A full-scale black soldier fly larvae (Hermetia illucens) bioconversion system for domestic biodegradable wastes to resource.

Domestic biodegradable wastes (DBW) pose a threat to environmental quality and human health. Bioconversion via black soldier fly larvae (BSFL; Hermitia illucens L.) is an expedient way for converting 'waste to resource' (insect protein and biofertilizer). Although researches abounded in laboratory-reared experiments and bioconversion mechanisms were pertinent, the void of data from actual and full-scale operation restricts the intensification of BSFL technology and its global adoption. Hence, a full-scale BSFL bioconversion system lasting 4 years in Hangzhou (China) was investigated, and the feasibility and efficiency of 15 tonnes of DBW per day were studied. Through continuous technical optimization, the average production of fresh larvae was increased from 8.5% in 2017 to 15.3% in 2020, along with bioconversion rate of final vermicompost decreased from 35.4% to 14.5%. The total biomass reduction rate in 2020 was 68.7 ± 17.4 kg/(m3 d), equivalent to 0.735 ± 0.215 kg/(kg d) in the form of fresh larvae. Crude fat in fresh larvae accounted for 13.4%, and crude protein accounted for 16.2% in which the determined amino acid profile bore a strong resemblance to fish meal only except histidine and tyrosine. Its economic benefits proved the feasibility of this technology, and the profit reached up to 35.9 US$ per tonne of DBW in 2019. In conclusion, BSFL bioconversion system under current 'insect-farm' operation was a promising solution for DBW treatment with value-added waste recycling.

Rumen fermentative metabolomic and blood insights into the effect of yeast culture supplement on growing bulls under heat stress conditions.

This study was conducted to investigate the effects of yeast culture supplements on the physiological state and growth performance of growing bulls under heat stress conditions and the underlying mechanism. A total of 14 (6.0 ± 1.0 months old) growing bulls with similar body weight were randomly assigned into the control group (YC0g/d ) and yeast culture supplement group (YC40g/d ). YC0g/d contained three replicates, with two bulls in each replicate, which were fed a basal diet. Meanwhile, the YC40g/d treatment contained four replicates, with two bulls in each replicate, which were fed a basal diet supplemented with 40 g/day of yeast culture per cattle. Growth performance, nutrient digestibility, rumen fermentable metabolites, serum immunity, serum hormones, and serum antioxidant parameters were measured. Results showed that the average daily gain significantly increased (P < 0.05), while the feed-to-gain ratio significantly decreased (P < 0.01) after YC supplementation compared with the YC0g/d . The digestibility of neutral detergent fiber (P < 0.05) was higher in YC40g/d . There were no significant differences in ruminal pH, NH3-N, butyrate, or acetate/propionate (P > 0.05). Besides, the rumen MCP, acetate, propionate, and total VFA content remarkably increased with the supplement of YC (P < 0.05). Yeast culture supplementation increased the concentration of nicotinamide riboside, neuromedin B, peptides, and formyl-5-hydroxykynurenamine. The YC40g/d group had a significantly (P < 0.05) higher serum triiodothyronine level, serum glutathione peroxidase levels, and total antioxidant capacity while having a lower serum malondialdehyde level than the YC0g/d group. In conclusion, the addition of yeast culture in the diet improves the growth performance of growing bulls under heat stress by increasing nutrient digestibility, rumen fermentation function, antioxidant capacity, and rumen metabolites.

Analysis of the risk factors for severe lung injury after radical surgery for tetralogy of fallot.

Objective: This study aimed to determine the risk factors for severe lung injury (SLI) (partial pressure of oxygen/fraction of inspired oxygen <150) after radical surgery for tetralogy of Fallot with pulmonary stenosis (TOF/PS) in children. Method: A retrospective analysis was conducted including a total of 287 children with TOF/PS aged below 10 years (including 166 males) who had undergone radical surgery at the Center of Pediatric Heart Disease of the Beijing Anzhen Hospital (China) from 2018 to 2020. Results: A total of 83 cases (28.9%) had SLI after surgery. Univariate analysis showed that age, weight, pulmonary artery index (PAI), cardiopulmonary bypass (CPB) time, and polymorphonuclear leukocyte (PMN) percentage on the first day after surgery were risk factors for postoperative SLI. Multivariate logistic regression analysis showed that PAI, PMN percentage on the first day postoperatively, and CPB time were independent risk factors for SLI after surgery. The prediction model was established as follows: Logit(P) = 2.236 + 0.009*CPB-0.008*PAI-0.035*PMN, area under the curve (AUC) = 0.683, P < 0.001, sensitivity 65.8%, and specificity 68.6%. Following surgery, static lung compliance was significantly lower in the SLI group compared with the routine group. Complication rates and mortality were significantly higher in the SLI than in the routine group. Ventilator support times, the length of intensive care unit stays, and the total lengths of hospital stay were significantly longer in the SLI than in the routine group. Conclusion: The occurrence of SLI following radical surgery for TOF in children significantly affected postoperative recovery, and PAI, PMN percentage on the first day postoperatively, and CPB time were independent risk factors for SLI.

