Four new earthworm species of the genera Amynthas and Metaphire (Oligochaeta, Megascolecidae) from Hunan and Anhui provinces, China.

This paper describes four new species earthworms from Hunan and Anhui provinces, China, Amynthasxiangtanensis Qiu & Jin, sp. nov., Amynthastaoyuanensis Qiu & Jin, sp. nov., Amynthasxuanchengensis Jin & Li, sp. nov. and Metaphiredonganensis Jin & Jiang, sp. nov. Amynthasxiangtanensis sp. nov., and A.taoyuanensis sp. nov. belong to the Amynthascorticis group. Both have four pairs of intersegmental spermathecal pores in 5/6-8/9; male pores in segment XVIII, separated by 1/3 of body circumference, each on top of a slightly raised porophore, surrounded by several tiny genital papillae. Amynthastaoyuanensis sp. nov. prostate glands are degenerated. Amynthasxuanchengensis sp. nov. belongs to the Amynthasmorrisi group, it has two pairs of spermathecal pores in 5/6 and 6/7; male pores in XVIII, separated by 1/3 of body circumference, each on top of a slightly raised, circular porophore. Metaphiredonganensis sp. nov. belongs to the Metaphirehoulleti group. It has three pairs of spermathecal pores in 6/7-8/9; male pores in XVIII, separated by 1/3 of body circumference, each on the bottom center of the longitudinal copulatory chamber.

Utilizing Liquid-liquid phase separation-related lncRNAs to predict the prognosis and treatment response of PCa.

BACKGROUND: Studies have indicated a close association between genes linked to liquid-liquid phase separation (LLPS) and the progression of prostate cancer (PCa). However, the interplay among long non-coding RNAs (lncRNAs) linked to LLPS in PCa remains elusive. Therefore, we constructed a prediction model based on LLPS-related LncRNA in PCa to explore its relationship with the prognosis and drug treatment of PCa. METHODS: We obtained clinical and sequencing data from TCGA and LLPS genes from the Phase Separation Protein Database. By analyzing the differential expression of LLPS-related genes and lncRNAs in prostate cancer, and using Poisson correlation, we identified LLPS-related lncRNAs. Prognostic LLPS-lncRNAs were found through prognostic correlation analysis and included in a Cox model to compute regression coefficients. Patients were scored and divided into high- and low-risk groups. Independent prognostic factors were integrated into a prognostic nomogram with risk and Gleason scores. We also conducted drug sensitivity analyses, GSEA, and validated the impact of key lncRNAs through functional experiments. RESULTS: Our study identified five LLPS-associated lncRNAs that are of prognostic importance. And found notable disparities in biochemical recurrence rates and survival outcomes between these risk groups, with the low-risk cohort exhibiting superior prognostic indicators. Moreover, our prediction nomogram demonstrated robust predictive accuracy and significant clinical utility. Furthermore, our model exhibited promising capabilities in forecasting patient sensitivity to various conventional therapeutic drugs, thereby highlighting its potential in personalized treatment strategies. GSEA showed that these lncRNAs may influence PCa prognosis and sensitivity to therapeutic agents by affecting pathways such as cell cycle. Knockdown of AC009812.4 could inhibit the ability of PCa cells to proliferate, migrate and invade, and compare to paracancerous tissue, AC009812.4 in PCa tissue has significantly higher expression. CONCLUSION: Our research uncovers the prognostic significance of lncRNAs associated with LLPS in PCa and established a model exhibiting excellent predictive accuracy for prognosis. Those lncRNAs may influence progress of PCa as well as sensitivity to therapy drugs through pathways such as cell cycle.

Mitochondrial DNA evidence reflects high genetic divergence of Amynthas aspergillum (Oligochaeta: Megascolecidae) in southern China.

