96-h LC50 Research Articles

Sublethal and transgenerational effects of broflanilide on Myzus persicae (Sulzer) (Hemiptera: Aphididae)

Myzus persicae (Sulzer) is an important agricultural pest worldwide that imparts substantial damage to various horticultural and agricultural crops. Broflanilide is a newly developed insecticide that exhibits high activity against crop pests such as M. persicae. Nevertheless, the effects of sublethal levels of broflanilide on M. persicae fitness and population growth remain unclear. We assessed the sublethal and transgenerational effects of broflanilide on M. persicae performance using an age-stage, two-sex life table. Our results reveal that broflanilide is highly toxic to M. persicae with a 72-h LC50 of 2.541 mg/L. The LC25 of broflanilide significantly reduced the longevity and fecundity of the F0 and F1 generations, and the LC10 and LC25 of broflanilide significantly prolonged the developmental period of the third instar nymph in the F1 generation. The net reproductive rate (R0), intrinsic rate of increase (r), and finite rate of increase (λ) were significantly lower in the LC25-treated aphids. LC25 broflanilide treatment significantly downregulated the reproduction-related genes ERR and Vg and significantly upregulated JHEH expression in the F0 generation. LC25 and LC10 broflanilide treatments significantly upregulated the detoxification-related genes CarE and CYP6CY3, respectively, which may be related to broflanilide resistance in M. persicae. These results showed that broflanilide LC25 exhibited a significant inhibitory effect on M. persicae populations, which can persist in the F1 generation. Our study facilitates the understanding of the sublethal effect of broflanilide on aphid performance and helps develop methods and programs for controlling M. persicae using low broflanilide doses.

Toxicity of triclosan, an antimicrobial agent, to a nontarget freshwater zooplankton species, Moina macrocopa.

The toxicity of triclosan (TCS) on the freshwater cladoceran Moina macrocopa was investigated by acute and chronic toxicity assessments followed by genotoxicity and oxidative stress response analyses. The 48-h LC50 of TCS for ≤24-h-old M. macrocopa was determined as 539 μg L-1 . Chronic exposure to TCS at concentrations ranging from 5 to 100 μg L-1 showed a stimulatory effect at low concentrations (≤10 μg L-1 ) and an inhibitory effect at high concentrations (≥50 μg L-1 ) on growth, reproduction, and population-growth-related parameters of M. macrocopa. The genotoxicity test results indicated that TCS concentrations ranging from 50 to 100 μg L-1 can alter individuals' DNA. Analysis of the antioxidant enzymes catalase (CAT) and glutathione s-transferase (GST) demonstrated increased levels of these enzymes at high TCS concentrations. Our results indicated that TCS concentrations found in the natural environment have minimal acute toxicity to M. macrocopa. However, TCS at even low concentrations can significantly affect its growth, reproduction, and population-growth-related characteristics. The observed responses suggest a hormetic dose-response pattern and imply a potential endocrine-disrupting effect of TCS. Our molecular and biochemical findings indicated that high concentrations of TCS have the potential to induce oxidative stress that may lead to DNA alterations in M. macrocopa.

Acute toxicity of palm oil mill effluent on zebrafish (Danio rerio Hamilton-Buchanan, 1822): Growth performance, behavioral responses and histopathological lesions

Evaluating the toxicity of Palm Oil Mill Effluent (POME) is critical as part of the effort to develop waste management regulations for the palm oil industry. In this study, we investigated the acute toxicity of POME on growth performance, behavioral response, and histopathology of gill and liver tissues of zebrafish (Danio rerio). In total, 550 adult male zebrafish were used for the toxicity experiment including range finding test, acute toxicity test, growth performance and behaviour test. Static non-renewal acute toxicity bioassays were conducted by exposing fish to POME (1.584–9.968 mL/L) for 96 h. Growth performance, behavior response, and histopathological lesions in untreated and POME treated (96-h LC50: 5.156 mL/L) fish were measured at 24, 48, 72 and 96 h. Time-dependent significant decline in body length and body weight of POME-exposed zebrafish was observed. Furthermore, several behavioral changes were recorded, including hyperactivity, loss of balance, excessive mucus secretion, and depigmentation. Decreasing operculum movement and oxygen consumption rate as well as alterations in gill tissues (i.e. hyperplasia, hypertrophy, hemorrhage, and necrosis) of POME-exposed zebrafish were observed, suggesting a dysfunction in respiratory performance. On the other hand, liver tissue alterations (congestion, hemorrhage, hyperplasia, shrinkage of hepatocytes, hydrophilic degeneration, and necrosis) indicated a disruption in detoxification performance. We conclude that exposure to POME at acute concentration caused histopathological lesions both in gill and liver tissue along with changes in fish behaviors which disrupted respiratory and detoxification performance, resulting in mortality and reduced growth of zebrafish. These findings might provide valuable information for guiding POME management and regulation.

