Calculated LC50 Values Research Articles

Investigation of the effect of water and feed sourced boron on the growth performance and blood parameters of Nile tilapia, Oreochromis niloticus

AbstractThis study investigated the effects of water and feed‐derived boron on the growth performance and blood parameters of Nile tilapia fingerlings. Two different experiments were designed for this purpose. The first phase of the study determined the LC50 (96‐h) value of boron for Nile tilapia. Fish were then fed in water containing boron at a ratio of 1:20 of the LC50 value. In the second experiment, feed containing boron at different rates (0.00%, 0.01%, 0.05%, and 0.10%) was fed for 40 days. At the end of the feeding period, growth performance, hematology, and blood biochemistry parameters were determined. The study concluded with a calculated LC50 value of 161.053 mg/L boron for Nile tilapia. The initial weight was 12.51 ± 0.79 g; at the end of feeding, the final weights were determined as 26.36 ± 0.15 g for the control and 28.07 ± 0.23, 32.28 ± 0.25 and 24.81 ± 0.48 g for 0.01%, 0.05% and 0.10% of boron feed treatments, respectively. At the end of feeding in water containing boron, the final weight of Nile tilapia was determined as 24.26 ± 0.26 g (LC50/20%). The results showed that feeding Nile tilapia with 0.05% boron‐supplemented feed stimulated growth and positively affected blood parameters, whereas waterborne boron inhibited Nile tilapia growth and negatively affected blood parameters.

Insecticidal and Repellent Activity of Essential Oils from Copaifera reticulata, Citrus paradisi, Lavandula hybrida and Salvia sclarea Against Immature and Adult Stages of Ctenocephalides felis felis.

The flea Ctenocephalides felis (Siphonaptera: Pulicidae), parasitizes dogs and cats globally, acting as a vector for various pathogens affecting both animals and humans. Growing interest in environmentally friendly, plant-based products prompted this study. The aim of the study was to determine the chemical composition of essential oils (EOs) from Copaifera reticulata, Citrus paradisi, Lavandula hybrida and Salvia sclarea, assessing their insecticidal and repellent properties, determining lethal concentrations (LC50 and LC90), and evaluating residual efficacy in vitro against Ctenocephalides felis felis. Gas Chromatography with Flame Ionization Detector analyzed EO composition. In vitro tests involved preparing EO solutions at various concentrations. Ten specimens from each life stage (egg, larva, pupa, adult) were used for insecticidal activity assessment. Adulticidal activity was assessed using 10cm2 filter paper strip, each treated with 0.200mL of the test solution. Immature stages activities were evaluated using 23.76cm2 discs of the same filter paper, each treated with 0.470mL of the test solution. Mortality percentage was calculated using (number of dead insects × 100) / number of incubated insects. Probit analysis calculated LC50 values with a 95% confidence interval. Major EO constituents were β-caryophyllene (EOCR), linalool (EOLH), linalyl acetate (EOSS), and limonene (EOCP). LC50 values were obtained for all stages except for the essential oil of C. paradisi. All oils showed repellent activity at 800μg/cm2. OECR exhibited greater residual efficacy. Each EO demonstrated superior insecticidal activity against specific C. felis felis stages.

Integrated Analysis of Pollution Characteristic and Ecotoxicological Effect Reveals the Fate of Lithium in Soil-Plant Systems: A Challenge to Global Sustainability.

Lithium, as an emerging contaminant, lacks sufficient information regarding its environmental and ecotoxicological implications within soil-plant systems. Employing maize, wheat, pea, and water spinach, we conducted a thorough investigation utilizing a multispecies, multiparameter, and multitechnique approach to assess the pollution characteristics and ecotoxicological effects of lithium. The findings suggested that lithium might persist in an amorphous state, altering surface functional groups and chemical bonds, although semiquantitative analysis was unattainable. Notably, lithium demonstrated high mobility, with a mild acid-soluble fraction accounting for 29.66-97.02% of the total, while a minor quantity of exogenous lithium tended to be a residual fraction. Plant analysis revealed that in 10-80 mg Li/kg soils lithium significantly enhanced certain growth parameters of maize and pea, and the calculated LC50 values for aerial part length across the four plant species varied from 173.58 to 315.63 mg Li/kg. Lithium accumulation in the leaves was up to 1127.61-4719.22 mg/kg, with its inorganic form accounting for 18.60-94.59%, and the cytoplasm fraction (38.24-89.70%) predominantly harbored lithium. Furthermore, the model displayed that growth stimulation might be attributed to the influence of lithium on phytohormone levels. Water spinach exhibited superior accumulation capacity and tolerance to lithium stress and was a promising candidate for phytoremediation strategies. Our findings contribute to a more comprehensive understanding of lithium's environmental behavior within soil-plant systems, particularly within the context of global initiatives toward carbon neutrality.

