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.

Insights into the reduction of antibiotic-resistant bacteria and mobile antibiotic resistance genes by black soldier fly larvae in chicken manure.

The increasing prevalence of antibiotic-resistant bacteria (ARB) from animal manure has raised concerns about the potential threats to public health. The bioconversion of animal manure with insect larvae, such as the black soldier fly larvae (BSFL, Hermetia illucens [L.]), is a promising technology for quickly attenuating ARB while also recycling waste. In this study, we investigated BSFL conversion systems for chicken manure. Using metagenomic analysis, we tracked ARB and evaluated the resistome dissemination risk by investigating the co-occurrence of antibiotic resistance genes (ARGs), mobile genetic elements (MGEs), and bacterial taxa in a genetic context. Our results indicated that BSFL treatment effectively mitigated the relative abundance of ARB, ARGs, and MGEs by 34.9%, 53.3%, and 37.9%, respectively, within 28 days. Notably, the transferable ARGs decreased by 30.9%, indicating that BSFL treatment could mitigate the likelihood of ARG horizontal transfer and thus reduce the risk of ARB occurrence. In addition, the significantly positive correlation links between antimicrobial concentration and relative abundance of ARB reduced by 44.4%. Moreover, using variance partition analysis (VPA), we identified other bacteria as the most important factor influencing ARB, explaining 20.6% of the ARB patterns. Further analysis suggested that antagonism of other bacteria on ARB increased by 1.4 times, while nutrient competition on both total nitrogen and crude fat increased by 2.8 times. Overall, these findings provide insight into the mechanistic understanding of ARB reduction during BSFL treatment of chicken manure and provide a strategy for rapidly mitigating ARB in animal manure.

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.

Isolated and identified pathogenic bacteria from black soldier fly larvae with "soft rot" reared in mass production facilities and its incidence characteristics.

The black soldier fly larvae (BSFL) can transform organic waste into high-end proteins, lipids, chitin, biodiesel, and melanin at an industrial scale. But scaling up of its production capacity has also posed health risks to the insect itself. In this investigation, larval "soft rot" which is occurring in mass production facilities that cause larval developmental inhibition and a certain degree of death was reported. Responsible pathogen GX6 was isolated from BSFL with "soft rot" and identified to be Paenibacillus thiaminolyticus. No obvious impact on larval growth was observed when treated with GX6 spores, whereas mortality of 6-day-old BSFL increased up to 29.33% ± 2.05% when GX6 vegetative cells (1 × 106 cfu/g) were inoculated into the medium. Moreover, higher temperature further enhanced the BSFL mortality and suppressed larval development, but increasing substrate moisture showed the opposite effect. The middle intestine of infected larvae became swollen and transparent after dissection and examination. Transmission electron microscopy (TEM) observation indicated that GX6 had destroyed the peritrophic matrix and intestinal microvilli and damaged epithelial cells of larval gut. Furthermore, 16S rRNA gene sequencing analysis of intestinal samples revealed that gut microflora composition was significantly altered by GX6 infection as well. It can be noticed that Dysgonomonas, Morganella, Myroides, and Providencia bacteria became more numerous in the intestines of GX6-infected BSFL as compared to controls. This study will lay foundations for efficient control of "soft rot" and promote healthy development of the BSFL industry to contribute to organic waste management and circular economy.

Mitigation of antibiotic resistome in swine manure by black soldier fly larval conversion combined with composting.

The increasing prevalence of antibiotic resistance genes (ARGs) in animal manure has attracted considerable attention because of their potential contribution to the development of multidrug resistance worldwide. Insect technology may be a promising alternative for the rapid attenuation of ARGs in manure; however, the underlying mechanism remains unclear. This study aimed to evaluate the effects of black soldier fly (BSF, Hermetia illucens [L.]) larvae conversion combined with composting on ARGs dynamics in swine manure and to uncover the mechanisms through metagenomic analysis. Compared to natural composting (i.e. without BSF), BSFL conversion combined with composting reduced the absolute abundance of ARGs by 93.2 % within 28 days. The rapid degradation of antibiotics and nutrient reformulation during BSFL conversion combined with composting indirectly altered manure bacterial communities, resulting in a lower abundance and richness of ARGs. The number of main antibiotic-resistant bacteria (e.g., Prevotella, Ruminococcus) decreased by 74.9 %, while their potential antagonistic bacteria (e.g., Bacillus, Pseudomonas) increased by 128.7 %. The number of antibiotic-resistant pathogenic bacteria (e.g., Selenomonas, Paenalcaligenes) decreased by 88.3 %, and the average number of ARGs carried by each human pathogenic bacterial genus declined by 55.8 %. BSF larvae gut microbiota (e.g., Clostridium butyricum, C. bornimense) could help reduce the risk of multidrug-resistant pathogens. These results provide insight into a novel approach to mitigate multidrug resistance from the animal industry in the environment by using insect technology combined with composting, in particular in light of the global "One Health" requirements.

