Toxic effects of microplastics and nitrite exposure on intestinal histology, digestion, immunity, and microbial community of shrimp Litopenaeus vannamei.

Nitrite and microplastics (MPs) are environmental pollutants that threaten intestinal integrity and affect immune function of shrimp. In this study, the shrimp Litopenaeus vannamei were exposed to the individual and combined stress of nitrite and microplastics for 14 days, and the changes of intestinal histology and physiological functions were investigated. After single and combined stress, affectations occurred in intestinal tissue; the antioxidant enzyme activities (MDA, H2O2, CAT increased) and gene expression levels (CAT, SOD, GPx, HSP70 up-regulated) changed. The expression levels of detoxification genes (CYP450, UGT down-regulated, GST up-regulated), apoptosis genes (CASP-3 up-regulated) and endoplasmic reticulum stress genes (Bip, GRP94 down-regulated) changed. Furthermore, the stress also increased intestinal microbial diversity, causing bacterial composition variation, especially beneficial bacteria and pathogenic bacteria. These results suggested that nitrite and microplastics stress had adverse effects on the intestinal health of L. vannamei by affecting intestinal tissue morphology, immune response and microbial community.

Chronic Ethanol Exposure Enhances Facial Stimulation-Evoked Mossy Fiber-Granule Cell Synaptic Transmission via GluN2A Receptors in the Mouse Cerebellar Cortex.

Sensory information is transferred to the cerebellar cortex via the mossy fiber-granule cell (MF-GC) pathway, which participates in motor coordination and motor learning. We previously reported that chronic ethanol exposure from adolescence facilitated the sensory-evoked molecular layer interneuron-Purkinje cell synaptic transmission in adult mice in vivo. Herein, we investigated the effect of chronic ethanol exposure from adolescence on facial stimulation-evoked MF-GC synaptic transmission in the adult mouse cerebellar cortex using electrophysiological recording techniques and pharmacological methods. Chronic ethanol exposure from adolescence induced an enhancement of facial stimulation-evoked MF-GC synaptic transmission in the cerebellar cortex of adult mice. The application of an N-methyl-D-aspartate receptor (NMDAR) antagonist, D-APV (250 µM), induced stronger depression of facial stimulation-evoked MF-GC synaptic transmission in chronic ethanol-exposed mice compared with that in control mice. Chronic ethanol exposure-induced facilitation of facial stimulation evoked by MF-GC synaptic transmission was abolished by a selective GluN2A antagonist, PEAQX (10 µM), but was unaffected by the application of a selective GluN2B antagonist, TCN-237 (10 µM), or a type 1 metabotropic glutamate receptor blocker, JNJ16259685 (10 µM). These results indicate that chronic ethanol exposure from adolescence enhances facial stimulation-evoked MF-GC synaptic transmission via GluN2A, which suggests that chronic ethanol exposure from adolescence impairs the high-fidelity transmission capability of sensory information in the cerebellar cortex by enhancing the NMDAR-mediated components of MF-GC synaptic transmission in adult mice in vivo.

Unveiling sugarcane defense response to Mythimna separata herbivory by a combination of transcriptome and metabolic analyses.

BACKGROUND: Sugarcane is the most important sugar crop in the world. Like other crops, sugarcane suffers from herbivorous insect attack. The oriental armyworm Mythimna separata is a devastating pest of various crops in northeast Asia and an outbreak of this pest can result in substantial yield loss for sugarcane. However, the plant defense response situation is widely acquisition in model crops, but there is little information about how sugarcane plants defend themselves against this herbivore at the molecular and biochemical levels. RESULTS: We combined transcriptome and metabolomic analysis to investigate the changes in gene expression and metabolic processes that occurred in sugarcane plants after continuous feeding by M. separata larvae for 12 and 24 h. We identified 13 662 genes and 55 metabolites that were differentially regulated in sugarcane plants fed on by M. separata. The genes involved in phytohormones, transcription factors, and kinase-related were activated and metabolism compounds such as carbohydrate, amino acid, ferulic substances and glutathione were detected regulated in sugarcane defense response. Comparable analyses showed a close correspondence relationship among pathways of phenylalanine metabolism, phenylpropanoid biosynthesis, and flavonoid biosynthesis in transcript and metabolite profiles. Furthermore, a bioassay experiment was conducted to test the influence of up-regulated metabolites on M. separata growth and found chlorogenic acid had a lethal effect. CONCLUSION: The results of our study greatly enhanced the understanding of the sugarcane-induced defense response mechanism against herbivore infestation at the transcriptional and metabolic levels. Also make contributions to provide clues for development of green pest control method. © 2021 Society of Chemical Industry.