Histopathological Alterations in Gills, Liver and Kidney of African Catfish (Clarias gariepinus, Burchell 1822) Exposed to Melaleuca cajuputi Extract.

This study evaluated the histopathological changes in the gill, liver and kidney of African catfish (Clarias gariepinus) intoxicated with a sub-lethal dose of Melaleuca cajuputi leaves extract (MCLE) for 96 h. The acute toxicity test has been determined previously with a value of 96-h LC50 = 127 mg/L, hence the selection of sub-lethal ranges from 60 mg/L to 160 mg/L of MCLE. Degenerative alterations were prominent in all tested organs, particularly after exposure to a high concentration of MCLE. Gill exhibited haemorrhage, epithelial lifting, lamellar disorganisation, and necrosis after exposure to a high MCLE concentration. Alterations in the liver include congestion, hydropic degeneration, and vacuolation, whereas lesions in the kidney were pyknosis, vacuolation, hydropic degeneration, and tubular necrosis. The obtained data showed that the organs experienced severe changes proportional to the increase in MCLE concentration. In addition, fish exposed to higher concentrations than the LC50 value experienced irreversible lesions. The present study suggests that the use of MCLE below the LC50 is recommended to avoid severe alterations to organs, particularly in African catfish. This study demonstrated that the use of MCLE above the LC50 promotes severe damage to the gills, liver and kidney of African catfish. However, further investigations are needed to define the causing-mechanisms underlying these effects.

Toxicity assessment of poultry-waste biosynthesized nanosilver in Anabas testudineus (Bloch, 1792) for responsible and sustainable aquaculture development-A multi-biomarker approach

The current study investigates the potential utilization of poultry intestines for the synthesis of stable silver nanoparticles (AgNPs) and their impact on fish physiology. The AgNPs were synthesized and characterized using various analytical techniques. The toxicity of AgNPs on Anabas testudineus was evaluated, determining a 96-h LC50 value of 25.46 mg l−1. Subsequently, fish were exposed to concentrations corresponding to 1/10th, 1/25th, 1/50th, and 1/100th of the estimated LC50 for a duration of 60 days in a sub-acute study. A comprehensive range of biomarkers, including haematological, serum, oxidative stress, and metabolizing markers, were analyzed to assess the physiological responses of the fish. Additionally, histopathological examinations were conducted, and the accumulation of silver in biomarker organs was measured. The results indicate that silver tends to bioaccumulate in all biomarker organs in a dose- and time-dependent manner, except for the muscle tissue, where accumulation initially increased and subsequently decreased, demonstrating the fish's inherent ability for natural attenuation. Analysis of physiological data and integrated biomarker responses reveal that concentrations of 1/10th, 1/25th, and 1/50th of the LC50 can induce stress in the fish, while exposure to 1/100th of the LC50 shows minimal to no stress response. Overall, this study provides valuable insights into the toxicity and physiological responses of fish exposed to poultry waste biosynthesized AgNPs, offering potential applications in aquaculture while harnessing their unique features.

Acute mancozeb-fungicide exposure induces neuro-ethology disruption, health disorders, and immune-oxidative dysfunction in Nile tilapia (Oreochromis niloticus)

