Persistent transmission of onchocerciasis in Kwanware-Ottou focus in Wenchi health district, Ghana.

BACKGROUND: This study examined onchocerciasis transmission in Kwanware and Ottou in the Wenchi Health District of Ghana, where persistent onchocercal microfilariae (mf) levels have been reported since 2012. METHODS: This study was conducted from 2019 to 2021 and involved the following: (i) reviewing past records of ivermectin mass drug administration (MDA); (ii) conducting a treatment coverage evaluation survey (CES); (iii) conducting key informant interviews; (iv) prospecting blackfly breeding sites; (v) collecting and dissecting blackflies; and (vi) conducting parasitological and serological surveys. RESULTS: (i) The review indicated ongoing MDA treatment for the past 27 years, with a reported coverage of over 65% in the last 17 yearly rounds; (ii) estimated treatment coverage by the CES in 2019 was 71.3%, with most of those not taking medicine stating that they were not offered; (iii) however, the key informant interviews revealed insufficiencies in reaching a significant number of people for treatment due to remote settlement, mobility, transport logistical issues, failure to register some people for treatment, leading to a false impression of good coverage, and a short distribution time; (iv) the most productive breeding was found within 5 km of Kwanware-Ottou; and (v) blackfly daily biting rates were highest in Kwanware and Ottou, with 199 and 160 bites per day, respectively. Infection in blackflies was found only in Kwanware and Ottou, with infectivity rates of 5.9‰ (per 1000) and 6.7‰, respectively. (vi) The mf prevalence in Ottou and Kwanware, respectively, was 40.0% and 30.0% among adults aged ≥ 20 years, and the anti-(Onchocerca volvulus) Ov16 IgG4 antibodies seroprevalence rates were 8.3% and 13.3% among children aged 5-9 years. These values were reduced to undetectable levels at a radius of 10 km from Ottou. CONCLUSIONS: This study confirms that active onchocerciasis transmission centres on Kwanware/Ottou and is confined to a 10 km radius despite 27 yearly treatment rounds. The main contributing factors are suboptimal coverage and high biting rates. Identifying and targeting such a focus with a combination of interventions will be cost-effective in accelerating onchocerciasis elimination in Ghana.

Vesicular Stomatitis Virus Detected in Biting Midges and Black Flies during the 2023 Outbreak in Southern California.

Vesicular stomatitis (VS) is a viral disease that affects horses, cattle, and swine that is transmitted by direct contact and hematophagous insects. In 2023, a multi-state outbreak of vesicular stomatitis New Jersey virus (VSNJV) occurred in California, Nevada, and Texas, infecting horses, cattle, and rhinoceros. To identify possible insect vectors, we conducted insect surveillance at various locations in San Diego County, CA, including at a wildlife park. CO2 baited traps set from mid-May to mid-August 2023 collected 2357 Culicoides biting midges and 1215 Simulium black flies, which are insect genera implicated in VSNJV transmission. Insects were pooled by species, location, and date, then tested for viral RNA. Nine RNA-positive pools of Culicoides spp. and sixteen RNA-positive pools of Simulium spp were detected. Infectious virus was detected by cytopathic effect in 96% of the RNA-positive pools. This is the first report of VSNJV in wild-caught C. bergi, C. freeborni, C. occidentalis, S. argus, S. hippovorum, and S. tescorum. The vector competency of these species for VSNJV has yet to be determined but warrants examination. Active vector surveillance and testing during disease outbreaks increases our understanding of the ecology and epidemiology of VS and informs vector control efforts.

Turning black soldier fly rearing by-products into valuable materials: Valorisation through chitin and chitin nanocrystals production.

The industry of insect-based proteins as feed and food products has been encountering a huge development since the last decade, and industrial-scale factories are now arising worldwide. Among all the species studied, Black Soldier Fly is one of the most promising and farmed. This rearing activity generates several by-products in the form of chitin-rich biomass that can be valorised to keep a virtuous production cycle embedded in the scope of the bioeconomy. Herein, we report the isolation of chitin and, for the first time, chitin nanocrystals (ChNCs) from all the BSF rearing by-products, i.e., moults (larval exuviae, puparium) and dead adults. Extraction yields, were dependent on the type of by-products and ranged from 5.8 % to 20.0 %, and the chemical structure of the extracts exhibited typical features of α-chitin, confirmed by FTIR, NMR, XRD and TGA analysis. Both STEM in SEM and AFM analysis confirmed the isolation of chitin nanocrystals presenting a rod-like morphology. The average nanocrystal height estimated by AFM ranged from 13 to 27 nm depending on the by-product sample. The following results highlighted the potential of BSF rearing by-products, promoting an approach to valorise those industrial waste and paving the way towards insect-based biorefinery.

