Polyurethane foam induces epigenetic modification of mitochondrial DNA during different metamorphic stages of Tenebrio molitor.

The present work investigated the changes in DNA methylation pattern of Tenebrio molitor mitochondria genome at different development stages, which was fed with polyurethane foam as a sole diet. Polyurethane foam could influence the global methylation levels in mitochondria DNA of Tenebrio molitor. Different leves of 5-methylcytosine appeared at CpG and non-CpG sites of Tenebrio molitor mtDNA while they were fed with polyurethane foam: 10 CpG and 49 non-CpG sites at larval stage, 4 CpG and 31 non-CpG sites at pupa stage, 7 CpG and 56 non-CpG sites at adult stage in general. Moreover, we observed the decreased levels of ATP generation with the mitochondria DNA methylation variation. The results demonstrated that mitochondria DNA gene could be methylated in response to environmental pollutants to modulate stage-specific functions. Moreover, mtDNA methylation of polyurethane-foam-feeding Tenebrio molitor existed discrepancy in the developmental stage. The tentative methylation mechanism of mtDNA might be that polyurethane foam induced oxidative stress and increased the permeability of mitochondrial membranes, which resulted in transmethylase entry into mitochondria.

Myclobutanil accumulation, transcriptional alteration, and tissue injury in lizards (Eremias argus) treated with myclobutanil enantiomers.

Enantioselective toxicokinetics, accumulation, and toxicity of myclobutanil were investigated by oral exposure of myclobutanil enantiomers to lizards. After a single oral administration, the absorption half-lives ( [Formula: see text] ) and elimination half-lives (t1/2k) were in the range of 0.133-14.828 and 3.641-17.682 h, respectively. The absorption and elimination half-lives of (+)-myclobutanil showed no significant differences from those of (-)-myclobutanil in lizard blood, whereas preferential enrichment of (-)-enantiomer was observed in the liver, fat, skin, intestine, lung and kidney. In the bioaccumulation experiments, the residue of (-)-myclobutanil was detected in most tissues at 7, 14, and 28 days, while (+)-myclobutanil was found only in lizard skin, at a concentration lower than that of (-)-myclobutanil. Thus, (-)-myclobutanil was preferentially accumulated in lizards. The transcriptional responses of metabolic enzyme genes indicated that cytochrome P450 1a1 (cyp1a1), cyp2d3, cyp2d6, cyp3a4 and cyp3a7 played a crucial role in the metabolism of (+)-myclobutanil, whereas cyp1a1, cyp2d3, cyp2d6, cyp2c8, and cyp3a4 contributed to the metabolism of (-)-myclobutanil. The difference in metabolism pathways may be a reason for the enantioselectivity of myclobutanil in lizard. Myclobutanil also affected the expression of antioxidant enzyme genes, and the (+)-myclobutanil treatment might produce higher oxidative stress in lizard liver when compared with its antipode. Hepatic histopathological changes such as hepatocellular hypertrophy, nuclear pyknosis, vacuolation, and non-zonal macrovesicular lipid accumulation were observed in the liver of lizards for both (+)-myclobutanil and (-)-myclobutanil treatments. Thus, myclobutanil could affect lizard liver upon multiple exposure. The findings of this study provide specific insights into the enantioselective metabolism and toxicity of chiral triazole fungicides in lizards.

Enantioselective Distribution, Degradation, and Metabolite Formation of Myclobutanil and Transcriptional Responses of Metabolic-Related Genes in Rats.

