Assessment of the in vitro acaricidal activity of Bravecto® (fluralaner) and a proposed orange oil-based formulation vehicle for the treatment of Sarcoptes scabiei.

BACKGROUND: Sarcoptic mange is a serious animal welfare concern in bare-nosed wombats (Vombatus ursinus). Fluralaner (Bravecto®) is a novel acaricide that has recently been utilised for treating mange in wombats. The topical 'spot-on' formulation of fluralaner can limit treatment delivery options in situ, but dilution to a volume for 'pour-on' delivery is one practicable solution. This study investigated the in vitro acaricidal activity of Bravecto, a proposed essential oil-based diluent (Orange Power®), and two of its active constituents, limonene and citral, against Sarcoptes scabiei. METHODS: Sarcoptes scabiei were sourced from experimentally infested pigs. In vitro assays were performed to determine the lethal concentration (LC50) and survival time of the mites when exposed to varying concentrations of the test solutions. RESULTS: All compounds were highly effective at killing mites in vitro. The LC50 values of Bravecto, Orange Power, limonene and citral at 1 h were 14.61 mg/ml, 4.50%, 26.53% and 0.76%, respectively. The median survival times of mites exposed to undiluted Bravecto, Orange Power and their combination were 15, 5 and 10 min, respectively. A pilot survival assay of mites collected from a mange-affected wombat showed survival times of < 10 min when exposed to Bravecto and Orange Power and 20 min when exposed to moxidectin. CONCLUSIONS: These results confirm the acaricidal properties of Bravecto, demonstrate acaricidal properties of Orange Power and support the potential suitability of Orange Power and its active constituents as a diluent for Bravecto. As well as killing mites via direct exposure, Orange Power could potentially enhance the topical delivery of Bravecto to wombats by increasing drug penetration in hyperkeratotic crusts. Further research evaluating the physiochemical properties and modes of action of Orange Power and its constituents as a formulation vehicle would be of value.

Evaluation of in silico model predictions for mammalian acute oral toxicity and regulatory application in pesticide hazard and risk assessment.

The United States Environmental Protection Agency (USEPA) uses the lethal dose 50% (LD50) value from in vivo rat acute oral toxicity studies for pesticide product label precautionary statements and environmental risk assessment (RA). The Collaborative Acute Toxicity Modeling Suite (CATMoS) is a quantitative structure-activity relationship (QSAR)-based in silico approach to predict rat acute oral toxicity that has the potential to reduce animal use when registering a new pesticide technical grade active ingredient (TGAI). This analysis compared LD50 values predicted by CATMoS to empirical values from in vivo studies for the TGAIs of 177 conventional pesticides. The accuracy and reliability of the model predictions were assessed relative to the empirical data in terms of USEPA acute oral toxicity categories and discrete LD50 values for each chemical. CATMoS was most reliable at placing pesticide TGAIs in acute toxicity categories III (>500-5000 mg/kg) and IV (>5000 mg/kg), with 88% categorical concordance for 165 chemicals with empirical in vivo LD50 values ≥ 500 mg/kg. When considering an LD50 for RA, CATMoS predictions of 2000 mg/kg and higher were found to agree with empirical values from limit tests (i.e., single, high-dose tests) or definitive results over 2000 mg/kg with few exceptions.

Oral acute toxicity study and in vivo antimalarial activity of Strychnos lucida R. Br. tablet.