Ongoing transmission of onchocerciasis in the Pru District of Ghana after two decades of mass drug administration with ivermectin and comparative identification of members of the Simulium damnosum complex using cytological and morphological techniques.

BACKGROUND: Human onchocerciasis remains a public health problem in Ghana. Mass drug administration (MDA) with ivermectin (IVM) has reduced disease morbidity and prevalence, but the transmission of onchocerciasis remains ongoing in several endemic foci. We investigated parasite transmission in some endemic communities in Ghana that had received > 18 rounds of annual MDA with IVM and determined the species composition of black fly (Simulium damnosum) vectors in these areas. METHODS: Adult female black flies were collected using human landing catches and identified as either forest or savanna species using morpho-taxonomic keys. The adult flies underwent dissection to determine their parity and detect any O. volvulus larvae, followed by the calculation of entomological indices. Simulium damnosum s.l. larvae were collected and preserved in freshly prepared Carnoy's fixative and were later used for cytotaxonomic studies. RESULTS: A total of 9,983 adult flies were caught: 6,569 and 3,414 in the rainy and dry seasons respectively. Black fly biting activities over the study period showed bimodal or trimodal patterns. The highest monthly biting rate (MBR) of 10,578.75 bites/person/month was recorded in July in Beposo, while the highest monthly transmission potential of 100.69 infective bites/person/month was recorded in Asubende in August. Morphological analysis of 2,032 flies showed that 99.8% (2,028) of the flies were savanna species, with only 4 (0.2%) adult flies being of the forest species. Cytogenetic studies on 114 black fly larvae revealed three cytospecies (Simulium damnosum s.s., S. sirbanum and S. sanctipauli) in the study area. CONCLUSIONS: The present studies confirmed an ongoing transmission of onchocerciasis in the study communities except Abua-1. It also provides further information on biting behaviors and onchocerciasis transmission indices in the study communities. Further, our data confirmed the savanna species (S. damnosum s.s. and S. sirbanum) of the S. damnosum s.l. to be the major vectors of onchocerciasis in the study areas, with only an occasional influx of forest cytotypes.

Morphological description and genetic analysis of a new black fly species (Diptera: Simuliidae) in the subgenus Asiosimulium from central Thailand.

BACKGROUND: Black flies are among the most medically and veterinary important insects, as adult females of certain species are the sole vector of Onchocerca volvulus. Here, a new black fly species belonging to the subgenus Asiosimulium Takaoka & Choochote, 2005, is described and formally named as Simulium (Asiosimulium) kittipati sp. nov. METHODS: Pupae and larvae of black flies were collected from available substrates in the stream from central Thailand. Pupae were individually separated in plastic tubes and maintained until adult flies emerged. The emerged adult flies associated with their pupal exuviae and cocoon as well as mature larvae preserved in 85% ethanol were used to describe the new species based on an integrated approach of morphological examination and molecular analysis of the COI gene. RESULTS: The new species is characterized in the female by the medium-long sensory vesicle with a medium-sized opening apically, scutum with three faint longitudinal vittae, and the ellipsoidal spermatheca; in the male by the number of upper-eye (large) facets in 20 vertical columns and 21 horizontal rows, hind basitarsus slender, nearly parallel-sided, and median sclerite much wider and upturned apically; in the pupa by the head and thoracic integument densely covered with tiny tubercles, and the pupal gill of arborescent type with 28-30 filaments; and in the larva by the postgenal cleft deep, nearly reaching the posterior margin of the hypostoma, and dark pigmented sheath of the subesophageal ganglion. The DNA barcode successfully differentiated the new species from its congeners with an interspecific genetic divergence of 1.74-18.72%, confirming the morphological identification that the species is a new member of the subgenus Asiosimulium. Phylogenetic analyses also indicated that the new species is genetically closely related to Simulium phurueaense Tangkawanit, Wongpakam & Pramual, 2018, further supporting its morphological classification. CONCLUSIONS: This is the ninth species assigned to the subgenus Asiosimulium within the genus Simulium Latreille, 1802. Taxonomic notes and identification keys are given to distinguish this new species from the eight known species members in its same subgenus. Additionally, a distribution map of all species members in this subgenus occurring in Thailand and other countries is provided.