Antifouling activities of proteinase K and α-amylase enzymes: Laboratory bioassays and in silico analysis

The application of enzymes as antifoulants is one of the environment-friendly strategies in biofouling management. In this study, antifouling activities of commercially available proteinase K and α-amylase enzymes were evaluated using barnacle larva and biofilm-forming bacteria as test organisms. The enzymes were also tested against barnacle cement protein through in silico analysis. The results showed that both enzymes inhibited the attachment of bacteria and settlement of barnacle larvae on the test surface. The lowest minimum inhibitory concentration of 0.312 mg ml−1 was exhibited by proteinase K against biofilm-forming bacteria. The calculated LC50 values for proteinase K and α-amylase against the barnacle nauplii were 91.8 and 230.96 mg ml−1 respectively. While α-amylase showed higher antibiofilm activity, proteinase K exhibited higher anti-larval settlement activity. Similarly, in silico analysis of the enzymes revealed promising anti-settlement activity, as the enzymes showed good binding scores with barnacle cement protein. Overall, the results suggested that the enzymes proteinase K and α-amylase could be used in antifouling coatings to reduce the settlement of biofouling on artificial materials in the marine environment.

Potential of Thuja occidentalis L. Essential Oil and Water Extracts against Field Crop Pests.

Thuja occidentalis L. essential oil (EOTO) and its compounds, such as terpinyl acetate, bornyl acetate, and β-thujone, are claimed to be highly effective against some storage pests, sanitary insects, or pests of fruit trees, while data about its use in protecting field crops are very scarce. There is also a lack of information in the literature about the insecticidal value of water extracts from T. occidentalis (WETOs). Both essential oils (EOs) and water extracts (WEs) from various plants have advantages and disadvantages in terms of their use as insecticides. EOs are generally more effective, but their preparation is more complicated and quite expensive. In turn, WEs are simple to prepare and cheap, but they often have limited effectiveness. Moreover, significant differences in responses exist depending on the species of the donor plant, the method of preparing the extract, its concentration, the species of the pest being controlled, the developmental stage, and even the gender of the pest. The goals of the research were to assess the effect of EOTO and WETOs prepared from dry and fresh matter on the mortality, feeding, and body mass changes of important crop pests, i.e., the black bean aphid, pea leaf weevil, and Colorado potato beetle (CPB), respectively, as well as on the mortality and voracity of non-target organism Asian lady beetle young larvae. EOTO showed significant aphicidal activity with LC50 = 0.8267% and 0.2453% after 42 h of the experiment for nymphs and wingless females of black bean aphid, respectively. Adults of CPB were more resistant to EOTO than aphids, with LC50 values for females equal to 1.5327% and 1.3113% after 48 h and after 72 h of the experiment. There was no significant effect of EOTO on CPB foraging. Calculated LC50 values for pea leaf weevil adults were lower than those for CPB (0.9638% and 0.8573% for males after 12 h and 24 h, respectively). In the case of this pest, a clear reduction in foraging was obtained, with higher concentrations of EOTO resulting in more pronounced reductions in foraging behavior. Concentrations of EOTO above 0.5%, which showed efficacy against the aphid, were lethal to 3-day-old larvae of the Asian lady beetle. WETOs, in turn, showed significant potential in inhibiting adult pea leaf weevil feeding, with very low or no effectiveness in reducing A. fabae and CPB, respectively.

