Strategic pathway analysis for dual management of epilepsy and comorbid depression: a systems biology perspective.

Depression is a common psychiatric comorbidity among patients with epilepsy (PWE), affecting more than a third of PWE. Management of depression may improve quality of life of epileptic patients. Unfortunately, available antidepressants worsen epilepsy by reducing the seizure threshold. This situation demands search of new safer target for combined directorate of epilepsy and comorbid depression. A system biology approach may be useful to find novel pathways/markers for the cure of both epilepsy and associated depression via analyzing available genomic and proteomic information. Hence, the system biology approach using curated 64 seed genes involved in temporal lobe epilepsy and mental depression was applied. The interplay of 600 potential proteins was revealed by the Disease Module Detection (DIAMOnD) Algorithm for the treatment of both epilepsy and comorbid depression using these seed genes. The gene enrichment analysis of seed and diamond genes through DAVID suggested 95 pathways. Selected pathways were refined based on their syn or anti role in epilepsy and depression. In conclusion, total 8 pathways and 27 DIAMOnD genes/proteins were finally deduced as potential new targets for modulation of selected pathways to manage epilepsy and comorbid depression. Supplementary Information: The online version contains supplementary material available at 10.1007/s40203-024-00208-1.

Dynamic alterations in physiological and biochemical indicators of Cirrhinus mrigala hatchlings: A sublethal exposure of triclosan.

Triclosan (TCS), a biocide used in various day-to-day products, has been associated with several toxic effects in aquatic organisms. In the present study, biochemical and hematological alterations were evaluated after 14 d (sublethal) exposure of tap water (control), acetone (solvent control), 5, 10, 20, and 50 µg/L (environmentally relevant concentrations) TCS to the embryos/hatchlings of Cirrhinus mrigala, a major freshwater carp distributed in tropic and sub-tropical areas of Asia. A concentration-dependent increase in the content of urea and protein carbonyl, while a decrease in the total protein, glucose, cholesterol, triglycerides, uric acid, and bilirubin was observed after the exposure. Hematological analysis revealed a decrease in the total erythrocyte count, hemoglobin, and partial pressure of oxygen, while there was an increase in the total leucocyte count, carbon dioxide, and partial pressure of carbon dioxide and serum electrolytes. Comet assay demonstrates a concentration-dependent increase in tail length, tail moment, olive tail moment, and percent tail DNA. An amino acid analyzer showed a TCS-dose-dependent increase in various amino acids. Sodium dodecyl sulphate polyacrylamide gel electrophoresis analysis revealed different proteins ranging from 6.5 to 200 kDa, demonstrating TCS-induced upregulation. Fourier transform infrared spectra analysis exhibited a decline in peak area percents with an increase in the concentration of TCS in water. Curve fitting of amide I (1,700-1600 cm-1) showed a decline in α-helix and turns and an increase in ß-sheets. Nuclear magnetic resonance study also revealed concentration-dependent alterations in the metabolites after 14 d exposure. TCS caused alterations in the biomolecules and heamatological parameters of fish, raising the possibility that small amounts of TCS may change the species richness in natural aquatic habitats. In addition, consuming TCS-contaminated fish may have detrimental effects on human health. Consequently, there is a need for the proper utilisation and disposal of this hazardous compound in legitimate quantities.

Pre-clinical and cellular safety assessment of oral administered DHA rich microalgae oil from Schizochytrium sp. (Strain ATCC-20889): acute, sub-chronic and genotoxicity.

The lack of toxicity data for DHA-rich oil from Schizochytrium sp. (Strain ATCC-20889) leads to its exclusion from the Qualified Presumption of Safety list. Therefore, present study addresses toxicity evaluation of DHA-rich microalgae oil using ex-vivo (cytotoxicity assay) and in-vivo methods (acute (OECD 423 guidelines), sub-chronic (OECD 452 guidelines), and genotoxicity assay). The ex-vivo results showed >90% cell viability of Caco-2 cells after 48 h of treatment (200 µg/mL of DHA). Additionally, the in-vivo acute toxicity study found that microalgae oil was nontoxic and classified under category 5 molecule according to OECD 423 guidelines with a highest degree of safety at 2000 mg/kg b.w. The in-vivo sub-chronic study revealed no significant mortality and changes in feed intake, body weight, haematological, biochemical, neurological, and urine parameters after repeated 180-days administration of DHA-rich microalgae oil at 250 mg/kg, 500 mg/kg, and 1000 mg/kg. Moreover, histopathology evaluation, comet assay, chromosomal aberration, and micronuclei assay also confirmed the nontoxic behavior of DHA-rich oil. Thus, the results from the ex-vivo and in-vivo studies indicate that DHA-rich oil from Schizochytrium sp. (Strain ATCC-20889) is safe for use as a novel food, and can be included in infants, adults, pregnant women, and children formula.

