The lipase inhibitory effect of mulberry leaf phenolic glycosides: The structure-activity relationship and mechanism of action.

The present study found for the first time that phenolic glycosides were an important material basis for mulberry leaves to inhibit lipase. The corresponding IC50 for hyperoside, rutin, astragalin and quercetin were 68, 252, 385 and 815 µg/mL respectively. The inhibitory effect was ranked as monoglycosides > phenolic hydroxyl groups > disaccharides on the benzone ring. Hyperoside bound to lipase in competitive inhibition type with one binding site, while the others bound to lipase in a mixed inhibition type by two similar sites. All four compounds altered the microenvironment and secondary conformation of lipase through static quenching. The docking score, stability, and binding energy were consistent with the compound inhibitory activity. The main binding between compounds and lipase amino acid residues were spontaneously though hydrophobic interactions and hydrogen bonding. The strong hydrogen bonds formed with SER-152 inside the lipase pocket, might be important for the strong inhibitory activity of hyperoside.

Stable isotope labeling and functional gene prediction elucidate the carbon metabolism in fermentative bacteria and microalgae coupling system.

The application of the fermentative bacteria and microalgae coupling system in the wastewater treatment has been studied, but there remains few knowledge regarding the organic and inorganic carbon metabolism within this system. In this study, the carbon metabolism of microalgae and fermentative bacteria was elucidated by 13C stable isotope labeling and functional gene prediction, respectively. The 13C glucose and 13C NaHCO3 were used as stable isotope tracers to clarify the organic and inorganic carbon metabolism of microalgae, indicating that approximately 71.5 % of the Acetyl-CoA in microalgae was synthesized from organic carbon sources, while 26.8 % was synthesized through the utilization of inorganic carbon sources. Inorganic carbon sources can enhance the activity of photosynthetic system and facilitate the Calvin cycle. Considering the adequate organic carbon sources and insufficient inorganic carbon sources in the fermentative bacteria and microalgae coupling system, NaHCO3 was added to improve carbon utilization of microalgae. The maximum microalgal lipid yield reached 1130.37 mg/L with 1000 mg/L NaHCO3 supplementation. Functional gene prediction was used to analysis the effect of various carbon composition on the bacterial carbon metabolism. Notably, the additional inorganic carbon sources increased the abundance of bacterial functional genes associated with the fermentation and acetic acids synthesis, which was advantageous for VFAs production and further promoted microalgae growth. This study can gain a deeper understanding of microbial metabolic mechanisms during the operation of fermentative bacteria and microalgae system, and improve its sustained operational stability.

Effects of mulberry leaf powder water extract supplementation on the growth performance, immunity, antioxidant, meat quality and intestinal microbiota of yellow feather broilers.

A 50-day feeding trial was conducted to evaluate the effects of mulberry leaf powder water extract (MLE) on the growth performance, immunity, antioxidant, meat quality and intestinal microbiota of yellow feather broilers. A total of 720 birds (initial body weight 40.07 ± 0.05 g) were randomly distributed into four groups with six replicates per group and 30 birds per replicate. Four diets were formulated with 0% (CON), 200 mg/kg MLE (MLE200), 400 mg/kg MLE (MLE400) and 600 mg/kg MLE (MLE600) supplementation. Results showed that the addition of 200-600 mg/kg MLE to the diet significantly increased the body weight (BW) and average daily weight gain (ADG), but feed to gain ratio (F/G) were linearly decreased (p = 0.045) as dietary MLE increased. Birds fed MLE400 had higher (p < 0.05) total antioxidant capacity (T-AOC), interleukin-10 (Il-10), secretory immunoglobulin A (SIgA) and complement 3 (C3) contents than those fed CON, whereas MLE400 had lower malondialdehyde (MDA) content than CON (p < 0.05). Analysis of 16 S rDNA indicated that supplementation with 200 mg/kg MLE increased the Shannon indices in the caecum (p < 0.05). Supplementation with MLE decreased the abundance of the phylum Proteobacteria and genus Helicobacter, and increased the abundance of the phylum Bacteroidetes in the caecum in broiler chickens (p < 0.05). The drip loss rate in the MLE600 was significantly diminished (p < 0.05), whereas the shear force was significantly elevated (p < 0.05). In summary, dietary supplementation with MLE can effectively improve growth performance, intestinal immunity, serum antioxidant capacity, meat quality and intestinal microbiota of yellow feather broilers. The most appropriate MLE supplementation level was 400 mg/kg. This study provides a practical strategy for the dietary application of MLE in yellow feather broilers.