Abstract 3333: Targeting the MDM2-p53 interaction: Time- and concentration-dependent studies in tumor and normal human bone marrow cells reveal strategies for an enhanced therapeutic index

Abstract Aim We aimed to design an MDM2-p53 antagonist with a differentiated tolerability profile that could be used to treat patients with wild-type TP53 malignancies. As part of an alliance between Newcastle University, Astex Pharmaceuticals, and Cancer Research Horizons, we discovered ASTX295, a potent inhibitor of the MDM2-p53 interaction that is currently under clinical investigation in patients with solid tumors (NCT03975387). We selected ASTX295 as a compound with a predicted short plasma half-life, which we hypothesised would help to mitigate the dose-limiting neutropenia and thrombocytopenia observed with earlier MDM2-p53 antagonists in clinical studies. To examine this hypothesis in vitro, we determined time- and concentration-dependent responses to ASTX295 treatment in healthy volunteer-derived human bone marrow cells, megakaryocytes, and in a panel of human tumor cell lines. Methods Samples containing bone marrow cells from healthy patients undergoing hip surgery were obtained under the ethical approval of the Newcastle Biobank (REC 12/NE/0395). Following Lymphoprep™ separation, cells were treated ex vivo with ASTX295 and seeded for Granulocyte-macrophage (GM) colony-forming assays in methylcellulose. Megakaryocytes were obtained from in vitro differentiation of CD34+ stem/progenitor cells. Human tumor cell lines (including MDM2-amplified SJSA-1) were treated with ASTX295 in vitro and seeded at low density for colony-forming assays. Exposures of 6, 12, or 24h were examined, and the data plotted to calculate LC50 values. Results The clonogenic survival of tumor cells was time-dependent, with LC50 values (mean ± SEM) in SJSA1 cells being 238 ± 46nM and 75 ± 7nM respectively (n = 3-4), following a 12h or 24h exposure to ASTX295. Time-dependent effects were also evident in five human bone marrow samples but with LC50 values of 1.9, >3, >10, >10, and >10uM being achieved at 12h, and 860 ± 268nM at 24h. Megakaryocytes showed similar time-dependent sensitivities in which daily treatment of 2 or 6h over three days did not induce apoptosis while significant cell death was observed when the treatment time was extended to 16-24h daily. In contrast, short, daily pulse treatment of 2-6h in cell lines (MV4-11, MOLM-13, SJSA-1) over three days was sufficient to induce cell death. Conclusions ASTX295 is a potent antagonist of the MDM2-p53 interaction. Collectively, our in vitro data suggest that a shorter exposure to ASTX295 (up to 12h), may help to spare healthy bone marrow cells whilst killing tumor cells. Hence, intermittent exposure to an MDM2-p53 antagonist could favourably modulate its therapeutic index. The predicted short plasma half-life of ASTX295 should provide flexibility in controlling the duration of exposure in vivo, potentially enabling a more bone-marrow sparing approach to MDM2-p53 antagonism to be utilised. Citation Format: Elaine Willmore, Maria Ahn, Suzanne Kyle, Yan Zhao, Huw Thomas, Kenneth S. Rankin, Luke Bevan, Lynsey Fazal, Keisha Hearn, Nicola Wilsher, Justyna Kucia-Tran, Nicola Ferrari, Nicola Wallis, Neil Thompson, John Lyons, Duncan Miller, Celine Cano, Martin E. Noble, Ian R. Hardcastle, Steven Howard, Gianni Chessari, John Lunec, David R. Newell, Steve R. Wedge. Targeting the MDM2-p53 interaction: Time- and concentration-dependent studies in tumor and normal human bone marrow cells reveal strategies for an enhanced therapeutic index [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3333.

Photocatalytic, Antibacterial, Cytotoxic and Bioimaging Applications of Fluorescent CdS Nanoparticles Prepared in DNA Biotemplate.

Synthesizing nanoparticles in biotemplates has been cited as one of the most promising way to obtain monodispersed inorganic nanoparticles. In this method, uniform voids in porous materials serve as hosts to confine the synthesized nanoparticles. DNA template can be described as a smart glue for assembling nanoscale building blocks. Here we investigate the photocatalytic, antibacterial, cytotoxic, and bioimaging applications of DNA capped CdS. XRD, SEM, TEM, UV-visible absorption, and photoluminescence spectra were used to study structural, morphological, and optical properties of CdS nanoparticles. Prepared CdS nanoparticles exhibit visible fluorescence. The photocatalytic activity of CdS towards Rhodamine 6G and Methylene blue are 64% and 91% respectively. A disc-diffusion method is used to demonstrate antibacterial screening. It was shown that CdS nanoparticles inhibit Gram-positive bacteria and Gram-negative bacteria effectively.DNA capped CdS shows higher activity than uncapped CdS nanoparticles. MTT cell viability assays were carried out in HeLa cells to investigate the cytotoxicity for 24h. At a concentration 2.5µg/ml, it shows 84% cell viability and 43% viability at12.5µg/ml. The calculated LC50 value is equal to 8µg/ml. These DNA capped CdS nanoparticles were taken for an in-vitro experiment with HeLa cells to exhibit the possibility of bioimaging applications. The present study suggests that the synthesized CdS nanoparticles could be a potential photocatalyst, antibacterial agent, and biocompatible nanoparticle for bioimaging applications.