Insights on dissolved organic matter and bacterial community succession during secondary composting in residue after black soldier fly larvae (Hermetia illucens L.) bioconversion for food waste treatment.

Black soldier fly larvae (Hermetia illucens L. BSFL) bioconversion is a promising biotechnology for food waste treatment. However, the separated residues still do not meet criteria for use as land application biofertilizers. In this work, we investigated a full-scale BSFL bioconversion project to explore features of dissolved organic matter (DOM) and its associated responses of bacterial community succession in residue during secondary composting. Data showed that the concentrations of total nitrogen and ammonium nitrogen decreased by 11.8% and 22.6% during the secondary composting, respectively, while the nitrate nitrogen concentration increased 18.7 times. The DOM concentration decreased by 69.1%, in which protein-like, alcohol-phenol, and biodegradable aliphatic substances were metabolized by bacteria during the thermophilic phase together with the accumulation of humus-like substances, resulting in an increase in the relative concentration of aromatic compounds. The structure of the bacterial community varied at different stages of the bioprocess, in which Bacteroidetes, Actinobacteria, Proteobacteria, and Firmicutes were the dominant bacterial phyla. Lysinibacillus, Pusillimonas, and Caldicoprobacter were found to be key contributors in the degradation and formation of DOM. The DOM concentration (33.4%) and temperature (17.7%) were the prime environmental factors that promoted succession of the bacterial community. Through bacterial metabolism, the structural stability of DOM components was improved during the composting process, and the degrees of humification and aromaticity were also increased. This study depicted the dynamic features of DOM and the associated bacterial community succession in residue during secondary composting, which is conducive with the reuse of BSFL residue as biofertilizer for agriculture.

Efficient Isolation and Characterization of a Cellulase Hyperproducing Mutant Strain of Trichoderma reesei.

A cellulase hyperproducing mutant strain, JNDY-13, was obtained using the ARTP mutation system and with Trichoderma reesei RUT-C30 as the parent strain. Whole-genome sequencing of JNDY-13 confirmed that 105 of the 653 SNPs were point mutations, 336 mutations were deletions and 165 were insertions. Moreover, 99 mutations were insertions and duplications. Among all the mutations, the one that occurred in the galactokinase gene might be related to the production of cellulases in T. reesei JNDY-13. Moreover, the up-regulation of cellulase and hemicellulase genes in JNDY-13 might contribute to higher cellulases production. Under optimal conditions, the highest cellulase activity by batch fermentation reached 4.35 U/ml, and the highest activity of fed-batch fermentation achieved was 5.40 U/ml.

[Optimization of coding sequences and expression of antimicrobial peptide magainin II in Escherichia coli and Pichia pastoris].

The antimicrobial peptide magainin II is expressed in the skin of the African clawed frog, Xenopus laevis, and exhibits a broad spectrum of antimicrobial activity as well as tumoricidal properties at low concentrations. In addition, magaininII plays a synergistic role during antimicrobial and tumoricidal processes with another antimicrobial peptide PGLa that is also expressed in Xenopus laevis. The optimized cDNA sequence of magainin II and magainin II-PGLa hybrid peptide according to E. coli or Pichia pastoris codon usage frequency were synthesized and sub-cloned into prokaryotic expression vector pGEX and Pichia pastoris secreted expression vector pPIC9k. The resulting recombinant plasmids were named as pGEX-magainin II and pPIC9k-magainin II-PGLa. The GST-magainin II fusion protein was highly expressed in E. coli. Furthermore, magainin II was successfully purified by digestion with PreScission Protease to cleave the GST tag. Additionally, our data obtained from the ELISA revealed that magainin II -PGLa hybrid peptide was successfully expressed in Pichia pastoris. These experiments establish a useful system for further studies of these antimicrobial peptides.