ETHNOPHARMACOLOGICAL RELEVANCE: Malaria eradication has been a major goal of the Indonesian government since 2020. Medicinal plants, such as Strychnos lucida R. Br., are empirically used to treat malaria through traditional preparation methods. However, the safety and efficacy of these plants have not yet been confirmed. Therefore, further investigations are necessary to confirm the safety and efficacy of S. lucida as an antimalarial agent. AIMS OF THE STUDY: To quantify the concentration of brucine in the S. lucida extract, determine the acute oral toxicity of the standardized extract, and evaluate the in vivo antimalarial potency of S. lucida tablet (SLT). MATERIALS AND METHODS: Acute oral toxicity of S.lucida extract was determined using the Organization for Economic Co-operation and Development 420 procedure, and the analytical method for brucine quantification was validated using high-performance liquid chromatography. In addition, antimalarial activity was determined using the Peter's four-day suppressive method. RESULTS: Acute toxicity analysis revealed S. lucida as a low-toxicity compound with a cut-off median lethal dose of 2000-5000 mg/kg body weight [BW], which was supported by the hematological and biochemical profiles of the kidneys, liver, and pancreas (p > 0.05). Extract standardization revealed that S. lucida contained 3.91 ± 0.074% w/w brucine, adhering to the limit specified in the Indonesian Herbal Pharmacopeia. Antimalarial test revealed that SLT inhibited the growth of Plasmodium berghei by 27.74-45.27%. Moreover, SLT improved the hemoglobin and hematocrit levels. White blood cell and lymphocyte counts were lower in the SLT-treated group than in the K (+) group (p < 0.05). CONCLUSION: Histopathological and biochemical evaluations revealed that S. lucida extract was safe at a dose of 2000 mg/kg BW with low toxicity. SLT inhibited Plasmodium growth and improved the hemoglobin, hematocrit, and red blood cell profiles. Additionally, SLT reduced the lymphocyte and WBC counts and increased the monocyte and thrombocyte counts as part of the immune system response against Plasmodium infection.

Multi-biomarker approach to evaluate the toxicity of chlorpyrifos (active ingredient and a commercial formulation) on different stages of Biomphalaria straminea.

Biomphalaria straminea is a freshwater gastropod native to South America and used in toxicological assessments. Our aim was to estimate 48 h-LC50 and sub-chronic effects after the exposure to low concentrations of chlorpyrifos as commercial formulation (CF) and active ingredient (AI) on B. straminea adult, embryos and juveniles. Concentrations between 1 and 5000 µg L-1 were chosen for acute exposures and 0.1 and 1 µg L-1 for the sub-chronic one. After 14 days biochemical parameters, viability and sub-populations of hemocytes, reproductive parameters, embryotoxicity and offspring' survival were studied. Egg masses laid between day 12 and 14 were separated to continue the exposure and the embryos were examined daily. Offspring' survival and morphological changes were registered for 14 days after hatching. 48 h-LC50, NOEC and LOEC were similar between CF and AI, however the CF caused more sub-lethal effects. CF but not the AI decreased carboxylesterases, catalase and the proportion of hyalinocytes with respect to the total hemocytes, and increased superoxide dismutase and the % of granulocytes with pseudopods. Also CF caused embryotoxicity probably due to the increase of embryos' membrane permeability. Acetylcholinesterase, superoxide dismutase, hemocytes sub-populations, the time and rate of hatching and juveniles' survival were the most sensitive biomarkers. We emphasize the importance of the assessment of a battery of biomarkers as a useful tool for toxicity studies including reproduction parameters and immunological responses. Also, we highlight the relevance of incorporating the evaluation of formulations in order to not underestimate the effects of pesticides on the environment.

Multilevel assessment of carbamazepine effects: An integrative approach using zebrafish early-life stages.

Carbamazepine (CBZ) is the most commonly used drug in epilepsy treatment, and its metabolites are commonly detected among persistent pharmaceuticals in the aquatic environment. This study aimed to investigate CBZ effects on early-life-stage zebrafish (Danio rerio) (from 2 to 168 hpf) by employing of an integrative approach linking endpoints from molecular to individual level: (i) development; (ii) locomotor activity; (iii) biochemical markers (lactate dehydrogenase, glutathione-S-transferase, acetylcholinesterase and catalase) and (iv) transcriptome analysis using microarray. A 168 h - LC50 of 73.4 mg L-1 and a 72 h - EC50 of 66.8 mg L-1 for hatching were calculated while developmental effects (oedemas and tail deformities) were observed at CBZ concentrations above 37.3 mg L-1. At the biochemical level, AChE activity proved to be the most sensitive parameter, as evidenced by its decrease across all concentrations tested (∼25% maximum reduction, LOEC (lowest observed effect concentration) < 0.6 µg L-1). Locomotor behaviour seemed to be depressed by CBZ although this effect was only evident at the highest concentration tested (50 mg L-1). Molecular analysis revealed a dose-dependent effect of CBZ on gene expression. Although only 25 genes were deregulated in organisms exposed to CBZ when compared to controls, both 0.6 and 2812 µg L-1 treatments impaired gene expression related to development (e.g. crygmxl1, org, klf2a, otos, stx16 and tob2) and the nervous system (e.g. Rtn3, Gdf10, Rtn3), while activated genes were associated with behavioural response (e.g. prlbr and taar). Altogether, our results indicate that environmentally relevant CBZ concentrations might affect biochemical and genetic traits of fish. Thus, the environmental risk of CBZ cannot be neglected, especially in a realistic scenario of constant input of domestic effluents into aquatic systems.