Multiple Receptors Contribute to the Attractive Response of Caenorhabditis elegans to Pathogenic Bacteria.

Nematodes feed mainly on bacteria and sense volatile signals through their chemosensory system to distinguish food from pathogens. Although nematodes recognizing bacteria by volatile metabolites are ubiquitous, little is known of the associated molecular mechanism. Here, we show that the antinematode bacterium Paenibacillus polymyxa KM2501-1 exhibits an attractive effect on Caenorhabditis elegans via volatile metabolites, of which furfural acetone (FAc) acts as a broad-spectrum nematode attractant. We show that the attractive response toward FAc requires both the G-protein-coupled receptors STR-2 in AWC neurons and SRA-13 in AWA and AWC neurons. In the downstream olfactory signaling cascades, both the transient receptor potential vanilloid channel and the cyclic nucleotide-gated channel are necessary for FAc sensation. These results indicate that multiple receptors and subsequent signaling cascades contribute to the attractive response of C. elegans to FAc, and FAc is the first reported ligand of SRA-13. Our current work discovers that P. polymyxa KM2501-1 exhibits an attractive effect on nematodes by secreting volatile metabolites, especially FAc and 2-heptanone, broadening our understanding of the interactions between bacterial pathogens and nematodes. IMPORTANCE Nematodes feed on nontoxic bacteria as a food resource and avoid toxic bacteria; they distinguish them through their volatile metabolites. However, the mechanism of how nematodes recognize bacteria by volatile metabolites is not fully understood. Here, the antinematode bacterium Paenibacillus polymyxa KM2501-1 is found to exhibit an attractive effect on Caenorhabditis elegans via volatile metabolites, including FAc. We further reveal that the attractive response of C. elegans toward FAc requires multiple G-protein-coupled receptors and downstream olfactory signaling cascades in AWA and AWC neurons. This study highlights the important role of volatile metabolites in the interaction between nematodes and bacteria and confirms that multiple G-protein-coupled receptors on different olfactory neurons of C. elegans can jointly sense bacterial volatile signals.

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.

Hermetia illucens L. larvae-associated intestinal microbes reduce the transmission risk of zoonotic pathogens in pig manure.

Black soldier fly (BSF) larvae are considered a promising biological reactor to convert organic waste and reduce the impact of zoonotic pathogens on the environment. We analysed the effects of BSF larvae on Staphylococcus aureus and Salmonella spp. populations in pig manure (PM), which showed that BSF larvae can significantly reduce the counts of the associated S. aureus and Salmonella spp. Then, using a sterile BSF larval system, we validated the function of BSF larval intestinal microbiota in vivo to suppress pathogens, and lastly, we isolated eight bacterial strains from the BSF larval gut that inhibit S. aureus. Results indicated that functional microbes are essential for BSF larvae to antagonise S. aureus. Moreover, the analysis results of the relationship between the intestinal microbiota and S. aureus and Salmonella spp. showed that Myroides, Tissierella, Oblitimonas, Paenalcalignes, Terrisporobacter, Clostridium, Fastidiosipila, Pseudomonas, Ignatzschineria, Savagea, Moheibacter and Sphingobacterium were negatively correlated with S. aureus and Salmonella. Overall, these results suggested that the potential ability of BSF larvae to inhibit S. aureus and Salmonella spp. present in PM is accomplished primarily by gut-associated microorganisms.

Reproductive Toxicity of Furfural Acetone in Meloidogyne incognita and Caenorhabditis elegans.

Furfural acetone (FAc) is a promising alternative to currently available nematicides, and it exhibits equivalent control efficiency on root-knot nematodes with avermectin in fields. However, its effect on the reproduction of root-knot nematode is poorly understood. In this study, the natural metabolite FAc was found to exhibit reproductive toxicity on Meloidogyne incognita and Caenorhabditis elegans. The number of germ cells of C. elegans was observed to decrease after exposure to FAc, with a reduction of 59.9% at a dose of 200 mg/L. FAc in various concentrations induced the germ-cell apoptosis of C. elegans, with an increase over six-fold in the number of apoptotic germ cells at 200 mg/L. These findings suggested that FAc decreased the brood size of nematode by inducing germ-cell apoptosis. Moreover, FAc-induced germ-cell apoptosis was suppressed by the mutation of gene hus-1, clk-2, cep-1, egl-1, ced-3, ced-4, or ced-9. The expression of genes spo-11, cep-1, and egl-1 in C. elegans was increased significantly after FAc treatment. Taken together, these results indicate that nematode exposure to FAc might inflict DNA damage through protein SPO-11, activate CEP-1 and EGL-1, and induce the core apoptosis pathway to cause germ-cell apoptosis, resulting in decreased brood size of C. elegans.