Bacterial consortia constructed for the decomposition of Agave biomass.

Research has shown that a greater variety of enzymes, as well as variety of microorganisms producing enzymes, can have an overall synergistic effect on the decomposition of lignocellulosic biomass for the production of value-added bio-products. Here, 8 cellulase-degrading bacterial isolates were selected to develop co-, tri-, and tetra-cultures for the decomposition of lignocellulosic biomass. Glucose and xylose equivalents released from imitation biomass media containing 0.5% (w/v) beechwood xylan and 0.5% (w/v) Avicel was measured using di-nitrosalicylic acid for all consortia, along with cell growth and survival. Thereafter, 6 co- and 2 tri-cultures with greatest decomposition were examined for ability to degrade Agave americana fiber. Interestingly, when strains were paired up in co-culture, four pairs: G+5, G+A, C+A1, and G+A1 produced high reducing sugars in 24 h: 6 µM, 8 µM, 8 µM, and finally, 6 µM, respectively. From 4 co-cultures with highest reducing sugar equivalents, tri- and tetra-cultures were produced. The bacterial consortia which had the highest reducing sugars detected were 2 tri-cultures: G + A1 + A4 and G + A1 + 5, displaying levels as high as 9 µM and 5 µM in day 1, respectively. All co- and tri-cultures maintained high cell survival for 14 days with 0.5 g ground Agave. Upon evaluating Agave dry weight after treatment, it was evident that almost half the biomass could be decomposed in 14 days. Scanning electron microscopy of treated Agave supported decomposition when compared with the control. These bacterial consortia have potential for further study of value-added by-product production during metabolism of lignocellulosic biomasses.

Optimization of technological conditions for one-pot synthesis of (S)-alpha-cyano-3-phenoxybenzyl acetate in organic media.

Optically active form of alpha-cyano-3-phenoxybenzyl (CPB) alcohol, building block of pyrethroid insecticides, was synthesized as its acetate by the combination of anion-exchange resin (D301)-catalyzed transcyanation between m-phenoxybenzaldehyde (m-PBA) and acetone cyanohydrin (AC), and lipase (from Alcaligenes sp.)-catalyzed enantioselective transesterification of the resulting cyanohydrin with vinyl acetate. Through optimizing technological conditions, the catalyzing efficiency was improved considerably compared to methods previously reported. Concentrations of CPB acetate were determined by gas chromatograph. The enantio excess (e.e.) values of CPB acetate were measured by NMR (nuclear magnetic resonance) method. Effects of solvents and temperatures on this reaction were studied. Cyclohexane was shown to be the best solvent among the three tested solvents. 55 degrees C was the optimal temperature for higher degree of conversion. External diffusion limitation was excluded by raising the rotational speed to 220 r/min. However, internal diffusion could not be ignored, since the catalyst (lipase) was an immobilized enzyme and its particle dimension was not made small enough. The reaction rate was substantially accelerated when the reactant (m-PBA) concentration was as high as 249 mmol/L, but decreased when the initial concentration of m-PBA reached to 277 mmol/L. It was also found that the catalyzing capability of recovered lipase was high enough to use several batches. Study of the mole ratio of AC to m-PBA showed that 2:1 was the best choice. The strategy of adding base catalyst D301 was found to be an important factor in improving the degree of conversion of the reaction from 20% to 80%. The highest degree of conversion of the reaction has reached up to 80%.

Decomposition dynamic of higher plant pigments by HPLC analysis.

The fate of the litter of dominant vegetation(willows and reeds) is one of the aspects studied in the frame of the project "Onderzoek Milieu Effecten Sigmaplan". One of the questions to be considered is how long the litter stays within the estuary. In this paper, the time the leaf litter (Salix triandra and Phragmites australis) stayed in the Schelde estuary was studied by using plant pigment as biomarkers with HPLC application. After analyzing the original data from the incubation experiment described by Dubuison and Geers (1999), the decomposition dynamics patterns of pigments were analyzed and described, and these decomposition dynamics patterns were used as calibration patterns. By using Spearman Rank Order Correlation, the calibration patterns of the pigments which were significant (p < 0.05) were grouped. In this way, several groups of the calibration patterns of pigment decomposition were achieved. The presence or absence of these groups of pigments (whether they can be detected or not from HPLC) was shown to be useful in determining the time the litter has stayed in the water. Combining data of DW and POC, more precise timing can be obtained.