Exploring honey bee toxicological data as a proxy for assessing dimethoate sensitivity in stingless bees.

The high diversity and distinctive characteristics of stingless bees pose challenges in utilizing toxicity test results for agrochemical registrations. Toxicity assessments were performed on 15 stingless bee species, along with the honey bee, using the insecticide dimethoate, following adapted OECD protocols. Median lethal doses over 24 h (24 h-LD50) were determined for exposure routes (acute oral or contact) and species. Species sensitivity distribution (SSD) curves were constructed and the 5% hazard doses (HD5) were estimated based on 24 h-LD50 values. The SSD curve was adjusted as the body weight and dimethoate response were correlated. Lighter bees (<10 mg) had lower 24 h-LD50 values. Contact exposure for adjusted HD5 suggested insufficient protection for Melipona mondury, whereas the oral exposure HD5 indicated no risks for the other 14 species. Comprehensive risk assessments are crucial for understanding the agrochemical impact on stingless bees, emphasizing the need for a broader species range in formulating conservation strategies.

Toxicity of minoxidil - Comprehensive in silico prediction of main toxicity endpoints: Acute toxicity, irritation of skin and eye, genetic toxicity, health effect, cardiotoxicity and endocrine system disruption.

This article focusses on elucidating the toxicological profile of minoxidil, a widely used pharmacological agent for alopecia, through the application of in silico methods (Percepta ACD/Labs software). This research is driven by the need to understand key toxicological endpoints: acute toxicity, skin and eye irritation, genetic toxicity, cardiotoxicity, disruption of the endocrine system, and estimation of various health effects due to the lack of experimental data for this drug. These parameters are critically evaluated to meet the stringent requirements of the pharmaceutical industry's safety assessments. The results obtained for acute toxicity (LD50 for rats and mouse) indicate that minoxidil exhibits a species-dependent acute toxicity profile e.g. 51 mg/kg bw for intravenous administration in mice. The predicted health effects indicate a 93% risk to the gastrointestinal system, 54% for the kidneys, 52% for the liver, 42% for the blood and lungs, and 39% for the cardiovascular system. The prediction of genotoxicity suggests a moderate probability (48%) of inducing a positive Ames test result. Furthermore, moderate inhibition of the hERG channel indicates potential cardiac risks of Minoxidil. Based on the information obtained, we propose subjecting minoxidil to additional toxicological assessments. The successful adoption of these in silico methodologies aligns with the 3 R s principle (replacement, reduction, and refinement) in the field of modern toxicological studies of minoxidil, all without the use of laboratory animals for the novelty of our toxicity assessment.

Development of a zebrafish model for toxicity evaluation of adulterated Apis mellifera honey.

