Assessment of radiosensitivity and enhancing key steviol glycosides in Stevia rebaudiana Bertoni through gamma radiation.

PURPOSE: Stevia rebaudiana Bertoni is a perennial herb, widely used as a natural sweetener around the globe. The key compounds responsible for its sweetness includes stevioside and rebaudioside-A. In order to improve these steviol glycosides, the present study was initiated to study the effect of induced mutagenesis on growth parameters, steviol glycosides and nuclear DNA content in Stevia rebaudiana Bertoni using ten doses of gamma-rays (5-100 kR). MATERIALS AND METHODS: Healthy seeds of 'Madhuguna' variety of Stevia rebaudiana Bertoni developed and maintained at stevia breeding farm, Agrotechnology division, CSIR-Institute of Himalayan Bioresource Technology, Palampur (HP), India were irradiated with ten doses of gamma rays (600 seeds each/dose) ranging from 5 kR to 100 kR (i.e., 5, 10, 15, 20, 30, 40, 50, 60, 80 and 100 kR) using Co60 gamma irradiation chamber at CCS Haryana Agricultural University, Hisar, (Haryana), India. RESULTS: Significant variations were recorded for all the seedling traits studied while major impact was noticed on the seedling after reaching the cotyledonary stage and doses above 40 kR showed absolute mortality of the seedlings. Based on probit analysis, the optimum LD50 dose lies in the range of 20-23 kR. Glycosidic profiling of 296 mutants using high-performance liquid chromatography showed decreased total steviol glycoside content with increased radiation dose. Doses 5 kR and 10 kR, were found to be effective in increasing the overall glycosidic content. A total of 72 promising mutants were also screened for increased rebaudioside-A stevioside ratio. Comparison of nuclear DNA content using flow cytometry revealed a similar decrease in the total nuclear DNA content with increase in dosage of gamma rays. The average genome size at 5, 10, 15, 20 and 30 kR treatments were 2.72, 2.69, 2.68, 2.70 and 2.66 pg as compared to 2.72 pg in control. CONCLUSIONS: Mild dose of gamma rays (5 and 10 kR) in stevia were found to be effective in improving the mean steviol glycoside content and may be used in future stevia mutation programmes.

Efficient imputation methods in case of measurement errors.

This manuscript develops few efficient difference and ratio kinds of imputations to handle the situation of missing observations given that these observations are polluted by the measurement errors (ME). The mean square errors of the developed imputations are studied to the primary degree approximation by adopting Taylor series expansion. The proposed imputations are equated with the latest existing imputations presented in the literature. The execution of the proposed imputations is assessed by utilizing a broad empirical study utilizing some real and hypothetically created populations. Appropriate remarks are made for sampling respondents regarding practical applications.

Design based synthetic imputation methods for domain mean.

In real life, situations may arise when the available data are insufficient to provide accurate estimates for the domain, the small area estimation (SAE) technique has been used to get accurate estimates for the variable under study. The problem of missing data is a serious problem that has an impact on sample surveys, but small area estimates are especially prone to it. This paper is a basic effort that suggests design based synthetic imputation methods for the domain mean estimation using simple random sampling in order to address the issue of missing data under SAE. The expression of the mean square error for the proposed imputation methods are obtained up to first order approximation. The efficiency conditions are determined and a thorough simulation study is carried out using artificially generated data sets. An application is included with real data that further supports this study.

Identification of potential breeding ground of flathead grey mullet, Mugil cephalus (Linnaeus, 1758), along the Mumbai coast, India, for ecological monitoring and conservation strategies.

Identifying the breeding grounds of fishes is crucial for the sustainable management of fisheries resources. The present study is aimed at identifying the potential breeding ground of Mugil cephalus along the estuary of the North Mumbai coast. A total of 1197 specimens of M. cephalus, including 546 eggs, 271 larvae, 235 juveniles, and 235 adults, were collected from four sampling stations in the Karanja estuary between January to October 2022. Water quality parameters, plankton dynamics in the estuary, and the reproductive and feeding biology of M. cephalus were also examined. The eggs, larvae, juveniles, and adults were identified using traditional morpho-meristic and DNA barcoding techniques. The results revealed a potential spawning ground of M. cephalus in the Karanja estuary. The results of reproductive biology also confirmed the occurrence of matured fishes during May-July. The abundance of eggs and larvae at the estuary's mouth and the presence of juveniles and mature individuals of M. cephalus dominantly in the Karanja estuary from May to July infer the presence of a spawning site. It is also recorded that M. cephalus spawn in higher salinity (35 ppt) and seawater temperature (33 °C) where the hatching of offspring takes place successfully. This study emphasizes the significance of DNA barcoding in guiding routine monitoring surveys and demonstrates its usefulness when combined with these techniques in identifying fish spawning grounds. The study findings will serve as baseline information to develop effective conservation and management strategies and protect the ideal spawning stock.

