Assessment of Lambda-Cyhalothrin and Spinetoram Toxicity and Their Effects on the Activities of Antioxidant Enzymes and Acetylcholinesterase in Honey Bee (Apis mellifera) Larvae.

Honeybees play a crucial role as agricultural pollinators and are frequently exposed to various pollutants, including pesticides. In this study, we aimed to evaluate the toxicity of lambda-cyhalothrin (LCY) and spinetoram (SPI) in honey bee larvae reared in vitro through single (acute) and repeated (chronic) exposure. The acute LD50 values for LCY and SPI were 0.058 (0.051-0.066) and 0.026 (0.01-0.045) µg a.i./larva, respectively. In chronic exposure, the LD50 values of LCY and SPI were 0.040 (0.033-0.046) and 0.017 (0.014-0.019) µg a.i./larva, respectively. The chronic no-observed-effect dose of LCY and SPI was 0.0125 µg a.i./larva. Adult deformation rates exceeded 30% in all LCY treatment groups, showing statistically significant differences compared to the solvent control group (SCG). Similarly, SPI-treated bees exhibited significantly more deformities than SCG. Furthermore, we examined the activities of several enzymes, namely, acetylcholinesterase (AChE), glutathione-S-transferase (GST), catalase (CAT), and superoxide dismutase (SOD), in larvae, pupae, and newly emerged bees after chronic exposure at the larval stage (honey bee larval chronic LD50, LD50/10 (1/10th of LD50), and LD50/20 (1/20th of LD50)). LCY and SPI induced significant changes in detoxification (GST), antioxidative (SOD and CAT), and signaling enzymes (AChE) during the developmental stages (larvae, pupae, and adults) of honey bees at sublethal and residue levels. Our results indicate that LCY and SPI may affect the development of honey bees and alter the activity of enzymes associated with oxidative stress, detoxification, and neurotransmission. These results highlight the potential risks that LCY and SPI may pose to the health and normal development of honey bees.

Transcriptome Profiling of Etridiazole-Exposed Zebrafish (Danio rerio) Embryos Reveals Pathways Associated with Cardiac and Ocular Toxicities.

Etridiazole (EDZ) is a thiadiazole-containing fungicide commonly used to control Pythium and Phytophthora spp. Although previous studies have shown that EDZ is teratogenic, the exact molecular mechanisms underlying its toxicity remain unknown. In this study, a zebrafish (Danio rerio; ZF) model was used to explore the molecular pathways associated with EDZ toxicity. The whole transcriptome of ZF embryos exposed to 96 h of EDZ was analyzed, along with developmental abnormalities. EDZ-induced malformations were primarily related to the eyes, heart, and growth of the ZF. Compared to untreated ZF, etridiazole-treated ZF had 2882 differentially expressed genes (DEGs), consisting of 1651 downregulated genes and 1231 upregulated genes. Gene ontology enrichment analysis showed that DEGs were involved in biological processes, such as sensory perception, visual perception, sensory organ development, and visual system development, and showed transmembrane transporter and peptidase regulator activities. Metabolism, phototransduction, aminoacyl-tRNA biosynthesis, MAPK signaling pathway, calcium signaling pathway, and vascular smooth muscle contraction were among the most enriched KEGG pathways. The qPCR analyses of the eight random genes were in good agreement with the transcriptome data. These results suggest several putative mechanisms underlying EDZ-induced developmental deformities in ZF.

Transcriptome Analysis of Thiram-Treated Zebrafish (Danio rerio) Embryos Reveals Disruption of Reproduction Signaling Pathways.