Effect of light intensity and photoperiod on high-value production and nutrient removal performance with bacterial-algal coupling system.

Direct discharge of mariculture wastewater can lead to eutrophication, posing a threat to aquatic ecosystems. A novel Bacteria-Algae Coupled Reactor (BACR) offers advantages in treating mariculture wastewater, which can effectively remove pollutants while simultaneously obtaining microalgal products. However, there is limited information available on how illumination affects the cultivation of mixotrophic microalgae in this bacteria-algae coupling system. Therefore, a combined strategy of photoperiod and light intensity regulation was employed to improve the biological mariculture wastewater remediation, promote microalgae biomass accumulation, and increase the high-value product yield in this study. Optimal light conditions could effectively enhance microalgal carbohydrate, protein, lipid accumulation and photosynthetic activity, with the carbohydrate, protein and lipid contents reached 44.11, 428.57 and 399.68 mg/L, respectively. Moreover, excellent removal rates were achieved for SCOD, NH4+-N and TP, reaching 86.68%, 87.35% and 95.13% respectively. This study proposes a comprehension of BACR processes in mariculture wastewater under different light conditions.

Ecdysteroid UDP-Glucosyltransferase Expression in Beauveria bassiana Increases Its Pathogenicity against Early Instar Silkworm Larvae.

Beauveria bassiana (B. bassiana) is a broad-spectrum entomopathogenic fungus that can control pests in agriculture and forestry. In this study, encoding ecdysteroid uridine diphosphate glucosyltransferase gene (egt) was successfully screened in B. bassiana on the medium containing 500µg/mL G418 sulfate solution through the protoplast transformation method. This enzyme has the function of 20E (20-hydroxyecdysone) inactivation, thus increasing the mortality of the early instar larvae infected with B. bassiana. In this study, we transformed B. bassiana with the egt gene, which deactivates 20-hydroxyecdysone, a key hormone in insect development. The results showed that transgenic B. bassiana killed more silkworms of the 2nd instar larvae than the wild-type with a shorter LT50 time, which was reduced by approximately 20% (day 1 of the 2nd instar silkworm infection of B. bassiana) and 26.4% (day 2 of the 2nd instar silkworm infection of B. bassiana) compared to the wild-type, and also showed a higher mortality number before molting. The transgenic B. bassiana had a higher coverage of the body surface of silkworms compared to the wild type on the 3rd instar. In summary, improving entomopathogenic fungi using biological methods such as genetic engineering is feasible.

Effects of Cinnamon Powder on Glucose Metabolism in Diabetic Mice and the Molecular Mechanisms.

The liver is the primary organ regulating glucose metabolism. In our recent study, cinnamon improved liver function in diabetic mice. However, it is not clear whether cinnamon can reduce the glycemia of diabetic animals by regulating liver glucose metabolism. The purpose of this study was to investigate the hypoglycemic mechanism of cinnamon powder (CP) from the perspective of regulating liver glucose metabolism. To achieve this, different doses of CP (200, 400, or 800 mg/kg body weight) were given to diabetic mice by gavage once per day for 8 weeks. These mice were compared with healthy controls, untreated diabetic mice, and diabetic mice treated with metformin (the main first-line drug for type 2 diabetes). CP treatment effectively reduced fasting blood glucose levels and food intake, improved glucose tolerance and fasting serum insulin levels, and decreased glycated serum protein levels in diabetic mice. Furthermore, treatment with CP increased liver glycogen content and reduced the level of the gluconeogenesis precursor pyruvate in the liver. Data obtained by qPCR and western blotting suggested that CP improved glucose metabolism disorders by regulating AMPKα/PGC1α-mediated hepatic gluconeogenesis and PI3K/AKT-mediated hepatic glycogen synthesis. CP exhibits good hypoglycemic effects by improving hepatic glycogen synthesis and controlling hepatic gluconeogenesis. Therefore, CP may be applied as a functional food to decrease blood glucose.