Since ancient times, honey has been used for medical purposes and the treatment of various disorders. As a high-quality food product, the honey industry is prone to fraud and adulteration. Moreover, limited experimental studies have investigated the impact of adulterated honey consumption using zebrafish as the animal model. The aims of this study were: (1) to calculate the lethal concentration (LC50) of acid-adulterated Apis mellifera honey on embryos, (2) to investigate the effect of pure and acid-adulterated A. mellifera honey on hatching rate (%) and heart rate of zebrafish (embryos and larvae), (3) to elucidate toxicology of selected adulterated honey based on lethal dose (LD50) using adult zebrafish and (4) to screen the metabolites profile of adulterated honey from blood serum of adult zebrafish. The result indicated the LC50 of 31.10 ± 1.63 (mg/ml) for pure A. mellifera honey, while acetic acid demonstrates the lowest LC50 (4.98 ± 0.06 mg/ml) among acid adulterants with the highest mortality rate at 96 hpf. The treatment of zebrafish embryos with adulterated A. mellifera honey significantly (p ≤ 0.05) increased the hatching rate (%) and decreased the heartbeat rate. Acute, prolong-acute, and sub-acute toxicology tests on adult zebrafish were conducted at a concentration of 7% w/w of acid adulterants. Furthermore, the blood serum metabolite profile of adulterated-honey-treated zebrafish was screened by LC-MS/MS analysis and three endogenous metabolites have been revealed: (1) Xanthotoxol or 8-Hydroxypsoralen, (2) 16-Oxoandrostenediol, and (3) 3,5-Dicaffeoyl-4-succinoylquinic acid. These results prove that employed honey adulterants cause mortality that contributes to higher toxicity. Moreover, this study introduces the zebrafish toxicity test as a new promising standard technique for the potential toxicity assessment of acid-adulterated honey in this study and hazardous food adulterants for future studies.

Transcriptome Analysis of Host Anti-Vibrio harveyi Infection Revealed the Pathogenicity of V. harveyi to American Eel (Anguilla rostrata).

Vibrio harveyi, a recently discovered pathogenic bacterium isolated from American eels (Anguilla rostrata), poses uncertainties regarding its pathogenesis in American eel and the molecular mechanisms underlying host defense against V. harveyi infection. This study aimed to determine the LD50 of V. harveyi in American eel and assess the bacterial load in the liver, spleen, and kidney post-infection with the LD50 dose. The results showed that the LD50 of V. harveyi via intraperitoneal injection in American eels over a 14d period was determined to be 1.24 × 103 cfu/g body weight (6.2 × 104 cfu/fish). The peak bacterial load occurred at 36 h post-infection (hpi) in all three organs examined. Histopathology analysis revealed hepatic vein congestion and thrombi, tubular vacuolar degeneration, and splenic bleeding. Moreover, quantitative reverse transcription polymerase chain reaction (qRT-PCR) results indicated significant up or downregulation of 18 host immune- or anti-infection-related genes post 12 to 60 hpi following the infection. Additionally, RNA sequencing (RNA-seq) unveiled 7 hub differentially expressed genes (DEGs) and 11 encoded proteins play crucial roles in the anti-V. harveyi response in American eels. This study firstly represents the comprehensive report on the pathogenicity of V. harveyi to American eels and RNA-seq of host's response to V. harveyi infection. These findings provide valuable insights into V. harveyi pathogenesis and the strategies employed by the host's immune system at the transcriptomic level to combat V. harveyi infection.

Determination of the median lethal dose of zinc gluconate in mice and safety evaluation.

BACKGROUND: Zinc Gluconate (ZG) is a safe and effective supplement for zinc. However, there is limited research on the optimal dosage for intravenous injection and the safety evaluation of animal models for ZG. This study aims to determine the safe dose range of ZG for intravenous injection in C57BL/6J mice. METHODS: A Dose titration experiment was conducted to determine the LD50 and 95% confidence interval (95%CI) of ZG in mice. Based on the LD50, four sub-lethal doses (SLD) of ZG were evaluated. Following three injections of each SLD and monitoring for seven days, serum zinc levels were measured, and pathological changes in the liver, kidney, and spleen tissues of mice were determined by histological staining. RESULTS: The dose titration experiment determined the LD50 of ZG in mice to be 39.6 mg/kg, with a 95%CI of 31.8-49.3 mg/kg. There was a statistically significant difference in the overall serum zinc levels (H = 36.912, P < 0.001) following SLD administration. Pairwise comparisons showed that the serum zinc levels of the 1/2 LD50 and 3/4 LD50 groups were significantly higher than those of the control group (P < 0.001); the serum zinc level of the 3/4 LD50 group was significantly higher than those of the 1/8 LD50 and 1/4 LD50 groups (P < 0.05). There was a positive correlation between the different SLDs of ZG and the serum zinc levels in mice (rs = 0.973, P < 0.001). H&E staining showed no significant histological abnormalities or lesions in the liver, kidney, and spleen tissues of mice in all experimental groups. CONCLUSION: The appropriate dose range of ZG for intravenous injection in C57BL/6J mice was clarified, providing a reference for future experimental research.