A linker protein from a red-type pyrenoid phase separates with Rubisco via oligomerizing sticker motifs.

The slow kinetics and poor substrate specificity of the key photosynthetic CO2-fixing enzyme Rubisco have prompted the repeated evolution of Rubisco-containing biomolecular condensates known as pyrenoids in the majority of eukaryotic microalgae. Diatoms dominate marine photosynthesis, but the interactions underlying their pyrenoids are unknown. Here, we identify and characterize the Rubisco linker protein PYCO1 from Phaeodactylum tricornutum. PYCO1 is a tandem repeat protein containing prion-like domains that localizes to the pyrenoid. It undergoes homotypic liquid-liquid phase separation (LLPS) to form condensates that specifically partition diatom Rubisco. Saturation of PYCO1 condensates with Rubisco greatly reduces the mobility of droplet components. Cryo-electron microscopy and mutagenesis data revealed the sticker motifs required for homotypic and heterotypic phase separation. Our data indicate that the PYCO1-Rubisco network is cross-linked by PYCO1 stickers that oligomerize to bind to the small subunits lining the central solvent channel of the Rubisco holoenzyme. A second sticker motif binds to the large subunit. Pyrenoidal Rubisco condensates are highly diverse and tractable models of functional LLPS.

A comparative analysis of growth kinetics, image analysis, and biofuel potential of different algal strains.

Due to the global population growth and economic development, energy demand has increased worldwide. Countries take steps to improve their alternative and renewable energy sources. Algae is one of the alternative energy sources and can be used to produce renewable biofuel. In this study, nondestructive, practical, and rapid image processing techniques were applied to determine the algal growth kinetics and biomass potential of four algal strains, including C. minutum, Chlorella sorokiniana, C. vulgaris, and S. obliquus. Laboratory experiments were conducted to determine different aspects of biomass and chlorophyll production of those algal strains. Suitable non-linear growth models, including Logistic, modified Logistic, Gompertz, and modified Gompertz models, were employed to determine the growth pattern of algae. Moreover, the methane potential of harvested biomass was calculated. The algal strains were incubated for 18 days, and the growth kinetics were determined. After the incubation, the biomass was harvested and assessed for its chemical oxygen demand content and biomethane potential. Among the tested strains, C. sorokiniana was the best in biomass productivity (111.97 ± 0.9 mg L-1d-1). The calculated vegetation indices, namely; colorimetric difference, color index vegetation, vegetative, excess green, excess green minus excess red, combination, and brown index values showed a significant correlation with biomass and chlorophyll content. Among the tested growth models, the modified Gompertz shows the best growth pattern. Further, the estimated theoretical CH4 yield was highest for C. minutum (0.98 mL g-1) compared to other tested strains. The present findings suggest that image analysis can be used as an alternative method to study the growth kinetics and biomass production potential of different algae during cultivation in wastewater.

Anomalous Oligomerization Behavior of E. coli Aquaporin Z in Detergent and in Nanodiscs.