Effects of heat treatment on rumen degradability and protein intestinal digestibility of black soldier fly (Hermetia illucens L.) in goat.

The black soldier fly larvae (BSF) are used as a substitute for soybean meal due to their high crude protein content. This experiment aims to assess the impact of heat treatment on the rumen degradability of BSF and protein digestion in the small intestine using the in situ nylon bag method and the three-step in vitro method. This study comprises a total of 8 groups (n = 6). The negative control group includes only full-fat soybeans (FFS) and BSF (FF group and BS group). The positive control groups consist of a 95% BSF or 95% FFS mixed with 5% cassava (FFC and BSC groups). The treatment groups involve adding 75% water to the positive control mixture, followed by vigorous kneading to achieve uniform mixing. The resulting mixture was then pressed to a thickness of approximately 5 cm, placed in an oven, and dried for 120 min at temperatures of 120 °C and 140 °C (12FFC, 14FFC, 12BSC, and 14BSC groups). Nylon bags will be incubated in the rumen for 0, 2, 4, 8, 12, 24, and 48 h, and the small intestine protein digestion rate will be analyzed at 16 h. Compared to the BS group, heat-treated BSF showed increased (P < 0.05) rumen DM degradability and effective degradability. The 14BSC group increased (P < 0.05) rumen CP degradability and degradation kinetic parameters, while the 12BSC group decreased (P < 0.05) these parameters. The CP degradability of BSF was significantly higher (P < 0.05) than that of full-fat soybeans. The Idg and IDCP of heat-treated full-fat soybeans were significantly higher (P < 0.05) than those of other treatment groups. At the same time, heat treatment was beneficial for increasing (P < 0.05) the Idg and IDCP of BSF, and the 14BSC treatment effect was significantly better (P < 0.05) than that of the 12BSC group. Therefore, based on the results of this experiment, it is recommended to supplement BSF with cassava and subject them to heat treatment at 140 °C.

Effects of black soldier fly larvae on biotransformation and residues of spent mushroom substrate and wet distiller's grains.

Black soldier fly larvae (BSFL) could convert a variety of organic wastes, including spent mushroom substrate (SMS) and wet distiller's grains (WDG). Nevertheless, little is known about the conversion of these wastes by BSFL. Thus, this study investigates the conversion of SMS and WDG in five different proportions by BSFL. This study demonstrates that BSFL can convert SMS, WDG, and their mixtures. It can also encourage the humification of the substrate, increasing the amount of element in the residues. It is evident that there were differences in the carbon and nitrogen element fractionation mode as well as the microbial community present in the residue. The microbial community of the substrate and the physiochemical parameters are intimately related to this. Although the mixture treated with BSFL helps to generate a residue with more humus, it might not be stable.

Breaking down barriers: live or dehydrated dietary whole black soldier fly larvae supplementation in slow growing chickens preserve meat quality and sensory traits.

This study investigated the effects of supplementing the diet of a slow-growing autochthonous chicken breed with dehydrated or live Black Soldier Fly Larvae (BSFL) on meat quality and sensory attributes. The research, conducted at the University of Turin, Italy, involved 144 male birds distributed in three experimental groups. The control group (C) was fed a basal diet in which soybean meal was completely substituted with alternative ingredients. The 2 experimental groups were administered a diet identical to the control group but supplemented with either whole dehydrated black soldier fly larvae (DL) or whole live black soldier fly larvae (LL) at a level equal to 5% expected daily feed intake of dry matter. We evaluated the following parameters: nutrient intake, slaughtering performance, physical and nutritional meat quality, fatty acid composition, proteomics, and sensory characteristics. The results demonstrated BSFL supplementation to have no detrimental effects on overall meat quality or sensory attributes. Specifically, there were no significant differences in physical meat quality parameters, nutritional composition, lipid oxidation, or protein digestibility between control and BSFL-fed groups. Fatty acid analysis revealed higher concentrations of lauric and myristic acids in BSFL-fed chicken breast (p < 0.005), suggesting potential nutritional benefits from the supplement. The proteomic analysis also showed no significant differences in the expression of abundant proteins in the breast meat between groups, indicating minimal physiological impact of BSFL supplementation. Overall, this study provides reassurance to consumers and industries about the suitability of BSFL as a sustainable feed supplement for poultry that also offers potential benefits in terms of optimizing the fatty acid profile of chicken meat.