Myclobutanil (MT), a chiral fungicide, can be metabolized enantioselectively in organisms. In this work, the associated absorption, distribution, metabolism and transcriptional responses of MT in rats were determined following a single-dose (10 mg·kg-1 body weight) exposure to rac-, (+)- or (-)-MT. The enantiomer fractions (EFs) were less than 0.5 with time in the liver, kidney, heart, lung, and testis, suggesting preferential enrichment of (-)-MT in these tissues. Furthermore, there was conversion of (+)-form to (-)-form in the liver and kidney after 6 h exposure to enantiopure (+)-MT. Enrichment and degradation of the two enantiomers differed between rac-MT and MT-enantiomers groups, suggesting that MT bioaccumulation is enantiomer-specific. Interestingly, the degradation half-life of MT in the liver with rac-MT treatment was shorter than that with both MT-enantiomer treatments. One reason may be that the gene expression levels of cytochrome P450 1a2 ( cyp1a2) and cyp3a2 genes in livers treated with rac-MT were the highest among the three exposure groups. In addition, a positive correlation between the expression of cyp2e1 and cyp3a2 genes and rac-MT concentration was found in livers exposed to rac-MT. Simultaneously, five chiral metabolites were detected, and the enantiomers of three metabolites, RH-9090, RH-9089, and M2, were separated. The detected enantiomers of (+)-MT metabolites were in complete contrast with those of (-)-MT metabolites. According to the results, a metabolic pathway of MT in male rats was proposed, which included the following five metabolites: RH-9089, RH-9090, RH-9090 Sulfate, M1, and M2. The possible metabolic enzymes were marked in the pathway. The findings of this study provide more specific insights into the enantioselective metabolic mechanism of chiral triazole fungicides.

The metabolism distribution and effect of imidacloprid in chinese lizards (Eremias argus) following oral exposure.

Systematically evaluation of the metabolism, distribution and effect of imidacloprid in Chinese lizards (Eremias argus) were carried out following oral exposure. Imidacloprid-olefin-guanidine was prone to accumulate in the brain and caused potential neurotoxicity. Percutaneous and excretory excretions were the primary ways for the elimination of imidacloprid and its metabolites. Liver was the main site for hydroxy reduction and nitro-reduction metabolism of imidacloprid. The metabolism of imidacloprid was a complex process in which many metabolic enzymes participated. Aldehyde oxidase and CYP2C9 were the key enzymes in nitro-reduction process. CYP3A4 dominated the process of hydroxylation and desaturation. The increase in Glutathione S-transferase expression may be related to the removal of imidacloprid, but also related to the oxidative stress reaction that imidacloprid may cause in tissues, especially in the kidney. The findings enrich and supplement the knowledge of the environmental fate of imidacloprid in reptiles.

Enantiomerization and stereoselectivity in bioaccumulation of furalaxyl in Tenebrio molitor larvae.

Furalaxyl is a chiral pesticide and widely used in modern agriculture as racemate mixture. The enantiomerization and enantioselecive bioaccumulation by a single dose of furalaxyl to Tenebrio molitor larvae under laboratory conditions were studied using a high-performance liquid chromatography tandem mass spectroscopy method based on a ChiralPAK IC column. Our results showed that a significant enantiomerization (interconversion between R-enantiomer and S-enantiomer) was observed in Tenebrio molitor larvae under R- or S-furalaxyl exposure. Though the two furalaxyl enantiomers exhibited low-capacity of bioaccumulation in Tenebrio molitor larvae, bioaccumulation of rac-furalaxyl was enantioselective with a preferential accumulation of S-furalaxyl at 10mg/kg dosage exposure. In addition, enantiomerization and enantioselective degradation of the two enantiomers was not observed in wheat bran. These results showed that enantioselectivtiy of furalaxyl enantiomers was an important process combined with degradation, metabolism and enatiomerization in organisms.

Tissue distribution and metabolism of triadimefon and triadimenol enantiomers in Chinese lizards (Eremias argus).

Triadimefon (TF, S-(+)-TF, R-(-)-TF) and its metabolite triadimenol (TN, TN-A1, A2 and TN-B1, B2) are two systemic fungicides and both of them are chiral pharmaceuticals which are widely used in agricultural industry. Many researches focused on the toxicity effects of triadimefon on mammals, while the ecotoxicological data of tiradimefon on reptiles is limited. In order to understand the toxicity mechanism of triadimefon in reptiles, the current study administrated S-(+)-TF or R-(-)-TF traidimefon (50mg/kgbw) to Chinese lizards (Eremias argus) respectively, the absorption, distribution of triadimefon and the formation of triadimenol were analysed at different sampling times. The metabolic pathways were demonstrated through relative gene expression using quantitative real-time PCR reaction. During the experiment time, triadimefon was quickly peaked to the maximum concentration within 12h in liver, brain, kidney, and plasma, eliminated slowly. The biotransformation in kidney was the lowest and fat possessed the worst degradation ability among others. The metabolite, triadimenol was detected in blood in 2h and reached to a plateau at about 12h in most organs (fat excepted), while the process of metabolism is stereoselective. The mainly metabolite in R-(-)-TF treated group was TN-B1, and TN-A2 in S-(+)-TF group which showed the selective metabolism to other species caused by environmental conditions, differences in the animal models and concentration of TF. The related gene expression of cyp1a1, cyp3a1 and hsd11ß mRNA level in lizards showed different metabolic pathways in the liver and brain. Both P450s enzymes and 11ß-hydroxysteroid dehydrogenase participated in metabolic reaction in liver, while no 11ß-hydroxysteroid dehydrogenase pathway observed in brain. This diversity in liver and brain may cause different degradation rate and ecotoxicological effect in different organs.