Amynthas aspergillum (Perrier, 1872), a natural resource used in traditional Chinese medicine (Guang-dilong) with high economic value, is widely distributed in forests and farmland habitats in the hilly areas of southern China. To investigate the extent of genetic differentiation and diversity in A. aspergillum, a population genetic structure study was performed on 157 samples from 75 locations in southern China using the mitochondrial genes COI, COII, 12S rRNA, 16S rRNA, and NDI. The results indicated that A. aspergillum had a high level of genetic diversity, and variation within populations was the main source of the total variation. Six deeply divergent mitochondrial clades (I-VI) were detected using both phylogenetic tree and haplotype network analyses. This finding was supported by the high Kimura two-parameter genetic distance and the pairwise fixation index value obtained based on the COI gene. No significant phylogeographic structures were observed. The widespread geographic distribution of clades II, IV, and VI suggested a recent demographic expansion based on multiple analysis results. These results include a high level of Hd and low π, star-shaped haplotype network structures with a high number of less frequent haplotypes, significantly negative neutrality test values, and a unimodal mismatch distribution pattern. The divergence time estimates and reconstruction of the ancestral area revealed that A. aspergillum originated in Guangxi Province and underwent initial intraspecific diversification in the early Pliocene to generate clade I. Then, it gradually dispersed eastward and rapidly differentiated into clades II-V during the Pleistocene. The Yunnan-Guizhou Plateau and Nanling and Wuyi Mountains might act as geographical barriers for the spread of A. aspergillum to the west and north.

Doxycycline decelerates aging in progeria mice.

Beyond the antimicrobial activity, doxycycline (DOX) exhibits longevity-promoting effect in nematodes, while its effect on mammals is unclear. Here, we applied a mouse model of Hutchinson-Gilford progeria syndrome (HGPS), Zmpste24 knockout (KO) mice, and analyzed the antiaging effect of DOX. We found that the DOX treatment prolongs lifespan and ameliorates progeroid features of Zmpste24 KO mice, including the decline of body and tissue weight, exercise capacity and cortical bone density, and the shortened colon length. DOX treatment alleviates the abnormal nuclear envelope in multiple tissues, and attenuates cellular senescence and cell death of Zmpste24 KO and HGPS fibroblasts. DOX downregulates the level of proinflammatory IL6 in both serum and tissues. Moreover, the elevated α-tubulin (K40) acetylation mediated by NAT10 in progeria, is rescued by DOX treatment in the aorta tissues in Zmpste24 KO mice and fibroblasts. Collectively, our study uncovers that DOX can decelerate aging in progeria mice via counteracting IL6 expression and NAT10-mediated acetylation of α-tubulin.

Three new species of the genus Amynthas (Oligochaeta: Megascolecidae) from Guangxi Zhuang Autonomous Region, China.

Three new earthworm species of the genus Amynthas Kinberg, 1867 were found in Guangxi Zhuang Autonomous Region, China: Amynthas huapingensis sp. nov., Amynthas retentus sp. nov. and Amynthas linguiensis sp. nov. Amynthas huapingensis sp. nov. belongs to Amynthas corticis-group characterized by having four pairs of spermathecal pores in 5/68/9; Amynthas retentus sp. nov., without spermathecal pores, belongs to Amynthas illotus-group; Amynthas huapingensis sp. nov. and Amynthas retentus sp. nov. have similar characteristics of male porophores medially surrounded by 23 genital papillae in a pulvinate pad within the male pore area. Amynthas linguiensis sp. nov. has variable spermathecal pores, three pairs in 6/78/9 or four pairs in 5/68/9. And its male pores each on the center of a large elliptical glandular porophore which extends from 1/4xvii to 1/4xix. The morphological comparison and COI gene data of these three new species and their similar species are given in this paper.

Dynamics of bacterial community in the foregut and hindgut of earthworms with the nutrition supplied by kitchen waste during vermicomposting.

Earthworm gut microbiota is vital in degrading bio-waste during vermicomposting. However, microbial dynamics in earthworm gut during this process are unclear. Thus, the aim is to firstly report the bacterial dynamics in both foregut and hindgut of earthworms over a 28 days' timeframe of vermicomposting by Eisenia foetida with the nutrition supplied by kitchen waste. Results showed that except the changing of the bacterial diversity, composition and structure, dynamics of the foregut and hindgut bacteria also differed during vermicomposting which related to the changes of nutrient provision. Day 3 was a turning point. The abundant bacteria of the top 20 % genera nearly did not overlap between the foregut and hindgut. In the end of vermicomposting, a remarkable stable bacterial structure appeared in the hindgut compared to somewhat muddled one in the foregut. Understanding the dynamics of earthworm gut microbiota enables the improvements to regulate the efficiency of organic waste vermicomposting.

Discovery and Mechanism of Action of a Novel Antimicrobial Peptide from an Earthworm.