Performance, egg quality and organ traits of laying hens fed black soldier fly larvae products.

Due to consumer demands and institutional pressure, the egg production sector, is looking for alternative protein sources for laying hen feed to support more sustainable, circular production. black soldier fly (BSF) larvae could be used as a protein source. In addition to protein the larvae contain large quantities of fat and can either be fed to laying hens unprocessed (alive) or processed (meal and oil). The current study was performed with 560 Brown Nick laying hens from 20 to 27 wk of age. The laying hens were divided over 5 treatments, each replicated 8 times. Treatments consisted of standard laying hen feed (control) and standard feed in which soybean meal was partly exchanged with live BSF larvae or BSF larvae meal and oil combined, at 2 inclusion levels. During the experiment production parameters, egg-quality, and length and weight of various organs were measured. Laying hens fed BSF larvae products consumed less feed compared to those of the control group. Most egg production parameters were similar, however laying hens fed diets with BSF larvae meal plus oil produced eggs with lower egg weight during the last 2 wk of the experiment, compared to the control group. All egg-quality characteristics remained the same across treatments, except for darker yolk colors when feeding BSF meal and oil and high inclusion of live BSF larvae. This is a favorable characteristic for European consumers. The weight of intestinal organs was largely unaffected by the treatments. The jejunum and ileum weight of laying hens fed live larvae was lower compared to the control group. As FCRs were similar or improved compared to the control group, we assume that nutrient utilization was not impaired. For most detected differences the type of BSF larvae product (live larvae or meal plus oil) rather than inclusion level was of significance.

Unraveling the role of black soldier fly larvae in chicken manure conversion: Facilitating maturation and enhancing humification.

Black soldier fly larvae (BSFL) have garnered considerable attention for their efficacy in mitigating waste management challenges. However, their potential in treating antibiotics contaminated chicken manure remains uncertain. This study investigates the physicochemical properties changes and nutrient dynamics during the composting of contaminated-chicken manure using BSFL. The results indicate that BSFL treatment reduces electrical conductivity (by 6.01-58.09 %), organic matter, and dissolved organic carbon content in chicken manure throughout the composting process, while maintaining a more stable pH value (pH âˆ¼ 6.0-8.0). This is attributed to the consumption of organic matter by BSFL and the subsequent promotion of organic acid formation. Additionally, BSFL treatment improves the degree of aromatization of dissolved organic matter (DOM) in chicken manure and increases the proportions of fulvic acid (up to 48.77 %) and humic acid (maximally 14.27 %) within the DOM. The germination index and pot experiments indicated improved compost maturity and plant growth in BSFL-treated composts. Furthermore, BSFL meal demonstrated high protein and essential fatty acid content, highlighting its potential as a protein supplement in animal feed. This study underscores the efficacy of BSFL in enhancing compost quality and nutrient availability, offering a sustainable solution for waste management and animal feed production.

Physical and chemical characterization of chitin and chitosan extracted under different treatments from black soldier fly.

