Occurrence, distribution, and health risk assessment of pyrethroid and neonicotinoid insecticides in aquatic products of China.

Synthetic pyrethroid insecticides (SPIs) and neonicotinoid insecticides (NEOs), now dominant in the insecticide market, are increasingly found in aquatic environments. This study focused on six SPIs and five NEOs in aquatic products from four Chinese provinces (Shandong, Hubei, Shanxi and Zhejiang) and the risk assessment of the safety for the residents was conducted. It revealed significantly higher residues of Σ6SPIs (6.27-117.19 µg/kg) compared to Σ5NEOs (0.30-14.05 µg/kg), with SPIs more prevalent in fish and NEOs in shellfish. Carnivorous fish showed higher pesticide levels. Residues of these two types of pesticides were higher in carnivorous fish than in fish with other feeding habits. In the four regions investigated, the hazard quotient and hazard index of SPIs and NEOs were all <1, indicating no immediate health risk to human from single and compound contamination of the two types of pesticides in aquatic products. The present study provides valuable information for aquaculture management, pollution control and safeguarding human health.

Endocytosis-mediated entry of a caterpillar effector into plants is countered by Jasmonate.

Insects and pathogens release effectors into plant cells to weaken the host defense or immune response. While the imports of some bacterial and fungal effectors into plants have been previously characterized, the mechanisms of how caterpillar effectors enter plant cells remain a mystery. Using live cell imaging and real-time protein tracking, we show that HARP1, an effector from the oral secretions of cotton bollworm (Helicoverpa armigera), enters plant cells via protein-mediated endocytosis. The entry of HARP1 into a plant cell depends on its interaction with vesicle trafficking components including CTL1, PATL2, and TET8. The plant defense hormone jasmonate (JA) restricts HARP1 import by inhibiting endocytosis and HARP1 loading into endosomes. Combined with the previous report that HARP1 inhibits JA signaling output in host plants, it unveils that the effector and JA establish a defense and counter-defense loop reflecting the robust arms race between plants and insects.

Kinetics of uptake and depuration of synthetic pyrethroid insecticides in manila clam (Ruditapes philippinarum).

Synthetic pyrethroid insecticides (SPIs) are frequently detected in water bodies and sediments, and they show high toxicity to aquatic organisms, but their toxicity kinetics remain unknown. In this work, the kinetics of uptake and depuration of three SPIs, fenpropathrin (FP), cypermethrin (CM) and deltamethrin (DM) were evaluated in manila clams (Ruditapes philippinarum) for the first time through a bioconcentration-semi-static test. Clams were exposed to three SPIs of different concentrations (2 ng/mL and 20 ng/mL) for 4 days, followed by a 10-day depuration stage. The results indicated that adult manila clams could absorb SPIs rapidly, and the bioconcentration factor (BCF) values of SPIs were different at high and low concentrations of contaminants. The depuration rate constants (k2) of SPIs in adult manila clams ranged from 0.024 h-1 to 0.037 h-1. The bioaccumulation factors ranged from 319.41 to 574.38. And the half-lives (t1/2) were in the range of 18.49 to 29.22 h. These results showed that manila clams have a high bioconcentration capacity, and SPIs have a high cumulative risk for bivalves. Moreover, after 10 days of elimination, SPIs can still be detected in manila clams at all concentrations, indicating that the complete elimination of SPIs required a longer time.

Pyrethroids contamination and health risk assessment in seafood collected from the coast of Shandong, China.

This study provides the first data on pyrethroid residues of seafood in China. A total of 192 seafood samples were randomly selected from four coastal cities of Shandong Province in 2020. The residues of fenpropathrin, cypermethrin and deltamethrin in seafood by GC-MS were ND to 26.82 µg/kg, ND to 19.18 µg/kg and ND to 15.56 µg/kg, respectively. The cumulative risk to general population of different age groups was assessed by the hazard quotient (HQ) and hazard index (HI) approaches, and showed that the maximum value of both HQ (1.81 × 10-3) and HI (2.9 × 10-3) were below the threshold 1. The present results indicated that the three main pyrethroids from the consumption of seafood is unlikely to pose a health risk to general populations in Shandong. The uncertainty analysis indicated that the monitoring study of pyrethroid residues in seafood is worthy of continuous attention to ensure food safety.

A highly accumulated secretory protein from cotton bollworm interacts with basic helix-loop-helix transcription factors to dampen plant defense.