Thiram, a dithiocarbamate fungicide, is used for the treatment of various fungal infections affecting crops and ornamentals. However, thiram-associated toxicity has been reported in animals, including fish, and the underlying molecular mechanisms are unclear. Herein, we employed zebrafish (ZF) to gain further insights into thiram toxicity-associated molecular mechanisms. We studied developmental abnormalities and performed whole-transcriptome analysis of ZF embryos exposed to thiram for 96 h. Embryos exposed to 4.0 µg/L thiram exhibited several phenotypic abnormalities, including bradycardia, spinal curvature, hatching arrest, and growth retardation. Whole-transcriptome analysis revealed 1754 differentially expressed genes (DEGs), with 512 upregulated and 1242 downregulated DEGs. The majority of biological processes affected by thiram were metabolic. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis yielded terms related to reproduction, such as steroid biosynthesis and steroid hormone biosynthesis. Quantitative real-time polymerase chain reaction validation results were in line with sequencing data for ten DEGs. The study results improve our current understanding of the effects of thiram exposure in ZF.

Assessment of acute and chronic toxicity of cyantraniliprole and sulfoxaflor on honey bee (Apis mellifera) larvae.

BACKGROUND: Recently, cyantraniliprole (CYA) and sulfoxaflor (SUL) have been considered as alternatives to neonicotinoid insecticides. In this study, we evaluated the acute and chronic toxicities of CYA and SUL on honey bee (Apis mellifera L.) larvae reared in vitro. RESULTS: In the acute toxicity test, the following test doses were used to determine the median lethal dose (LD50 ): CYA 0.007, 0.014, 0.028, 0.056 and 0.112 µg larva-1 ; SUL 2.5, 5, 10, 20 and 40 µg larva-1 . In the chronic toxicity test, the following test doses were used to determine the LD50 : CYA 0.00512, 0.0128, 0.032, 0.08 and 0.2 µg larva-1 ; SUL 0.0625, 0.125, 0.25, 0.5 and 1.0 µg larva-1 . The acute LD50 values of CYA and SUL were 0.047 and 11.404 µg larva-1 , respectively. Larvae acutely exposed to SUL had significantly lower body weight than controls, but those exposed to CYA showed no difference. The no observed adverse effect level (NOAEL) and LD50 values of the chronic toxicity tests for each insecticide were 0.00512 and 0.064 µg larva-1 for CYA, and 0.0625 µg larva-1 and 0.212 µg larva-1 for SUL, respectively. Larvae chronically exposed to SUL emerged as bees with deformed wings, reaching adult deformation rates of over 50%; however, CYA had no effect on adult deformation. CONCLUSION: Exposure to CYA increased larval mortality but did not cause any adult deformation, whereas SUL exposure increased pupal mortality and caused wing deformation in newly emerged bees. Our study may be useful for the assessment of pesticide toxicity by providing valuable findings on the effects of these insecticides on honey bee larvae. © 2022 Society of Chemical Industry.

Developmental Toxic Effects of Thiram on Developing Zebrafish (Danio rerio) Embryos.

Thiram, an oxidized dimer of dithiocarbamate, has fungicidal and ectoparasiticidal roles. This study aimed to determine the effects of thiram on the development of zebrafish (ZF) embryos. The developmental toxicity test was performed in accordance with the OECD 236 test guidelines, and ZF embryos were subjected to several thiram concentrations and a DMSO (0.01%) control. Subsequently, embryo mortalities and developmental anomalies were evaluated at different hours post fertilization (hpf). Thiram was highly toxic to ZF, with calculated median lethal concentrations (LC50) of thiram at 48 and 96 h as 13.10 ± 2.17 and 8.87 ± 2.09 µg/L, respectively. Thiram-treated embryos/larvae exhibited a variety of deformities, such as abnormal somites, reduced eye pigment, abnormal tail shape, yolk sac edema, hatching defects, and curved spines, with a median effective concentration (EC50) of 3.88 ± 1.23, 5.04 ± 1.82, 6.23 ± 0.92, 5.24 ± 2.22, 1.39 ± 0.25, and 2.60 ± 0.82 µg/L, respectively. Teratogenic index (TI) values ranged from 1.42 to 6.66 for the scored deformities. At 48 hpf, the average heartbeat of the control group was 177.20 ± 5.63 per minute, while the highest thiram-treated group (40 µg/L) was 99.50 ± 18.12 per minute. In addition, cardiac-related issues, such as pericardial edema and abnormal blood flow, were observed in thiram-treated ZF embryos. Overall, these findings suggest that thiram is teratogenic to ZF.