The shell of Hermetia illucens L. contains considerable amounts of chitin, which has various biological activities. So far, few studies have focused on chitin of Hermetia illucens L. as a source of chitosan and oligosaccharides. There is great potential for utilizing Hermetia illucens L. chitin to produce chitosan films in biomaterials. We studied different extraction conditions for chitin and extracted it from black soldier fly (BSF) (Hermetia illucens L.). Three processing steps were adopted: (1) demineralization, (2) deproteinization, and (3) decolorization. The chemical components (moisture, ash, protein, fat, residual protein, and residual mineral contents) and physicochemical characteristics of the chitin and chitosan extracted under these three conditions were determined. In addition, Fourier transform infrared spectroscopy and X-ray diffraction were used to analyze the extracted chitin and commercial samples, and the results showed that demineralization-deproteinization-decolorization treatments could achieve the highest chitin yield (7.18 ± 0.11 %), chitosan yield (64.22 ± 0.79 %), and the best purity (residual protein 0.56 ± 0.01 % and residual ash 0.58 ± 0.04 %), making it the best treatment method. Using this method, the residues produced from farmed BSF can be recycled and used as a new source of chitin.

Genetic relationships of three species of black flies (Diptera: Simuliidae) in Taiwan.

As an island about 150 km from the mainland, Taiwan would be expected to have endemic species. About 64 % of its 36 species of black flies are considered endemic, more than twice the level of endemicity that has been recorded for all insects on the island. To begin assessing the validity of the high level of endemism for the Simuliidae, we used giant chromosome banding patterns and cytochrome c oxidase I (COI) sequences, against a well-defined morphological backdrop, to evaluate three of Taiwan's black flies, Simulium chungi Takaoka & Huang, S. pingtungense Huang & Takaoka, and S. sakishimaense Takaoka. Molecular data revealed high similarity of populations of S. sakishimaense in Taiwan and at the type locality on Ishigaki Island, Japan, about 180 km to the east. Thus, populations referred to as S. sakishimaense in Taiwan are conspecific with typical S. sakishimaense in Japan, confirming their non-endemicity in Taiwan. Simulium sakishimaense might have reached Ishigaki by island hopping via Taiwan from the Chinese mainland. Chromosomes and the COI gene agree with morphology that S. sakishimaense is a member of the S. multistriatum species group although the chromosomal banding patterns do not indicate that it is distinct from S. fenestratum Edwards on the mainland. Although molecular sequences indicate S. sakishimaense is monophyletic, this taxon falls within the same Operational Taxonomic Unit as nine other members of the S. multistriatum group, including S. fenestratum. Simulium pingtungense, in agreement with morphology, is molecularly distinct from the 10 other analyzed members of the S. striatum species group, tentatively suggesting that it is endemic to Taiwan, pending analysis of samples from mainland China. Simulium chungi in Taiwan is chromosomally and molecularly unique, with larvae resembling those of S. saskishimaense. It is not, however, a member of either the S. multistriatum or S. striatum species groups. For now, the S. chungi species group remains a legitimate taxon consisting of two species. Strengthening the case for endemic taxa in Taiwan awaits analysis of key samples from the Chinese mainland.

Effects of brewery by-products on growth performance, bioconversion efficiency, nutritional profile, and microbiota and mycobiota of black soldier fly larvae.

Brewery by-products are recognised as suitable rearing substrates for Hermetia illucens, better known as black soldier fly (BSF) but information about the impact of different ratios of brewer's spent grains (BSG) and brewer's spent yeast (BSY) are still scarce. This study evaluated the effects of BSG-BSY-based diets on BSF larval growth, survival, bioconversion efficiency, nutritional profile, and microbiota and mycobiota. A total of 3 000 6-day-old BSF larvae were allotted to five dietary treatments (six replicate boxes/diet, 100 larvae/box): (i) BSY2.5 (25 g/kg of BSY+975 g/kg of BSG), (ii) BSY5 (50 g/kg of BSY+950 g/kg of BSG), (iii) BSY7.5 (75 g/kg of BSY+925 g/kg of BSG), (iv) BSY10 (100 g/kg of BSY+900 g/kg of BSG), and (v) control (Gainesville diet). Larval weight and substrate pH were recorded every 4 days. At the end of the trial (5% of prepupae), bioconversion efficiency corrected for residue (BER), reduction rate (RR), and waste reduction index (WRI) were calculated, and the larval proximate composition, microbiota and mycobiota characterised. At 10 and 14 days of age, BSY7.5 and BSY10 larvae displayed higher weight than BSY2.5 and BSY5 (P < 0.05), with BSY10 larvae showing the highest weight among the BSG-BSY-based diets at the end of the trial (P < 0.05). The BSY7.5 and BSY10 larvae also displayed a better BER than BSY2.5 and BSY5 (P < 0.01), whereas similar RR, WRI, survival and development time, as well as pH, were, however, observed among the BSG-BSY-based diets (P > 0.05). The BSY10 larvae displayed lower ether extract content than the other BSG-BSY-based diets (P > 0.001). The use of BSG-BSY-based diets did not influence the alpha diversity of larval microbiota and mycobiota (P > 0.05), but a specific microbial signature was identified per each dietary treatment (Porphyromonadaceae [BSY5], Sphingomonas [BSY7.5], Bacillus [BSY10] and Ruminococcus and Myroides [BSG-BSY-based diets]; P < 0.05). Co-occurrence and co-exclusion analysis also showed that Saccharomyces cerevisiae and Pichia excluded and favoured, respectively, the presence of Streptomyces and Fluviicola, while Clavispora lusitaniae was associated with Myroides (P < 0.05). In conclusion, BSG-BSY-based diets are suitable for rearing HI in terms of larval performance, nutritional profile, and microbiota and mycobiota, with 7.5 and 10% of BSY inclusion levels being able to improve larval growth and bioconversion efficiency.