Inhibition of Zoonotic Pathogens Naturally Found in Pig Manure by Black Soldier Fly Larvae and Their Intestine Bacteria.

Black soldier fly (BSF) larvae are often exposed to organic waste which harbors abundant zoonotic pathogens. We investigated the ability of BSF larvae to inhibit the zoonotic pathogens naturally found in pig manure. The zoonotic pathogens populations were detected by using selective medium during the conversion. Results showed that the viability of the zoonotic pathogens in pig manure was significantly affected. After eight days of conversion, the Coliform populations were undetected, and Staphylococcus aureus and Salmonella spp. decreased significantly on the eighth day. Antimicrobial assays of the purified recombinant defensin-like peptide 4 (DLP4) showed that this peptide exhibits inhibitory activity against S. aureus, Salmonella enterica serovar typhimurium, and Escherichia coli in vitro. Bacteria BSF-CL and BSF-F were isolated from the larvae gut, and both inhibited the growth of S. aureus and E. coli, but Salmonella spp. was sensitive to the BSF-CL strain (but not to the BSF-F strain). The results from our experiments indicate that BSF larvae are capable of functionally inhibiting potential zoonotic pathogens in pig manure through a variety of mechanisms including antimicrobial peptides expression and the gut associate microorganisms. This study provides a theoretical basis for further study on the combined mechanism of BSF larvae immunity and its gut microbes against the zoonotic pathogens in pig manure.

Characteristics and mechanisms of ciprofloxacin degradation by black soldier fly larvae combined with associated intestinal microorganisms.

Antibiotics are challenging to degrade and are excreted by livestock which results in environmental pollution. In this paper, we demonstrated that environmentally friendly manure bioremediation performed by black soldier fly larvae (BSFL) is a wise alternative, which could effectively degrade ciprofloxacin (CIP) by approached 85.48% in artificial diet and 84.22% in poultry manure within 12 days. They are up to 2.5-4.0 fold more than that achieved by natural fermentation. The five CIP-degrading strains were isolated from the larval gut, two of which, named by Klebsiella pneumoniae BSFLG-CIP1 and Proteus mirabilis BSFLG-CIP5, could degraded CIP by nearly 98.22% and 97.83% in vitro, respectively. When the intestinal isolates were re-inoculated to sterile BSFL system, the degradation level significantly increased up to 82.38%, comparing with the sterile BSFL system (21.76%). It is proved that the larvae intestinal microbiota might carry out this highly-efficient CIP-degradation. Furthermore, seven possible metabolites were identified for CIP-degradation in vitro, and they were referring three main potential degrading mechanisms of hydroxylize, piperazine ring substitute and cleavage, and quinoline ring cleavage. In conclusion, the present study may provide a strategy to reduce antibiotics pollution in animal waste through bioremediation with BSFL and adjusted intestinal microbes.

Structural and functional characterizations and heterogenous expression of the antimicrobial peptides, Hidefensins, from black soldier fly, Hermetia illucens (L.).

Insect defensins are effector components of the innate defense system. Defensins, which are widely distributed among insects, are a type of small cysteine-rich plant antimicrobial peptides with broad-spectrum antimicrobial activity. Here, the cDNAs of the black soldier fly, Hermetia illucens (L.), encoding six defensins, designated herein as Hidefensin1-1, 2, 3, 4, 5, 6. Moreover, Hidefensin1-1, 2, and 5 were identified for the first time by genome-targeted analysis. These Hidefensins were found to mainly adopt α-helix and ß-sheet conformation homology as modeled by PRABI, Swiss-Model and ProFunc server. Six conserved cysteine residues that contribute to three disulfide bonds formed the spacing pattern "C-X12-C-X3-C-X9-C-X5-C-X-C", which play a vital role in the molecular stability of Hidefensins. Phylogenetic analysis revealed that the homology of five Hidefensins (except Hidefensin4) was about 59%-92% compared with other insect defensins, indicating that they are novel antimicrobial peptides genes in black soldier fly. Furthermore, the Hidefensin1-1 was expressed in the Escherichia coli strain BL21(DE3) as a fusion protein with thioredoxin. Results showed that the purified TRX-Hidefensin1-1 exerted strong inhibitory effects against the Gram-positive bacteria Staphylococcus aureus and the Gram-negative bacteria Escherichia coli. The inhibitory efficacy of TRX-Hidefensin1-1 against Gram-positive bacteria was better than that against Gram-negative bacteria. These results indicated that Hidefensin1-1 has potent antimicrobial activities against test pathogens.