Evaluation of the Toxicity of Microcyclamide Produced by Microcystis aeruginosa in Danio rerio Embryos.

The genus of cyanobacteria Microcystis is one of the most recurrent in blooms and is associated with the hepatotoxin microcystin production. In addition to cyanotoxins, these bacteria produce a wide range of secondary metabolites with a wide repertoire of activities. The co-occurrence of cyanotoxins and other cyanopeptides during blooming is quite common, and the negative effects are not always limited to one class of toxins, which makes it essential to investigate the toxicity of the other compounds individually. The objective of this study was to isolate the cyanopeptide microcyclamide produced by the strain Microcystis aeruginosa LTPNA 08 by liquid chromatography coupled to high-resolution mass spectrometry with a quadrupole-time-of-flight analyzer (LC-HR-QTOF-MS/MS) and to evaluate its acute toxicity in embryos of Danio rerio through the Fish Embryo Acute Toxicity (FET) assay. The fraction containing microcyclamide (95% purity) caused lethality in 62% of the embryos after 96 h exposure (50 µg mL-1), with evidence of cardiotoxicity (cardiac edema). The calculated LC50 value was 42.98 µg mL-1 (with a concentration range of 37.79-48.89 µg mL-1). The characterization of the secondary metabolites produced by cyanobacteria and the investigation of the toxicity of these compounds individually are essential for the identification of the substances responsible for negative effects on living organisms and on the ecosystem, in addition to assisting in the development of risk management policies.

EVALUATION OF ACHE ACTIVITY AND LIPID PEROXIDATION IN FRESHWATER TELEOST UPON EXPOSURE TO HERBICIDE

In this study, the freshwater fish Channa punctatus was used to study the harmful effects of clomazone on AChE and LPO at three different doses and for four different exposure times. To determine the sublethal concentrations of clomazone, the LC50 of the herbicide was calculated. Based on the calculated LC50 value, three different concentrations were determined: high dose- 0.5 ppm, mild dose- 0.25 ppm, and low dose- 0.08 ppm. The fish were treated with these concentrations and the exposure period was 7, 14, 21, and 28 days respectively. The acetylcholinesterase activity and concentration of MDA were significantly decreased (p<0.05) and increased (p<0.05), respectively, upon higher concentration and also with the increase in exposure periods. The highest reduction of AChE was recorded in the liver, followed by the kidney and gill in all three concentrations. On the other hand, the MDA concentration was recorded highest in the kidney, followed by the liver and gills. Though, in the liver, the concentration of MDA is almost the same when exposed to concentrations of 0.5 and 0.25 ppm clomazone. Thus, our findings imply that AChE activity and lipid peroxidation may be potential indicators of environmental pollutants in aquatic systems.

Pathogenicity of an Entomopathogenic Nematode, Steinernema carpocapsae on Alphitobius diaperinus (Coleoptera: Tenebrionidae) Strains From Turkey.

Alphitobius diaperinus Panzer (Coleoptera: Tenebrionidae) is an insect pest in poultry production systems. This insect has developed resistance to many chemical insecticides. As an alternative to chemicals, entomopathogenic nematodes (EPNs) are one of the most commonly used agents against several pest arthropods. The pathogenicity of the EPN Steinernema carpocapsae Weiser on four field strains of lesser mealworm, A. diaperinus, from Turkey was studied. In the experiments, larvae of A. diaperinus were found to be more sensitive than adults to S. carpocapsae infection. In terms of LC50 values in both larval and adult insects, Balikesir strain was found to be the most susceptible, Manisa strain the most resistant. The calculated LC50 values for adults were 85.9, 205.6, 135.4, and 418.8 IJs/ml, and for larvae 31.2, 39.8, 34.8, and 70.9 IJs/ml for the Balikesir, Canakkale, İzmir, and Manisa strains, respectively. This is the first report about the pathogenicity of EPNs against larvae and adults of lesser mealworm A. diaperinus strains from Turkey. We conclude that S. carpocapsae can be used as a control agent for lesser mealworms.