Recombinant expression and characterization of the endochitinase Chit36-TA from Trichoderma asperellum in Komagataella phaffii for chitin degradation of black soldier fly exuviae.

The natural polymer chitin is an abundant source for valuable N-acetylchitooligosaccharides and N-acetylglucosamine applicable in several industries. The endochitinase Chit36-TA from Trichoderma asperellum was recombinantly expressed in Komagataella phaffii for the enzymatic degradation of chitin from unused insect exuviae into N-acetylchitooligosaccharides. Chit36-TA was purified by Ni-NTA affinity chromatography and subsequently biochemically characterized. After deglycosylation, the endochitinase had a molecular weight of 36 kDa. The optimum pH for Chit36-TA was 4.5. The temperature maximum of Chit36-TA was determined to be 50 °C, while it maintained > 93% activity up to 60 °C. The chitinase was thermostable up to 45 °C and exhibited ~ 50% activity after a 15 min incubation at 57 °C. Chit36-TA had a maximum specific enzyme activity of 50 nkat/mg with a Km value of 289 µM with 4-methylumbelliferyl-N,N',N″-triacetyl-ß-chitotrioside as substrate. Most tested cations, organic solvents and reagents were well-tolerated by the endochitinase, except for SDS (1 mM), Cu2+ (10 mM) and Mn2+ (10 mM), which had stronger inhibitory effects with residual activities of 3, 41 and 28%, respectively. With a degree of hydrolysis of 32% applying colloidal shrimp chitin (1% (w/v)) and 12% on insect larvae (1% (w/v)) after 24 h, the endochitinase was found to be suitable for the conversion of colloidal chitin as well as chitin from black soldier fly larvae into water-soluble N-acetylchitooligosaccharides. To prove scalability, a bioreactor process was developed in which a 55-fold higher enzyme activity of 49 µkat/l and a tenfold higher protein expression of 1258 mg/l were achieved.

Strategic approach for converting fat-rich food waste into high-quality biodiesel using black soldier fly larvae for sustainable bioenergy.

Food waste (FW) comprises carbohydrates, proteins, lipids, and water, posing technical challenges for effective treatment and valorisation. This study addresses these challenges by using black soldier fly larvae (BSFL) as a bioconversion medium to transform FW into biodiesel (BD). BSFL predominantly consumed the carbohydrates and proteins in FW (81 wt%), while showing a lower preference for lipids (<50 wt% consumed). Notwithstanding the lower consumption of lipids in the FW than that of carbohydrates and proteins, BSFL had a high lipid content (48.3 wt%). The subsequent conversion of the lipids extracted from BSFL into BD was tested via catalytic (acid/alkali) and non-catalytic transesterification processes. The BD yield from catalytic transesterification was lower than that from non-catalytic transesterification because of the low tolerance against free fatty acids (FFAs). BD was also produced from the lipid-concentrated residual FW through non-catalytic transesterification. Although the FW residue extracts contained high amounts of FFAs (49.9 wt%), non-catalytic transesterification displayed a high BD yield (92.4 wt%; yields from catalytic transesterification: < 80.0 wt%). Moreover, blending the BD derived from the BSFL and FW residue extracts enhanced the fuel properties. The BSFL-assisted FW management efficiently reduced FW by 90 wt% while producing a high-quality BD.