An acute exposure study of mancozeb (MAZ) fungicide was applied on Oreochromis niloticus for 96-h duration. Three hundred fish (20.50 ± 1.60 g) were assigned into six groups (50 fish/ group; 10 fish/replicate) and exposed to different six concentrations (0, 4, 8, 12, 16, and 20 mg L−1) of MAZ for 96-h. The Probit analysis program was used to compute the 96-h lethal concentration 50 (96-h LC50) of MAZ. During the exposure duration, the fish's behavior, clinical symptoms, and mortalities were recorded daily. After the exposure period was ended, the hematological, biochemical, immunological, and oxidant/antioxidant parameters were evaluated. The results of this study recorded the 96-h LC50 of MAZ for O. niloticus to be 11.49 mg L−1. Acute MAZ exposure badly affected the fish's behavior in the form of increased the breath gasping and swimming activity with aggressive mode. The exposed fish showed excessive body hemorrhages and fin rot. The survival rate of the exposed fish to MAZ was 100, 80, 66, 50, 38, and 30% in 0, 4, 8, 12, 16, and 20 mg L−1 MAZ, respectively. The hematological indices (red blood cell count, hemoglobin, packed cell volume%, and white blood cell count) were significantly decreased by increasing the MAZ exposure concentration (8–20 mg L−1). The acetylcholine esterase activity and immune indices (lysozyme, nitric oxide, immunoglobulin M, complement 3) were decreased by MAZ exposure (4–20 mg L−1). Acute MAZ exposure induced hepato-renal dysfunction and elevated stress-related parameter (cortisol) by increasing the MAZ concentration. A significant reduction in the antioxidant parameters (total antioxidant activity, catalase, and superoxide dismutase) with increasing the lipid peroxidation marker (malondialdehyde) was noticed by acute MAZ exposure (4 –20 mg L−1) in O. niloticus. Based on these outcomes, the MAZ exposure induced toxicity to the fish evident in changes in fish behavior, neurological activity, hepato-renal functioning, and immune-antioxidant responses which suggest physiological disruption.

Removal of the red tide dinoflagellate Cochlodinium polykrikoides using chemical disinfectants

For decades, red tide control has been recognized as necessary for mitigating financial damage to fish farms. Chemical disinfectants, frequently used for water disinfection, can reduce the risk of red tides on inland fish farms. This study systematically evaluated four different chemical disinfectants (ozone (O3), permanganate (MnO4−), sodium hypochlorite (NaOCl), and hydrogen peroxide (H2O2)) for their potential use in inland fish farms to control red tides by investigating their (i) inactivation efficacy regarding C. polykrikoides, (ii) total residual oxidant and byproduct formation, and (iii) toxicity to fish. The inactivation efficacy of C. polykrikoides cells by chemical disinfectants from highest to lowest followed the order of O3 > MnO4− > NaOCl > H2O2 for different cell density conditions and disinfectant doses. The O3 and NaOCl treatments generated bromate as an oxidation byproduct by reacting with bromide ions in seawater. The acute toxicity tests of the disinfectants for juvenile red sea bream (Pagrus major) showed that 72-h LC50 values were 1.35 (estimated), 0.39, 1.32, and 102.61 mg/L for O3, MnO4−, NaOCl, and H2O2, respectively. Considering the inactivation efficacy, exposure time of residual oxidants, byproduct formation, and toxicity toward fish, H2O2 is suggested as the most practical disinfectant for controlling red tides in inland fish farms.

2,4-Dinitrotoluene (2,4-DNT) exposure induces liver developmental damage and perturbs lipid metabolism and oxygen transport gene expression in zebrafish (Danio rerio).

2,4-Dinitrotoluene (2, 4-DNT) is a common environmental pollutant. The toxic effect on mammals of 2,4-DNT has been well studied, but its toxicity on aquatic organisms is little known. In this study, 126 healthy female zebrafish (Danio rerio) were exposed to different concentrations of 2,4-DNT (0, 2, 4, 8, 12 and 16mg/L) to determine 96-h semi-lethal concentrations (LC50). And then, 90 female zebrafish were exposed to 0, 2, 4 and 8mg/L 2,4-DNT for 5days to study liver toxicity. Exposed zebrafish developed hypoxia features, such as floating head and breathing rapidly, and then died. 96-h LC50 of 2,4-DNT in zebrafish was 9.36mg/L. Histological data revealed that 2,4-DNT severely damaged the liver tissues, following with the round nucleus, dense interstitial tissue, dense arranged hepatocyte cords and more inflammatory cells. Additionally, the further result showed that the lower levels of lipid transport and metabolism (apoα2, mtp, ppar-α and acox) were noticed. But, exposed to 2,4-DNT for 5days significantly upregulated the expression levels of genes involved in respiration (hif1a, tfa and ho1, p < 0.05). These results indicated that 2,4-DNT exposure disturbed lipid transport and metabolism and oxygen supply in zebrafish, which could contribute to severe damage in liver and death.