Оцінка потенційної токсичності мийно-дезінфекційних засобів на культурі клітин in vitro

Detergents with disinfectants properties today are widely used in various fields of human activity for cleaning surfaces, equipment, tools while destroying infectious agents. An active substance, as disinfectants, it contains various chemical compounds with antimicrobial, antiviral and antifungal properties. To establish the safety of these substances, along with toxicological studies on animals, experiments are conducted on alternative in vitro test systems, in particular on cell culture, which allows to quickly assess and predict the potential toxicity of detergents to humans and other living organisms. The purpouse of the study was assessment of the potential danger of detergents and disinfectants for humans by determining their cytotoxic effects in experiments on cell culture in vitro. Material and methods. Studies were performed to evaluate the cytotoxic effect of detergents with disinfectants properties : "Bioblysk", "Blysk Plus" and "Blysk" (manufactured by PE "HYGIENICS", Poland) on the lung cancer cells (line A-549) using cytochemical methods: MTT- with methyltetrazolium blue and CRB - test with sulforodamine B. Results. It was found that all detergents have the greatest cytotoxic effect on A-549 cells at a concentration of 1.0% of the native solution. The lowest concentrations 0.03% and 0.015% have no toxic effect on cells. With the help of probit analysis based on the obtained results, LC50 values were calculated: for Bioblysk - 0.15% (MTT test) and 0.15% (CRP test); for Blysk Plus LC50 - 0.25% (MTT test) and 0.2% (CRP test); for Blysk - 0.2% (MTT test and CRP test). LC50 values for detergents were 10 times lower than the recommended minimum concentrations of working solutions Conclusions. The obtained data indicate that the citotoxicity effect of detergents on A-549 cells was manifested by the effect on mitochondrial function and protein synthesis. The severity of studied drugs toxic effect depended on its chemical composition and pH, concentration in the incubation medium. The calculated LC50 values indicate their possible danger to the respiratory system, so they require appropriate protection.

Identification of Volatile Compounds and Insecticidal Activity of Essential Oils from Origanum compactum Benth. and Rosmarinus officinalis L. against Callosobruchus maculatus (Fab.)

This work was undertaken to investigate the volatile compounds and insecticidal activity of essential oils (EOs) from Origanum compactum Benth. and Rosmarinus officinalis L. against the crop pest Callosobruchus maculatus (Fab.). Essential oils of Origanum compactum (EOC) and Rosmarinus officinalis L. (EOR) were extracted by use of hydrodistillation, and their volatile compounds were profiled by gas chromatography-mass spectrometry (GC-MS). The insecticidal activity of extracted EOC and EOR was evaluated against C. maculatus. GC-MS analysis revealed that carvacrol (70.88%) and 1,8-cineole (62.35%) were the major constituents of EOC and EOR, respectively. EOC exhibited a potent insecticidal activity with calculated LC50 values of 6.77 and 3.57 μL/L air, 24 and 48 h posttreatment, respectively. Comparable LC50 values were obtained for EOR recording 6.25 and 3.82 μL/L, 48 h posttreatment. The effects of fumigation by the tested EOs on fertility (egg hatching) and the emergence of adult C. maculatus were also investigated. Notably, EOC completely abolished egg fertility judged by the abrogation of emergence of adults, regardless of the tested dose. By contrast, EOR completely inhibited the fertility and the emergence of C. maculatus adults at the dose of 16 μL/10 g. The outcome of the present study highlights the utility of the EOs from O. compactum Benth. and R. officinalis L. as natural sources of effective and ecofriendly pest-control agents.

A comparative investigation of toxicity of three polymer nanoparticles on acorn barnacle (Amphibalanus amphitrite)

Pollution from plastic waste is increasingly prevalent in the environment and beginning to generate significant adverse impact on the health of living organisms. In this study, we investigate the toxicity of polymer nanoparticles exposed to Acorn Barnacle (Amphibalanus amphitrite) nauplii, as an animal model. Highly stable aqueous dispersion of luminescent nanoparticles from three common polymers: polymethylmethacrylate (PMMA), polystyrene (PS), and polyvinylchloride (PVC), were prepared via nanoprecipitation and fully characterised. Exposure studies of these polymer particles to freshly spawned barnacle nauplii were performed within a concentration range from 1 to 25 mg/L under laboratory-controlled conditions. The exposure to PMMA and PS nanoparticles did not show detrimental toxicity and did not cause sufficient mortality to compute a LC50 value. However, PVC nanoparticles were significantly toxic with a mortality rate of up to 99% at 25 mg/L, and the calculated LC50 value for PVC nanoparticles was 7.66 ± 0.03 mg/L, 95% CI. Interestingly, PVC nanoparticle aggregates were observed to adhere to the naupliar carapace and appendages at higher concentrations and could not be easily removed by washings. To explore the possibility of chemical toxicity of polymer nanoparticles, analysis of the polymer powders which was used to prepare the nanoparticles was conducted. The presence of low molecular weight oligomers such as dimers, trimers and tetramers were observed in all polymer samples. The chemical nature and concentration of such compounds are likely responsible for the observed toxicity to the barnacle nauplii. Overall, our study shows that care should be exercised in generalising the findings of exposure studies performed using one type of plastic particles, as the use of different plastic particles may elicit different responses inside a living organism.