Adjunct antiseizure effect of clotrimazole in a rotenone corneal kindling mouse model of mitochondrial drug-resistant epilepsy.

This study aimed to investigate the therapeutic potential of clotrimazole, an inhibitor of the transient receptor potential cation channel, for treating mitochondrial drug-resistant epilepsy and to understand its underlying neurochemical mechanisms. Adult albino mice underwent rotenone-corneal kindling, receiving daily electric shocks (15 mA, 20 V, 6-Hz for 3 s) through a corneal electrode, to induce mitochondrial drug-resistant epilepsy. The onset of drug resistance was confirmed by the significant (p < 0.05) lack of seizure control with standard antiseizure medications including levetiracetam (40 mg/kg), valproate (250 mg/kg), phenytoin (35 mg/kg), lamotrigine (15 mg/kg), and carbamazepine (40 mg/kg). Drug-resistant mice were then classified into one vehicle-treated group and three groups treated with varying doses of clotrimazole (40, 80, and 160 mg/kg orally). Neurochemical analysis of the seizurogenic hippocampus and cerebral cortex was conducted using high-performance liquid chromatography with an electrochemical detector. Administration of clotrimazole alongside standard antiseizure medications led to a significant decrease (p < 0.05) in seizure scores suggesting the restoration of antiseizure effects. Neurochemicals, including tryptophan, serotonin, kynurenine, serine, taurine, gamma-aminobutyric acid, and glutamate, were significantly restored post-clotrimazole treatment. Overall, the present study underscores the adjunct antiseizure effect of clotrimazole in a rotenone corneal kindling mouse model of mitochondrial drug-resistant epilepsy, emphasising its role in neurochemical restoration.

A network pharmacology approach to explore pharmacological mechanisms of Asparagus racemosus for ameliorative effect in epilepsy and comorbid progressive memory dysfunction.

Background: Network pharmacology approach has been observed a powerful tool to predict underlying complex pharmacological mechanism of herbs. Asparagus racemosus has been reported to show ameliorative effects in treating epilepsy and comorbid memory dysfunction but mechanism of this amelioration is elusive. Hence a network pharmacology approach was employed to investigate the plausible mechanism of A. recemosus. Methodology: : Bioactive compounds of A. racemosus were extracted based on the TCMSP, PCIDB, and BATMAN-TCM database. The potential targets of bioactive compounds were collected using target fishing. Epilepsy and comorbid dementia genes were collected from DISGENET. A PPI network among these targets was constructed using the intersecting key targets between herb targets and disease targets. Besides, DAVID bioinformatics resource was utilized for the pathway enrichment analysis on GO and KEGG. Ultimately, phytochemical compound-target genes-Pathways network has been assembled utilizing Cytoscape to decipher the mechanism of the herb. Results: The network analysis revealed that 5 targets (CASP3, TNF, VEGFA, PTGS2 and CNR1) might be the key therapeutic targets of asparagus on Epilepsy comorbid Alzheimer's disease. Based on high connectivity, four hub compounds with the highest connectivity were noted and it includes Shatavarin V, Sarsasapogenin, Shatavarin IX, and Shatavarin VI. A total of 19 KEGG terms were enriched as the potential pathways of A. racemosus in Epilepsy comorbid Alzheimer's disease. Conclusion: This study envisaged the pharmacological and molecular mechanism of A. racemosus against epilepsy comorbid Alzheimer's disease and put forward a strategy to uncover the mechanisms of Traditional Indian Medicine based on network pharmacology. Supplementary Information: The online version contains supplementary material available at 10.1007/s40203-023-00169-x.

Physiological, biochemical and structural changes in tomato plants by vermicompost application in different exposure periods under glass house conditions.