Transcriptome-Based Identification of Genes Responding to the Organophosphate Pesticide Phosmet in Danio rerio.

Organophosphate pesticides (OPPs) are one of the most widely used insecticides. OPPs exert their neurotoxic effects by inhibiting acetylcholine esterase (AChE). Most of the gross developmental abnormalities observed in OPP-treated fish, on the other hand, may not be explained solely by AChE inhibition. To understand the overall molecular mechanisms involved in OPP toxicity, we used the zebrafish (ZF) model. We exposed ZF embryos to an OPP, phosmet, for 96 h, and then analyzed developmental abnormalities and performed whole transcriptome analysis. Phenotypic abnormalities, such as bradycardia, spine curvature, and growth retardation, were observed in phosmet-treated ZF (PTZF). Whole transcriptome analysis revealed 2190 differentially expressed genes (DEGs), with 822 and 1368 significantly up-and downregulated genes, respectively. System process and sensory and visual perception were among the top biological pathways affected by phosmet toxicity. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed significant enrichment of metabolic pathways, calcium signaling pathway, regulation of actin cytoskeleton, cardiac muscle contraction, drug metabolism-other enzymes, and phototransduction. Quantitative real-time PCR results of six DEGs agreed with the sequencing data expression profile trend. Our findings provide insights into the consequences of phosmet exposure in ZF, as well as an estimate of the potential risk of OPPs to off-target species.

Horizontal Honey-Bee Larvae Rearing Plates Can Increase the Deformation Rate of Newly Emerged Adult Honey Bees.

Rearing honey bee larvae in vitro is an ideal method to study honey bee larval diseases or the toxicity of pesticides on honey bee larvae under standardized conditions. However, recent studies reported that a horizontal position may cause the deformation of emerged bees. Accordingly, the purpose of this study was to evaluate the emergence and deformation rates of honey bee (Apis mellifera ligustica) larvae reared in horizontal and vertical positions. The study was conducted under the same laboratory conditions with three experimental groups, non-capped or capped horizontal plates and capped vertical plates. However, our results demonstrated that the exhibited adult deformation rates of the horizontal plates were significantly higher (27.8% and 26.1%) than those of the vertical plates (11.9%). In particular, the most common symptoms were deformed wings and an abnormal abdomen in the horizontal plates. Additionally, adults reared on horizontal plates were substantially smaller (10.88 and 10.82 mm) than those on vertical plates (11.55 mm). Considering these conclusions, we suggest that a vertical rearing method is more suitable when considering the deformation rates of the control groups to verify the sublethal effects of pesticides on honey bees.

Humification of effluent organic matters through a surface-flow constructed wetland.

Dominant fractions of wastewater effluent organic matter (EfOM) were changed from polysaccharides (PS) to polyhydroxyaromatics (PHA), throughout the constructed treatment wetland connected to a wastewater treatment plant, as measured using pyrolysis gas chromatography-mass spectrometry (Py-GC/MS). The changes in the fractions were also identified, with respect to molecular weight (MW) distributions of the effluent organic matters, as measured using high performance size exclusion chromatography equipped with both UV and fluorescence detectors, for aromatic/hydrophobic and protein-like organic substances, respectively; organic matter, with MWs of approximately 2,500 and 20,000 Da, and approximately 38,000 Da, as measured by the UV and fluorescence detectors, respectively, were newly formed after the wetlands, especially for the samples of the Typha wetland in June and August against in December. Thus, with the above two different analyses, the humification type of transformation of EfOM through the treatment wetland, was believed to occur, probably due to biological transformation (from the comparison of results in June and August with those in December). It was anticipated that the humification of EfOM could reduce biodegradable organic portions of wastewater effluents even though total organic carbon levels were not reduced that much after the treatment wetland.