Effect of 2 Naphthalene Sulfonate on Biochemical Stress Markers as well as Structural Integrity of DNA in Liver and Kidney Tissue of Channa punctatus after Acute Exposure

2Naphthalene Sulfonate (2NS) is an intermediate compound used in textile industries but being nonbiodegradable, the solicitude regarding its ecotoxicity has risen. Thus, an inquisition was undertaken with the objective of evaluating the oxidative stress and genotoxicity of 2NS in fresh water fish, Channa punctatus. Based upon calculated LC50 value, two sublethal doses were selected i.e. 2.38g/L and 4.77g/L for further investigation. In order to study acute effect of 2NS, liver and kidney samples were collected after 24h, 48h, 72h and 96h of exposure. Symbolic elevation in oxidative stress biomarkers and DNA damage was observed revealing the toxic impact of 2NS. The study would be helpful in assessing the risk impose by 2NS and calls for urgency in application of stringent policies against the indiscriminate use of such toxic compounds.

An integrated biomarker approach to assess the welfare status of Ompok bimaculatus (Pabda) in biofloc system with altered C/N ratio and subjected to acute ammonia stress

Biofloc aquaculture is emerging fast as a sustainable technology; however, the system faces challenges from ammonia spikes that affect the physiology of the fishes. Besides, the immuno-stimulatory role of the produced floc in encountering the induced stressor is poorly understood. In this context, the study investigates the effect of different carbon to nitrogen (C/N) ratio on the welfare status of Ompok bimaculatus subjected to acute ammonia stress using an integrated biomarker response (IBR) approach. Four treatments with different C/N ratio of 10, 15, 20 & 25 were designated as C/N10, C/N15, C/N20 and C/N25, alongside a control group with a provision for 80–90% daily water exchange. Twenty-day old O. bimaculatus (average weight = 0.082 ± 0.001 g) larvae were uniformly distributed in each tank (25 l) at a stocking density of 160 nos./tank. At the end of 45-days rearing, fishes were exposed to an acute 96-h ammonia challenge, using a calculated LC50 value of 12.19 mg l−1. Pre- and post-exposure, blood indices like RBC, PVC and Hb were found highest in C/N25, while WBC was highest in C/N20. Plasma total protein, albumin and globulin levels were also elevated in C/N20 group, before and after exposure. Immune scores like lysozyme and respiratory burst activity were found highest in C/N20, whereas the control group showed the lowest activity. Stress biomarkers (glucose and cortisol) showed no significant change (p > 0.05) before exposure, however, levels were elevated in all groups with the highest observed in control and lowest in C/N15 and 20. Before exposure, catalase (CAT) activity was highest in C/N20 in all tissues (liver, gut, muscle); whereas, levels decreased compared to control after exposure. Post exposure, superoxide dismutase (SOD) activity was highest in C/N20 in all tissues (p < 0.05). Before and after ammonia exposure, glutathione peroxidase (GPx) activity in liver was highest in C/N20, whereas, its activity in gut was highest in C/N15. IBR analysis reveals higher values in liver and gut for biomarkers like MDA and GPx, while, SOD showed a plausible level in muscle tissue. Semi-quantitative histological analysis of the gill tissue showed a distinctive pathological damage in all the groups, post-exposure, with the control receiving highest level of alterations, however, not significant with C/N 15 & 25. Probability of survival among groups was highest in C/N20. Overall results from this study indicates a varied immune-stimulatory effect of C/N manipulation with C/N20 delivering highest protection to O. bimaculatus fry against acute ammonia challenge. Further, the IBR approach delineates the probable use of the tissue-level biomarkers for ascertaining the welfare status of O. bimaculatus in the biofloc system.