In modern era, various inorganic fertilizers and pesticides are used as plant growth supplements in a variety of crop in order to gain maximum output and also reported as hazardous to mankind as well as environment. In addition, some of the plants died in initial phase of the growth after germination due to poor nutrient content of the soil or exposure to biotic stresses. In the beginning of sustainable agriculture, these chemical fertilizers were replaced with some alternative growth boosters such as organic fertilizers. In the present study, vermicompost was prepared using garden waste and cattle dung followed by analysis of various physico-chemical properties. Then tomato seeds were allowed to germinate in soil and supplemented with different doses of vermicompost (0-100%). The plants were harvested after 10 and 45 days of their germination and tissues were subjected to analysis of various morphological and biochemical parameters. Morphological parameters included root length, shoot length, root fresh weight, shoot fresh weight and number of leaves. Whereas biochemical parameters such as protein content, antioxidative enzymes (catalase, superoxide dismutase, ascorbate peroxidase, polyphenol oxidase), non-enzymatic antioxidants (ascorbic acid, glutathione, tocopherol), osmolytes (proline, carbohydrate), photosynthetic pigments (chlorophyll, carotenoid) and secondary metabolites (phenol, flavonoid, anthocyanin) were estimated on UV-visible spectrophotometer using standard protocols. Further, structural analysis of plant tissue was done using fourier transform infrared spectroscopy spectra (FTIR) and carbon hydrogen nitrogen (CHN) elemental analyzer. Results obtained from the present study revealed significant difference in all morphological and biochemical markers at both 10 and 45 days intervals of time. Further, growth of all plants was found to be directly proportional to the concentration of vermicompost and exposure duration. FTIR spectra and CHN analyses reveal the breakdown of various complex compounds and their transformation from Vcom amended soil to roots of plants. This is the first study in which significant changes were observed in growth, physiology and structural composition of tomato plants at two different exposure periods (10 and 45 days) under glass house conditions which further concluded that vermicompost has a significant potential for increasing plant growth.

Genotoxic and mutagenic potential of 7-methylxanthine: an investigational drug molecule for the treatment of myopia.

7-Methylxanthine (7-MX, CAS No. 552-62-5, purity 99.46%) is the first orally administered drug candidate, which showed anti-myopic activity in different pre-clinical studies. In the present study, we investigated the in-vivo genotoxic and mutagenic toxicity of 7-MX in Wistar rats using comet/single-cell gel electrophoresis, chromosomal aberration and micronucleus assays after oral administration. For the single-dose study (72 h), two doses of 7-MX 300 and 2000 mg/kg body weight were selected. For a repeated dose 28 d study, three doses (250, 500, and 1000 mg/kg) of 7-MX were selected. The doses were administered via oral gavage in the suspension form. Blood and major vital organs such as bone marrow, lung and liver were used to perform comet/single cell gel electrophoresis, chromosomal aberration, and micronucleus assays. The in-vitro Ames test was performed on TA98 and TA100 strains. In the chromosomal aberration study, a non-significant increase in deformities such as stickiness, ring chromosome, and endoreduplication was observed in bone marrow cells of 7-MX treated groups. These chromosomal alterations were observed upon treatment with doses of 2000 mg/kg single dose for 72 h and 1000 mg/kg repeated dose for 28 d. At a dose of 500 mg/kg, DNA damage in terms of tail length, tail moment, % tail DNA and the olive tail moment was also found to be non-significant in 7-MX treated groups. The Ames test showed the non-mutagenic nature of 7-MX in both strains of TA98 and TA100 of Salmonella typhimurium with or without metabolic activation. Thus, the present work is interesting in view of the non- genotoxicity and non-mutagenicity of repeated doses of 7-MX.

Biopreservative efficacy of Enterococcus faecium-immobilised film and its enterocin against Salmonella enterica.

The growing awareness about the adverse health effects of artificial synthetic preservatives has led to a rapid increase in the demand for safe food preservation techniques and bio preservatives. Thus, in this study, the biopreservatives efficacy of enterocin-producing Enterococcus faecium Smr18 and its enterocin, ESmr18 was evaluated against Salmonella enterica contamination in chicken samples. E. faecium Smr18 is susceptible to the antibiotics penicillin-G, ampicillin, vancomycin, and erythromycin, thereby indicating that it is a nonpathogenic strain. Further, the enterocin ESmr18 was purified and characterised as a 3.8 kDa peptide. It possessed broad spectrum antibacterial activity against both Gram-positive and Gram-negative pathogens including S. enterica serotypes Typhi and Typhimurium. Purified ESmr18 disrupted the cell membrane permeability of the target cell thereby causing rapid efflux of potassium ions from L. monocytogenes and S. enterica. Chicken samples inoculated with S. enterica and packaged in alginate films containing immobilised viable E. faecium resulted in 3 log10 colony forming units (CFU) reduction in the counts of S. enterica after 34 days of storage at 7-8 °C. The crude preparation of ESmr18 also significantly (p < 0.05) reduced the CFU counts of salmonella-inoculated chicken meat model. Purified ESmr18 at the concentration upto 4.98 µg/ml had no cytolytic effect against human red blood cells. Crude preparation of ESmr18 when orally administered in fish did not cause any significant (p < 0.05) change in the biochemical parameters of sera samples. Nonsignificant changes in the parameters of comet and micronucleus assays were observed between the treated and untreated groups of fishes that further indicated the safety profile of the enterocin ESmr18.