The robust innate immune system of the earthworm provides a potential source of natural antimicrobial peptides (AMPs). However, the cost and high rediscovery rate of direct separation and purification limits their discovery. Genome sequencing of numerous earthworm species facilitates the discovery of new antimicrobial peptides. Through predicting potential antimicrobial peptides in the open reading frames of the Eisenia andrei genome and sequence optimization, a novel antimicrobial peptide, named EWAMP-R (RIWWSGGWRRWRW), was identified. EWAMP-R demonstrated good activity against various bacteria, including drug-resistant strains. The antibacterial mechanisms of EWAMP-R were explored through molecular simulation and wet-laboratory experiments. These experiments demonstrated that the bacterial membrane may be one of the targets of EWAMP-R but that there may be different interactions with Gram-negative and Gram-positive bacterial membranes. EWAMP-R can disrupt bacterial membrane integrity; however, at low concentrations, it appears that EWAMP-R may get through the membrane of Escherichia coli instead of damaging it directly, implying the existence of a secondary response. Gene expression studies identified that in E. coli, only the apoptosis-like cell death (ALD) pathway was activated, while in Staphylococcus aureus, the MazEF pathway was also upregulated, limiting the influence of the ALD pathway. The different antimicrobial actions against Gram-positive and -negative bacteria can provide important information on the structure-activity relationship of AMPs and facilitate AMP design with higher specificity. This study identified a new source of antibacterial agents that has the potential to address the increasingly serious issue of antibiotic resistance. IMPORTANCE Drug-resistant bacteria are a great threat to public health and drive the search for new antibacterial agents. The living environment of earthworms necessitates a strong immune system, and therefore, they are potentially a rich resource of novel antibiotics. A novel AMP, EWAMP-R, with high antibacterial activity was found through in silico analysis of the Eisenia andrei genome. Molecular analysis investigating the interactions between EWAMP-R and the cell membrane demonstrated the importance of tryptophan and arginine residues to EWAMP-R activity. Additionally, the different secondary responses found between E. coli and S. aureus were in accordance with a common phenomenon where some antibacterial agents only target specific species of bacteria. These results provided useful molecular information to support further AMP research and design. Our study expands the sources of antimicrobial peptides and also helps to explain the adaptability of earthworms to their environment.

Physical and hydraulic properties of bioretention substrate using hexadecyl trimethyl ammonium bromide (HDTMA) modified zeolite.

This study using hexadecyl trimethyl ammonium bromide (HDTMA) modified zeolite as a component of bioretention substrate, to investigate the effect of HDTMA modification on the basic physical and hydraulic properties of substrate layer. Two different levels of HDTMA modified zeolite (ZHD10 and ZHD50) were mixed with a mixture consists of peat soil, river sand and compost (fixed volumetric proportion at 5:4:1) with varying volumetric percentage (25%, 50%, and 75%) to form substrate media. The modification only changes the physical properties of zeolite and media with zeolite slightly, while significant changes in surface hydrophobicity and hydraulic properties were observed. A distinct decline of saturated hydraulic conductivity (Ks) values of zeolite can be observed after the modification, Ks values drop 36.5% for ZHD10 and 55.1% for ZHD50. In contrast, Ks values of substrate media using zeolite increase after the modification at the same volumetric ratio of zeolite. When 50% of zeolite (v/v%) was used in substrate, Ks for natural zeolite, ZHD10 and ZHD50 was 0.024, 0.038 and 0.075 cm/s, respectively. Such alterations in Ks are associated with the changes of surface hydrophobicity after the modification and ion exchange between modified zeolite and other materials after soaking into water. Changes in water retention characteristics (WRC) curves were in good accord with the variations in Ks, and can be interpreted by the changed Ks of tested materials. The orientations of HDTMA molecules loaded on zeolite surface were suggested to play crucial roles in altering the hydraulic properties of zeolite added substrate.

Eco-risk management of tylosin fermentation residues using vermicomposting.

Tylosin fermentation residues (TFR) pose an ecotoxicological risk through antibiotic resistant bacteria (ARBs) and their corresponding genes (ARGs). This study evaluated the ecotoxicity of TFR to soil biological activity, and further explored the mechanisms of vermicomposting to reduce the toxicological risk. The results showed that tylosin (TYL) was moderately degradable with a half-life (t1/2) of 37.5 d, inducing 28-44% inhibition rate of nitrogen transformation in soil, and the EC50 of earthworm avoidance was 880 mg/kg. The 30-d vermicomposting reduced the pH and OM content, while increased the EC and TN content, accelerated compost maturation (C/N ratio up to 20), and enriched the microbial community. ARGs were reduced by earthworm through removal of TYL (>70% degradation, t1/2 of <20 d), inhibiting abundance of intI1 and ARBs. We conclude that vermicomposting is an efficient method for TFR treatment and its eco-risk management.

