and their sublethal effects on growth and oxidative stress enzymes in benthic oligochaete worm, Tubifex tubifex.

The present study was aimed to assess the acute toxicity of organophosphate pesticide, profenofos; synthetic pyrethroid pesticide, λ cyhalothrin and biopesticide, azadirachtin and their sublethal effects on growth rate and oxidative stress biomarkers in Tubifex tubifex in vivo. The results showed that 96 h LC50 value of profenofos, λ cyhalothrin and azadirachtin to Tubifex tubifex are 0.59, 0.13 and 82.15 mg L-1 respectively. Pesticide treated worms showed several behavioral abnormalities including increased mucus secretion, erratic movements, wrinkling activity and decreased clumping tendency during acute exposure. The percentage of autotomy increased significantly (p < 0.05) with the increasing concentration of the pesticides at 96 h of exposure. Sublethal concentrations of profenofos (0.059 and 0.118 mg L-1), λ cyhalothrin (0.013 and 0.026 mg L-1) and azadirachtin (8.2 and 16.4 mg L-1) caused significant alterations in growth rate and oxidative stress enzymes in T. tubifex during 14 days exposure period. The growth rate of the pesticide exposed worms decreased significantly (P < 0.05) in a concentration and duration-dependent manner. Superoxide dismutase (SOD), reduced glutathione (GSH), glutathione-s-transferase (GST) and glutathione peroxidase (GPx) demonstrated a noteworthy (p < 0.05) initial induction followed by a subsequent reduction, while catalase (CAT) and malondialdehyde (MDA) exhibited noteworthy induction (p < 0.05) all through the exposure time. Through principal component analysis, correlation matrix, and integrated biomarker response, the effects of profenofos, λ cyhalothrin and azadirachtin on T. tubifex were distinguished. These results indicate that exposure to profenofos, λ cyhalothrin and azadirachtin affect survivability, change the behavioral responses, reduce the growth rate and induce oxidative stress enzymes in T. tubifex.

De novo transcriptome analysis of halotolerant bacterium Staphylococcus sp. strain P-TSB-70 isolated from East coast of India: In search of salt stress tolerant genes.

In the present study, we identified salt stress tolerant genes from the marine bacterium Staphylococcus sp. strain P-TSB-70 through transcriptome sequencing. In favour of whole-genome transcriptome profiling of Staphylococcus sp. strain P-TSB-70 (GenBank Accn. No. KP117091) which tolerated upto 20% NaCl stress, the strain was cultured in the laboratory condition with 20% NaCl stress. Transcriptome analyses were performed by SOLiD4.0 sequencing technology from which 10280 and 9612 transcripts for control and treated, respectively, were obtained. The coverage per base (CPB) statistics were analyzed for both the samples. Gene ontology (GO) analysis has been categorized at varied graph levels based on three primary ontology studies viz. cellular components, biological processes, and molecular functions. The KEGG analysis of the assembled transcripts using KAAS showed presumed components of metabolic pathways which perhaps implicated in diverse metabolic pathways responsible for salt tolerance viz. glycolysis/gluconeogenesis, oxidative phosphorylation, glutathione metabolism, etc. further involving in salt tolerance. Overall, 90 salt stress tolerant genes were identified as of 186 salt-related transcripts. Several genes have been found executing normally in the TCA cycle pathway, integral membrane proteins, generation of the osmoprotectants, enzymatic pathway associated with salt tolerance. Recognized genes fit diverse groups of salt stress genes viz. abc transporter, betaine, sodium antiporter, sodium symporter, trehalose, ectoine, and choline, that belong to different families of genes involved in the pathway of salt stress. The control sample of the bacterium showed elevated high proportion of transcript contigs (29%) while upto 20% salt stress treated sample of the bacterium showed a higher percentage of transcript contigs (31.28%). A total of 1,288 and 1,133 transcript contigs were measured entirely as novel transcript contigs in both control and treated samples, respectively. The structure and function of 10 significant salt stress tolerant genes of Staphylococcus sp. have been analyzed in this study. The information acquired in the present study possibly used to recognize and clone the salt stress tolerant genes and support in developing the salt stress-tolerant plant varieties to expand the agricultural productivity in the saline system.