Enantioselective acute toxicity effects and bioaccumulation of furalaxyl in the earthworm (Eisenia foetida).

The enantioselectivities of individual enantiomers of furalaxyl in acute toxicity and bioaccumulation in the earthworm (Eisenia foetida) were studied. The acute toxicity was tested by filter paper contact test. After 48 h of exposure, the calculated LC50 values of the R-form, rac-form, and S-form were 2.27, 2.08, and 1.22 µg cm(-2), respectively. After 72 h of exposure, the calculated LC50 values were 1.90, 1.54, and 1.00 µg cm(-2), respectively. Therefore, the acute toxicity of furalaxyl enantiomers was enantioselective. During the bioaccumulation experiment, the enantiomer fraction of furalaxyl in earthworm tissue was observed to deviate from 0.50 and maintained a range of 0.55-0.60; in other words, the bioaccumulation of furalaxyl was enantioselective in earthworm tissue with a preferential accumulation of S-furalaxyl. The uptake kinetic of furalaxyl enantiomers fitted the first-order kinetics well and the calculated kinetic parameters were consistent with the low accumulation efficiency.

Enantiomerization and enantioselective bioaccumulation of metalaxyl in Tenebrio molitor larvae.

The enantiomerization and enantioselective bioaccumulation of metalaxyl by a single dose of exposure to Tenebrio molitor larvae under laboratory condition were studied by high-performance liquid chromatography tandem mass spectroscopy (HPLC-MS/MS) based on a ChiralcelOD-3R [cellulosetris-tris-(3, 5-dichlorophenyl-carbamate)] column. Exposure of enantiopure R-metalaxyl and S-metalaxyl in Tenebrio molitor larvae exhibited significant enantiomerization, with formation of the R enantiomers from the S enantiomers, and vice versa, which might be attributed to the chiral pesticide catalyzed by a certain enzyme in Tenebrio molitor larvae. Enantiomerization was not observed in wheat bran during the period of 21 d. In addition, bioaccumulation of rac-metalaxyl in Tenebrio molitor larvae was enantioselective with a preferential accumulation of S-metalaxyl. These results showed that enantioselectivity was caused not only by actual degradation and metabolism but also by enantiomerization, which was an important process in the environmental fate and behavior of metalaxyl enantiomers.

Enantioseletive bioaccumulation and metabolization of diniconazole in earthworms (Eiseniafetida) in an artificial soil.

Degradation and enantioselective bioaccumulation of diniconazole in earthworms (Eiseniafetida) in artificial soil was investigated using liquid chromatography-tandem mass spectrometry (LC-MS/MS) method under laboratory condition. Three exposure concentrations (1 mg/kg, 10 mg/kg and 25 mg/kg) of diniconazole in soil (dry weight) to earthworms were used. The uptake kinetics fitted the first-order kinetics well. The bioaccumulation factors (BAF) of R, S isomers were 6.6046 and 8.5115 in 25 mg/kg dose exposure, 2.6409 and 2.9835 in 10mg/kg dose exposure, 1.7784 and 2.0437 in 1 mg/kg dose exposure, respectively. Bioaccumulation of diniconazole in earthworm tissues was enantioselective with a preferential accumulation of S-diniconazole and the enantiomer fractions were about 0.45-0.50 in all three level dose exposures. In addition, it was obvious that both R-diniconazole and S-diniconazole had bioaccumulation effect in earthworm. Diniconazole was metabolized to 1,2,4-triazole, (E)-3-(1H-1,2,4-triazol-1-yl) acrylaldehyde, (E, S)-4-(2, 4-dichlorophenyl)-2, 2-dimethyl-5-(1H-1,2,4-triazol-1-yl)pent-4-ene-1,3-diol, and (E)-4-(2, 4-dichlorophenyl)-3-hydroxy-2,2-dimethyl-5-(1H-1,2,4-triazol-1-yl) pent-4-enoic acid in earthworms; the metabolites of 1,2,4-triazole and (E)-3-(1H-1,2,4-triazol-1-yl)acrylaldehyde could be detected in soil as well.