Effect of mariculture wastewater concentrations on high-value production and pollutants removal with bacterial-algal coupling reactor (BACR).

To achieve the goal of cost-effective mariculture wastewater treatment, a novel Bacteria-Algae Coupling Reactor (BACR) integrating acidogenic fermentation with microalgae cultivation was applied for the mariculture wastewater treatment. Currently, there is limited research on the impact of different concentrations of mariculture wastewater on the pollutant removal and the high-value products recovery. In this study, different concentrations (4, 6, 8, and 10 g/L) of mariculture wastewater were treated with BACR. The results showed thatoptimalMW concentrations of 8 g/L improved the growth viability and biochemical components synthetic of Chlorella vulgaris, which increased the potential for high-value products recovery. The BACR exhibited the excellent removal efficiency of chemical oxygen demand, ammonia-nitrogen and total phosphorus with 82.30%, 81.12% and 96.40%, respectively. This study offers an ecological and economic approach to improve the MW treatment through the utilization of a novel bacterial-algal coupling system.

Predatory stink bug, Eocanthecona furcellata (Wolff) responses to oral exposure route of λ-cyhalothrin via sex-specific modulation manner.

The toxic effects of insecticides on predatory arthropods have closely related to their exposure routes. However, little is known about the effects of insecticide on reproductive parameters when the route of exposure occurs at a trophic level via prey intake. We therefore conducted current studies assessing whether Eocanthecona furcellata adults would be affected by feeding with λ-cyhalothrin-contaminated prey. Reproductive parameters, i.e. prolonged premating and preoviposition durations, reduced number of egg batches and egg amount, disturbed ovarian development, and suppressed expression of reproductive related genes were observed in E. furcellata females by feeding with treated prey. Moreover, reduced survival rate and altered carbohydrate metabolism parameters were detected in male bugs. Biochemical parameters, including MDA content, the activities of three antioxidant enzymes and three detoxification enzymes exhibited sex-specific responses after oral-exposure to λ-cyhalothrin in E. furcellata. The results indicate that the insecticide affects the fitness and leads to impairing reproductive potential via sex-specific modulation manner in predator insects. Taken together, our results provide a comprehensive assessment about detrimental impacts of λ-cyhalothrin-exposure on predators via prey intake, as well as a solid basis for further research to protect the predators from hazardous impacts of insecticides.

Mechanism of lead adsorption by a Bacillus cereus strain with indole-3-acetic acid secretion and inorganic phosphorus dissolution functions.

BACKGROUND: Heavy metal pollution has become a major source of environmental pollution because of increasing industrialization. Microbial remediation is a promising approach to remediate lead-contaminated environments owing to its cost-effective, environment-friendly, ecologically sustainable, and highly efficient properties. In this study, the growth-promoting functions and lead-adsorption ability of Bacillus cereus SEM-15 were examined, and the functional mechanism of the strain was preliminarily identified using scanning electron microscopy, energy spectrum, infrared spectrum, and genome analyses, providing theoretical support for utilization of B. cereus SEM-15 in heavy metals remediation. RESULTS: B. cereus SEM-15 showed strong ability to dissolve inorganic phosphorus and secrete indole-3-acetic acid. The lead adsorption efficiency of the strain at lead ion concentration of 150 mg/L was more than 93%. Single factor analysis revealed the optimal conditions for heavy metal adsorption by B. cereus SEM-15 (adsorption time, initial lead ion concentration, pH, and inoculum amount were 10 min, 50-150 mg/L, 6-7, and 5 g/L, respectively) in nutrient-free environment, with the lead adsorption rate reaching 96.58%. Scanning electron microscopy of B. cereus SEM-15 cells before and after lead adsorption showed adherence of a large number of granular precipitates to the cell surface after lead adsorption. X-Ray photoelectron spectroscopy and Fourier transform infrared spectroscopy results indicated the characteristic peaks of Pb-O, Pb-O-R (R = functional group), and Pb-S bonds after lead adsorption, and a shift in the characteristic peaks of bonds and groups related to C, N, and O. Genome annotation results showed the presence of genes related to heavy metals tolerance and plant growth promotion in B. cereus SEM-15, providing a molecular basis for the strain's heavy metals tolerance and plant growth promotion functions. CONCLUSIONS: This study analyzed the lead adsorption characteristics of B. cereus SEM-15 and the associated influencing factors, and discussed the adsorption mechanism and related functional genes, providing a basis for clarifying the underlying molecular mechanism and offering a reference for further research on plant-microorganisms combined remediation of heavy metals polluted environments.