Deeper insight into the storage time of food waste on black soldier fly larvae growth and nutritive value: Interactions of substrate and gut microorganisms.

Biological treatment of food waste (FW) by black soldier fly larvae (BSFL) is considered as an effective management strategy. The composition and concentrations of nutrients in FW change during its storage and transport period, which potentially affect the FW conversion and BSFL growth. The present study systematically investigated the effect of different storage times (i.e., 0-15 d) on FW characteristics and its substantial influence on the BSFL growth. Results showed that the highest larvae weight of 282 mg and the shortest growth time of 14 days were achieved at the group of FW stored for 15 days, but shorter storage time (i.e., 2-7 d) had adverse effect on BSFL growth. Short storage time (i.e., 2-4 d) improved protein content of BSFL biomass and prolonged storage time (i.e., 7-10 d) led to the accumulation of fat content. The changes of substrate characteristics and indigenous microorganisms via FW storage time were the main reasons for BSFL growth difference. Lactic acid (LA) accumulation (i.e., 19.84 g/L) in FW storage for 7 days significantly limited the BSFL growth, leading to lowest larvae weight. Both the substrate and BSFL gut contained same bacterial communities (e.g., Klebsiella and Proteus), which exhibited similar change trend with the prolonged storage time. The transfer of Clostridioides from substrate to BSFL gut promoted nutrients digestion and intestinal flora balance with the FW stored for 15 days. Pathogens (e.g., Acinetobacter) in BSFL gut feeding with FW storage time of 7 days led to the decreased digestive function, consistent with the lowest larvae weight. Overall, shorter storage time (i.e., 2-7 d) inhibited the BSFL digestive function and growth performance, while the balance of the substrate nutrients and intestinal flora promoted the BSFL growth when using the FW stored for 15 days.

Wild-caught adult black flies (Diptera: Simuliidae) from various ecological landscapes in Malaysia.

Most studies on black flies focus on the taxonomy and ecology of their aquatic stages. Despite posing a public health threat, the adults remain poorly studied in many countries, including Malaysia. The present study represents the first investigation of the distribution of wild-caught black flies from various ecological landscapes and climatic conditions in Malaysia. CO2-baited Malaise traps were set randomly at 41 sampling sites across Peninsular Malaysia from 2020 to 2023. In total, 532 black flies belonging to 14 species of four subgenera were captured. To ensure taxonomic rigor, specimens were identified to species morphologically and molecularly. The subgenus Gomphostilbia was the most abundant (71.43 %), followed by Simulium (14.28 %) and Davieselleum and Nevermannia each representing 7.14 % of the total captures. These species represented 14.74 % of the total species recorded from Malaysia. The most frequently collected species were Simulium roslihashimi (24.39 %), followed by S. aureohirtum, S. vanluni, and S. (Gomphostilbia) sp. 1 with 7.32 % each. The highest relative abundance was found for S. vanluni (86.09 %) and S. roslihashimi (7.14 %). Most species were found at elevation below 300 m (78.57 %); fewer were at elevation higher than 1,000 m (21.43 %). Two principal components accounted for 85.3 % of the total intersite variance. Simulium roslihashimi was found at almost every site, with a maximum relative humidity of 90 %. Simulium aureohirtum and S. vanluni were found at sites with relative humidity up to 73 %, but S. aureohirtum was found at higher temperatures (31 °C) compared with S. roslihashimi (28 °C) and S. vanluni (29 °C). The present study establishes the groundwork for further studies of wild adults in Malaysia and identifies the need to use more traps over the range of seasons and environmental conditions, particularly near breeding sites.

Effects of ultrasonic treatment on the structure and functional characteristics of myofibrillar proteins from black soldier fly.