Life stage-specific effects of tire particle leachates on the cosmopolitan planktonic copepod Acartia tonsa.

Tire wear particles (TWP) are a major source of microplastics in the aquatic environment and the ecological impacts of their leachates are of major environmental concern. Among marine biota, copepods are the most abundant animals in the ocean and a main link between primary producers and higher trophic levels in the marine food webs. In this study, we determined the acute lethal and sublethal effects of tire particle leachates on different life stages of the cosmopolitan planktonic copepod Acartia tonsa. Median lethal concentration (LC50, 48 h) ranged from 0.4 to 0.6 g L-1 depending on the life stages, being nauplii and copepodites more sensitive to tire particle leachates than adults. The median effective concentration (EC50, 48 h) for hatching was higher than 1 g L-1, indicating a relatively low sensitivity of hatching to tire particle leachates. However, metamorphosis (from nauplius VI to copepodite I) was notably reduced by tire particle leachates with an EC50 (48 h) of 0.23 g L-1 and the absence of metamorphosis at 1 g L-1, suggesting a strong developmental delay or endocrine disruption. Leachates also caused a significant decrease (10-22%) in the body length of nauplii and copepodites after exposure to TWP leachates (0.25 and 0.5 g L-1). We tested a battery of enzymatic biomarkers in A. tonsa adult stages, but a sublethal concentration of 50 mg L-1 of tire particle leachates did not cause a statistically significant effect on the measured enzymatic activities. Our results show that tire particle leachates can negatively impact the development, metamorphosis, and survival of planktonic copepods. More field data on concentrations of TWPs and the fate and persistence of their leached additives is needed for a better assessment of the risk of tire particle pollution on marine food webs.

Towards safer pesticide management: A quantitative structure-activity relationship based hazard prediction model.

Pesticides are recognized as common environmental contaminants. The potential pesticide hazard to non-target organisms, including various mammal species, is a global concern. The global problem requires a comprehensive risk assessment. To assess the toxic effects of pesticides at the early stage, a toxicological risk analysis is conducted to determine pesticide hazard levels. World Health Organization (WHO) has established five pesticide hazard classes based on lethal dose (LD50) values to perform these assessments. In this paper, we have developed one-vs-all quantitative structure-activity relationship (OvA-QSAR) models using five machine-learning techniques with the selected optimum molecular descriptors. Descriptor selection was conducted based on correlation to evaluate the relevance and significance of individual features in our dataset. Our OvA-QSAR model was built using a dataset obtained from the WHO, covering a wide range of chemical pesticides. These models can predict the hazard category for a pesticide within the five available categories. Notably, our experiments demonstrate the outstanding performance and robustness of the Random Forest (RF) model in addressing the challenge of multi-class classification with the selected descriptors.

Alteration of haematological and biochemical biomarkers after sub-lethal chronic malathion (Elathion®) intoxication in freshwater fish, Labeo rohita (Hamilton, 1822).

The present investigation aimed to evaluate the long-term effects of malathion (Elathion®) at two sub-lethal concentrations (0.36 and 1.84 mgL-1) for 45 days after the determination of 96 h-LC50 value (18.35 mgL-1) in a commercially important aquaculture species, Labeo rohita by assaying multiple biomarker approaches. Total erythrocyte count (TEC), and haemoglobulin count (Hb) were found to be decreased while total leucocyte counts (TLC) were increased (p < 0.05) in malathion-intoxicated fish. Malathion exposure significantly reduced (p < 0.05) serum protein levels while significantly increased (p < 0.05) blood glucose levels. RNA activity in muscle was reduced (p < 0.05) while DNA activity increased (p < 0.05) in malathion-intoxicated fish. Acid phosphatase (ACP) activities in the brain; lacate dehydrogenase (LDH) activities in brain and liver were increased (p < 0.05), while alkaline phosphatase (ALP) activities in the brain; succinate dehydrogenase (SDH) activities in the brain, liver and kidney; acetylcholine esterase (AChE) activity in the brain; and ATPase activities in the brain, liver and kidney were reduced (p < 0.05) in comparison to control. Thus, the alteration in studied biomarkers was in a concentation-time dependent manner; however, it was more pronounced at the higher concentration at 45 days of exposure. The alteration in biomarker activity is probably a defensive mechanism/ adaptive response of fish to overcome the stress induced by malathion, which is a novel insight and possible impact on L.rohita. Our findings suggest malathion-induced stress, therefore, the use of malathion needs to be regulated to safeguard aquatic animals including fish and human health.