Aquaporins are tetrameric integral membrane proteins that act as water channels, and can also permeabilize membranes to other solutes. The monomer appears to be the functional form despite all aquaporins being organized as tetramers, which therefore must provide a clear functional advantage. In addition to this quaternary organization, some aquaporins can act as adhesion molecules in membrane junctions, when tetramers located in opposing membranes interact via their extracellular domains. These stacked forms have been observed in a range of aquaporins, whether using lipidic membrane environments, in electron crystallography, or using detergent micelles, in single-particle cryo-electron microscopy (cryo-EM). In the latter technique, structural studies can be performed when the aquaporin is reconstituted into nanodiscs of lipids that are surrounded by a protein scaffold. During attempts to study E. coli Aquaporin Z (AqpZ), we have found that in some conditions these nanodiscs tend to form filaments that appear to be either thicker head-to-tail or thinner side-to-side stacks of nanodiscs. Nanodisc oligomerization was observed using orthogonal analytical techniques analytical ultra-centrifugation and mass photometry, although the nature of the oligomers (head-to-tail or side-to-side) could not be determined. Using the latter technique, the AqpZ tetramer itself formed oligomers of increasing size when solubilized only in detergent, which is consistent with multiple stacking of AqpZ tetramers. We observed images consistent with both of these filaments in negative staining EM conditions, but only thicker filaments in cryo-EM conditions. We hypothesize that the apparent nanodisc side-to-side arrangement that can only be visualized in negative staining conditions is related to artifacts due to the sample preparation. Filaments of any kind were not observed in EM when nanodiscs did not contain AqpZ, or after addition of detergent into the nanodisc cryo-EM preparation, at concentrations that did not disrupt nanodisc formation. To our knowledge, these filaments have not been observed in nanodiscs preparations of other membrane proteins. AqpZ, like other aquaporins has a charge asymmetry between the cytoplasmic (more positive) and the extracellular sides, which may explain the likely head-to-tail stacking observed, both in nanodisc preparations and also in detergent micelles.

Study of feeding biology and diet-associated microplastic contamination in selected creek fishes of northeastern Arabian Sea: A multi-species approach.

This study investigated the diet composition and microplastic contamination in six fish species collected from the creek area of northeastern Arabian Sea. The results show that the diet of the fish is mainly composed of shrimps, algae, fish, and zooplankton, with microplastics constituting up to 4.83 % (Index of Preponderance) of their diet. The average abundance of microplastics ranges from 5.82 to 7.69 items per fish, and their ingestion is influenced by seasonal variation, gut fullness, and trophic level. Microplastic contamination has no significant effect on the condition factor and hepatosomatic index of the fish species. However, polymer hazard index indicates that microplastic pollution in fish is associated with a low to high risk factor, which might cause potential harm to aquatic lives and higher vertebrates via food chain. Therefore, this study highlights the need for immediate attention and effective regulations to reduce microplastic pollution to protect marine life.

Persistence of parental age effect on somatic mutation rates across generations in Arabidopsis.

In the model plant Arabidopsis thaliana, parental age is known to affect somatic mutation rates in their immediate progeny and here we show that this age dependent effect persists across successive generations. Using a set of detector lines carrying the mutated uidA gene, we examined if a particular parental age maintained across five consecutive generations affected the rates of base substitution (BSR), intrachromosomal recombination (ICR), frameshift mutation (FS), and transposition. The frequency of functional GUS reversions were assessed in seedlings as a function of identical/different parental ages across generations. In the context of a fixed parental age, BSR/ICR rates were unaffected in the first three generations, then dropped significantly in the 4th and increased in most instances in the 5th generation (e.g. BSR (F1 38 = 0.9, F2 38 = 1.14, F3 38 = 1.02, F4 38 = 0.5, F5 38 = 0.76)). On the other hand, with advancing parental ages, BSR/ICR rates remained high in the first two/three generations, with a striking resemblance in the pattern of mutation rates (BSR (F1 38 = 0.9, F1 43 = 0.53, F1 48 = 0.79, F1 53 = 0.83 and F2 38 = 1.14, F2 43 = 0.57, F2 48 = 0.64, F2 53 = 0.94). We adopted a novel approach of identifying and tagging flowers pollinated on a particular day, thereby avoiding biases due to potential emasculation induced stress responses. Our results suggest a time component in counting the number of generations a plant has passed through self-fertilization at a particular age in determining the somatic mutation rates.

Impacts of marine debris on coral reef ecosystem: A review for conservation and ecological monitoring of the coral reef ecosystem.

Coral reefs are the most spectacular underwater creation of nature. It enhances ecosystem functioning and marine biodiversity while also ensuring the livelihood of millions of coastal communities worldwide. Unfortunately, marine debris poses a serious threat to ecologically sensitive reef habitats and their associated organisms. Over the past decade, marine debris has been regarded as a major anthropogenic threat to marine ecosystems and gained scientific attention around the globe. However, the sources, types, abundance, distribution, and potential consequences of marine debris on reef ecosystems are hardly known. The goal of this review is to provide an overview of the current status of marine debris in various reef ecosystems across the world, with special emphasis on its sources, abundance, distribution, species impacted, major categories, potential impacts and management strategies. Furthermore, the adhesion mechanisms of microplastics to coral polyps, diseases caused by microplastics and are also highlighted.

