Larval biomass production from the co-digestion of mushroom root waste and soybean curd residues by black soldier fly larvae (Hermetia illucens L.).

People are increasingly using black soldier fly larvae (BSFL) as a sustainable waste management solution. They are high in protein and other essential nutrients, making them an ideal food source for livestock, poultry, and fish. Prior laboratory studies with BSFL developed on pure mushroom root waste (MRW) showed poor conversion efficiency compared to a regular artificial diet. Therefore, we mixed the nutrient-rich soybean curd residues (SCR) with MRW in different ratios (M2-M5). Pure mushroom root waste (M1, MRW 100%) had the lowest survival rate (86.2%), but it increased up to 96.9% with the SCR percentage increasing. M1 had the longest developmental period (31.1 days) and the lowest BSFL weight (7.4 g). However, the addition of SCR reduced the development time to 22.0 and 21.5 days in M4 (MRW 40%, SCR 60%) and M5 (MRW 20%, SCR 80%), respectively, and improved the larval weight to 10.9 g in M4 and 11.8 g in M5. Other groups did not have as much feed conversion ratio (FCR) (8.4 for M4 and M5), bioconversion (M4 5.4%; M5 5.9%), or lipid content (M4 25.2%; M5 24.3%). These mixtures did. Compare this to M1. We observed better results, with no significant differences between the M4 and M5 groups and their parameters. In the present study, our main target was to utilize more MRW. Therefore, we preferred the M4 group in our nutritional and safety investigation and further compared it with the artificial diet (M7). The heavy metals and essential amino acids (histidine 3.6%, methionine 2.7%, and threonine 3.8%) required for human consumption compared to WHO/FAO levels showed satisfactory levels. Furthermore, fatty acids (capric acid 1.9%, palmitic acid 15.3%, oleic acid 17.3%, and arachidonic acid 0.3%) also showed higher levels in M4 than M7. The SEM images and FT-IR spectra from the residues showed that the BSFL in group M4 changed the structure of the compact fiber to crack and remove fibers, which made the co-conversion mixture better.

Cellulose-degrading bacteria improve conversion efficiency in the co-digestion of dairy and chicken manure by black soldier fly larvae.

Black soldier fly larvae (BSFL) have potential utility in converting livestock manure into larval biomass as a protein source for livestock feed. However, BSFL have limited ability to convert dairy manure (DM) rich in lignocellulose. Our previous research demonstrated that feeding BSFL with mixtures of 40% dairy manure and 60% chicken manure (DM40) provides a novel strategy for significantly improving their efficiency in converting DM. However, the mechanisms underlying the efficient conversion of DM40 by BSFL are unclear. In this study, we conducted a holistic study on the taxonomic stucture and potential functions of microbiota in the larval gut and manure during the DM and DM40 conversion by BSFL, as well as the effects of BSFL on cellulosic biodegradation and biomass production. Results showed that BSFL can consume cellulose and other nutrients more effectively and harvest more biomass in a shorter conversion cycle in the DM40 system. The larval gut in the DM40 system yielded a higher microbiota complexity. Bacillus and Amphibacillus in the BSFL gut were strongly correlated with the larval cellulose degradation capacity. Furthermore, in vitro screening results for culturable cellulolytic microbes from the larval guts showed that the DM40 system isolated more cellulolytic microbes. A key bacterial strain (DM40L-LB110; Bacillus subtilis) with high cellulase activity from the larval gut of DM40 was validated for potential industrial applications. Therefore, mixing an appropriate proportion of chicken manure into DM increased the abundance of intestinal bacteria (Bacillus and Amphibacillus) producing cellulase and improved the digestion ability (particularly cellulose degradation) of BSFL to cellulose-rich manure through changes in microbial communities composition in intestine. This study reveals the microecological mechanisms underlying the high-efficiency conversion of cellulose-rich manure by BSFL and provide potential applications for the large-scale cellulose-rich wastes conversion by intestinal microbes combined with BSFL.

Black soldier fly, Hermetia illucens as a potential innovative and environmentally friendly tool for organic waste management: A mini-review.