Decreased cuticular penetration minimizes the impact of the pyrethroid insecticide λ-cyhalothrin on the insect predator Eocanthecona furcellata.

The use of broad-spectrum pesticides may reduce the biological control efficacy of predatory arthropods. Hence, the risks of pesticides to predators need to be evaluated. Here, we assessed the effects of a broad spectrum pyrethroid λ-cyhalothrin on a polyphagous predatory insect Eocanthecona furcellata via contact exposure route. The recommended application rate of λ-cyhalothrin was lower than the LR50 and HQ (in-field) was equal to 0.57, indicating the risk of λ-cyhalothrin to E. furcellata was low. Dried λ-cyhalothrin residue had no effect on the mortality, body weight, protein content of cuticle, or activities of major detoxification enzymes in E. furcellata. Residual of λ-cyhalothrin was only detected in the cuticle and legs of E. furcellata with a decreasing trend as time went by and no λ-cyhalothrin was detected inside the body. Additionally, a comparative transcriptome analysis was conducted to study global changes in gene expression in E. furcellata at different time points following exposure to λ-cyhalothrin-contaminated environment. A total of 57,839 unigenes with an average length of 1044 bp and an N50 of 1820 bp were obtained. In total, 118 and 109 differentially expressed genes (DEGs) at 12 h, and 60 h were identified between two groups. The DEGs were largely enriched in functional categories related to the structural constituent of cuticle. Accordingly, multiple cuticle protein-coding genes were up-regulated at 12 h after pesticide exposure. The present study stressed the importance of evaluating the compatibility between a specific pesticide (λ-cyhalothrin) and E. furcellata via simulating the releasing predators after insecticide application. The data could help optimize the pesticide use, optimizing the ecological services of E. furcellata as a BCA, and expanding its use into more areas of agriculture.

Pupal Diapause Termination and Transcriptional Response of Antheraea pernyi (Lepidoptera: Saturniidae) Triggered by 20-Hydroxyecdysone.

The pupal diapause of univoltine Antheraea pernyi hampers sericultural and biotechnological applications, which requires a high eclosion incidence after artificial diapause termination to ensure production of enough eggs. The effect of pupal diapause termination using 20-hydroxyecdysone (20E) on the eclosion incidence has not been well-documented in A. pernyi. Here, the dosage of injected 20E was optimized to efficiently terminate pupal diapause of A. pernyi, showing that inappropriate dosage of 20E can cause pupal lethality and a low eclosion incidence. The optimal ratio of 20E to 1-month-old pupae was determined as 6 µg/g. Morphological changes showed visible tissue dissociation at 3 days post-injection (dpi) and eye pigmentation at 5 dpi. Comprehensive transcriptome analysis identified 1,355/1,592, 494/203, 584/297, and 1,238/1,404 upregulated and downregulated genes at 1, 3, 6, and 9 dpi, respectively. The 117 genes enriched in the information processing pathways of "signal transduction" and "signaling molecules and interaction" were upregulated at 1 and 3 dpi, including the genes involved in FOXO signaling pathway. One chitinase, three trehalase, and five cathepsin genes related to energy metabolism and tissue dissociation showed high expression levels at the early stage, which were different from the upregulated expression of four other chitinase genes at the later stage. Simultaneously, the expression of several genes involved in molting hormone biosynthesis was also activated between 1 and 3 dpi. qRT-PCR further verified the expression patterns of two ecdysone receptor genes (EcRB1 and USP) and four downstream response genes (E93, Br-C, ßFTZ-F1, and cathepsin L) at the pupal and pharate stages, respectively. Taken together, these genes serve as a resource for unraveling the mechanism underlying pupal-adult transition; these findings facilitate rearing of larvae more than once a year and biotechnological development through efficient termination of pupal diapause in A. pernyi in approximately half a month.