Correction to: The effect of teprenone on the intestinal morphology and microbial community of Chinese sea bass (Lateolabrax maculatus) under intermittent hypoxic stress.

In the end of the Discussion section, following information should be included.

The effect of teprenone on the intestinal morphology and microbial community of Chinese sea bass (Lateolabrax maculatus) under intermittent hypoxic stress.

Hypoxia stress may affect the fish intestine and thereby threaten the growth and survival of the fish. Teprenone is a clinically effective agent in protecting gastrointestinal mucosa. This study aims to assess the effect of teprenone in the intestine of Chinese sea bass Lateolabrax maculatus under intermittent hypoxic stress. L. maculatus juveniles were either raised under intermittent hypoxic condition or normal condition (NC). Part of the hypoxic-intervened fish were treated with teprenone at different concentrations (HTs), and the rest were regarded as hypoxic control (HC). Histological analysis was performed on the epithelial tissue of the fish intestine. High-throughput sequencing technology was used to analyze the diversity and composition of the microbial community in L. maculatus intestine. Reduced villi length and goblet cell, exfoliated enterocyte, and improper arrangement of villi were observed in HC compared with NC and HTs. Proteobacteria, Firmicutes, and Bacteroidetes represented the most abundant phyla in each sample. Significantly higher microbial diversity was detected in HC compared with NC (P < 0.05). At the phylum level, HC presented significantly decreased relative abundance of Proteobacteria, and significantly increased relative abundance of Bacteroidetes, Chloroflex, and Cyanobacteria compared with NC (P < 0.05). At the class level, HC showed significantly reduced relative abundance of Alphaproteobacteria and Bacilli, and significantly increased relative abundance of Clostridia, Gammaproteobacteria, and Bacteroides (P < 0.05). Teprenone protects the intestine from epithelial damages and maintains the microbial harmony in L. maculatus under intermittent hypoxic stress.

Molecular cloning, characterization, and expression profiling analysis of Cry toxin receptor genes from sugarcane shoot borer Chilo infuscatellus (Snellen).

The sugarcane shoot borer Chilo infuscatellus (Snellen) is known for causing severe damage to sugarcane yield in China. Methods have been developed to control this pest, including Cry toxin pesticide and transgenic Bt plants. In order to investigate the molecular mechanism of the Cry toxin binding process and provide a basis for understanding the insect's resistance mechanism, we used a high throughput sequencing platform to perform a de novo transcriptome assembly across different larval developmental stages and analyzed Cry toxin receptors based on our assembled transcripts. We cloned twelve Cry toxin receptor genes including 1 cadherin (Cad), 7 aminopeptidase-Ns (APNs), 3 alkaline phosphatases (ALPs), and 1 ATP-binding cassette transporter subfamily C2 (ABCC2), and three of them with full length. The sublethal dosage of Cry1Ac toxin was applied to sugarcane shoot borer and identified some Cry toxin receptor genes that were significantly induced after 48 h of exposure. Furthermore, quantitative RT-PCR was conducted to detect the expression profiles of these genes. Our transcriptome sequence data provided a valuable molecular resource for further study and the identified Cry toxin receptor data gave insights for improved research into the mechanism of Bt resistance.

Molecular cloning, characterization and functional analysis of GluCl from the oriental armyworm, Mythimna separata Walker.