Health risk assessment and metal contamination in fish, water and soil sediments in the East Kolkata Wetlands, India, Ramsar site.

East Kolkata Wetlands (EKW) is an important site for fish culture in sewage-fed areas, which are major receivers of pollutants and wastages from Kolkata. EKW is internationally important as the Ramsar site was declared on Aug 2002 with an area of 125 km2. EKW is a natural water body where wastewater-fed natural aquaculture has been practiced for more than 70 years. It is ecologically vulnerable due to the discharge of toxic waste through sewage canals from cities. Assessing the EKW to understand the inflow and load of the toxic metal (s) in fish, water, and sediments samples is essential. The field (samples collection from 13 sites) and lab (determination of toxic level of metals) based research were carried out to assess metal toxicity and health risk assessment in EKW. The levels of eighteen metals (18), namely Chromium, Vanadium, Cobalt, Manganese, Copper, Nickel, Zinc, Silver, Molybdenum, Arsenic, Selenium, Tin, Gallium, Germanium, Strontium, Cadmium, Mercury, and Lead, were determined using Inductively coupled plasma mass spectrometry (ICP-MS) in five fish tissues viz. muscle, liver, kidney, gill and brain, along with the water samples and soil sediments in 13 sampling sites. The bioaccumulation and concentration of metals in fish tissues, soil sediments, and water samples were well within the safe level concerning the recommendation of different national and international agencies except for a few metals in a few sampling sites like Cd, As, and Pb. The geoaccumulation index (Igeo) was also determined in the soil sediments, indicating moderate arsenic, selenium, and mercury contamination in a few sites. The contamination index in water was also determined in 13 sampling sites. The estimated daily intake (EDI), reference dose (RfD), target hazard quotient (THQ), slope factor and cancer risk of Cr, Mn, Co, Ni, Cu, Zn, As, Se, Cd, Pb and Hg from fish muscle were determined. Based on the results of the present investigation, it is concluded that fish consumption in the East Kolkata Wetland (EKW) is safe. The effects of bioaccumulation of metals in muscle tissue were well within the safe level for consumption as recommended by WHO/FAO.

Evaluating the performance of memory type logarithmic estimators using simple random sampling.

In survey research, various types of estimators have been suggested that consider only the current sample information to compute the unknown population parameters. Therefore, we utilize the past sample information along with the current sample information in the form of hybrid exponentially weighted moving averages to suggest the memory type logarithmic estimators for time-based surveys. The expression of the mean square error of the suggested estimators is determined to the first order of approximation. A relative comparison of the suggested estimators with the existing estimators is performed and efficiency conditions are obtained. Further, a simulation study is accomplished using a hypothetically rendered population and a real data illustration to improve the theoretical results. The results of the simulation study and the real data application exhibit that the consideration of past and current sample information meliorates the efficiency of the suggested estimators.

Impact of key parameters involved with plant-microbe interaction in context to global climate change.

Anthropogenic activities have a critical influence on climate change that directly or indirectly impacts plant and microbial diversity on our planet. Due to climate change, there is an increase in the intensity and frequency of extreme environmental events such as temperature rise, drought, and precipitation. The increase in greenhouse gas emissions such as CO2, CH4, NOx, water vapor, increase in global temperature, and change in rainfall patterns have impacted soil-plant-microbe interactions, which poses a serious threat to food security. Microbes in the soil play an essential role in plants' resilience to abiotic and biotic stressors. The soil microbial communities are sensitive and responsive to these stressors. Therefore, a systemic approach to climate adaptation will be needed which acknowledges the multidimensional nature of plant-microbe-environment interactions. In the last two scores of years, there has been an enhancement in the understanding of plant's response to microbes at physiological, biochemical, and molecular levels due to the availability of techniques and tools. This review highlights some of the critical factors influencing plant-microbe interactions under stress. The association and response of microbe and plants as a result of several stresses such as temperature, salinity, metal toxicity, and greenhouse gases are also depicted. New tools to study the molecular complexity of these interactions, such as genomic and sequencing approaches, which provide researchers greater accuracy, reproducibility, and flexibility for exploring plant-microbe-environment interactions under a changing climate, are also discussed in the review, which will be helpful in the development of resistant crops/plants in present and future.

Blockchain Powered Vaccine Efficacy for Pharma Sector.