Chemical composition, mosquito larvicidal and molluscicidal activities of Magnolia foveolata leaf essential oil

Viral disease-causing pathogens transmitted by mosquito vectors are a global health problem, causing economic burden in many underdeveloped countries. Searching for pesticides from plant sources that are readily available in areas with mosquito-borne diseases to control mosquitoes may be an economical and timely solution to prevent and respond to pandemic outbreaks. Magnolia essential oils have shown a myriad of biological activities and we hypothesize that Magnolia foveolata essential oil may show pesticidal activities. The purpose of this investigation, therefore, was to characterize the essential oil of M. foveolata and to screen the essential oil for mosquito larvicidal and molluscicidal activities. Essential oil from the leaves of M. foveolata was obtained by hydrodistillation using a Clevenger type apparatus for 5 h. Chemical composition was analyzed by gas chromatography–mass spectrometry (GC-MS), gas chromatography with flame ionization detection (GC-FID), chiral gas chromatography - mass spectrometry. Pesticide activities of the essential oil were tested following WHO guidelines with modifications. The leaf essential oil of M. foveolata was dominated by the monoterpene hydrocarbons (+)-α-thujene (22.5%), (+)-α-pinene (6.9%) and (−)-α-pinene (9.0%), and p-cymene (9.5%), as well as the oxygenated monoterpenoid (−)-terpinen-4-ol (9.7%). The M. foveolata leaf essential oil was very active against the larvae of four mosquitoes Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, Culex fuscocephala with 24-h LC50 less than 40 μg/mL. The leaf essential oil of M. foveolata was also active against the fresh water snails, Pomacea canaliculata and Gyraulus convexiusculus, with LC90 values of 10.5 μg/mL and 23.6 μg/mL, respectively. Unfortunately, this essential oil also showed strong toxicity against non-target organisms Diplonychus rusticus, Anisops sp., Oreochromis sp., and Fejervarya limnocharis tadpoles. With appropriate care, the essential oil of M. foveolata may be considered as a pesticide applied to control mosquito species such as Ae. aegypti or Cx. quinquefasciatus living in urban areas.

Bifenthrin disrupts cytochrome c oxidase activity and reduces mitochondrial DNA copy number through oxidative damage in pool barb (Puntius sophore)

Bifenthrin (BF), a synthetic pyrethroid is used worldwide for both agricultural and non-agricultural purposes due to its high insecticidal activity and low toxicity in mammals. However, its improper usage implies a possible risk to aquatic life. The study was aimed to correlate the association of BF toxicity with mitochondrial DNA copy number variation in edible fish Punitus sophore. The 96-h LC50 of BF in P. sophore was 3.4 μg/L, fish was treated with sub-lethal doses ((⅒ and ⅕ of LC50;0.34 μg/L, 0.68 μg/L) of BF for 15 days. The activity and expression level of cytochrome c oxidase (Mt-COI) were measured to assess mitochondrial dysfunction caused by BF. Results showed BF reduced the level of Mt-COI mRNA in treated groups, hindered complex IV activity and increased ROS generation leading to oxidative damage. mtDNAcn was decreased in the muscle, brain and liver after BF treatment. Furthermore, BF induced neurotoxicity in brain and muscle cells through the inhibition of AchE activity. The treated groups showed elevated level of malondialdehyde (MDA) and an imbalance of antioxidant enzymes activity. Molecular docking and simulation analysis also predicted that BF binds to the active sites of the enzyme and restricts the fluctuation of its residues. Hence, outcome of the study suggests reduction of mtDNAcn could be a potential biomarker to assess Bifenthrin induced toxicity in aquatic ecosystem.

Phytochemical Compounds ofEuphorbia bivonaeExtract and Their Cytotoxicity Effects on the Lethality of Brine ShrimpArtemia salinaand Embryonic Kidney (HEK293) Cells.

In this research paper, we investigated the effect of Euphorbia bivonae extract compounds on the lethality of brine shrimp Artemia salina and on the embryonic cell line (HEK293) proliferation. Our GC/MS analysis revealed that the E. bivonae ethanolic extract contained essentially sitsoterol, euphol and lupeol. The 24-h LC50 was determined using the probit analysis method (LC50= 357.11mg l-1). Depending on this cytotoxicity test result, E. bivona extract induced a significant increase of Superoxide Dismutase (SOD), Catalase (CAT), Glutathione-Peroxidase (GPx) activities, and lipid peroxidation (LPO) in A. salina larvae. In addition, the cytotoxicity effect of this extract had proved against the HEK293 cell line in vitro. We suggest that the three compounds of E. bivonae extract (sitsoterol, euphol and lupeol) are the most responsible for this cytotoxicity. The possible application of this extract as an alternative natural antiproliferative is considered.