Stereoselective metabolism, distribution, and bioaccumulation brof triadimefon and triadimenol in lizards.

In this research, Chinese lizards (Eremias argus) were chosen as laboratory animal to evaluate the stereoselectivity in the processes of metabolism, distribution, and bioaccumulation of triadimefon. A validated chiral high-performance liquid chromatography coupled with triple quadruple mass spectrometry (HPLC-MS/MS) method was developed for determining enantiomers' residues of parent compound triadimefon and its metabolite triadimenol in lizard blood and tissues. Pharmacokinetic results of single-does exposure suggested that S-(+)-triadimefon was metabolized easier than R-(-)-triadimefon, and RR-(+)-triadimenol was the main metabolic product of triadimefon. During the continuous exposure of two dose (40mg/kg(bw)·d and 200mg/kg(bw)·d), enantiomers of triadimefon and triadimenol were detected in all body compartments, with the highest triadimefon concentrations in brain. However, the triadimenol concentrations were not significantly different among the compartments. The concentrations of RS-(+)-triadimenol were negative correlated with concentrations of RR-(+)-triadimenol both in blood (r=-0.775, p=0.024) and liver (r=-0.834, p=0.02) in 200mg/kg(bw)·d group, which indicates that chiral conversion between enantiomers of triadimenol might exist in the metabolic process of triadimefon. In all the processes, the enantiomer fractions (EFs) of R-(-)-triadimefon and RR-(+)-triadimenol were significantly different from their natural ratios, 0.5 and 0.1, respectively, which proved that metabolism, bioaccumulation, and distribution of triadimefon and triadimenol in lizards were enantioselective. These results help enrich and supplement the knowledge of the stereoselective behaviour of triadimefon and triadimenol in reptile.

Stereoselectivity in bioaccumulation and excretion of epoxiconazole by mealworm beetle (Tenebrio molitor) larvae.

Stereoselectivity in bioaccumulation and excretion of stereoisomers of epoxiconazole by mealworm beetle (Tenebrio molitor) larvae through dietary exposure was investigated. Liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method that use a ChiralcelOD-3R[cellulosetris-Tris-(3, 5-dichlorophenyl-carbamate)] chromatography column was applied to carry out chiral separation of the stereoisomers. Wheat bran was spiked with racemic epoxiconazole at two dose levels of 20mg/kg and 2mg/kg (dry weight) to feed T. molitor larvae. The results showed that both the doses of epoxiconazole were taken up by Tenebrio molitor larvae rapidly at the initial stages. There was a significant trend of stereoselective bioaccumulation in the larvae with a preferential accumulation of (-)-epoxiconazole in the 20mg/kg dose. The stereoselectivity in bioaccumulation in the 2mg/kg dosage was not obvious compared to the 20mg/kg group. Results of excretion indicated an active excretion is an important pathway for the larvae to eliminate epoxiconazole which was a passive transport process with non stereoselectivity. The faster elimination might be the reason for the low accumulation of epoxiconazole, as measured by bioaccumulation factor (BAF).

Stereoselective degradation and toxic effects of benalaxyl on blood and liver of the Chinese lizard Eremias argus.