Glutamate-gated chloride channels (GluCls) mediate inhibitory synaptic transmission in invertebrate nervous systems, and only one GluCl gene has been found in insects. Therefore, insect GluCls are one of the major targets of insecticides including avermectins. In the present study, a 1347 bp full-length cDNA encoding a 449-amino acid protein (named MsGluCl, GenBank ID: MK336885) was cloned from the oriental armyworm, Mythimna separata, and characterized two alternative splicing variants of MsGluCl. The protein shares 76.9-98.6% identity with other insect GluCl isoforms. Spatial and temporal expression analysis revealed that MsGluCl was highly expressed in the 3rd instar and adult head. Dietary ingestion of dsMsGluCl significantly reduced the mRNA level of MsGluCl and decreased abamectin mortality. Thus, our results reveal that MsGluCl could be the molecular target of abamectin and provide the basis for further understanding the resistance mechanism to abamectin in arthropods.

Assessment of the role of brine shrimp Artemia in white spot syndrome virus (WSSV) transmission.

Challenge tests with Artemia four different development stages (nauplii, metanauplii, pseudoadults and adults) to white spot syndrome virus was carried out by immersion challenge and virus-phytoplankton adhesion route in order to asses the possibility of Artemia acting as a vector of WSSV to penaeid shrimp Litopenaeus vannamei postlarvae. The WSSV succeeded in infecting four stages Artemia, and nested-PCR detection for WSSV revealed positive results to virus-phytoplankton adhesion route. No mass mortalities were observed in penaeid shrimp postlarvae fed with WSSV-positive Artemia which exposed to WSSV by virus-phytoplankton adhesion route, whereas WSSV DNA detected in penaeid shrimp postlarvae by nested-PCR. By contrary, no WSSV-positive was detected in any animal fed with WSSV-negative Artemia. These results indicated that Artemia could serve as a vector in WSSV transmission.

Bioassay evidence for the transmission of WSSV by the harpacticoid copepod Nitocra sp.

White Spot Syndrome Virus (WSSV) is now one of the most devastating and virulent viral agents threatening the penaeid shrimp culture industry and has been responsible for serious economic losses for shrimp farms worldwide. One remarkable characteristic of WSSV is its wide reservoir range, which contributes to its wide geographical distribution. Among epizootiological surveys, there is substantial evidence for WSSV-positive copepods found in shrimp farming ponds. Therefore, copepods are suspected to be the vector of WSSV. In the present study, nested-PCR analysis showed positive results in the harpacticoid copepod Nitocra sp. exposed to WSSV by virus-phytoplankton adhesion route. Oral route and intramuscular injection were used to test the pathogenicity of WSSV isolated from the WSSV-positive Nitocra sp. For the oral route of infection, Marsupenaeus japonicus postlarvae were fed with WSSV-positive copepods. The shrimp postlarvae in the infected treatment became WSSV-positive and occurred 52.50+/-5.00% mortality which was significant higher (P <0.05) than that in the control treatment (20.00+/-0.00%) when postlarvae were fed with WSSV free copepods. In the intramuscular injection challenge, M. japonicus juveniles were injected with the copepods inoculum extracted from the WSSV-positive Nitocra sp., and showed 72.50+/-9.57% mortality which was also significant higher (P <0.05) than that in the control treatment (22.50+/-5.00%) when juveniles were received mock injection of a tissue homogenate prepared from WSSV-negative Nitocra sp. Based on these laboratory challenge studies, it was confirmed that the copepods can serve as a vector in WSSV transmission.

White spot syndrome virus (WSSV) transmission from rotifer inoculum to crayfish.

To test the possibility that shrimp pond rotifer resting eggs and hatched rotifers could transmit white spot syndrome virus (WSSV) to crayfish (Procambarus clarkii), we injected crayfish with rotifer and resting egg inocula that were WSSV-positive only by dot-blot analysis of PCR products. No crayfish became WSSV-positive after challenge with the resting egg inoculum. However, 1/15 crayfish became WSSV-positive after challenge with the rotifer inoculum. The results demonstrated that rotifers constitute a potential risk for WSSV transmission to crayfish and other cultivated crustaceans. However, the actual quantitative risk of transmission in an aquaculture setting depends on many variables that remain untested.