Acute toxicity and histopathological effects of Malathion on shrimp Macrobrachium nipponense (De Haan, 1849) (Caridea: Palaemonidae)

In the south of Iraq, Malathion is frequently used in agricultural activities and over-fishing, significantly harming the environment and animals that live in the region's waterways.The 96-h LC50 toxicity of malathion was determined in Macrobrachium nipponense and the valuewas 61.27 µg/L. Based on the 96-h LC50 value, two concentrations (1:10 and 1:5 of the 96-h LC50)were selected as sublethal concentrations for 1 week exposure for histopathology analysis. Themost histopathological alterations in hepatopancreas were abnormal lumen ALU after 1 week ofexposure to Malathion with the sublethal concentration 6.12 µg/L. In the other sublethal concen-tration of Malathion 12.25 µg/L for 1 week exposure, abnormal lumen ALU; necrosis of epithelialcells NC; and necrosis of tubules were recorded.

Relationship closeness of tolerance to two neonicotinoids with their internal body burden in two estuarine resident marine crustaceans

The estuarine resident crustacean sand shrimp, Crangon uritai, has a higher tolerance to neonicotinoid insecticides than that of the kuruma prawns, Penaeus japonicus. However, the reason for the differential sensitivities between the two marine crustaceans remains to be understood. This study explored the mechanism underlying differential sensitivities based on insecticide body residues after exposing both said crustaceans to two insecticides (acetamiprid and clothianidin) with or without oxygenase inhibitor piperonyl butoxide (PBO) for 96 h. Two graded-concentration groups were formed; group H (1/15–1 times the 96-h LC50 values) and L (one-tenth the concentration of group H). Results showed that the internal concentration in survived specimens tended to be lower in sand shrimp than in kuruma prawns. Co-treatment of PBO with two neonicotinoids not only increased sand shrimp mortality in the H group, but also altered metabolism of acetamiprid into its metabolite, N-desmethyl acetamiprid. Furthermore, molting during the exposure period enhanced bioconcentration of insecticides, but not affects survival. Collectively, the higher tolerance of sand shrimp than that of kuruma prawns to the two neonicotinoids can be explained by lower bioconcentration potential and more involvement of oxygenase in their alleviating lethal toxicity.

Mortality and histopathology in sheepshead minnow (Cyprinodon variegatus) larvae exposed to pectenotoxin-2 and Dinophysis acuminata

Toxic species of the dinoflagellate genus Dinophysis can produce diarrheic toxins including okadaic acid (OA) and dinophysistoxins (DTXs), and the non-diarrheic pectenotoxins (PTXs). Okadaic acid and DTXs cause diarrheic shellfish poisoning (DSP) in human consumers, and also cause cytotoxic, immunotoxic and genotoxic effects in a variety of mollusks and fishes at different life stages in vitro. The possible effects of co-produced PTXs or live cells of Dinophysis to aquatic organisms, however, are less understood. Effects on an early life stage of sheepshead minnow (Cyprinodon variegatus), a common finfish in eastern USA estuaries, were evaluated using a 96-h toxicity bioassay. Three-week old larvae were exposed to PTX2 concentrations from 50 to 4000 nM, live Dinophysis acuminata culture (strain DAVA01), live cells resuspended in clean medium or culture filtrate. This D. acuminata strain produced mainly intracellular PTX2 (≈ 21 pg cell−1), with much lower levels of OA and dinophysistoxin-1.No mortality or gill damages were observed in larvae exposed to D. acuminata (from 5 to 5500 cells mL−1), resuspended cells and culture filtrate. However, exposure to purified PTX2 at intermediate to high concentrations (from 250 to 4000 nM) resulted in 8 to 100% mortality after 96 h (24-h LC50 of 1231 nM). Histopathology and transmission electron microscopy of fish exposed to intermediate to high PTX2 concentrations revealed important gill damage, including intercellular edema, necrosis and sloughing of gill respiratory epithelia, and damage to the osmoregulatory epithelium, including hypertrophy, proliferation, redistribution and necrosis of chloride cells. Tissue damage in gills is likely caused by the interaction of PTX2 with the actin cytoskeleton of the affected gill epithelia. Overall, the severe gill pathology observed following the PTX2 exposure suggested death was due to loss of respiratory and osmoregulatory functions in C. variegatus larvae.