The application of black soldier fly (BSF), Hermetia illucens based technology to process organic wastes presents a practical option for organic waste management by producing feed materials (protein, fat), biodiesel, chitin and biofertilizer. Therefore, BSF organic wastes recycling is a sustainable and cost-effective process that promotes resource recovery, and generates valuable products, thereby creating new economic opportunities for the industrial sector and entrepreneurs. Specifically, we discussed the significance of BSF larvae (BSFL) in the recycling of biowaste. Despite the fact that BSFL may consume a variety of wastes materials, whereas, certain lignocellulosic wastes, such as dairy manure, are deficient in nutrients, which might slow BSFL development. The nutritional value of larval feeding substrates may be improved by mixing in nutrient-rich substrates like chicken manure or soybean curd residue, for instance. Similarly, microbial fermentation may be used to digest lignocellulosic waste, releasing nutrients that are needed for the BSFL. In this mini-review, a thorough discussion has been conducted on the various waste biodegraded by the BSFL, their co-digestion and microbial fermentation of BSFL substrate, as well as the prospective applications and safety of the possible by-products that may be generated at the completion of the treatment process. Furthermore, this study examines the present gaps and challenges on the direction to the efficient application of BSF for waste management and the commercialization of its by-products.

Synergistic bioconversion of organic waste by black soldier fly (Hermetia illucens) larvae and thermophilic cellulose-degrading bacteria.

Introduction: This study examines the optimum conversion of Wuzhishan pig manure by Black Soldier Fly Larvae (BSFL) at various phases of development, as well as the impact of gut microbiota on conversion efficiency. Method and results: In terms of conversion efficiency, BSFL outperformed the growing pig stage (GP) group, with significantly higher survival rates (96.75%), fresh weight (0.23 g), and larval conversion rate (19.96%) compared to the other groups. Notably, the GP group showed significant dry matter reductions (43.27%) and improved feed conversion rates (2.17). Nutritional composition varied, with the GP group having a lower organic carbon content. High throughput 16S rRNA sequencing revealed unique profiles, with the GP group exhibiting an excess of Lactobacillus and Clostridium. Promising cellulose-degrading bacteria in pig manure and BSFL intestines, including Bacillus cereus and Bacillus subtilis, showed superior cellulose degradation capabilities. The synergy of these thermophilic bacteria with BSFL greatly increased conversion efficiency. The BSFL1-10 group demonstrated high growth and conversion efficiency under specific conditions, with remarkable larval moisture content (71.11%), residual moisture content (63.20%), and waste reduction rate (42.28%). Discussion: This study sheds light on the optimal stages for BSFL conversion of pig manure, gut microbiota dynamics, promising thermophilic cellulose-degrading bacteria, and the significant enhancement of efficiency through synergistic interactions. These findings hold great potential for sustainable waste management and efficient biomass conversion, contributing to environmental preservation and resource recovery.

Pharmacological Validation for the Folklore Use of Ipomoea nil against Asthma: In Vivo and In Vitro Evaluation.

Oxidative stress is the key factor that strengthens free radical generation which stimulates lung inflammation. The aim was to explore antioxidant, bronchodilatory along with anti-asthmatic potential of folkloric plants and the aqueous methanolic crude extract of Ipomoea nil (In.Cr) seeds which may demonstrate as more potent, economically affordable, having an improved antioxidant profile and providing evidence as exclusive therapeutic agents in respiratory pharmacology. In vitro antioxidant temperament was executed by DPPH, TFC, TPC and HPLC in addition to enzyme inhibition (cholinesterase) analysis; a bronchodilator assay on rabbit's trachea as well as in vivo OVA-induced allergic asthmatic activity was performed on mice. In vitro analysis of 1,1-Diphenyl-2-picrylhydrazyl radical (DPPH) expressed as % inhibition 86.28 ± 0.25 with IC50 17.22 ± 0.56 mol/L, TPC 115.5 ± 1.02 mg GAE/g of dry sample, TFC 50.44 ± 1.06 mg QE/g dry weight of sample, inhibition in cholinesterase levels for acetyl and butyryl with IC50 (0.60 ± 0.67 and 1.5 ± 0.04 mol/L) in comparison with standard 0.06 ± 0.002 and 0.30 ± 0.003, respectively, while HPLC characterization of In.Cr confirmed the existence with identification as well as quantification of various polyphenolics and flavonoids i.e., gallic acid, vanillic acid, chlorogenic acid, quercetin, kaempferol and others. However, oral gavage of In.Cr at different doses in rabbits showed a better brochodilation profile as compared to carbachol and K+-induced bronchospasm. More significant (p < 0.01) reduction in OVA-induced allergic hyper-responses i.e., inflammatory cells grade, antibody IgE as well as altered IFN-α in airways were observed at three different doses of In.Cr. It can be concluded that sound mechanistic basis i.e., the existence of antioxidants: various phenolic and flavonoids, calcium antagonist(s) as well as enzymes' inhibition profile, validates folkloric consumptions of this traditionally used plant to treat ailments of respiration.