In the process of utilizing black soldier fly larvae (BSFL) lipids to develop biodiesel, many by-products will be produced, especially the underutilized protein components. These proteins can be recycled through appropriate treatment and technology, such as the preparation of feed, biofertilizers or other kinds of bio-products, so as to achieve the efficient use of resources and reduce the generation of waste. Myofibrillar protein (MP), as the most important component of protein, is highly susceptible to environmental influences, leading to oxidation and deterioration, which ultimately affects the overall performance of the protein and product quality. For it to be high-quality and fully exploited, in this study, black soldier fly myofibrillar protein (BMP) was extracted and primarily subjected to ultrasonic treatment to investigate the impact of varying ultrasonic powers (300, 500, 700, 900 W) on the structure and functional properties of BMP. The results indicated that as ultrasonic power increased, the sulfhydryl content and turbidity of BMP decreased, leading to a notable improvement in the stability of the protein emulsion system. SEM images corroborated the changes in the microstructure of BMP. Moreover, the enhancement of ultrasound power induced modifications in the intrinsic fluorescence spectra and FTIR spectra of BMP. Additionally, ultrasonic treatment resulted in an increase in carbonyl content and emulsifying activity of BMP, with both peaking at 500 W. It was noteworthy that BMP treated with ultrasound exhibited stronger digestibility compared to the untreated. In summary, 500 W was determined as the optimal ultrasound parameter for this study. Overall, ultrasound modification of insect MPs emerges as a dependable technique capable of altering the structure and functionality of BMP.

Unlocking the potential of black soldier fly frass as a sustainable organic fertilizer: A review of recent studies.

Using Hermetia illucens, or Black Soldier Fly (BSF) frass as an organic fertilizer is becoming increasingly popular in many countries. As a byproduct derived from BSF larvae that feed on organic waste, BSF frass has tremendous potential for preserving the environment and promoting the circular economy. Since it has diverse biochemical properties influenced by various production and environmental factors, further research is needed to evaluate its potential for extensive use in crop production and agriculture. Our review summarizes recent findings in BSF frass research by describing its composition and biochemical properties derived from various studies, including nutrient contents, biostimulant compounds, and microbial profiles. We also discuss BSF frass fertilizers' effectiveness on plant growth and contribution to environmental sustainability. Great compositions of BSF frass increase the quality of plants/crops by establishing healthy soil and improving the plants' immune systems. Special emphasis is given to potentially replacing conventional fertilizer to create a more sustainable cropping system via organic farming. Besides, we discuss the capability of BSF bioconversion to reduce greenhouse gas emissions and improve the socioeconomic aspect. The prospects of BSF frass in promoting a healthy environment by reducing greenhouse gas emissions and improving the socioeconomic aspects of communities have also been highlighted. Overall, BSF frass offers an alternative approach that can be integrated with conventional fertilizers to optimize the cropping system. Further studies are needed to fully explore its potential in establishing sustainable system that can enhance socioeconomic benefits in the future.

Exploring the role of RNASET2 in the immune response of black soldier fly larvae.

T2 RNases are transferase-type enzymes distributed across phyla, crucial for breaking down single-stranded RNA molecules. In addition to their canonical function, several T2 enzymes exhibit pleiotropic roles, contributing to various biological processes, such as the immune response in invertebrates and vertebrates. This study aims at characterizing RNASET2 in the larvae of black soldier fly (BSF), Hermetia illucens, which are used for organic waste reduction and the production of valuable insect biomolecules for feed formulation and other applications. Given the exposure of BSF larvae to pathogens present in the feeding substrate, it is likely that the mechanisms of their immune response have undergone significant evolution and increased complexity. After in silico characterization of HiRNASET2, demonstrating the high conservation of this T2 homolog, we investigated the expression pattern of the enzyme in the fat body and hemocytes, two districts mainly involved in the insect immune response, in larvae challenged with bacterial infection. While no variation in HiRNASET2 expression was observed in the fat body following infection, a significant upregulation of HiRNASET2 synthesis occurred in hemocytes shortly after the injection of bacteria in the larva. The intracellular localization of HiRNASET2 in lysosomes of plasmatocytes, its extracellular association with bacteria, and the presence of a putative antimicrobial domain in the molecule, suggest its potential role in RNA clean-up and as an alarm molecule promoting phagocytosis activation by hemocytes. These insights contribute to the characterization of the immune response of Hermetia illucens larvae and may facilitate the development of animal feedstuff enriched with highly valuable BSF bioactive compounds.