Spider mite resistance to miticides in South Carolina strawberry and implications for improved integrated pest management.

Tetranychus urticae Koch (Acari: Tetranychidae), twospotted spider mite, is a major secondary pest of strawberry and can cause significant yield loss. Tetranychus urticae is typically controlled using miticides, which has led to rapid resistance development. In South Carolina (USA), extension agents and growers have reported field failures of miticides (inadequate pest suppression), but resistance has not been quantitatively determined. In 2018, we determined the level of miticide resistance of six T. urticae populations found on strawberry across South Carolina. We examined efficacy of all miticides registered for use on US strawberry by conducting an initial diagnostic bioassay at 20% of the maximum labeled field rate. Any population × active ingredient combination resulting in < 55% mortality was identified as 'potentially resistant' and concentration-response bioassays were then conducted to calculate LC50 values for an individual population. These values were compared with those of a known-susceptible laboratory population to calculate resistance ratios (RR). Our results indicate that examined South Carolina populations of T. urticae from strawberry were highly resistant to bifenthrin (RR = 100-60,000) and there was reduced susceptibility to fenbutatin-oxide (RR = 25-123). The 'Sardinia' population had decreased abamectin susceptibility (RR = 25). No resistance to hexythiazox, etoxazole, acequinocyl, bifenazate, fenpyroximate, spiromesifen, or cyflumetofen was found. Based on available data, it appears that miticide resistance is not a likely cause of field failures and issues related to application error and coverage should be investigated. Overall, this work supports the need to reduce the use of broad-spectrum pesticides and older products, in favor of newer miticide chemistries due to resistance issues.

Bioactivities of rose-scented geranium nanoemulsions against the larvae of Anopheles stephensi and their gut bacteria.

Anopheles stephensi with three different biotypes is a major vector of malaria in Asia. It breeds in a wide range of habitats. Therefore, safer and more sustainable methods are needed to control its immature stages rather than chemical pesticides. The larvicidal and antibacterial properties of the Pelargonium roseum essential oil (PREO) formulations were investigated against mysorensis and intermediate forms of An. stephensi in laboratory conditions. A series of nanoemulsions containing different amounts of PREO, equivalent to the calculated LC50 values for each An. stephensi form, and various quantities of surfactants and co-surfactants were developed. The physical and morphological properties of the most lethal formulations were also determined. PREO and its major components, i.e. citronellol (21.34%), L-menthone (6.41%), linalool (4.214%), and geraniol (2.19%), showed potent larvicidal activity against the studied mosquitoes. The LC50/90 values for mysorensis and intermediate forms were computed as 11.44/42.42 ppm and 12.55/47.69 ppm, respectively. The F48/F44 nanoformulations with 94% and 88% lethality for the mysorensis and intermediate forms were designated as optimized formulations. The droplet size, polydispersity index, and zeta-potential for F48/F44 were determined as 172.8/90.95 nm, 0.123/0.183, and -1.08/-2.08 mV, respectively. These results were also confirmed by TEM analysis. Prepared formulations displayed antibacterial activity against larval gut bacteria in the following order of decreasing inhibitory: LC90, optimized nanoemulsions, and LC50. PREO-based formulations were more effective against mysorensis than intermediate. Compared to the crude PREO, the overall larvicidal activity of all nanoformulations boosted by 20% and the optimized formulations by 50%. The sensitivity of insect gut bacteria may be a crucial factor in determining the outcome of the effect of toxins on target insects. The formulations designed in the present study may be a good option as a potent and selective larvicide for An. stephensi.

Metarhizium Entomopathogenic Fungi Against the Beetle Brontispa longissima (Coleoptera: Hispidae): Isolation and Species Identification.