Benalaxyl as a xylem-systemic fungicide is usually direct sprayed on the soil surface, which is potential harm to the animals lived in the soil. However, the stereoselectivity of benalaxyl in reptiles have rarely been studied. In this study, Chinese lizards (Eremias argus) were firstly used to evaluate the stereoselectivity in biodegradation and toxicity of racemate and individual enantiomers of benalaxyl. A method for determining residues of the two enantiomers of benalaxyl in lizard blood and liver by high-performance liquid chromatography coupled with triple quadrupole mass spectrometry (HPLC-MS/MS) was developed. The degradation followed pseudo first-order kinetics and the degradation of the (S)-(+)-benalaxyl was faster than its antipode in blood and liver (Half-time t1/2 of (R)-(-)-benalaxyl and (S)-(+)-benalaxyl were 5.08 h and 3.75 h in blood, 6.21 h and 4.45 h in liver, separately). Moreover, antioxidant defenses consisting of activities of superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST) and lipid peroxide malondialdehyde (MDA) were determined in 24h acute exposure. Enantioselectivity of acute toxicity depended on the concentration and form of benalaxyl. In addition, cellular degeneration, decrease of cell number, clustering phenomena of cell nuclei and preliminary liver fibrosis were observed in pathological detection at the termination of 21-d subchronic exposure (20 mg/kg(-bw) of racemate and individual enantiomers of benalaxyl). The enantiomer fractions (EFs) in racemate and individual enantiomer groups were approached both in blood and liver caused by the chiral conversion. The chiral conversion from (R)-(-)-benalaxyl to (S)-(+)-benalaxyl and (S)-(+)-benalaxyl to (R)-(-)-benalaxyl were the primary cause for no remarkable differences in toxicity between the enantiomers of benalaxyl.

Determination of bromothalonil residues and degradation in apple and soil by QuEChERS and GC-MS/MS.

The dissipation and residues of bromothalonil in apple and soil under field condition were analyzed by QuEChERS (quick, easy, cheap, effective, rugged and safe) combined with gas chromatography-mass spectrometer method. The recoveries were ranged from 80.8 % to 106 % with coefficient variation for repeatability ranged from 3.08 % to 7.09 % at fortification levels of 0.02, 0.04 and 0.2 mg/kg in apple and soil. The limit of quantification of the method was 0.31 µg/kg. The dissipation rates of bromothalonil followed the first-order kinetics and the half-lives were from 3.61 to 3.98 days in apple and from 4.65 to 9.29 days in soil. In apple, the terminal residues of bromothalonil were below the China maximum residue limit (0.2 mg/kg) after 7 days of application. This work contributed to provide the basic information for a safe usage of bromothalonil in apple orchard and preventing health problem from consumers in China.

A Magnetic Beads-Based Sandwich Chemiluminescence Enzyme Immunoassay for the Rapid and Automatic Detection of Lactoferrin in Milk.

Lactoferrin (LF), an iron-binding glycoprotein with immunological properties and a high nutritional value, has emerged as a prominent research focus in the field of food nutrition. Lactoferrin is widely distributed in raw milk and milk that has undergone low-temperature heat treatment during pasteurization, making its rapid and accurate detection crucial for ensuring the quality control of dairy products. An enzyme-linked immunosorbent assay-based analytical protocol has often been referred to for the detection of LF in real samples. Signal amplification was accomplished using the streptavidin-biotin system. Here, an automated magnetic beads-based sandwich chemiluminescence enzyme immunoassay (MBs-sCLEIA) system was developed for the quantification of lactoferrin in pasteurized milk. The MBs-sCLEIA system consists of an automated chemiluminescence-based analyzer and a lactoferrin MBs-sCLEIA assay kit. Notably, our proposed method eliminates the need for pretreatment procedures and enables the direct addition of milk samples, allowing for the automatic quantitative detection of lactoferrin within a rapid 17 min timeframe for up to eight samples simultaneously. The MBs-sCLEIA was linear over the range of 7.24-800 ng/mL and displayed a limit of detection (LOD) of 2.85 ng/mL. As its good recovery and CV values indicate, the method exhibited high precision and accuracy. Furthermore, it was verified that it was selective towards five additional common milk proteins. A good correlation was observed between the results from the MBs-sCLEIA and heparin affinity column-HPLC (r2 = 0.99042), which proves to be a useful and practicable way of conducting an accurate analysis of lactoferrin in dairy products.