Infectious and contagious diseases exist in humanity for many centuries which causes a curb in the growth of the population. Immunization plays a vital role to prevent mortality and morbidity against infectious diseases. COVID-19 pandemic continues to rage the urgency of developing a vaccine that should ensure the safety, efficacy, swift and fair deployment, implementation, and monitoring of vaccines across the globe. In the present context, the vaccine production to immunization campaign is a critical challenge. Therefore, an effective vaccine supply chain mechanism is required to address issues such as counterfeit vaccines, reduce vaccine wastages, and vaccine record fraud. In this paper, a blockchain-enabled vaccine supply chain is proposed to ensure the correctness, transparency, trust, and immutable log and improve the efficiency of vaccine distribution in the cold chain. The uniqueness of the proposed system is to provide distributed system to verify the reliability and efficacy of the vaccine from production to end beneficiaries' feedback about the vaccine. Our proposed system gives a clear view to the users as well as to the healthcare provider about the vaccination and ensures the anticounterfeit vaccine. The proposed system minimizes counterfeit vaccines and records, provides transparent communication between stakeholders in the supply chain, and improves the security of the vaccine supply chain and immutable feedback system about the vaccine.

Correction: Chrysomycins A-C, antileukemic naphthocoumarins from Streptomyces sporoverrucosus.

[This corrects the article DOI: 10.1039/C3RA42884B.].

Plant probiotics - Endophytes pivotal to plant health.

Endophytes as a ubiquitous associate of the plant are considered as a promising candidate for sustainable agriculture. These organisms play a pivotal role in the regulation of the primary and secondary metabolism of their host plant. The direct and long-lasting interaction of endophytes with the host enables them to escape from harsh environmental conditions. Especially, their endophytic nature makes them better candidates over epiphytes and rhizospheric microbes in interaction with plants. Current research findings revealed that the endophytes help plants in making nutrient acquisition from the soil, nitrogen fixation, phosphate availability, phytohormone and antimicrobial production. There is a huge potential for developing novel products like endophytes-based microbial formulations and elicitors to improve plant health, ameliorating stress tolerance in plants and source of therapeutically important secondary metabolites. The present review specifically dealt with attributes such as host-tissue specificity of endophytes, the importance of seed-associated endophytes, endophyte-parasite plant-host plant interaction as well as their applications in plant in-vitro systems and as microbial consortium. In addition, the conserved endophytic microbial communities in different plants are also looked upon possibly to understand the plant-endophytic microbiome on similar lines of the animal-gut microbiome. Primarily, the purpose of this review is to implicate the endophytic flora as probiotics influencing overall plant health and their survival under extreme environmental conditions.

Mechanisms Underlying C-type Inactivation in Kv Channels: Lessons From Structures of Human Kv1.3 and Fly Shaker-IR Channels.

Voltage-gated potassium (Kv) channels modulate the function of electrically-excitable and non-excitable cells by using several types of "gates" to regulate ion flow through the channels. An important gating mechanism, C-type inactivation, limits ion flow by transitioning Kv channels into a non-conducting inactivated state. Here, we highlight two recent papers, one on the human Kv1.3 channel and the second on the Drosophila Shaker Kv channel, that combined cryogenic electron microscopy and molecular dynamics simulation to define mechanisms underlying C-type inactivation. In both channels, the transition to the non-conducting inactivated conformation begins with the rupture of an intra-subunit hydrogen bond that fastens the selectivity filter to the pore helix. The freed filter swings outwards and gets tethered to an external residue. As a result, the extracellular end of the selectivity filter dilates and K+ permeation through the pore is impaired. Recovery from inactivation may entail a reversal of this process. Such a reversal, at least partially, is induced by the peptide dalazatide. Binding of dalazatide to external residues in Kv1.3 frees the filter to swing inwards. The extracellular end of the selectivity filter narrows allowing K+ to move in single file through the pore typical of conventional knock-on conduction. Inter-subunit hydrogen bonds that stabilize the outer pore in the dalazatide-bound structure are equivalent to those in open-conducting conformations of Kv channels. However, the intra-subunit bond that fastens the filter to the pore-helix is absent, suggesting an incomplete reversal of the process. These mechanisms define how Kv channels self-regulate the flow of K+ by changing the conformation of the selectivity filter.

Impact of COVID-19 lockdown on aquatic environment and fishing community: Boon or bane?