The Spasmolytic, Bronchodilator, and Vasodilator Activities of Parmotrema perlatum Are Explained by Anti-Muscarinic and Calcium Antagonistic Mechanisms.

Parmotremaperlatum is traditionally used in different areas of Pakistan to treat gastrointestinal, respiratory, and vascular diseases. This study evaluates the underlying mechanisms for traditional uses of P. perlatum in diarrhea, asthma, and hypertension. In vitro pharmacological studies were conducted using isolated jejunum, trachea, and aortic preparations, while the cytotoxic study was conducted in mice. Crude extract of P. perlatum(Pp.Cr), comprising appreciable quantities of alkaloids and flavonoids, relaxed spontaneously contracting jejunum preparation, K+ (80 mM)-induced, and carbachol (1 µM)-induced jejunum contractions in a concentration-dependent manner similar to dicyclomine and dantrolene. Pp.Cr showed a rightward parallel shift of concentration-response curves (CRCs) of Cch after a non-parallel shift similarto dicyclomine and shifted CRCs of Ca+2 to rightward much likeverapamil and dantrolene, demonstrating the coexistence of antimuscarinic and Ca+2 antagonistic mechanism. Furthermore, Pp.Cr, dicyclomine, and dantrolene relaxed K+ (80 mM)-induced and Cch (1 µM)-induced tracheal contractions and shifted rightward CRCs of Cch similar to dicyclomine, signifying the dual blockade. Additionally, Pp.Cr also relaxed the K+ (80 mM)-induced and phenylephrine (1 µM)-induced aortic contraction, similarly to verapamil and dantrolene, suggesting Ca+2 channel antagonism. Here, we explored for the first time thespasmolytic and bronchodilator effects of Pp.Crand whether they maybe due to the dual blockade of Ca+2 channels and muscarinic receptors, while the vasodilator effect might be owing to Ca+2 antagonism. Our results provide the pharmacological evidence that P. perlatum could be a new potential therapeutic option to treat gastrointestinal, respiratory, and vascular diseases. Hence, there is a need for further research to explore bioactive constituent of P. perlatum as well as further investigation by suitable experimental models are required to further confirm the importance and usefulness of P. perlatum in diarrhea, asthma, and hypertension treatment.

In silico bimolecular characterization of anticancer phytochemicals from Fagonia indica.

The in silico molecular dynamics and structure-based site-specific drug design of indigenous plant biomolecules and selected proteins have remarkable potential for cancer therapy. A set of five proteins included for this research were epidermal growth factor protein (PDB ID; 1M17), crystal structure of mutated EGFR kinase (PDB ID; 2EB3), crystal structure of Bcl-xl (PDB ID; 2YXJ), apoptosis regulator protein MCL-1 BH3 (PDB ID; 3MK8) and apoptosis proteins (PDB ID; 5C3H). The present study on in silico investigation of fifteen indigenous medicinal plants were selected there one hundred thirty four ligands available literature were docked against five proteins involved in carcinogenesis. The highest scoring in silico plant, Fagonia indica was subjected to in vitro cytotoxic effects on HCT116, HepG-2 and HeLa human carcinoma cell lines. Molecular dynamics showed best ligand-protein inhibition interaction between Coumarin-2xyj and Kaempferol-2eb3 with promising binding affinities. Whereas, on HeLa human cervical cancer cell line IC50 was 28.3±0.102/ml. Fagonia indica could be potential source from natural products that have cytotoxic properties against cervical cancer cells by blocking mutant epidermal growth factor tyrosine or peroxisome proliferators activated receptor proteins.