One-pot green synthesis of ZIF-8/IgG composite for the precise orientation and protection of antibody and its application in purification and detection of aflatoxins in peanut oil.

This study presents a novel approach toward the one-pot green synthesis of ZIF-8/IgG composite, focusing on its precise orientation and protection of the anti-aflatoxins antibody. The antibody orientation is achieved through the specific binding of IgG to the Fc region of the antibody, while the antibody protection is accomplished by the structural change restriction of ZIF-8 framework to the antibody. Consequently, the antibody exhibits enhanced target capability and significantly improved tolerance to organic solvents. The ZIF-8/IgG/anti-AFT was employed for the purification and detection of AFTs by coupling with UPLC. Under optimized conditions, the recoveries of spiked AFTs in peanut oils are between 86.1% and 106.4%, with relative standard deviations (RSDs) ranging from 0.8% to 8.8%. The linearity range is 0.5-20.0 ng for AFB1 and AFG1, 0.125-5.0 ng for AFB2 and AFG2, the limit of detection is 0.1 ng for AFB1 and AFG1, 0.03 ng for AFB2 and AFG2.

[Simultaneous determination of 21 perfluorinated and polyfluoroalkyl substances in plant oils by ultra-high performance liquid chromatography-triple quadrupole mass spectrometry].

Edible plant oils are a key component of the daily human diet, and the quality and safety of plant oils are related to human health. Perfluorinated and polyfluoroalkyl substances (PFASs) are pollutants that can contaminate plant oil through the processing of raw materials or exposure to materials containing these substances. Thus, establishing a sensitive and accurate analytical method for the determination of PFASs is critical for ensuring the safety of plant oils. In this study, a method based on acetonitrile extraction and solid phase extraction purification combined with ultra-high performance liquid chromatography-triple quadrupole mass spectrometry (UHPLC-MS/MS) was developed for the simultaneous determination of 21 PFASs, including perfluorocarboxylic acids, perfluoroalkyl sulfonic acids, and fluorotelomer sulfonic acids, in edible plant oils. The chromatographic conditions and MS parameters were optimized, and the influences of the extraction solvents and purification method were systematically studied. Plant oil samples were directly extracted with acetonitrile and purified using a weak anion-exchange (WAX) column. The 21 target PFASs were separated on a reversed-phase C18 chromatographic column and detected using a triple quadrupole mass spectrometer with an electrospray ionization source. The mass spectrometer was operated in negative-ion mode. The target compounds were analyzed in multiple reaction monitoring (MRM) mode and quantified using an internal standard method. The results demonstrated that the severe interference observed during the detection of PFASs in the co-extracted substances was completely eliminated after the extraction mixture was purified using a WAX column. The 21 target PFASs showed good linearity in their corresponding ranges, with correlation coefficients greater than 0.995. The limits of detection (LODs) and limits of quantification (LOQs) of the method were in the range of 0.004-0.015 and 0.015-0.050 µg/kg, respectively. The recoveries ranged from 95.6% to 115.8%, with relative standard deviations (RSDs) in the range of 0.3%-10.9% (n=9). The established method is characterized by simple sample pretreatment, good sensitivity, high immunity to interferences, and good stability, rendering it suitable for the rapid analysis and accurate determination of typical PFASs in edible plant oils.

Bioaccumulation and excretion of enantiomers of myclobutanil in Tenebrio molitor larvae through dietary exposure.

The bioaccumulation and excretion of enantiomers of myclobutanil in Tenebrio molitor larvae through dietary exposure under laboratory conditions were investigated using high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) based on a ChiralcelOD-3R [cellulosetris-tris-(3, 5-dichlorophenyl-carbamate)] column. The wheat bran fed to Tenebrio molitor larvae was spiked with racemic myclobutanil at two dose levels of 20 mg/kg and 2 mg/kg (dry weight). The results showed that there was a significant trend of enantioselective bioaccumulation in the larvae with a preferential accumulation of (-)-myclobutanil in 20 mg/kg dose exposure, but it was not obviously observed in the 2 mg/kg dose group. A kinetic model considering enantiomerization between the two enantiomers based on first-order reactions was built and the rate constants were estimated to discuss the kinetic reason for the different concentrations of individual enantiomers in the larvae. The approximations implied an inversion between the two enantiomers with a relatively higher rate of the inversion from (-)-myclobutanil to (+)-myclobutanil. Meanwhile, analysis of data of excretion samples suggested the active excretion is probably an important pathway for the insect to eliminate myclobutanil rapidly with nonenantioselectivity as a passive transport process, which was consistent with the low accumulation efficiency of myclobutanil measured by BAF (bioaccumulation factor).