Physiological and molecular responses in the silver carp (Hypophthalmichthys molitrix) larvae after acute mercury exposure.

The current study was performed to examine the acute toxicity of mercuric chloride (HgCl2) on the silver carp (Hypophthalmichthys molitrix) larvae. Probit analysis was used to determine the median lethal concentration (LC50). The LC50 values of Hg2+ for the fish larvae at 24, 48, 72, and 96h were 267.72, 252.97, 225.57, and 97.80μg/L, respectively. The safe concentration of Hg was 9.78μg/L for fish larvae. Based on the 96h LC50, fish were exposed to four different groups including 0, 6.11, 12.23, and 24.45μg/L for 96h to assess the effects of different concentrations of Hg2+ on antioxidant capacity, energy metabolism parameters, and related gene expression. The findings revealed that there were no significant differences in the activities of superoxide dismutase (SOD) and total antioxidant capacity (T-AOC) in fish larvae among all the groups (P < 0.05). In the 12.23μg/L group, fish larvae had a maximum in catalase (CAT) activity. The creatine kinase (CK) activities of fish larvae in control and 6.11μg/L groups were significantly lower than those groups (P < 0.05). A high concentration of Hg2+ significantly upregulated the mRNA levels of heat shock protein 70 (HSP70) and metallothionein (MT) genes in fish larvae. Furthermore, the IBR index value showed the highest value in the 24.45μg/L group. Overall, this study provides an increased understanding of the effects of Hg-acute toxicity on silver carp larvae.

Evaluation of Toxicity Induced by Engineered CuO Nanoparticles in Freshwater Fish, Labeo rohita

With the fast development of industries relevant to nanotechnology, the inappropriate disposal of nanoproducts may initiate a new source of pollution in aquatic ecosystems, thus posing a possible danger to aquatic life. This study evaluated the eco-toxicological effects of waterborne copper oxide nanoparticles (CuO-NPs) having a 32.84nm size and rod shape on a freshwater fish, Labeo rohita. 96-h LC50 of CuO-NPs was 353.98mg/L. Two sub-lethal concentrations equivalent to 1/3rd and 1/5th LC50/96h (70.79 and 117.99 mg/L) of CuO-NPs were selected for 15, 30, and 45-day exposure tests. Bioaccumulation for the 1/3rd 96h LC50 was significantly higher compared to 1/5th of 96-h LC50 of CuO-NPs. There was a sharp decrease in the CAT activity and this decline ultimately increased the TBARS contents. The highest percentage of damaged nuclei and genetic damage index in fish erythrocytes was recorded at the highest concentration and after 45 days of treatment. The adverse effects of CuO-NPs were examined to be dose and duration dependent with increasing extent during all studied time intervals. Summarizing, exposure to sublethal concentrations of CuO-NPs is sufficient to cause alterations in ecotoxicological endpoints such as metal overload, oxidative stress and genotoxicity after chronic exposure.

Combined toxic effects of T-2 toxin and propiconazole on the early life stages of zebrafish (Danio rerio)

BackgroundAgricultural products are frequently co-contaminated by mycotoxins and pesticides. Most studies have only focused on the single toxicities of these chemicals, while their combined effects are neglected. Therefore, we investigated the combined toxic effects of T-2 toxin (T-2) and propiconazole (PRO) on zebrafish (Danio rerio).ResultsOur data exhibited that T-2 had higher toxicity to embryonic fish (96-h LC50 value of 0.39 mg a.i. L−1) than PRO (96-h LC50 value of 17.16 mg a.i. L−1). The mixture of T-2 and PRO showed an acute synergistic effect on zebrafish. Meanwhile, indicators associated with oxidative stress (SOD, Mn-sod, and cat) displayed significant variations in most exposures to T-2 and PRO mixtures (MTP) compared with the single exposures. The expressions of apoptosis-related genes cas3 and cas9 were also substantially elevated in the high-dose MTP exposure compared with the corresponding T-2 exposure. Besides, the expressions of endocrine system-related genes (TRβ, tsh, crh, cyp19a, and vtg1) were markedly varied in most MTP exposures compared with the corresponding single exposures. Our present results suggested that the mixture of T-2 and PRO could cause enormous effects on oxidative stress, cellular apoptosis, and the hypothalamic–pituitary–thyroid/hypothalamic–pituitary–gonadal (HPG/HPT) axis of zebrafish.ConclusionsOur results provided new insights into the development of combined pollution standards for agricultural products. Taken together, the impact of the combined effects could be considered and regulated as priorities.