Physicochemical, antidiarrheal and antidiabetic potential of super food (Moringa oleifera Lam.).

There is a long history of natural products for the treatment of infections and diseases. The objective of present study was to investigate the organoleptic, microscopic, physico-chemical, phytochemical, antidiarrheal and antidiabetic potential of leaf, flowering bud and stem bark of Moringa oleifera L. Macroscopic, microscopic, physico-chemical parameters and phytochemical screening were carried out. Diarrhea was induced with castor oil (10ml/kg), verapamil (3, 10 and 30mg/kg) were used as standard antidiarrheal drug and extract of Moringa oleifera at (100, 300 and 1000mg/kg) was used for treatment. Alpha glucosidase inhibitory assay was carried out by using acarbose (0.5mM) and extracts (5.0 mg/Ml). Diabetes was induced by alloxan (150mg/kg), while glibenclamide (10mg/kg) was used as standard drug, and extracts (at the doses of 500mg/kg) were used to determine the antidiabetic activity. Results showed the presence of primary and secondary metabolites, treatment at the dose of 1.0g/kg of leaf, flowering bud and stem bark showed 94 ±2.527, 85.42±5.460 and 84.58±6.138% protection respectively whereas verapamil (10mg/kg) showed 94.84±3.27% protection. Alpha glucosidase inhibition of stem bark (0.5mg/ml) was 95.43±1.47 and flowering bud 94.78±1.25 whereas acarbose (5mM) inhibition was 92.23±0.14%. Stem bark and flowering bud extract (500mg/kg) decreases the blood glucose level from 388.5±35.83 to 226.3±47.10 and 322.5±48.35 to 173.8±29.5 respectively whereas glibenclamide (10 mg/kg) decreases the blood glucose level from 320.7±22.9 to 146.3±17.7 and increases the body weight of the experimental animal. It was concluded from the results that stem bark has strong antidiabetic potential while leaves of the plant have promising antidiarrheal effect.

Response of lymphatic tissues to natural feed additives, curcumin (Curcuma longa) and black cumin seeds (Nigella sativa), in broilers against Pasteurella multocida.

The antibiotic residues and pathogenic resistance against the drug are very common in poultry because of antibiotics used in their feed. It is necessary to use natural feed additives as effective alternatives instead of a synthetic antibiotic. This study aimed to investigate the immune response of Nigella sativa and Curcuma longa in broilers under biological stress against Pasteurella multocida. The total 100, one-day-old chicks were divided into 5 groups. Groups 1 and 2 served as control negative and control positive. Both control groups were receiving simple diet without any natural feed additives, but the infection was given in group 2 at day 28 with the dose of 5.14 × 107 CFU by IV. Groups 3A and 3B were offered 2% seed powder of Nigella sativa, groups 4A and 4B were offered C. longa 1% in powdered form, and group 5A and 5B were offered both C. longa 1% and N. sativa 2% in the feed from day 1 and groups 3B, 4B, and 5B were challenged with P. multocida. The haemagglutination inhibition titter against Newcastle Disease virus (NDV), feed conversion ratio, mortality, gross, and histopathology were studied. The results of this study revealed that hemagglutination inhibition titers against NDV were highly significant (P < 0.05) in treated groups, highest titers (3A, 6.8; 3B, 6.4; and 5A, 7.2) were obtained from treated Groups. The feed conversion ratio of N. sativa + C. longa treated groups (5A, 1.57, and 3A, 1.76) were higher than that of other nontreated groups. The gross and histopathological changes were much severe in control positive, but fewer changes were seen in treated groups. Therefore, we recommend that natural feed additives, black cumin (N. sativa) and turmeric (C. longa), act as an immune enhancer in broilers against P. multocida.