Environmentally relevant concentrations of Triclosan cause transcriptomic and biomolecular alterations in the hatchlings of Labeo rohita.

Suppression (p ≤ 0.05) of antioxidative/detoxification (except GPx and CYP3a) and cytoskeletal (except DHPR) genes but induction of metabolic (except for AST and TRY) and heat shock (except HSP60) genes of Labeo rohita hatchlings after 14 days of exposure to environmentally relevant concentrations of Triclosan (0.0063, 0.0126, 0.0252 and 0.06 mg/L) was followed by an increase (p ≤ 0.05) for most of the genes after 10 days recovery period. After recovery, LDH, ALT, CK, CHY, PA, HSP47 and DHPR declined, while SOD, CAT, GST, GR, GPx, CYP1a, CYP3a, AST, AChE, TRY, HSP60, HSP70, HSc71, HSP90 MLP-3, α-tropomyosin, desmin b and lamin b1 increased over exposure. Peak area of biomolecules (except 3290-3296, 2924-2925 and 2852-2855 cm-1) declined (p ≤ 0.01) more after recovery [except for an increase (p ≤ 0.01) at 1398-1401 cm-1]. CYP3a, CK, HSP90, MLP-3 and secondary structure of amide A are the most sensitive markers for the environmentally relevant concentrations of Triclosan.

Combined effects of vermicompost and vermicompost leachate on the early growth of Meloidogyne incognita stressed Withania somnifera (L.) Dunal.

Roots of Withania somnifera, an important medicinal herb, are prone to the infection of Meloidogyne incognita (a root parasitic nematode). The infection lowers the quality and quantity of plant material and poses a challenge in crop cultivation and obtaining desirable yield. In the present study, in vitro inhibitory activity of vermicompost leachate (Vcom-L) was assessed against % hatch and survival of M. incognita in a 96 h assay. Then, Vcom-L was used as soil supplement in combination with vermicompost (Vcom) to evaluate their nematode inhibitory and stress alleviating effect in W. somnifera, in a pot experiment. Root galling intensity and growth performance of nematode-stressed W. somnifera raised from seeds pre-soaked in distilled water (DW), Vcom-L, vermicompost tea (Vcom-T) and different dilutions of Vcom-L were assessed. We observed 79% suppression of egg hatching and 89% juvenile (J2) mortality after 96 h compared to control, at 100% concentration of Vcom-L. Significant reduction in gall formation with increase in growth parameters of seedlings was observed after combined application of Vcom (60% or 100%) + Vcom-L and was evident as enhancement in seedling biomass and contents of chlorophyll and protein. However, proline, total phenolics and malondialdehyde (MDA) content declined significantly in these combinations compared to the control (0% Vcom). Activity of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidise (APX), guaiacol peroxidise (GPX) and glutathione reductase (GR) declined with Vcom as well as Vcom + Vcom-L and corresponded with decline in the accumulation of reactive oxygen species in leaves. Further, 1:5 and 1:10 dilutions of Vcom-L in combination with Vcom (60%) proved beneficial in mitigating the nematode-induced stress in W. somnifera. Present results showed the potential of Vcom and Vcom-L in standardised combination as an effective strategy in controlling the pathogenicity of M. incognita in medicinal plants such as W. somnifera.

Potential of vermicompost extract in enhancing the biomass and bioactive components along with mitigation of Meloidogyne incognita-induced stress in tomato.