Chemical, ecotoxicological, cytotoxic, and mutagenic evaluation of gelling agents used in the production of 70% alcohol gel.

With COVID-19, there has been an increase in the use of gelling agents for hand sanitizer production, and as a result, the release of this product into wastewater could induce impacts and adverse reactions in living organisms. Thus, ecotoxicological and cytotoxicological assessments of gelling agents with test organisms from different trophic levels are necessary to assess their environmental safety. For this, seven cellulose-based gelling agents and a polyacrylic acid derivative (C940) were selected for tests with Artemia salina. The most toxic agent was tested on Allium cepa to assess cytotoxicity. The volatile compounds of the gelling agents were analyzed. Cellulose-based gelling agents were not considered toxic according to their LC50, but C940 presented moderate toxicity to A. salina and cytotoxicity to Allium cepa, but without mutagenicity. In addition, C940 contained cyclohexane as a volatile compound. Thus, cellulose-based gelling agents are better environmental options than carbomer for 70% alcohol gel sanitizer.

Assessing the disparity: comparative toxicity of Copper in zebrafish larvae exposes alarming consequences of permissible concentrations in Brazil.

Copper (Cu) is a naturally occurring metal with essential micronutrient properties. However, this metal might also pose increased adverse environmental and health risks due to industrial and agricultural activities. In Brazil, the maximum allowable concentration of Cu in drinking water is 2 mg/L. Despite this standard, the impact of such concentrations on aquatic organisms remains unexplored. This study aimed to evaluate the toxicity of CuSO4 using larval zebrafish at environmentally relevant concentrations. Zebrafish (Danio rerio) larvae at 72 hr post-fertilization (hpf) were exposed to nominal CuSO4 concentrations ranging from 0.16 to 48 mg/L to determine the median lethal concentration (LC50), established at 8.4 mg/L. Subsequently, non-lethal concentrations of 0.16, 0.32, or 1.6 mg/L were selected for assessing CuSO4 -induced toxicity. Morphological parameters, including body length, yolk sac area, and swim bladder area, were adversely affected by CuSO4 exposure, particularly at 1.6 mg/L (3.31 mm ±0.1, 0.192 mm2 ±0.01, and 0.01 mm2 ±0.05, respectively). In contrast, the control group exhibited values of 3.62 mm ±0.09, 0.136 mm2 ±0.013, and 0.3 mm2 ±0.06, respectively. Behavioral assays demonstrated impairments in escape response and swimming capacity, accompanied by increased levels of reactive oxygen species (ROS) and lipid peroxidation. In addition, decreased levels of non-protein thiols and reduced cellular viability were noted. Data demonstrated that exposure to CuSO4 at similar concentrations as those permitted in Brazil for Cu adversely altered morphological, biochemical, and behavioral endpoints in zebrafish larvae. This study suggests that the permissible Cu concentrations in Brazil need to be reevaluated, given the potential enhanced adverse health risks of exposure to environmental metal contamination.

Susceptibility of the cultured Amazonian fish, Colossoma macropomum, to experimental infection with Aeromonas species from ornamental fish.