Correction: Saeed, A., et al. Incidence of Vancomycin-Resistant Phenotype of the Methicillin-Resistant Staphylococcus aureus Isolated from a Tertiary Care Hospital in Lahore. Antibiotics 2020, 9, 3.

Staphylococcus aureus (S. aureus)-associated infections are one of the major threats to public health. The aim of the present study was to determine the antibiotic resistance pattern, as well as the genetic characterization, of methicillin and vancomycin-resistant S. aureus (VRSA) isolated from a tertiary care hospital in Lahore. The S. aureus isolates were isolated from different clinical samples, identified by biochemical testing, and subjected to antibiotic susceptibility testing via the disc diffusion method or broth microdilution method. The methicillin resistance gene (mecA) and vancomycin resistance gene (vanA) were amplified by the polymerase chain reaction. The S. aureus isolates showed high incidences of resistance against methicillin (76%) and moderate incidences of resistance to vancomycin (14%). Isolates were also resistant to several other drugs, such as cefoxitin (76%), ertapenem (83%), ampicillin (81%), tobramycin (78%), moxifloxacin (76%), and tetracycline (74%). An encouraging finding was that 98% of isolates were susceptible to tigecycline, indicating its possible role in the treatment of methicillin-resistant Staphylococcus aureus (MRSA) and VRSA, as well as the multi-drug resistant S. aureus. The mecA gene was detected in 33.3% of tested isolates (10/30), while the vanA gene was also detected in 30% (9/30) of the tested isolates. In conclusion, the frequent presence of methicillin and vancomycin resistance in S. aureus appraises the cautious use of these antibiotics in clinical practices. Furthermore, it is suggested that there should be continuous monitoring of tigecycline treatments in clinical setups in order to delay the development of resistance against it.

Physicochemical structure of chitin in the developing stages of black soldier fly.

Black soldier fly can convert organic wastes into their own functional biological macromolecules i.e. chitin that has great potential for biotechnological, biomedical and cosmetic application. The variation in the physiochemical structure of chitin was proved for several insects during metamorphosis stages whereas, it remains unknown for black soldier fly (BSF), a recognized resource insect for industrial production and organic waste management. The current work noted results on how the chitin matrix can undergo physicochemical changes during the developmental phases (larvae, prepupa, puparium, and adults) of BSF. Chitin content was determined around 3.6%, 3.1%, 14.1% and 2.9%, respectively. Fourier transform infrared spectroscopy, thermogravimetric analysis and x-ray diffraction analysis showed that chitin from BSF different stages was all α-chitin with similar thermal stability. The chitin crystalline index increased gradually with development from larvae to adult, 33.09%, 35.14%, 68.44% and 87.92%, respectively. Moreover, it was observed by scan electron microscopy that surface morphology characteristics of chitin vary significantly in developmental phases. These results confirmed that Hermetia illucens is promising for converting organic wastes into valuable biopolymers i.e. chitin and its physiochemical properties in various developmental stages help to determine the related biomedical, biotechnological, cosmetic and functional food utilization potential.

Incidence of Vancomycin Resistant Phenotype of the Methicillin Resistant Staphylococcus aureus Isolated from a Tertiary Care Hospital in Lahore.

Staphylococcus aureus (S. aureus) associated infections are one of the major threats to public health. The aim of the present study was to determine the antibiotic resistance pattern as well as the genetic characterization of methicillin and vancomycin resistant S. aureus (VRSA) isolated from a tertiary care hospital in Lahore. The S. aureus were isolated from different clinical samples, identified by biochemical testing, and subjected to antibiotic susceptibility testing by disc diffusion method or broth microdilution method. Methicillin resistance gene (mecA) and vancomycin resistance gene (vanA) were amplified by polymerase chain reaction. The S. aureus isolates showed high incidence of resistance against methicillin (76%) and moderate incidence of resistance to vancomycin (44%). Isolates were also resistant to several other drugs such as cefoxitin (76%), ertapenem (83%), ampicillin (81%), tobramycin (78%), moxifloxacin (76%), and tetracycline (74%). An encouraging finding was that 98% of isolates were susceptible to tigecycline, indicating its possible role in the treatment of MRSA, VRSA, as well as multi-drug resistant S. aureus. The mecA gene was detected from 33.3% (10/30) while vanA gene was detected from 46.6% (14/30) of the tested isolates. In conclusion, frequent presence of methicillin and vancomycin resistance in S. aureus appraises the cautious use of these antibiotics in clinical practices. Furthermore, it is suggested that there should be continuous monitoring of tigecycline treatments in clinical setups in order to delay the development of resistance against it.