Enhanced protein degradation by black soldier fly larvae (Hermetia illucens L.) and its gut microbes.

Black soldier fly larvae (BSFL) can convert a variety of organic wastes into biomass, and its gut microbiota are involved in this process. However, the role of gut microbes in the nutrient metabolism of BSFL is unclear. In this study, germ-free BSFL (GF) and gnotobiotic BSFL (GB) were evaluated in a high-protein artificial diet model. We used 16S rDNA sequencing, ITS1 sequencing, and network analysis to study gut microbiota in BSFL that degrade proteins. The protein reduction rate of the GB BSFL group was significantly higher (increased by 73.44%) than that of the GF BSFL group. The activity of gut proteinases, such as trypsin and peptidase, in the GB group was significantly higher than the GF group. The abundances of different gut microbes, including Pseudomonas spp., Orbus spp. and Campylobacter spp., were strongly correlated with amino acid metabolic pathways. Dysgonomonas spp. were strongly correlated with protein digestion and absorption. Issatchenkia spp. had a strong correlation with pepsin activity. Campylobacter spp., Pediococcus spp. and Lactobacillus spp. were strongly correlated with trypsin activity. Lactobacillus spp. and Bacillus spp. were strongly correlated with peptidase activity. Gut microbes such as Issatchenkia spp. may promote the gut proteolytic enzyme activity of BSFL and improve the degradation rate of proteins. BSFL protein digestion and absorption involves gut microbiota that have a variety of functions. In BSFL the core gut microbiota help complete protein degradation. These results demonstrate that core gut microbes in BSFL are important in protein degradation.

Volatile Organic Compounds from Bacillus aryabhattai MCCC 1K02966 with Multiple Modes against Meloidogyne incognita.

Plant-parasitic nematodes cause severe losses to crop production and economies all over the world. Bacillus aryabhattai MCCC 1K02966, a deep-sea bacterium, was obtained from the Southwest Indian Ocean and showed nematicidal and fumigant activities against Meloidogyne incognita in vitro. The nematicidal volatile organic compounds (VOCs) from the fermentation broth of B. aryabhattai MCCC 1K02966 were investigated further using solid-phase microextraction gas chromatography-mass spectrometry. Four VOCs, namely, pentane, 1-butanol, methyl thioacetate, and dimethyl disulfide, were identified in the fermentation broth. Among these VOCs, methyl thioacetate exhibited multiple nematicidal activities, including contact nematicidal, fumigant, and repellent activities against M. incognita. Methyl thioacetate showed a significant contact nematicidal activity with 87.90% mortality at 0.01 mg/mL by 72 h, fumigant activity in mortality 91.10% at 1 mg/mL by 48 h, and repellent activity at 0.01-10 mg/mL. In addition, methyl thioacetate exhibited 80-100% egg-hatching inhibition on the 7th day over the range of 0.5 mg/mL to 5 mg/mL. These results showed that methyl thioacetate from MCCC 1K02966 control M. incognita with multiple nematicidal modes and can be used as a potential biological control agent.

Control of Meloidogyne incognita in Three-Dimensional Model Systems and Pot Experiments by the Attract-and-Kill Effect of Furfural Acetone.

Meloidogyne incognita causes large-scale losses of agricultural crops worldwide. The natural metabolite furfural acetone has been reported to attract and kill M. incognita, but whether the attractant and nematicidal activities of furfural acetone on M. incognita function simultaneously in the same system, especially in three-dimensional spaces or in soil, is still unknown. Here, we used 23% Pluronic F-127 gel and a soil simulation device to demonstrate that furfural acetone has a significant attract-and-kill effect on M. incognita in both three-dimensional model systems. At 24 h, the chemotaxis index and the corrected mortality of nematodes exposed to 60 mg/ml of furfural acetone in 23% Pluronic F-127 gel were as high as 0.82 and 74.44%, respectively. Soil simulation experiments in moist sand showed that at 48 h, the chemotaxis index and the corrected mortality of the nematode toward furfural acetone reached 0.63 and 82.12%, respectively, and the effect persisted in the presence of tomato plants. In choice experiments, nematodes selected furfural acetone over plant roots and were subsequently killed. In pot studies, furfural acetone had a control rate of 82.80% against M. incognita. Collectively, these results provide compelling evidence for further investigation of furfural acetone as a novel nematode control agent.