Increasing inorganic fertilizer and pesticide use has been linked to increased health risks for humans and cattle, as well as substantial water and soil contamination. In recent years, vermicomposting has shown to be a viable alternative to chemical pesticides. Vermicompost and vermicompost products such as extract and leachate assist plants in a number of ways. According to recent studies, vermicompost extract (VCE), when used as a supplement, is thought to work as a growth and stress tolerance booster for plants. These liquid supplements also help to suppress a range of pests, such as root knot nematodes. In the present study, neem- and cattle dung-based vermicompost extracts of different concentrations (0, 20, 40, 60, 80 and 100%) were prepared and used for their application against nematode infection in tomato seedlings under laboratory conditions. Apart from its antagonistic action against Meloidogyne incognita, the influence of VCE on plant growth was investigated by analyzing its morphological characteristics in tomato seedlings infected and uninfected with M. incognita. Seeds were pre-soaked in VCE for the seed priming process before being allowed for germination. After 10 days of nematode inoculation, biochemical parameters like protein content, activity of antioxidative enzymes, non-enzymatic antioxidants, stress indices, photosynthetic pigments, proline content and secondary metabolites were also analyzed. The results revealed that neem-based VCE was fatal to second-stage juveniles, with an 82% mortality rate following exposure to the highest dose. When eggs were exposed to 100% VCE, 33.8% of hatching was suppressed, indicating that VCE had an antagonistic effect on nematode egg hatching. Further, all the morphological and biochemical parameters were significantly enhanced in VCE-treated tomato seedlings as compared to untreated seedlings. Stress indices were also found to be significantly lowered by the VCE treatments in the infected plants. The effect of VCE on seedling growth and physiology was shown to be concentration dependent. As a result, the current findings show that VCE has the potential to be used as a plant growth accelerator as well as an environmentally friendly biocontrol agent against nematode pathogenesis in tomato plants.

Biomolecular alterations in the early life stages of four food fish following acute exposure of Triclosan.

We investigated the effect of acute concentrations of triclosan (TCS; 96 h exposure and 10d post exposure) on the free amino acid, primary (SDS-PAGE) and secondary (FT-IR) structure of proteins in the embryos/larvae of Cyprinus carpio, Ctenopharyngodon idella, Labeo rohita and Cirrhinus mrigala. A concentration dependent increase in free amino acids, upregulation of polypeptides (100 and 70 kDa in C. carpio, C. idella and L. rohita, 55, 45, 36 kda in C. idella and L. rohita and 22 kDa in all the fish) and a decline in percent area of all the selected peaks of the FT-IR spectra was observed after exposure and recovery period. The decline in percent area was greatest for L. rohita at peak 1080 - 1088 cm-1 (-75.99%) after exposure and at peak 2854 - 2855 cm-1 (-53.59%) after recovery. Curve fitting analysis revealed a decrease in α-helices and increase in ß-sheets in all fish after exposure and recovery period. The results suggest that TCS elicits alterations in biomolecules of fish embryos.

Supplementing hand washing with proper use of alcoholic hand rub in a special neonatal care unit in a large academic public health institute at Jabalpur, Madhya Pradesh, India.

OBJECTIVE: The purpose was to increase use of alcoholic hand rub (AHR) in specialised newborn care unit (SNCU) to improve hand hygiene in order to reduce neonatal sepsis and mortality at Netaji Subhash Chandra Bose Medical College and Hospital, Jabalpur. DESIGN: A prospective interventional and observational study. METHODOLOGY: We formed a quality improvement (QI) team in our SNCU consisting of doctors, nurses, auxiliary staff and parents (a floating member) to improve proper use of AHR. To identify the barriers to the problem, we used fishbone analysis tool. The barriers which were not allowing the health providers to use AHR properly identified were amount of AHR in millilitres to be used per day per baby, how much and when the amount of AHR to be indented from the main store and what is the proper site to place the bottle. We used plan-do-study-act cycles to test and adapt solutions to these problems. Within 5-6 weeks of starting our project, AHR use increased from 44 mL to 92 mL per baby per day and this is sustained around 100 mL per baby per day for over 2 years now. RESULTS: Significant decrease in neonatal mortality was observed (reduced from median of 41.0 between August 2016 and April 2018 to 24.0 between May 2018 and December 2019). The neonates discharged alive improved from 41.2 to 52.3 as a median percentage value. The percentage of babies who were referred out and went Left Against Medical Advice (LAMA) deceased too. CONCLUSION: Multiple factors can lead to neonatal deaths, but the important factors are always contextual to facilities. QI methodology provides health workers with the skills to identify the major factors contributing to mortality and develop strategies to deal with them. Improving processes of care can lead to improved hand hygiene and saves lives.