Development, characterization and evaluation of ginger extract loaded microemulsion: In vitro and Ex vivo release studies.

Zingeber officinale (ginger) has been used for a long time in conventional medicine for the management of many diseases most important of which is inflammatory diseases. The aim of this study was formulation of topical microemulsion system to enhance the solubility, stability and release profile of ginger extract, as it is unstable in the presence of light, air, heat and long term storage. The solubility of ginger extract in different oils, surfactants, and cosurfactants was determined in order to find the optimal components for microemulsion. Isopropyl myristate (IPM) was selected as oil phase, tween 80 and PEG 400 were selected as surfactant and co-surfactant respectively based on highest solubility values. Pseudo-ternary phase diagram was constructed in order to find out the microemulsion region. The prepared microemulsions were evaluated for pH, viscosity, conductivity, refractive index, globular size, zeta potential, polydispersity index, ginger extract content, in-vitro and ex-vivo release profiles. The formulation GE1 showed best physicochemical properties with smallest globular size (19.75nm), highest release rate and flux value. It also showed significant (p<0.05) anti-inflammatory effect as compared to reference piroxicam drug solution. It is concluded that ginger extract can be used to develop stable microemulsion system with better skin permeation and promising antiinflammatory activity.

De novo transcriptome sequencing and analysis revealed the molecular basis of rapid fat accumulation by black soldier fly (Hermetia illucens, L.) for development of insectival biodiesel.

BACKGROUND: Black soldier fly (BSF, Hermetia illucens L.) can efficiently degrade organic wastes and transform into a high fat containing insect biomass that could be used as feedstock for biodiesel production. Meanwhile, the molecular regulatory basis of fat accumulation by BSF is still unclear; it is necessary to identify vital genes and regulators that are involved in fat accumulation. RESULTS: This study analyzed the dynamic state of fat content and fatty-acid composition of BSF larvae in eight different stages. The late prepupa stage exhibited the highest crude fat, with lauric acid being the main component. Therefore, to provide insight into this unexplained phenomenon, the molecular regulation of rapid fat accumulation by BSF larvae was investigated. The twelve developmental stages of BSF were selected for transcriptome analysis, including the eight stages used for investigation of fat content and fatty-acid composition. By Illumina sequencing, 218,295,450,000 nt were generated. Through assembly by Trinity, 70,475 unigenes were obtained with an average length of 1064 nt and an N50 of 1749 nt. The differentially expressed unigenes were identified by DESeq, with 9159 of them being up-regulated and 10,101 of them were down-regulated. The several putative genes that are involved in the formation of pyruvate, acetyl-CoA biosynthesis, acetyl-CoA transcription, fatty-acid biosynthesis, and triacylglycerol biosynthesis were identified. The four vital metabolic genes that are associated with fat accumulation were validated by quantitative real-time PCR (qRT-PCR). The molecular mechanism of fat accumulation in BSF was clarified in this investigation through the construction of a detailed fat accumulation model from our results. CONCLUSION: The study provides an unprecedented level of insight from transcriptome sequencing to reveal the crude fat accumulation mechanism in developing BSF. The finding holds considerable promise for insectival biodiesel production, and the fat content and fatty-acid composition can be altered by genetic engineering approaches in the future for the insect production industry.

Enhanced bioconversion of dairy and chicken manure by the interaction of exogenous bacteria and black soldier fly larvae.