Evaluation of Multiple Impacts of Furfural Acetone on Nematodes In Vitro and Control Efficiency against Root-Knot Nematodes in Pots and Fields.

Root-knot nematodes (RKNs) seriously endanger agricultural development and cause great economic losses worldwide. Natural product furfural acetone (FAc) is a promising nematicide with strong attractant and nematicidal activities, but baseline information about the impact of FAc on the reproduction, egg hatching, feeding, and growth of nematodes and its pest control efficiency in field are lacking. Here, the inhibition effects of FAc on nematodes in vitro and its RKN control efficiency in pot and field were investigated. FAc inhibited the egg hatching of Meloidogyne incognita by 91.7% at 200 mg/L after 2 days and suppressed the reproduction, feeding, and growth of Caenorhabditis elegans in vitro. In pot experiments, FAc in various dosages reduced the disease index of plant root significantly. In field experiments, FAc exhibited control effect on RKNs equivalent to commercial nematicides avermectin and metam sodium, with a reduction in disease index by 36.9% at a dose of 50 mg/plant. FAc also reduced the population density of RKNs in soil, with a reduction rate of 75.3% at the dose of 750 mg/m2. No adverse effect was detected on plant growth after FAc application. These results provide compelling evidence for development of FAc as an appropriate alternative for current nematicides.

Dual oxidase Duox and Toll-like receptor 3 TLR3 in the Toll pathway suppress zoonotic pathogens through regulating the intestinal bacterial community homeostasis in Hermetia illucens L.

Black soldier fly (BSF; Hermetia illucens L.) larvae can convert fresh pig manure into protein and fat-rich biomass, which can then be used as aquafeed for select species. Currently, BSF is the only approved insect for such purposes in Canada, USA, and the European Union. Pig manure could serve as a feed substrate for BSF; however, it is contaminated with zoonotic pathogens (e.g., Staphylococcus aureus and Salmonella spp.). Fortunately, BSF larvae inhibit many of these zoonotic pathogens; however, the mechanisms employed are unclear. We employed RNAi, qRT-PCR, and Illumina MiSeq 16S rDNA high-throughput sequencing to examine the interaction between two immune genes (Duox in Duox-reactive oxygen species [ROS] immune system and TLR3 in the Toll signaling pathway) and select pathogens common in pig manure to decipher the mechanisms resulting in pathogen suppression. Results indicate Bsf Duox-TLR3 RNAi increased bacterial load but decreased relative abundance of Providencia and Dysgonomonas, which are thought to be commensals in the BSF larval gut. Bsf Duox-TLR3 RNAi also inactivated the NF-κB signaling pathway, downregulated the expression of antimicrobial peptides, and diminished inhibitory effects on zoonotic pathogen. The resulting dysbiosis stimulated an immune response by activating BsfDuox and promoting ROS, which regulated the composition and structure of the gut bacterial community. Thus, BsfDuox and BsfTLR3 are important factors in regulating these key gut microbes, while inhibiting target zoonotic pathogens.

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.

Identification and Characterization of Nematicidal Volatile Organic Compounds from Deep-Sea Virgibacillus dokdonensis MCCC 1A00493.

Root-knot nematode diseases cause severe yield and economic losses each year in global agricultural production. Virgibacillus dokdonensis MCCC 1A00493, a deep-sea bacterium, shows a significant nematicidal activity against Meloidogyne incognita in vitro. However, information about the active substances of V. dokdonensis MCCC 1A00493 is limited. In this study, volatile organic compounds (VOCs) from V. dokdonensis MCCC 1A00493 were isolated and analyzed through solid-phase microextraction and gas chromatography-mass spectrometry. Four VOCs, namely, acetaldehyde, dimethyl disulfide, ethylbenzene, and 2-butanone, were identified, and their nematicidal activities were evaluated. The four VOCs had a variety of active modes on M. incognita juveniles. Acetaldehyde had direct contact killing, fumigation, and attraction activities; dimethyl disulfide had direct contact killing and attraction activities; ethylbenzene had an attraction activity; and 2-butanone had a repellent activity. Only acetaldehyde had a fumigant activity to inhibit egg hatching. Combining this fumigant activity against eggs and juveniles could be an effective strategy to control the different developmental stages of M. incognita. The combination of direct contact and attraction activities could also establish trapping and killing strategies against root-knot nematodes. Considering all nematicidal modes or strategies, we could use V. dokdonensis MCCC 1A00493 to set up an integrated strategy to control root-knot nematodes.