Triclosan elicited biochemical and transcriptomic alterations in Labeo rohita larvae.

In the current study, Triclosan (TCS, a commonly used antimicrobial agent) induced alterations in biochemical parameters and gene expression were recorded in the larvae of Labeo rohita after 96 h exposure and 10 days recovery period to find out health status biomarkers. 96 h exposure to 0.06, 0.067 and 0.097 mg/L TCS significantly declined the levels of glucose, triglycerides, urea and uric acid and activity of alkaline phosphatase (ALP), glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT). There was a non-significant decline in the levels of cholesterol and total protein but albumin and total bilirubin showed no change. After 10 days of recovery period, trend was opposite for glucose, urea and ALP only. Decline in the expression of trypsin and pancreatic amylase and elevation in creatine kinase during exposure to TCS showed a reverse trend after recovery period. However, concentration dependent elevation of chymotrypsin persisted till the end of recovery period. Principal Component Analysis (PCA) showed association of total protein, ALP, GOT, creatine kinase and pancreatic amylase with PC1 after exposure as well as recovery period. Therefore, these can be considered as important biomolecules for identification of health status of TCS stressed fish.

Genomic markers for the biological responses of Triclosan stressed hatchlings of Labeo rohita.

Triclosan (TCS) used commonly in pharmaceuticals and personal care products has become the most common pollutant in water. Three-day-old hatchlings of an indigenous fish, Labeo rohita, were given 96h exposure to a nonlethal (60 µg L-1) and two moderately lethal concentrations (67 and 97 µg L-1) of TCS and kept for 10 days of recovery for recording transcriptomic alterations in antioxidant/detoxification (SOD, GST, CAT, GPx, GR, CYP1a and CYP3a), metabolic (LDH, ALT and AST) and neurological (AchE) genes and DNA damage. The data were subjected to principal component analysis (PCA) for obtaining biomarkers for the toxicity of TCS. Hatchlings were highly sensitive to TCS (96h LC50 = 126 µg L-1 and risk quotient = 40.95), 96h exposure caused significant induction of CYP3a, AChE and ALT but suppression of all other genes. However, expression of all the genes increased significantly (except for a significant decline in ALT) after recovery. Concentration-dependent increase was also observed in DNA damage [Tail Length (TL), Tail Moment (TM), Olive Tail Moment (OTM) and Percent Tail DNA (TDNA)] after 96 h. The damage declined significantly over 96h values at 60 and 67 µg L-1 after recovery, but was still several times more than control. TCS elicited genomic alterations resulted in 5-11% mortality of exposed hatchlings during the recovery period. It is evident that hatchlings of L. rohita are a potential model and PCA shows that OTM, TL, TM, TDNA, SOD and GR (association with PC1 during exposure and recovery) are the biomarkers for the toxicity of TCS. Graphical abstract.

Biomarkers for the toxicity of sublethal concentrations of triclosan to the early life stages of carps.

Accumulation, contents of protein, non-enzymatic antioxidant glutathione (GSH and GSSG), lipid peroxidation product (melondialdehyde-MDA) and organic acids (fumarate, succinate, malate and citrate), and activities of neurological (acetylcholinesterase-AChE), detoxification (glutathione S-transferase-GST) and metabolic (lactate dehydrogenase-LDH, aspartate transaminase-AST and alanine transaminase-ALT) enzymes were recorded in the hatchlings of Cyprinus carpio, Ctenopharyngodon idella, Labeo rohita and Cirrhinus mrigala after 7 and 14 days exposure and 10 days post exposure (recovery period) to sublethal concentrations (0.005, 0.01, 0.02 and 0.05 mg/L) of triclosan, a highly toxic and persistent biocide used in personal care products. Accumulation was maximum between 7-14 days at 0.01 mg/L for C. carpio and L. rohita but at 0.005 mg/L for C. idella and C. mrigala. No triclosan was observed at 0.005 mg/L in C. carpio and C. mrigala after recovery. Significant decline in protein, glutathione and acetylcholinesterase but increase in glutathione S-transferase, lactate dehydrogenase, aspartate transaminase, alanine transaminase, melondialdehyde and organic acids over control during exposure continued till the end of recovery period. Integrated biomarker response (IBR) analysis depicted higher star plot area for glutathione and glutathione S-transferase during initial 7 days of exposure, thereafter, during 7-14 days of exposure and the recovery period, higher star plot area was observed for acetylcholinesterase, aspartate transaminase, alanine transaminase and organic acids. Higher star plot area was observed for protein in all the species throughout the study. The study shows that L. rohita is most sensitive and glutathione, acetylcholinesterase, aspartate transaminase and alanine transaminase are the biomarkers for the toxicity of sublethal concentrations of TCS.