Generation of insects' biomass from lignocellulose rich organic wastes is of significant challenges in reducing the environmental impact of wastes and in sustaining feed and food security. This research looked at the effects of lignocellulotic exogenous bacteria in the black soldier fly (BSF) organic waste conversion system for biomass production and lignocellulose biodegradation of dairy and chicken manures. Six exogenous bacteria were investigated for cellulolytic activity with carboxymethyl cellulose and found that these tested bacterial strains degrade the cellulose. In this study; a co-conversion process using Hermetia illucens larvae to convert the previously studied best mixing ratio of dairy manure (DM) and chicken manure (CHM) (2:3) and cellulose degrading bacteria was established to enhance the larval biomass production, waste reduction and manure nutrient degradation. BSF larvae assisted by MRO2 (R5) has the best outcome measures: survival rate (99.1%), development time (19.0 d), manure reduction rate (48.7%), bioconversion rate (10.8%), food conversion ratio (4.5), efficiency of conversion of ingestion (22.3), cellulose (72.9%), hemicellulose (68.5%), lignin (32.8%), and nutrient utilization (protein, 71.2% and fat, 67.8%). By analyzing the fiber structural changes by scanning electron microscopy and Fourier-transformed infrared spectroscopy (FT-IR), we assume that exogenous bacteria assist the BSF larvae that trigger lead to structural and chemical modification of fibers. We hypothesized that these surface and textural changes are beneficial to the associated gut bacteria, thereby helping to larval growth and reduce waste. The finding of the investigation showed that enhanced conversion of DM and CHM by BSF larvae assisted with lignocellulotic exogenous bacteria could play key role in the manure management.

Dynamic Effects of Initial pH of Substrate on Biological Growth and Metamorphosis of Black Soldier Fly (Diptera: Stratiomyidae).

Edible insects have become a recognized alternative and sustainable source of high-quality proteins and fats for livestock or human consumption. In the production process of black soldier fly (BSF), (Hermetia illucens L. [Diptera: Stratiomyidae]), initial substrate pH is a critical parameter to ensure the best value of insect biomass, life history traits, and quality bio-fertilizer. This study examined the impact of initial pH values on BSF larvae production, development time, and adult longevity. The BSF were reared on artificial diet with initial pH of 2.0, 4.0, 6.0, 8.0, and 10.0; the control was set at 7.0. Final BSF larval weight was significantly greater in substrates having initial pH 6.0 (0.21 g), control 7.0 (0.20 g), and 10.0 (0.20 g) with no significant difference among them, whereas larval weight reared with initial pH 2.0 and 4.0 were lowest at 0.16 g (-23%). Prepupal weight was greatest when larvae were reared on substrates with initial pH 6.0 (0.18 g), control 7.0 (0.19 g), 8.0 (0.18 g), and 10.0 (0.18 g). In contrast, the prepupal weight of larvae reared on diets with initial pH 2.0 was lowest at 0.15 g (-22%). Larval development time was 21.19 d at pH 8.0, about 3 d (12.5%) shorter than that of those reared on diets with initial pH 6.0, 7.0 control, and 10.0. In all treatments, pH shifted to 5.7 after 3-4 d and 8.5 after 16-17 d except for two groups (2.0 and 4.0) where the pH remained slightly acidic 5.0 and 6.5, respectively.

Insect biorefinery: a green approach for conversion of crop residues into biodiesel and protein.

BACKGROUND: As a major lignocellulosic biomass, which represented more than half of the world's agricultural phytomass, crop residues have been considered as feedstock for biofuel production. However, large-scale application of this conventional biofuel process has been facing obstacles from cost efficiency, pretreatment procedure, and secondary pollution. To meet the growing demands for food, feed, and energy as the global population continues to grow, certain kinds of insects, many of which are voracious feeders of organic wastes that may help address environmental, economic, and health issues, have been highlighted as a source of protein and fat. RESULTS: The biorefinery studied includes initial corn stover degradation by yellow mealworm (Tenebrio molitor L.), followed by a second stage that employs black soldier fly (Hermetia illucens L.), to utilize the residues produced during the first stage. These two insect-based biorefinery yielded 8.50 g of insect biomass with a waste dry mass reduction rate of 51.32%, which resulted in 1.95 g crude grease from larval biomass that produced 1.76 g biodiesel, 6.55 g protein, and 111.59 g biofertilizer. The conversion rate of free fatty acids of crude grease into biodiesel reached 90%. The components of cellulose, hemicellulose, and lignin contained in corn stover hydrolyzed harmoniously, resulting in declines of 45.69, 51.85, and 58.35%, respectively. Moreover, fluctuations in lipid, protein, and reducing sugar were also analyzed. CONCLUSION: The investigation findings demonstrated that successive co-conversion of corn stover by insects possessing different feeding habits could be an attractive option for efficient utilization of lignocellulosic resources, and represents a potentially valuable solution to crop residues management, rise of global liquid energy, and animal feed demand.