Mitochondrial bioenergetic, oxidative stress and burrowing responses in earthworm exposed to roxarsone in soil.

The eco-risk of roxarsone (ROX) was evaluated using multiple responses of earthworm biomarkers under different ROX concentrations for 28 d. With the increasing total arsenic accumulation (TAs-E), biological responses in earthworm generally showed a two-stage changes of homeostasis dysregulation and dose-dependent alterations. At the early periods, ROX stress increased the reactive oxygen species (ROS) and lipid peroxidation (LPO) in a similar manner, and apparently disrupted mitochondrial calcium ([Ca2+]m). But earthworms regulated their mitochondrial and redox homeostasis through stable mitochondrial membrane potential (MMP) and increase of ATP level, superoxide dismutase (SOD) and catalase (CAT). After 14 d, the positively correlated mitochondrial effects of ROS, [Ca2+]m, MMP and ATP were related to the behavioral inhibition of burrow length, depth and reuse rate as well as antioxidant up-regulation of Nrf2, HO-1, sod1 and cat. These results contributed possible biomarkers from the dose-dependent relationship between mitochondrial, antioxidant and behavioral responses. Multiple biological detection in earthworms can better reflect the sub-chronic ecotoxicity of phenylarsenic pollutants in soil.

Toxicity assessment of earthworm exposed to arsenate using oxidative stress and burrowing behavior responses and an integrated biomarker index.

Arsenate (As-V) is a ubiquitous contaminant in soil as a result of excessive use of veterinary drugs and pesticides, causing enormous environmental risks. Multiple biomarkers have been used to assess the ecotoxicity of arsenic, however, the mechanisms of toxicity remain unclear. This paper describes the exposure of the earthworm (Eisenia fetida) to natural soil with different As-V concentrations for 28 days, then biomarkers from oxidative stress and burrowing behavior were quantified to evaluate As-V stress. Dynamic changes in reactive oxygen species (ROS), lipid peroxidation (MDA), adenosine triphosphate (ATP) content and antioxidant enzymes activity (Gpx, SOD, CAT) implied two stages of intensified stress responses and physiological adaptability. The transcriptional expression and regulation of antioxidant enzymes showed different responses. The mRNA expression of sod1 was up-regulated, while that of cat showed no significant change. The related regulators, nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1), showed dose-dependent activation, suggesting antioxidant defense induced by Nrf2 signaling. The burrowing behavior after 14-day exposure indicated that As-V inhibited burrowing activity, especially the burrow length and maximum burrow depth. These multiple biomarkers were integrated using a biomarker response index (BRI) model, which showed significant dose-effect relationship especially on day 28, and suggested that ATP was a sensitive and representative biomarker. This study provided evidence that burrowing activity, Nrf2 and HO-1 were useful biomarkers warranting inclusion into the BRI model. Arsenic toxicity was comprehensively understood through redox homeostasis regulation, biochemical and behavioral changes, and these results suggested new strategies for soil pollutants diagnosis.

Three new widely distributed and polymorphic species of Amynthas earthworms (Oligochaeta, Clitellata, Megascolecidae) from South China.

Three new earthworm species are described from South China, namely Amynthas rusticanus sp. nov., Amynthas scaberulus sp. nov., and Amynthas yuanjiangensis sp. nov. They are widely distributed in South China, and be polymorphic in reproductive organs. Of three new species, A. rusticanus sp. nov. is a small species with three pairs of spermathecal pores in 5/6-7/8 or two pairs of spermathecal pores in 5/6-6/7; it has developed prostate glands in XVI-XX. A. scaberulus sp. nov. is a medium-sized species with two pairs of spermathecal pores in 7/8-8/9; the lower edge of each spermathecal pore forms a tiny papilla, and the prostate glands are either degenerated or developed. A. yuanjiangensis sp. nov. is a medium-sized species with four pairs of spermathecal pores in 5/6-8/9 and has degenerated or developed prostate glands. Barcode data for each species (partial sequence of mitochondrial cytochrome c oxidase subunit I) are provided as well.

Population Genetic Structure Reveals Two Lineages of Amynthas triastriatus (Oligochaeta: Megascolecidae) in China, with Notes on a New Subspecies of Amynthas triastriatus.