Acute aquatic toxicity of two commonly used fungicides to midwestern amphibian larvae.

Fungicide usage has increased globally in response to the rise in fungal pathogens, especially in the agricultural sector. However, research examining the toxicity of fungicides is still limited for many aquatic species. In this study, we examined the acute toxicity of two widely used fungicides, chlorothalonil and pyraclostrobin, on six North American larval amphibian species across multiple families using 96-h LC50 tests. We found that pyraclostrobin was approximately 3.5x more toxic than chlorothalonil; estimated LC50 values ranged from 5-18 µg/L for pyraclostrobin and 15-50 µg/L for chlorothalonil. Comparing across amphibian groups, we found that salamanders were 3x more sensitive to pyraclostrobin than anuran species and equally as sensitive to chlorothalonil. Notably, our estimated LC50 values within the range of the expected environmental concentration for these fungicides suggesting environmental exposures could lead to direct mortality in these species. Given the widespread and increasing usage of fungicides, additional work should be conducted to assess the general risk posed by these chemicals to amphibian and their associated aquatic habitats.

Mixture toxic effects of thiacloprid and cyproconazole on honey bees (Apis mellifera L.)

Pesticide exposure remains one of the main factors in the population decline of insect pollinators. It is urgently necessary to assess the effects of mixtures on pollinator risk assessments because they are often exposed to numerous agrochemicals. In the present study, we explored the mixture toxic effects of thiacloprid (THI) and cyproconazole (CYP) on honey bees (Apis mellifera L.). Our findings revealed that THI possessed higher acute toxicity to A. mellifera (96-h LC50 value of 216.3 mg a.i. L−1) than CYP (96-h LC50 value of 601.4 mg a.i. L−1). It's worth noting that the mixture of THI and CYP exerted an acute synergistic effect on honey bees. At the same time, the activities of detoxification enzyme cytochrome P450s (CYP450s) and neuro target enzyme Acetylcholinesterase (AChE), as well as the expressions of seven genes (CRBXase, CYP306A1, CYP6AS14, apidaecin, defensing-2, vtg, and gp-93) associated with detoxification metabolism, immune response, development, and endoplasmic reticulum stress, were significantly altered in the combined treatment compared with the corresponding individual exposures of THI or CYP. These data indicated that a mixture of THI and CYP could disturb the physiological homeostasis of honey bees. Our study provides a theoretical basis for in-depth studies on the impacts of pesticide mixtures on the health of honey bees. Our study also provides important guidance for the rational application of pesticide mixtures to protect pollinators in agricultural production effectively.

Mixture toxicities of tetrachlorantraniliprole and tebuconazole to honey bees (Apis mellifera L.) and the potential mechanism

The extensive use of pesticides has negative effects on the health of insect pollinators. Although pollinators in the field are seldom exposed to individual pesticides, few reports have assessed the toxic impacts of pesticide combinations on them. In this work, we purposed to reveal the combined impacts of tetrachlorantraniliprole (TET) and tebuconazole (TEB) on honey bees (Apis mellifera L.). Our data exhibited that TET had greater toxicity to A. mellifera (96-h LC50 value of 298.2 mg a.i. L-1) than TEB (96-h LC50 value of 1,841 mg a.i. L-1). The mixture of TET and TEB displayed acute synergistic toxicity to the pollinators. Meanwhile, the activities of CarE, CYP450, trypsin, and sucrase, as well as the expressions of five genes (ppo, abaecin, cat, CYP4G11, and CYP6AS14) associated with immune response, oxidative stress, and detoxification metabolism, were conspicuously altered when exposed to the mixture relative to the individual exposures. These results provided an overall comprehension of honey bees upon the challenge of sublethal toxicity between neonicotinoid insecticides and triazole fungicides and could be used to assess the intricate toxic mechanisms in honey bees when exposed to pesticide mixtures. Additionally, these results might guide pesticide regulation strategies to enhance the honey bee populations.