Biochemical markers for prolongation of the acute stress of triclosan in the early life stages of four food fishes.

In the present study, embryos of four food fishes viz. Cyprinus carpio, Ctenopharyngodon idella, Labeo rohita and Cirrhinus mrigala were given acute (96 h) exposure to their respective LC0, LC10 and LC30 (causing 0, 10 and 30% mortality, respectively) concentrations of triclosan [TCS, 5-chloro-2-(2,4-dichlorophenoxy) phenol], a broad spectrum biocide. Bioaccumulation, contents of protein, non-enzymatic antioxidants (GSH and GSSG), MDA (lipid peroxidation product) and organic acids (fumarate, succinate, malate and citrate) along with the activities of AChE (neurological enzyme), GST (detoxification enzyme) and three metabolic enzymes (LDH, AST and ALT) were estimated after 48 and 96 h exposure and 10 days post exposure. Around 1/10 of the TCS in water got accumulated in the hatchlings after 96 h, increase over 48 h values was maximum at LC0 (+195.30, +143.23 and + 140.75%) but minimum at LC30 (+89.62, +84.26 and + 126.72%) for C. idella, L. rohita and C. mrigala, respectively. In C. carpio, TCS got accumulated only at LC30 after 48 h but at all the concentrations after 96 h exposure. Contents of protein, GSH, GSSG and activity of AChE decreased but activities of GSH, LDH, AST and ALT and contents of MDA and organic acids increased concentration dependently in all the fishes. TCS declined by 85-90% but its toxic effects on biomolecules prolonged till the end of the recovery period. Such acute exposures are accidental but there is a need to evaluate biomarkers for prolongation of the stress of small concentrations especially LC0 and LC10 (causing negligible mortality) of lipophilic pollutants like TCS.

Azolla pinnata, Aspergillus terreus, and Eisenia fetida for fasterrecycling of nutrients from wheat straw.

A vast amount of surplus wheat straw/stubble (a carbon-rich bioresource) is wasted every year by burning. Harmful gases and residue matter released due to burning cause harmful effects on the environment and human health. Therefore, there is a strong need to recycle this bioresource in a sustainable manner. In the present study, wheat straw (W) was spiked with cattle dung (C), Azolla pinnata (A), and Aspergillus terreus (F) to make eight different treatments (1 kg each), viz. W (1 kg), WC (666 g + 334 g), WA (980 g + 20 g), WF (980 g + 20 ml), WCF (666 g + 314 g + 20 ml), WCA (666 g + 314 g + 20 g), WFA (960 g+ 20 ml + 20 g), and WCFA(666 g + 294 g + 20 ml + 20 g), and subjected to vermicomposting (Vcom) and aerobic composting (Acom). A comparison was made for the time required for degradation and nutrient profile of the products. The fastest recycling of wheat straw/stubble (120 days) was observed in WCA and WCFA, but the nutrient quality of WCA was better (N 18.67, P 3.88, K 38.84 g/kg). In the Acom group, longer time was required for degradation of various mixtures, but in this group also, WCA was degraded first of all (138 days) and yielded a product with the best nutrient quality (N 14.77, P 2.56, K 28.80 g/kg). Maximum growth of E. fetida and maximum number of hatchlings were observed in WCA while the highest cocoon production was observed in WCFA. It was observed that azolla enhanced conversion of wheat straw into a nutrient-rich product for agronomic use. Thus its use will reduce the amount of cattle dung in the mixture and the bulk to be handled by the farmers for ecosafe disposal of surplus straw/stubble. Therefore, this technology can be adopted as an alternative to burning.

Teratogenicity and accumulation of triclosan in the early life stages of four food fish during the bioassay.