Amynthas triastriatus (Oligochaete: Megascolecidae) is a widely distributed endemic species in Southern China. To shed light on the population genetic diversity and to elucidate the population differentiation and dispersal of A.triastriatus, a population genetic structure study was undertaken based on samples from 35 locations collected from 2010 to 2016. Two exclusive lineages within A. triastriatus-lineage A and lineage B-were revealed. Lineage A was mainly distributed at high altitudes while lineage B was mainly distributed at low altitudes in Southeast China. The genetic diversity indices indicated that the populations of A. triastriatus had a strong genetic structure and distinct dispersal histories underlying the haplogroups observed in this study. Combined with morphological differences, these results indicated a new cryptic subspecies of A. triastriatus. Lineage A was almost degenerated to parthenogenesis and lineage B had a trend to parthenogenesis, which suggested that parthenogenesis could be an internal factor that influenced the differentiation and dispersal of A. triastriatus. The divergence time estimates showed that A.triastriatus originated around Guangxi and Guangdong provinces and generated into two main lineages 2.97 Ma (95%: 2.17-3.15 Ma) at the time of Quaternary glaciation (2.58 Ma), which suggested that the Quaternary glaciation may have been one of main factors that promoted the colonization of A.triastriatus.

Comparative study of vermicomposting of garden waste and cow dung using Eisenia fetida.

Vermicomposting is the process of composting using worms and is applied in waste management to produce high-quality organic fertilizer. Garden waste (GW) is often mixed with other raw materials for vermicomposting. In the present study, the feasibility of vermicomposting using only GW was investigated in comparison with cow dung (CD). The total nitrogen (TN), total phosphorus (TP), and total potassium (TK) contents and the electrical conductivity increased, while total organic carbon (TOC) and the C/N ratio decreased in both substrates after vermicomposting. The nutrient content (TN, TP, and TK) of the GW vermicompost was promoted less than that in CD. Scanning electron microscopy images and specific surface area analysis showed that the vermicompost was strongly disaggregated and became more compacted and fragmented compared with the raw substrates. No mortality of earthworms was observed in GW; however, the earthworms had a higher mean body weight and reproduction rate in CD than that in GW. There were higher bacterial community richness and diversity in the vermicompost than that in the raw materials, and the dominant phylum species were Proteobacteria, Actinobacteria, and Bacteroidetes. Redundancy analysis demonstrated that TN, C/N ratio, and TOC play an important role in bacterial community dynamics. These data indicate that vermicomposting is a robust process that is suitable for the management of GW.

Soil Behaviour of the Veterinary Drugs Lincomycin, Monensin, and Roxarsone and Their Toxicity on Environmental Organisms.

Lincomycin, monensin, and roxarsone are commonly used veterinary drugs. This study investigated their behaviours in different soils and their toxic effects on environmental organisms. Sorption and mobility analyses were performed to detect the migration capacity of drugs in soils. Toxic effects were evaluated by inhibition or acute toxicity tests on six organism species: algae, plants, daphnia, fish, earthworms and quails. The log Kd values (Freundlich model) of drugs were: lincomycin in laterite soil was 1.82; monensin in laterite soil was 2.76; and roxarsone in black soil was 1.29. The Rf value of lincomycin, roxarsone, monensin were 0.4995, 0.4493 and 0.8348 in laterite soil, and 0.5258, 0.5835 and 0.8033 in black soil, respectively. The EC50 for Scenedesmus obliquus, Arabidopsis thaliana, Daphnia magna and LC50/LD50 for Eisenia fetida, Danio rerio, and Coturnix coturnix were: 13.15 mg/L,32.18 mg/kg dry soil,292.6 mg/L,452.7 mg/L,5.74 g/kg dry soil and 103.9 mg/kg (roxarsone); 1.085 mg/L, 25 mg/kg dry soil, 21.1 mg/L, 4.76 mg/L, 0.346 g/kg dry soil and 672.8 mg/kg (monensin); 0.813 mg/L, 35.40 mg/kg dry soil, >400 mg/L, >2800 mg/L, >15 g/kg dry soil, >2000 mg/kg (lincomycin). These results showed that the environmental effects of veterinary drug residues should not be neglected, due to their mobility in environmental media and potential toxic effects on environmental organisms.

Effects of Salinity on Earthworms and the Product During Vermicomposting of Kitchen Wastes.