Complete genome sequence of Brucella pseudintermedia ASAG-D25, isolated from the wheat ear.

Brucella pseudintermedia ASAG-D25 was isolated from the wheat ear sample in Xuzhou City, Jiangsu Province, China. The complete genome sequence of B. pseudintermedia will provide an important resource for better understanding of the genetic features of the species within the family of Brucellaceae.

Biotransformation of Deoxynivalenol by a Dual-Member Bacterial Consortium Isolated from Tenebrio molitor Larval Feces.

In this study, a dual-member bacterial consortium with the ability to oxidize deoxynivalenol (DON) to 3-keto-DON, designated SD, was first screened from the feces of Tenebrio molitor larvae. This consortium consisted of Pseudomonas sp. SD17-1 and Devosia sp. SD17-2, as determined by 16S rRNA-based phylogenetic analysis. A temperature of 30 °C, a pH of 8.0-9.0, and an initial inoculum concentration ratio of Devosia to Pseudomonas of 0.1 were optimal single-factor parameters for the DON oxidation activity of the bacterial consortium SD. Genome-based bioinformatics analysis revealed the presence of an intact PQQ biosynthesis operon (pqqFABCDEG) and four putative pyrroloquinoline quinone (PQQ)-dependent alcohol dehydrogenase (ADH) genes in the genomes of Pseudomonas strain SD17-1 and Devosia strain SD17-2, respectively. Biochemical analyses further confirmed the PQQ-producing phenotype of Pseudomonas and the DON-oxidizing enzymatic activities of two of four PQQ-dependent ADHs in Devosia. The addition of PQQ-containing a cell-free fermentation supernatant from Pseudomonas activated DON-oxidizing activity of Devosia. In summary, as members of the bacterial consortium SD, Pseudomonas and Devosia play indispensable and complementary roles in SD's oxidation of DON. Specifically, Pseudomonas is responsible for producing the necessary PQQ cofactor, whereas Devosia expresses the PQQ-dependent DON dehydrogenase, together facilitating the oxidation of DON.

Four PQQ-Dependent Alcohol Dehydrogenases Responsible for the Oxidative Detoxification of Deoxynivalenol in a Novel Bacterium Ketogulonicigenium vulgare D3_3 Originated from the Feces of Tenebrio molitor Larvae.

Deoxynivalenol (DON) is frequently detected in cereals and cereal-based products and has a negative impact on human and animal health. In this study, an unprecedented DON-degrading bacterial isolate D3_3 was isolated from a sample of Tenebrio molitor larva feces. A 16S rRNA-based phylogenetic analysis and genome-based average nucleotide identity comparison clearly revealed that strain D3_3 belonged to the species Ketogulonicigenium vulgare. This isolate D3_3 could efficiently degrade 50 mg/L of DON under a broad range of conditions, such as pHs of 7.0-9.0 and temperatures of 18-30 °C, as well as during aerobic or anaerobic cultivation. 3-keto-DON was identified as the sole and finished DON metabolite using mass spectrometry. In vitro toxicity tests revealed that 3-keto-DON had lower cytotoxicity to human gastric epithelial cells and higher phytotoxicity to Lemna minor than its parent mycotoxin DON. Additionally, four genes encoding pyrroloquinoline quinone (PQQ)-dependent alcohol dehydrogenases in the genome of isolate D3_3 were identified as being responsible for the DON oxidation reaction. Overall, as a highly potent DON-degrading microbe, a member of the genus Ketogulonicigenium is reported for the first time in this study. The discovery of this DON-degrading isolate D3_3 and its four dehydrogenases will allow microbial strains and enzyme resources to become available for the future development of DON-detoxifying agents for food and animal feed.