Caterpillar oral secretion (OS) contains active molecules that modulate plant defense signaling. We isolated an effector-like protein (Highly Accumulated Secretory Protein 1, HAS1) from cotton bollworm (Helicoverpa armigera) that is the most highly accumulated secretory protein of the nondigestive components in OS and belongs to venom R-like protein. Elimination of HAS1 by plant-mediated RNA interference reduced the suppression of OS on the defense response in plants. Plants expressing HAS1 are more susceptible to insect herbivory accompanied by the reduced expressions of multiple defense genes. HAS1 binds to the basic helix-loop-helix (bHLH) transcription factors, including GoPGF involved in pigmented gland formation and defense compounds biosynthesis in cotton and MYC3/MYC4 the main regulators in jasmonate (JA) signaling in Arabidopsis. The binding activity is required for HAS1 to inhibit the activation of bHLHs on plant defense gene expressions. Together with our previous study that another venom R-like protein HARP1 in cotton bollworm OS blocks JA signaling by interacting with JASMONATE-ZIM-domain repressors, we conclude that the venom R-like proteins in OS interfere with plant defense in a dual suppression manner. Considering the venom proteins in parasitic wasp assault the immune system of its host animal, our investigation reveals their conserved function in carnivorous and herbivorous insects.

Use of GAL4 factor-based yeast assay to quantify the effects of xenobiotics on RXR homodimer and RXR/PPAR heterodimer in scallop Chlamys farreri.

Retinoid X receptor (RXR) and peroxisome proliferators-activated receptors (PPAR) have been shown as important targets of endocrine disrupting effects caused by organotin compounds (OTCs). In vitro methods for non-model species are instrumental in revealing not only mechanism of toxicity but also basic biology. In the present study, we constructed the GAL4 factor-based recombinant yeast systems of RXRα/RXRα (RR), RXRα/PPARα (RPα) and RXRα/PPARγ (RPγ) of the scallop Chlamys farreri to investigate their transcriptional activity under the induction of OTCs (tributyltin chloride, triphenyltin chloride, tripropyltin chloride and bis(tributyltin)oxide), their spiked sediments and five other non­tin compounds (Wy14643, rosiglitazone, benzyl butyl phthalate, dicyclohexyl phthalate and bis(2-ethylhexyl) phthalate). The results showed that the natural ligand of RXR, 9-cis-retinoic acid (9cRA), induces transcriptional activity in all three systems, while four OTCs induced the transcriptional activity of the RR and RPα systems. None of the five potential non­tin endocrine disruptors induced effects on the RPα and RPγ systems. The spiked sediment experiment demonstrated the feasibility of the recombinant yeast systems constructed in this study for environmental sample detection. These results suggest that OTCs pose a threat to affect function of RXRα and PPARα of bivalve mollusks. The newly developed GAL4 factor-based yeast two-hybrid system can be used as a valuable tool for identification and quantification of compounds active in disturbing RXR and PPAR of bivalves.

Impact of P-Chloroaniline on Oxidative Stress and Biomacromolecules Damage in the Clam Ruditapes philippinarums: A Simulate Toxicity Test of Spill Incident.

As a hazardous chemical, p-chloroaniline (PCA) shows intensive adsorption and accumulation after entering the aquatic ecosystem, which can be enriched in organisms and cause damage. With the objective of achieving an integrated and mechanistic view of the toxic effects of PCA in the marine sentinel organism Ruditapes philippinarum, Manila clams were exposed to different concentration of PCA (0.5, 2 and 5 mg/L) for 15 days. Focusing on the gills, first targeting the toxic and digestive gland, the metabolic detoxification organ, we detected dose- and time-related changes inantioxidase activities and biomacromolecular damages in treated clams. Glutathione S-transferase (GST) activity and glutathione (GSH) contents were significantly induced, and superoxide dismutase (SOD) activity increased at the beginning of exposure and then decreased. The malondialdehyde (MDA) and protein methylation (PC) contents which represent lipid peroxidation and carbonylation of proteins, increased first with exposure time and then decreased in the digestive gland. DNA strand break levels were consistently higher than those in the control group. The digestive gland showed more sensitivity to the stress of PCA than the gills. GST and MDA in the gill and GST, GSH, SOD, DNA strand break level in the digestive gland showed significant correlation with PCA exposure, which indicated that these parameters can be used as sensitive biomarkers to indicate toxic effects from chloraniline leakage.

Effects of crustacean hyperglycaemic hormone RNA interference on regulation of glucose metabolism in Litopenaeus vannamei after ammonia-nitrogen exposure.