Mechanism of carbendazim in treating pebrine disease of Bombyx mori based on GC/MS-based metabonomics.

Pebrine disease is caused by microporidia (Nosema bombycis) and is destructive to sericulture production. A carbendazim-based drug FangWeiLing (FWL) has a significant control effect on the disease, which is a successful example of drug treatment of microsporidia. In this study, the therapeutic effect and critical action time of FWL were investigated by silkworm rearing biological test. Besides, the hemolymph samples from silkworms in the control group, model group, and FWL group were analyzed by metabonomics based on gas chromatography-mass spectrometry (GC/MS). The results showed that FWL had a significant therapeutic effect on pebrine disease, and the critical action time was 24 ~ 48 h post inoculation. Forty-seven different metabolites related to pebrine disease were screened out, and correlated with starch and sucrose metabolism; aminoacyl-tRNA biosynthesis; arginine biosynthesis; glycine, serine, and threonine metabolism; and phenylalanine, tyrosine, and tryptophan biosynthesis. After pretreatment with FWL, the metabolites were all effectively regulated, indicating productive intervention. Principal component analysis (PCA) also showed that the overall metabolic profile of the FWL group tended toward the control group. Compared with the control group, 16 different metabolites were obtained from the hemolymph of B.mori in FWL group, mainly involving aminoacyl-tRNA biosynthesis and taurine and hypotaurine metabolism. It indicated that FWL had some effect on silkworm metabolism, which might be related to the decrease in cocoon quality. In conclusion, combined with the life cycle of N. bombycis, the mechanism of carbendazim in the treatment of pebrine disease can be fully revealed. Carbendazim can effectively reduce the destruction of amino acid metabolism and carbohydrate metabolism by N. Bombycis infection by inhibiting the proliferation of the meronts in silkworms, thus maintaining the normal physiological state of B. mori and achieve therapeutic effects. GC/MS-based metabonomics is a valuable and promising strategy to understand the disease mechanism and drug treatment of pebrine disease.

Pb Content, Risk Level and Primary-Source Apportionment in Wheat and Rice Grains in the Lihe River Watershed, Taihu Region, Eastern China.

This study detailed a complete research from Lead (Pb) content level to ecological and health risk to direct- and primary-sources apportionment arising from wheat and rice grains, in the Lihe River Watershed of the Taihu region, East China. Ecological and health risk assessment were based on the pollution index and US Environmental Protection Agency (EPA) health risk assessment model. A three-stage quantitative analysis program based on Pb isotope analysis to determine the relative contributions of primary sources involving (1) direct-source apportionment in grains with a two-end-member model, (2) apportionment of soil and dustfall sources using the IsoSource model, and (3) the integration of results of (1) and (2) was notedly first proposed. The results indicated that mean contents of Pb in wheat and rice grains were 0.54 and 0.45 mg/kg and both the bio-concentration factors (BCF) were <<1; the ecological risk pollution indices were 1.35 for wheat grains and 1.11 for rice grains; hazard quotient (HQ) values for adult and child indicating health risks through ingestion of grains were all <1; Coal-fired industrial sources account for up to 60% of Pb in the grains. This study provides insights into the management of grain Pb pollution and a new method for its source apportionment.

Effects of Certain cis-Regulatory Elements on Stage-Specific vitellogenin Expression in the Bombyx mori (Lepidoptera: Bombycidae).