TCS [5-chloro-2-(2,4-dichlorophenoxy)phenol] caused a concentration dependent delay in embryonic development, delay and decline in hatching and reduction in length and weight of hatchlings along with an increase in abnormal/deformed embryos and larvae and percent mortality. These parameters varied in a species specific manner and increased with TCS residue in body. The 96 h LC50 values of TCS for Cyprinus carpio, Ctenopharyngodon idella, Labeo rohita and Cirrhinus mrigala were estimated at 0.315, 0.116, 0.096 and 0.131 mg/L, respectively. Hatching got delayed by 16.33 h for C. carpio (0.47 and 0.50 mg/L TCS) and C. idella (0.20 mg/L TCS) but by 18.07 h for L. rohita (0.15 mg/L TCS) and by 19.33 h for C. mrigala (0.18 mg/L TCS). Spine malformations, oedema (yolk sac and cardiac) and deflated swim bladder were present in 100% larvae of C. carpio, C. idella, L. rohita and C. mrigala at 0.30, 0.08, 0.13 and 0.14 mg/L TCS, respectively. TCS also caused hemorrhage (all but C. idella, only 3.33%), albinism and deformed caudal fin (C. idella), hypopigmentation and rupturing of yolk sac (C. mrigala), gas bubble disease (C. mrigala and L. rohita), fusion of eyes (C. carpio) and degeneration of digestive tract (L. rohita) in 10-40% hatchlings. Exposed hatchlings were very weak and paralyzed, could not swim and remained settled at the bottom of jars. Embryonic development was observed to be an early indicator of the toxicity of TCS as oedema and bubbles in yolk were observed in 40-100% embryos/hatchlings at 0.08 mg/L TCS while 100% mortality was observed between 0.15 and 0.50 mg/L TCS. L. rohita was most sensitive and C. carpio was least sensitive to the stress of TCS. Accumulation of TCS in the hatchlings (1/10 of TCS in water) after 96 h exposure hints that even small quantities of TCS may change species diversity in natural waters.

Ultra-morphology of the scale as an indicator of the stress of Acid Black-1 (AB-1, CI: 20470) and zinc (Zn).

High contents of azo dyes and heavy metals enter surface waters with the wastewater from dying and dye-manufacturing industries and pose serious threat to fish. In the present study, changes in the ultra-morphological features of the scale have been evaluated as indicators of the stress of lethal and sublethal concentrations of an azo dye Acid Black-1 (AB-1, CI 20470), zinc (Zn), and their mixture AB-1 + Zn to Labeo rohita. Fish were exposed for 96 h to lethal concentration (LC) causing 0-70% mortality, i.e., LC0, LC20, LC50, and LC70 of AB-1 (4, 6, 8, and 10 mg/L respectively) and Zn (25, 50, 55, and 60 mg/L respectively) and LC0, LC50, and LC70 of AB-1 + Zn (2 + 15, 2 + 20, and 2 + 25 mg/L respectively). Subchronic exposures of 150 days were given to 1/12, 1/6, and 1/3 of 96 h LC50 values of AB-1, Zn, and AB-1 + Zn. After each exposure, the fish were kept for a recovery period of 90 days. Breakage of circuli, erosion and breakage of lepidonts, uprooting of tubercles, and disappearance of intercircular teeth were observed in all the fish, after 96 h exposure to AB-1, Zn, and AB-1 + Zn. However, damage to focus and holes were common on the scales of Zn-exposed fish. The mixture AB-1 + Zn was more toxic than either of the two as loss of circuli, lepidonts, and intercircular teeth, and sloughing of surface were observed in the scales after 96 h exposure to 2 + 25 mg/L. Damage at this concentration was more than the damage at 8/10 mg/L AB-1 and 55/60 mg/L Zn. After 150 days of exposure, damage due to sublethal concentrations was more than the damage due to all the concentrations of 96 h exposure. Irregular and sloughed circuli were common at 2 + 1.79 and 2 + 3.59 mg/L AB-1 + Zn. Cracks all over the surface (2 + 7.18 mg/L), calcium projections (2 + 3.59 and 2 + 7.18 mg/L), and holes (2 + 7.18 mg/L) hint towards synergistic toxicity of the mixture. It seems that the present dye and metal formed complexes with collagen and osteoblastic cells of the scale that caused an increase in damage during the post-exposure period. Scales of 2 + 7.18 mg/L AB-1 + Zn exposed fish were completely devoid of the normal architectural pattern on the 90th day of the recovery period. Changes in the ultra-morphology of scales at LC0 (0% mortality) and sublethal concentrations show that these are early indicators of the stress of minute quantities of dyes and metals in water. This is a first report on the cumulative toxicity of the two most abundant components of textile industry effluents.