To unveil the adaptation of Litopenaeus vannamei to elevated ambient ammonia-N, crustacean hyperglycaemic hormone (CHH) was knocked down to investigate its function in glucose metabolism pathway under ammonia-N exposure. When CHH was silenced, haemolymph glucose increased significantly during 3-6 h, decreased significantly during 12-48 h and recovered to the control groups' level at 72 h. After CHH knock-down, dopamine (DA) contents reduced significantly during 3-24 h, which recovered after 48 h. Besides, the expressions of guanylyl cyclase (GC) and DA1R in the hepatopancreas decreased significantly, while DA4R increased significantly. Correspondingly, the contents of cyclic AMP (cAMP), cyclic GMP (cGMP) and diacylglycerol (DAG) and the expressions of protein kinase A (PKA), protein kinase G (PKG), AMP active protein kinase α (AMPKα) and AMPKγ were significantly down-regulated, while the levels of protein kinase C (PKC) and AMPKß were significantly up-regulated. The expressions of cyclic AMP response element-binding protein (CREB) and GLUT2 decreased significantly, while GLUT1 increased significantly. Moreover, glycogen content, glycogen synthase and glycogen phosphorylase activities in hepatopancreas and muscle were significantly increased. Furthermore, the levels of key enzymes hexokinase, pyruvate kinase and phosphofructokinase in glycolysis (GLY), rate-limiting enzymes citrate synthase in tricarboxylic acid and critical enzymes phosphoenolpyruvate carboxykinase, fructose diphosphate and glucose-6-phosphatase in gluconeogenesis (GNG) were significantly decreased in hepatopancreas. These results suggest that CHH affects DA and then they affect their receptors to transmit glucose metabolism signals into the hepatopancreas of L. vannamei under ammonia-N stress. CHH acts on the cGMP-PKG-AMPKα-CREB pathway through GC, and CHH affects DA to influence cAMP-PKA-AMPKγ-CREB and DAG-PKC-AMPKß-CREB pathways, thereby regulating GLUT, inhibiting glycogen metabolism and promoting GLY and GNG. This study contributes to further understand glucose metabolism mechanism of crustacean in response to environmental stress.

Induction of time- and dose-dependent oxidative stress of triazophos to brain and liver in zebrafish (Danio rerio).

Extensive use of triazophos for the chemical control of pests in agriculture or aquaculture might strongly disturb the aquatic environment due to residue accumulation through various routes like surface run-off, spray-drift and effluent from factories, which have potential negative effects to non-target aquatic organisms. Previous studies have documented the antioxidative effects of triazophos to mammals, however, the oxidative toxicity of triazophos to fish has not been adequately studied to date. Thus, an acute exposure (96 h) to triazophos at different concentrations of 0.06, 0.3 and 1.5 mg/L (corresponding to 1/50th, 1/10th and 1/2th of 96 h-LC50, respectively), was conducted to investigate the triazophos-induced oxidative stress in adult zebrafish (Danio rerio). The results showed that the time- and dose-dependent induction of oxidative stress except for the ROS level in liver after 24 and 48 h exposure, as indicated by increased reactive oxygen species (ROS), malondialdehyde (MDA) level and a compromised antioxidant defense system, including increased superoxide dismutase (SOD) activity, catalase (CAT) activity, glutathione (GSH) content as well as the increased at first and decreased afterwards genes expression (Sod1, Sod2, Cat and Gpx) in brain. Simultaneously, ROS and MDA showed an increased trend, SOD activity, CAT activity and GSH content showed a trend of increasing at 24 h and decreasing at 48 h, and then increasing at 96 h and Sod1, Sod2, Cat and Gpx gene showed decreasing at first and then increasing in liver tissue. The present study concluded that the damage of the antioxidant system by triazophos induced oxidative stress in the brain and liver of zebrafish with concomitant lipid peroxidation, which is an important mechanism underlying the triazophos-induced acute toxicity.

Effect of acute exposure of triazophos on histological structure and apoptosis of the brain and liver of zebrafish (Danio rerio).

Triazophos (TAP) has become a part of widespread pollutant of the aquatic environment due to its residue. Current study was designed to investigate the toxic effect of TAP at different doses (0.06, 0.3 and 1.5 mg/L) to the model organism of zebrafish (Danio rerio) by using multi-endpoint analysis in a 96 h acute exposure test. The direct observation that histological and ultrastructural alteration of zebrafish brain and liver were carried out via paraffin section in hematoxylin and eosin (H&E) staining and transmission electron microscopy (TEM), respectively. In addition, a series of methods were applied for exploring the physiological parameters related to cellular apoptosis. Results indicated that vacuolar structure after 96 h treatment with TAP were appeared in the molecular and granular layers of cerebellum. A large number of nuclear retraction, tissues vacuolation and cytoplasmic loss were observed in liver at histological level. From the fine structural level, the mitochondrial vacuolation and membrane damage of brain cells were found and the cristae of mitochondria disintegrated partly in hepatocytes. Onset of such histological structure alterations were one of the most intuitive reflection to TAP exposure, which needs to analyze biochemical alterations for further study. The mitochondrial membrane potential (MMP) showed a downward trend in the brain and liver of zebrafish. Simultaneously, the activity of caspase-3 and caspase-9 increased after 96 h exposure with a concentration-dependent manner, which could be served as a suitable indicator of cellular apoptosis. Furthermore, apoptosis-related genes (Apaf-1, p53, Bax, Bcl-2, caspase-3 and caspase-9) transcription showed different alterations in response to the TAP treatment. These results indicated that TAP exposure led to apoptosis in zebrafish brain and liver and it was speculated that the apoptosis may occur through mitochondrial pathway. The present study demonstrated that the exposure of zebrafish to the insecticide TAP led to observe its effects at both histological structure and apoptosis level in liver and brain.