The global ornamental fish trade carries important risk factors for spreading pathogens between different countries and regions, not only for ornamental fish but also for cultured fish and even other animal species. In the current study, we reported the capacity of Aeromonas veronii and A. hydrophila isolated from ornamental fish to experimentally infect the reared Amazonian fish Colossoma macropomum. For this, those bacteria were identified, and a primary characterization was performed. Fish were inoculated with 0.1 mL of increasing concentrations of A. hydrophila or A. veronii (C1 = 1 × 102; C2 = 1.8 × 104; C3 = 2.1 × 106; C4 = 2.4 × 108 bacterial cells per mL) in the coelomic cavity. In the control group, fish received the same volume of sterile saline solution (0.9 %). Fish presented petechiae, skin suffusions, and mortality rates up to 100 % according to the inoculum concentration. Histopathologically, fish presented necrosis with karyolysis, loss of the cytoplasmic delimitation of cells of the renal tubules and hepatocytes, hemorrhage, cellular edema, and the presence of bacterial cells. The LD50-96h of A. veronii on C. macropomum was estimated at 2.4 × 106 CFU mL-1 and of A. hydrophila at 1.408 × 105 CFU mL-1. The results demonstrated that it is possible that Aeromonas species isolated from ornamental fish affect C. macropomum, causing similar clinical signs and lesions. This shows the importance of promoting risk control measures worldwide regarding the trade of ornamental fish.

Deleterious effects of benzotriazoles on zebrafish development and neurotransmission: 5-Chloro-benzotriazole versus 1H-benzotriazole.

Benzotriazoles are heterocyclic compounds typically presenting a benzene ring fused with a triazole molecule. The industry uses these compounds as anti-corrosion agents and recently, they have been employed in the pharmaceutical industry and in detergent formulations. Benzotriazoles persist in the environment, and water treatment plants cannot degrade them completely. Consequently, these compounds have been detected in rivers, lakes, and drinking water, which makes assessing their safety for the human and aquatic animal populations crucial. Here, we have evaluated and compared how exposure to 1H-benzotriazole or 5-chloro-benzotriazole affect the zebrafish embryo-larval stages. We have determined the acute toxicity, morphometric alterations, and acetylcholinesterase activity on zebrafish embryos, as well as behavioral endpoints using the tail coiling assay. The estimated LC50 of 5-chloro-benzotriazole was 19 mg/L, whereas 1H-benzotriazole caused no mortality. The zebrafish embryos exposed to 20 and 25 mg/L 5-chloro-benzotriazole had decreased hatching rate and exhibited pericardial and yolk sac edemas. Furthermore, the embryo length and eye area were decreased, in contrast with an increased yolk sac after exposure to 20 mg/L 5-chloro-benzotriazole. In turn, 1H-benzotriazole also decreased the eye area of zebrafish embryos, but no other significant morphological alterations were observed. The tail coiling assay showed that the zebrafish embryos increased the percentage of time moving and the number of embryonic movements per minute after exposure to 1H-benzotriazole (15 mg/L) or 5-chloro-benzotriazole (20 and 25 mg/L), indicating that these compounds were potentially neurotoxic. However, acetylcholinesterase activity was not significantly altered in embryos exposed to 1H-benzotriazole, but significantly decreased when exposed to 0.05 mg/L 5-chloro benzotriazole confirming its neurotoxicity at a much lower concentration. Our findings showed that 5-chloro-benzotriazole seems to induce more harmful alterations to zebrafish embryos than 1H-benzotriazole. Nevertheless, 1H-benzotriazole seems to induce a direct effect on eye development for concentrations lower than the ones of 5-chloro-benzotriazole affecting zebrafish embryos.

Design, Characterization and SAR Studies of Novel Bioactive Benzylideneacetophenone Derivatives as Insecticidal Agents against Spodoptera frugiperda (Lepidoptera: Noctuidae).

Unintentional environmental effects brought on by insecticides encourage the creation of safer substitutes. A very polyphagous migrating lepidopteran pest species in Africa called S. Frugiperda causes terrible damage. In the current paper, treatment of 4-acetylphenyl 4-methylbenzenesulfonate with different aromatic aldehydes in the presence of NaOH afforded benzylideneacetophenones. The structure of the newly prepared compounds were proved by different spectroscopic techniques such as IR, 1 H-NMR, 13 C NMR, and elemental analysis. We looked at the association between contact with S. frugiperda and stricture reaction to examine their harmful effect. Additionally, S. frugiperda was used for testing the newly created compounds for their ability to kill insects. The majority of substances have been proven to be effective and promising. It has been found that 4-[3-(4-Methylphenyl)prop-2-enoyl]phenyl-4-methyl benzenesulfonate (4) was the most active with an LC50 =3.46 mg/L of 2nd instar larvae and LC50 =9.45 mg/L of 4th instar larvae. Moreover, some of biological and histopathological aspects of the synthesized products were investigated under laboratory conditions.