Bombyx mori vitellogenin (BmVg) is highly upregulated during pupation, and the 20-hydroxyecdysone and amino acids may regulate stage-specific BmVg expression. However, previous studies showed that other factors may also affect stage-specific BmVg expression. Here, we characterized effective BmVg transcription factors by identifying the corresponding cis-regulatory elements (CREs). We prepared transgenic B. mori, in which DsRed was driven by various lengths of BmVg promoter. qRT-PCR analysis showed that DsRed expression driven by a 1.0-kb BmVg promoter (VgP1.0K) was consistent with endogenous BmVg. VgP1.0K specificity was closer to the endogenous BmVg promoter than that of VgP0.8K. These results suggest that CREs affecting stage-specific BmVg expression were localized to the 1.0-kb BmVg promoter. We investigated the effects of certain CREs that could influence the stage specificity of BmVg promoter on BmVg expression in transgenic B. mori. The relative DsRed expression was significantly reduced in transgenic female B. mori and the peak in DsRed expression was delayed after E-box CRE mutation. These results demonstrate that the E-box element enhanced BmVg expression and also affected stage-specific BmVg expression. Moreover, the relative DsRed expression was significantly increased in transgenic female of B. mori after 3×BD CRE mutation in BmVg promoter. However, the stage specificity of the mutated promoter was consistent with that of the endogenous BmVg promoter. The 3×BD element downregulated BmVg but had no effect on stage-specific BmVg expression. The present study promoted the process of elucidating the regulatory network for stage-specific BmVg expression and furnished a theoretical basis for the application of BmVg promoter.

Isolation of Bacillus subtilis strain SEM-2 from silkworm excrement and characterisation of its antagonistic effect against Fusarium spp.

Fusarium wilt is a devastating soil-borne disease mainly caused by highly host-specific formae speciales of Fusarium spp. Antagonistic microorganisms play a very important role in Fusarium wilt control. Isolation of potential biocontrol strains has become increasingly important. Bacterial strain SEM-2 was isolated from the high-temperature stage of silkworm excrement composting. SEM-2 exhibited a considerable antagonistic effect against Fusarium graminearum mycelial growth and spore germination. The results of pot experiments suggested that SEM-2 has a better inhibitory effect on the early stage of disease occurrence. The green fluorescent protein labelled SEM-2 coated on the surface of tomato seeds colonised the roots of tomato plants in 15 days. Genome sequencing identified SEM-2 as a new strain of Bacillus subtilis, and genome annotation and analysis determined gene clusters related to the biosynthesis of antimicrobials, such as bacillaene, fengycin, bacillibactin, subtilosin A, surfactin, and bacilysin. Interestingly, liquid chromatography - quadrupole time-of-flight mass spectrometry revealed that metabolites in pathways associated with the synthesis of secondary metabolites and antibiotics were highly differentially expressed. These findings may help to explain the mode of action of B. subtilis SEM-2 against Fusarium spp.

bmo-miR-2739 and the novel microRNA miR-167 coordinately regulate the expression of the vitellogenin receptor in Bombyx mori oogenesis.

Vitellogenin receptors (VgRs) play crucial roles in oogenesis by mediating endocytosis of vitellogenin and other nutrients in ovipara. We conducted small RNA sequencing and screening with a luciferase reporter system, and found that bmo-miR-2739 and a novel miRNA (novel-miR-167) coordinately regulate the expression of VgR in Bombyx mori (BmVgR). Further analyses suggested that these two miRNAs direct target repression by binding directly to the BmVgR 3' untranslated region. Forced expression of either miRNA using the piggyBac system blocked vitellogenin (Vg) transport and retarded ovariole development. Antagomir silencing of bmo-miR-2739 or novel-miR-167 resulted in increased amounts of BmVgR protein in the ovaries and BmVgR mRNA in the fat body. This evidence, combined with spatiotemporal expression profiles, revealed that these two miRNAs function together to fine-tune the amount of BmVgR protein for ovarian development. Additionally, novel-miR-167 was mainly responsible for the post-transcriptional repression of BmVgR in non-ovarian tissues. The results of this study contribute to our understanding of the function of miRNAs during ovarian development of a lepidopteran and suggest a new strategy for controlling insect reproduction.

Phylogenetic Analysis of the Complete rRNA Gene Sequence of Nosema sp. SE Isolated from the Beet Armyworm Spodoptera exigua.

The microsporidium Nosema sp. SE is a pathogen that infects the beet armyworm Spodoptera exigua. The complete sequence of its 4,302-base pair (bp) ribosomal ribonucleic acid (rRNA) gene region was obtained by polymerase chain reaction amplification and sequencing. The rRNA organization of Nosema sp. SE was 5'-large subunit (LSU) rRNA-internal transcribed spacer-small subunit (SSU) rRNA-intergenic spacer-5S-3', which corresponded to the pattern of Nosema bombycis. Phylogenetic analysis based on LSU rRNA and SSU rRNA both indicated that the parasite had a close relationship with other true Nosema species, confirming that Nosema sp. SE belongs to true Nosema group of the genus Nosema.