Population growth and social changes have recently contributed to an exaggerated increase in kitchen wastes in China. Vermicomposting has recently been recognized as an effective and eco-friendly method of organic waste treatment through the combination of earthworms and microbes. However, the influence of salt in kitchen wastes on vermicomposting have been unknown. The goal of this study was to analyze the influence of different salinities on earthworms (Eisenia fetida) and the products during the vermicomposting of kitchen wastes. In our research, kitchen wastes were divided into four different salinities: 0% (A), 0.1% (B), 0.2% (C) and 0.3% (D). The chemical characters of substrates and earthworm growth were measured on the 14th day and the 28th day of composting. Our results show that the high salinity (measured >0.2%) prevented earthworms from properly growing and had negative effects on quality of products in composting. T2 (measured salinity = 0.2%) had the highest average body weight, nitrate nitrogen, and available phosphorus. Thus, the salinity of kitchen wastes should be pretreated to less than 0.2% before vermicomposting.

Three new species and one subspecies of the Amynthas corticis-group from Guangxi Zhuang Autonomous Region, China (Oligochaeta, Megascolecidae).

Three new species and one subspecies of the genus Amynthas are described from Guangxi Zhuang Autonomous Region, China: The new species are: Amynthas maximus Qiu & Dong, sp. nov. and Amynthas tortuosus Qiu & Dong, sp. nov., and Amynthas shengtangmontis Dong & Jiang, sp. nov., the subspecies is Amynthas shengtangmontis minusculus subsp. nov. All have four pairs of spermathecal pores in 5/6-8/9, which indicates that they should belong to the corticis-group. Their morphological characteristics are compared to other similar species in the corticis-group from China and other Asian countries, such as Amynthas pulvinus Sun & Jiang, 2013, Amynthas homosetus (Chen, 1938), Amynthas corticis (Kinberg, 1867), Amynthas dorsualis Sun & Qiu, 2013, and Amynthas carnosus (Goto & Hatai, 1899). In addition, the results presented are confirmed by the pairwise comparison of COI barcode sequences. The pairwise distances between each new species and the other eighteen corticis-group species are greater than 14.7% on average. Furthermore, the pairwise distance between A. shengtangmontis shengtangmontis and A. shengtangmontis minusculus is 10.7-11.4%.

Three new species of earthworms (Oligochaeta: Megascolecidae) from Yunnan Province, China.

Earthworm specimens collected from Yunnan Province, China were found to represent three new species of Megascolecidae: Amynthas demptus sp. nov., Amynthas lacustris sp. nov. and Metaphire reclusa sp. nov. A. demptus belongs to Amynthas hawayanus-group and has three pairs of spermathecal pores in 5/6-7/8. A. lacustris belongs to Amynthas morrisi-group and has two pairs of spermathecal pores in 5/6-6/7. M. reclusa has no spermathecal pores. A. demptus has no male pores. The male pores of A. lacustris are about 1/3 body circumference ventrally apart, with a presetal and a postsetal papilla medial to each pore, surrounded with two circular folds. The male pores of M. reclusa are in copulatory chambers at XVIII, with openings of copulatory chambers about 1/3 body circumference ventrally apart. In this paper, the morphological characters of the three new species are described. We also sequenced COI and 12S gene fragments of the holotypes and selected paratype specimens to provide molecular data.

Hypoxic environment protects cowpea bruchid (Callosobruchus maculatus) from electron beam irradiation damage.

BACKGROUND: Electron beam (eBeam) irradiation and hermetic storage are safe and effective technologies to protect stored products. Although hypoxic environment improves performance of some irradiated insects, whether hypoxia affects irradiation of storage insects and impacts pest control efficacy remains to be investigated. RESULTS: Using cowpea bruchid (Callosobruchus maculatus) larvae, we showed that, relative to eBeam irradiation under normoxia, the adult emergence rate increased substantially if they were exposed to hypoxia prior to and during eBeam treatment. Conversely, exposure to hypoxia only after eBeam irradiation did not have this protective effect. eBeam irradiation caused an increase in reactive oxygen species (ROS) in normoxic larvae but not in hypoxic larvae. The activity of citrate synthase, a pace-making enzyme in the citric acid cycle, was suppressed under hypoxia but resumed normal function within hours of reoxygenation, suggesting that reduced mitochondrial activity, and thus less ROS production under hypoxia increased insect tolerance to irradiation. Furthermore, reoxygenation accelerated eBeam-induced glutathione-S-transferase activation and potentiated eBeam-enhanced catalase activities. Faster and stronger detoxification capacity in eBeam-irradiated, hypoxic larvae may have protected them from oxidative damage. CONCLUSION: Hypoxic environment enhanced radiotolerance of bruchid larvae, presumably due to limited ROS production and elevated antioxidant enzymatic activities after reoxygenation. © 2018 Society of Chemical Industry.