Seven entomopathogenic fungi strains (M1-7) were isolated from field-obtained dead coconut hispine beetles Brontispa longissima (Gestro), identified to species, and bioassayed for their pathogenicity. According to ITS sequences, all isolates belong in the genus Metarhizium, mainly M. flavoviride and M. anisopliae. Measured median lethal times (LT50) of 1×107 conidia/ml of M1-7 against fourth-instar B. longissima larvae within 15 d following exposure were, respectively: 5.43, 10.64, 11.26, 10.93, 6.62, 4.73, and 5.95 d. The isolate M6 yielded the highest mortality to fourth-instar larvae, and was thus selected to be tested against other larval instars and adults of B. longissima, after Time-Dose-Mortality (TDM) models. M6 proved more pathogenic against larvae than adults. The obtained bioassays data produced a good fit to the TDM models, yielding estimated LC50 and LT50 for each of the tested developmental stages of B. longissima. Both the obtained dose (β) and time effect (ri) parameters from TDM models suggest that first-instar larvae are the most susceptible life stage of the pest insect, while adults are more resistant to M6 infection. Calculated LC50 values were, respectively, 1.23×103 and 1.15×106 conidia/ml for first-instar larvae and adults, on the 15th day following M6 inoculation. Estimated LT50 were 3.3 and 5.9 d for first-instar larvae and adults, respectively, at 1×108 conidia/ml. Taken together, these results would suggest Metarhizium M6 as an option for the biological control of B. longissima in the field.

Occurrence of chemical pollutants in major e-waste sites in West Africa and usefulness of cytotoxicity and induction of ethoxyresorufin-O-deethylase (EROD) in determining the effects of some detected brominated flame retardants and e-waste soil-derived extracts.

We investigated the occurrence of chemical pollutants in major e-waste sites in West Africa and usefulness of cytotoxicity and induction of ethoxyresorufin-O-deethylase (EROD) in determining the effects of some detected brominated flame retardants (BFRs) and e-waste soil-derived extracts. Analysis of the e-waste site samples using AAS and GC-MS techniques revealed the presence of a range of toxic metals as well as persistent and toxic organic pollutants, respectively, in the vicinity of the e-waste sites. As expected, the occurrence (%) of all the detected chemical pollutants in experimental soils significantly (P < 0.05) differs from occurrence (%) in control soil. The calculated LC50 values on RBL-2H3 cells of the detected tetrabromobisphenol A (TBBPA) and hexabromocyclododecane (HBCD) were 3.75 μM and 4.2 μM, respectively. Tribromophenol (TBP), dibromobiphenyl (DBB), and decabromodiphenyl ether (DBDE) were remarkably less toxic on RBL-2H3 cells compared with TBBPA and HBCD as they did not reduce RBL-2H3 cell viability below 50% in the tested concentration range (0-20 μM). The study revealed that TBBPA and HBCD could induce significant RBL-2H3 cell death through caspase-dependent apoptosis. The study further shows that the cytotoxicity of some of these BFRs could increase synergistically when in mixtures and potentially activate inflammation through the stimulation of mast cell degranulation. The e-waste soil-derived extracts induced a concentration-dependent increase in EROD activity in the exposed RTG-W1 cells. Ultimately, nonpolar extracts had higher EROD-inducing potency compared with polar extracts and hence suggesting the presence in higher amounts of AhR agonists in nonpolar e-waste soil-derived extracts than polar extracts. Overall, there is urgent need for actions in order to improve the environmental quality of the e-waste sites.

Evaluation of pyrethroids and organophosphates in insecticide mixtures for management of western corn rootworm larvae.

The western corn rootworm is an economically important pest of corn. Management tactics include pyrethroid and organophosphate insecticides, which may be applied as a mixture to protect corn roots. The goal of our study was to characterize the effects of pyrethroids and organophosphates alone and in combination on larval corn rootworm mortality and injury to corn roots. We evaluated two insecticide combinations: tebupirimphos with β-cyfluthrin and chlorethoxyfos with bifenthrin. Using a soil-based, laboratory bioassay, we exposed larvae to five concentrations of the pyrethroid alone, the organophosphate alone, the combined formulation, and a water control. We calculated LC50 values and co-toxicity factors to determine synergism or antagonism between organophosphates and pyrethroids. We also measured adult emergence and root injury in a field experiment that tested tebupirimphos alone, β-cyfluthrin alone, the combined formulation, and an untreated control. Bioassay results indicated antagonism between the pyrethroid and organophosphate at most concentrations for both insecticide combinations. In the field experiment, tebupirimphos alone or in combination with β-cyfluthrin significantly reduced adult emergence and root injury compared to the untreated controls, but β-cyfluthrin alone did not differ from the untreated control for either metric. These results suggest that, at the concentrations tested, the pyrethroid component of pyrethroid-organophosphate mixtures may not contribute to a reduction of rootworm emergence or root injury. While these pyrethroids may confer a management benefit for other pests, such as seedcorn maggot, the concentrations of pyrethroids present in current formulations of these mixtures are likely too low for effective rootworm management. © 2020 Society of Chemical Industry.