Unveiling first report on in silico modeling of aquatic toxicity of organic chemicals to Labeo rohita (Rohu) employing QSAR and q-RASAR.

Labeo rohita, a fish species within the Carp family, holds significant dietary and aquacultural importance in South Asian countries. However, the habitats of L. rohita often face exposure to various harmful pesticides and organic compounds originating from industrial and agricultural runoff. It is challenging to individually investigate the effects of each potentially harmful compound. In such cases, in silico techniques like Quantitative Structure-Activity Relationship (QSAR) and quantitative Read-Across Structure-Activity Relationship (q-RASAR) can be employed to construct algorithmic models capable of simultaneously assessing the toxicity of numerous compounds. We utilized the US EPA's ToxValDB database to curate data regarding acute median lethal concentration (LC50) toxicity for L. rohita. The experimental variables included study type (mortality), study duration (ranging from 0.25 h to 4 h), exposure route (static, flowthrough, and renewal), exposure method (drinking water), and types of chemicals (industrial chemicals and pharmaceuticals). Using this dataset, we developed regression-based QSAR and q-RASAR models to predict chemical toxicity to L. rohita based on chemical descriptors. The key descriptors for predicting the toxicity of L. rohita in the regression-based QSAR model include F05[S-Cl], SpMax_EA(ri), s4_relPathLength_2, and SpDiam_AEA(ed). These descriptors can be employed to estimate the toxicity of untested compounds and aid in the development of compounds with lower toxicity based on the presence or absence of these descriptors. Both the QSAR and q-RASAR models serve as valuable tools for understanding the chemicals' structural features responsible for toxicity and for filling gaps in aquatic toxicity data by predicting the toxicity of newly untested compounds in relation to L. rohita. Finally, the developed best model was employed to predict 297 external chemicals, the most toxic substances to L. rohita were identified as cyhalothrin, isobornyl thiocyanatoacetate, and paclobutrzol, while the least toxic ones included ethyl acetate, ethylthiourea, and n-butyric acid.

Propidium monoazide PCR, a method to determine OsHV-1 undamaged capsids and to estimate virus Lethal Dose 50.

Ostreid herpes virus 1 (OsHV-1) has been classified within the Malacoherpesviridae family from the Herpesvirales order. OsHV-1 is the etiological agent of a contagious viral disease of Pacific oysters, C. gigas, affecting also other bivalve species. Mortality rates reported associated with the viral infection vary considerably between sites and countries and depend on the age of affected stocks. A variant called µVar has been reported since 2008 in Europe and other variants in Australia and in New Zealand last decade. These variants are considered as the main causative agents of mass mortality events affecting C. gigas. Presently there is no established cell line that allows for the detection of infectious OsHV-1. In this context, a technique of propidium monoazide (PMA) PCR was developed in order to quantify "undamaged" capsids. This methodology is of interest to explore the virus infectivity. Being able to quantify viral particles getting an undamaged capsid (not only an amount of viral DNA) in tissue homogenates prepared from infected oysters or in seawater samples can assist in the definition of a Lethal Dose (LD) 50 and gain information in the experiments conducted to reproduce the viral infection. The main objectives of the present study were (i) the development/optimization of a PMA PCR technique for OsHV-1 detection using the best quantity of PMA and verifying its effectiveness through heat treatment, (ii) the definition of the percentage of undamaged capsids in four different tissue homogenates prepared from infected Pacific oysters and (iii) the approach of a LD50 during experimental viral infection assays on the basis of a number of undamaged capsids. Although the developped PMA PCR technique was unable to determine OsHV-1 infectivity in viral supensions, it could greatly improve interpretation of virus positive results obtained by qPCR. This technique is not intended to replace the quantification of viral DNA by qPCR, but it does make it possible to give a form of biological meaning to the detection of this DNA.