Quality Formation Mechanism of Stiff Silkworm, Bombyx batryticatus Using UPLC-Q-TOF-MS-Based Metabolomics.

Bombyx batryticatus is a well-known animal in traditional Chinese medicine. The aim of the research was to reveal the quality formation mechanism of B. batryticatus and to screen out the characteristic component used for the quality control. The anticonvulsant effects of B. batryticatus with a stiff time of one, five, and nine days (D1, D5 and D9, respectively) and healthy silkworm of the same developmental stage (SW) were determined by animal experiment. The dynamic changes in chemical composition were analyzed using UPLC-Q-TOF-MS-based metabolomics. D5 and D9 B. batryticatus exhibited significant anticonvulsant effects (p < 0.05 and p < 0.01, respectively). Accordingly, principal component analysis (PCA) and partial least squares discrimination analysis (PLS-DA) indicated that the chemical composition of D5 and D9 B. batryticatus changed significantly. The different metabolites mainly consisted of primary metabolites such as lipids and amino acids and secondary metabolites such as flavonoids, beauvericin, and glycolipids. Interestingly, the relative abundance of quercetin-7-O-ß-d-4-O-methylglucoside, the characteristic component of B. batryticatus, increased with stiff time and was promised to be used as an index component of quality control. The results expand our understanding of the quality formation mechanism of B. batryticatus. In addition, it highlights the potential of UPLC-Q-TOF-MS-based metabolomics for the quality control purpose of TCMs.

Characterization and sequence analysis of potential biofertilizer and biocontrol agent Bacillus subtilis strain SEM-9 from silkworm excrement.

Fusarium wilt is a devastating soil-borne disease caused mainly by highly host-specific formae speciales of Fusarium oxysporum. Antagonistic microorganisms play a very important role in Fusarium wilt control, and the isolation of potential biocontrol strains is becoming more and more important. We isolated a bacterial strain (SEM-9) from the high-temperature stage of silkworm excrement composting, which had a marked ability to solubilize phosphorus, promote the growth and increase the yield of the small Chinese cabbage, and which also exhibited considerable antagonistic effect towards Fusarium sambucinum and other fungi. The result of physiological and biochemical analyses, as well as genome sequencing, showed that SEM-9 was a strain of Bacillus subtilis. Through genome annotation and analysis, it was found that SEM-9 contained genes related to the regulation of biofilm formation, which may play an important role in colonization, and gene clusters encoding the biosynthesis of antimicrobials, such as surfactin, bacilysin, fengycin, and subtilosin-A. The production of such antifungal compounds may constitute the basis of the mode-of-action of SEM-9 against Fusarium spp. These data suggested that the SEM-9 strain has potential as both a biofertilizer and a biocontrol agent, with the potential to manage Fusarium wilt disease in crops.

Morphological and phylogenetic analysis of a microsporidium (Nosema sp.) isolated from rice stem borer, Chilo suppressalis (Walker) (Lepidoptera: Pyralidae).

A new microsporidium was isolated from Chilo suppressalis (Walker) (Lepidoptera: Pyralidae), one of the most important rice pests in China. The morphology and molecular systematics of this novel microsporidium were described in this study. The spores were long oval and measured 3.17 × 1.64 µm on fresh smears. Ultrastructure of the spores was characteristic for the genus Nosema: a diplokaryon, 10-12 polar filament coils of the same type, and posterior vacuole. Small subunit rRNA gene sequence data and phylogenetic analysis further confirmed that the microsporidian species from C. suppressalis belong to the true Nosema sub-group of the genus Nosema. Besides, the microsporidium Nosema sp. CS could cause systemic infection of Bombyx mori and infect silkworms through vertical transmission. Therefore, mulberry field pest control should be carefully monitored, and sanitation of mulberry leaves is essential to control the pebrine disease in sericulture.