Cellulose decomposition and larval biomass production from the co-digestion of dairy manure and chicken manure by mini-livestock (Hermetia illucens L.).

World trends toward the modern dairies intensification on large production units cause massive animal manure production and accumulation. Improper handling of manure produced by industrial farm operation greatly deteriorates the major environmental media including air, water and soil. The black soldier fly utilizes organic waste and converts it into larvae biomass to be used as livestock feed and into residues to be used as bio-fertilizer. However, due to the high ratio of cellulose, hemicellulose and lignin in dairy manure, this conversion is difficult. Therefore, dairy manure treated with chicken manure was digested by Hermetia illucens. In this paper, we found that the co-digestion process significantly enhanced the larval production, waste mass reduction, rate of larvae conversion, feed conversion ratio, nutrient reduction and fibers utilization. Whereas 40% dairy manure and 60% chicken manure group show better results than other manure mixtures and had a significantly increased the cellulose consumption by 61.19%, hemicellulose consumption by 53.22% and lignin consumption by 42.23% compared with 49.89%, 49.77% and 31.95%, respectively, in the dairy-only manure group. Finally, scanning electron microscopy was used to analyze the structural changes of dairy manure, chicken manure and their co-digestion mixtures. The scan electron microscopy showed the deterioration in the structure of dairy and chicken manure fibers by Hermetia illucens. Moreover, the carbon-nitrogen ratio was decreased in all end products of post vermicomposting. The results suggest that the co-digestion of 40% dairy manure with 60% chicken manure is an appropriate proportion for dairy manure management with the black soldier fly.

Allele frequency distribution of CYP2C19*2 allelic variants associated with clopidogrel resistance in cardiac patients.

Drug resistance is a phenomenon that has become a critical issue in medical practice. Such is the case in the response to clopidogrel treatment, which is variable inter-individually and inter-ethnically due to genetic polymorphisms in the cytochrome P40 (CYP) gene. Clopidogrel is an anti-platelet agent administered to cardiac patients in the form of a prodrug, which is further metabolized into an active form by CYP enzymes. There are many allelic variants of the CYP gene that are involved in clopidogrel resistance, of which CYP2C19*2 has been demonstrated to be one of the most significant loss-of-function alleles. In the present study, 100 cardiac patients with percutaneous coronary intervention (PCI) or acute coronary syndrome (ACS) who were undergoing treatment with clopidogrel were selected and the patients were analyzed for CYP2C19*2 allelic variants using an allele-specific primer extension polymerase chain reaction method. The variant amplicons were visualized on gel and validated by Sanger sequencing. The observed allelic frequency distribution of CYP2C19*2 variants was 18% heterozygous for CYP2C19*2 A/C/G variants, 35% heterozygous for A/G variants, 13% heterozygous for C/G variants, 6% heterozygous for A/C variants, 7% homozygous for A variant, 5% homozygous for C variant and 16% homozygous for G wild-type. Furthermore, tri-allelic single nucleotide polymorphisms (SNPs) were identified in the CYP2C19*2 allele in cardiac patients for the first time, to the best of our knowledge; these were CYP2C19*2 A/C/G SNPs (18%). The overall frequency observed for new allelic variant C of CYP2C19*2 was 42%. These results suggested that there are significant inter-ethnic variations in the allelic frequencies of CYP2C19*2, which may be responsible for the variable clopidogrel response in cardiac patients.