COVID-19 pandemic is a serious threat for mankind having an extensive socio-economic impact. However, it is considered as an unfortunate event with some positive environmental effects where nature is retrieving itself. The water quality index in different places of the world was reported to be improved during the lockdown, which in turn whipped up the regenerative process of fishes, sea turtles, marine mammals, and aquatic birds. Additionally, ecologically sensitive areas such as mangroves and coral reefs were also seen rejuvenating during COVID-19 seal off. But these favourable implications are temporary as there is an unexpected surge in plastic waste generation in the form of PPE kits, face masks, gloves, and other healthcare equipment. Moreover, the outbreak of the pandemic resulted in the complete closure of fishing activities, decline in fish catch, market disruption, and change in consumer preference. To address these multidimensional effects of the COVID-19 pandemic, government organizations, NGOs, and other concerned authorities should extend their support to amplify the positive impacts of the lockdown and reduce the subsequent pollution level while encouraging the fisheries sector.

Efficacy and Safety of a Phytopharmaceutical Drug Derived from Cocculus hirsutus in Adults with Moderate COVID-19: a Phase 2, Open-label, Multicenter, Randomized Controlled Trial.

INTRODUCTION: There is an urgent need for an effective, oral therapy for COVID-19. Purified aqueous extract of Cocculus hirsutus (AQCH) has shown robust antiviral activity in in vitro studies. We aimed to evaluate the efficacy and safety of AQCH plus standard of care in hospitalized patients with moderate COVID-19. METHODS: In an open-label, multicenter, randomized controlled trial conducted in India, eligible patients (aged 18-75 years) were randomized (1:1) to receive AQCH 400 mg orally three times a day plus standard of care (AQCH group) or standard of care alone (control group) for 10 days. Primary endpoint was the proportion of patients showing clinical improvement by day 14. Time to clinical improvement, time to viral clearance, and duration of hospitalization were secondary endpoints. RESULTS: A total of 210 patients were randomized. By day 14 most patients in both groups showed clinical improvement [difference - 0.01 (95% CI - 0.07 to 0.05); p = 1.0]. Median time to clinical improvement was 8 days (IQR 8-11) in the AQCH group versus 11 days (IQR 8-11) in the control group [HR 1.27 (95% CI 0.95-1.71); p = 0.032]. Time to viral clearance and duration of hospitalization were also significantly shorter in the AQCH group (p = 0.0002 and p = 0.016, respectively). AQCH was well tolerated, with no safety concerns identified. CONCLUSIONS: AQCH significantly reduced time to clinical improvement, time to viral clearance, and duration of hospitalization. In a pandemic, this has significant potential to decrease healthcare resource utilization and increase hospital bed availability. Further investigation of the therapeutic potential of AQCH in patients with COVID-19 is warranted. TRIAL REGISTRATION: Clinical Trials Registry - India (CTRI/2020/05/025397).

Multi-biomarker approach to assess chromium, pH and temperature toxicity in fish.

Chromium (Cr) is considered as the most common ubiquitous pollutant for aquatic animals including fish. An experiment was conducted to determine the acute and chronic toxicity of Cr, pH and high temperature in Anabas testudineus. Lethal concentration (LC50) of Cr alone was determined as 55.02 mg L-1, Cr and low pH 48.19 mg L-1 and Cr, low pH and high temperature 47.16 mg L-1. The chronic toxicity of low dose of Cr, pH and high temperature (1/10th and 1/20th of LC50) was designed to execute the experiment for 72 days. The stress enzymes and biomarkers were determined viz. superoxide dismutase, catalase, glutathione peroxidase, glutathione-s-transferase, lipid peroxide, acetylcholine esterase, cortisol, HSP-70, blood glucose, aspartate amino transferase, alanine amino transferase and malate dehydrogenase, lactate dehydrogenase, ATPase and genotoxicity in this study. We had also studied the integrated biomarker response (IBR), which revealed that Cr toxicity enhanced with concurrent exposure to pH and high temperature. All the biochemical attributes were significantly altered with exposure to Cr alone and with low pH and high temperature except gill SOD. Further, thermal tolerance was also determined, and results revealed that thermal tolerance was significantly reduced with exposure to Cr alone and Cr and low pH exposure in A. testudineus. The present study concluded that, the chronic toxicity of Cr is enhanced with low pH and high temperature and it has led to understanding the multi-approach of Cr toxicity which affect, stress biomarkers, cellular metabolic stress and thermal tolerance of A. testudineus.