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.

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.

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.

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.

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.

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.

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.

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.

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.

Diverting organic waste from landfills via insect biomanufacturing using engineered black soldier flies (Hermetia illucens).

A major roadblock towards the realisation of a circular economy are the lack of high-value products that can be generated from waste. Black soldier flies (BSF; Hermetia illucens) are gaining traction for their ability to rapidly consume large quantities of organic wastes. However, these are primarily used to produce a small variety of products, such as animal feed ingredients and fertiliser. Using synthetic biology, BSF could be developed into a novel sustainable biomanufacturing platform to valorise a broader variety of organic waste feedstocks into enhanced animal feeds, a large variety of high-value biomolecules including industrial enzymes and lipids, and improved fertiliser.

Safety and transfer of veterinary drugs from substrate to black soldier fly larvae.

There is an increasing interest in edible insects in Europe for feed and food purposes. Quantitative information on the transfer of chemical hazards from substrates to larvae is needed to evaluate food and feed safety aspects. This evaluation is especially needed when organic substrates or residual streams such as manure will be applied as substrate, contributing to a circular food system. This study investigated the transfer of veterinary drugs from spiked substrate to black soldier fly larvae (Hermetia illucens). Veterinary drugs that are commonly administered to chicken, fattening pigs, and cattle and regularly detected in manure were included: three different antibiotics (enrofloxacin, oxytetracycline, sulfamethoxazole), three coccidiostats (narasin, salinomycin, toltrazuril) and one antiparasitic drug (eprinomectin). The chemicals were spiked to insect substrate to reach final concentrations of 0.5 and 5 mg/kg for the antibiotics and the antiparasitic drug, and 5 and 50 mg/kg for the coccidiostats. Black soldier fly larvae were reared for 1 week on the spiked substrates, and the transfer of the veterinary drugs to the larvae and frass was quantified using liquid chromatography coupled with tandem mass spectrometry. Only oxytetracycline and eprinomectin reduced the average weight and/or survival of the black soldier fly larvae. The transfer of the veterinary drugs to the larvae was on average 19.2% for oxytetracycline, 12% for enrofloxacin, 9.5% for narasin, 8.1% for eprinomectin, 3.9% for salinomycin, 4.2% for toltrazuril, and 0.2% for sulfamethoxazole, relative to concentrations in the substrate. Mass-balance calculations revealed that the larvae seem to metabolise veterinary drugs, and indeed, metabolites of enrofloxacin, sulfamethoxazole, and toltrazuril were detected in the larvae and frass. In conclusion, insect-rearing substrates should be evaluated for the presence of veterinary drug residues to ensure feed (and food) safety, as well as because of possible effects on insect growth.

Modelling onchocerciasis-associated epilepsy and the impact of ivermectin treatment on its prevalence and incidence.

Retrospective cohort studies in Cameroon found an association between Onchocerca volvulus microfilarial load in childhood (measured in 1991-1993) and risk of developing epilepsy later in life (measured in 2017). We parameterised and integrated this relationship (across children aged 3-15 years) into the previously published, stochastic transmission model, EPIONCHO-IBM, for Simulium damnosum sensu lato-transmitted onchocerciasis. We simulated 19 years (1998-2017) of annual ivermectin mass drug administration (MDA) reflecting coverage in the study area, and modelled epilepsy prevalence and incidence. Scenario-based simulations of 25 years of (annual and biannual) MDA in hyper- and holoendemic settings, with 65% and 80% therapeutic coverage, were also conducted. EPIONCHO-IBM predicted 7.6% epilepsy prevalence (compared to 8.2% in the Cameroon study) and incidence of 317 cases/100,000 person-years (compared to 350). In hyperendemic areas, 25 years of biannual MDA (80% coverage) eliminated onchocerciasis-associated epilepsy (OAE) and protected untreated under-fives from its development. Strengthening onchocerciasis programmes, implementing alternative strategies, and evaluating treatment for under-fives and school-age children are crucial to prevent OAE in highly-endemic settings.

Dehydrated and live black soldier fly larvae as environmental enrichment in indigenous slow-growing chickens: performance, gut health, and chitinolytic enzyme activity.

The demand for sustainable and ethically farmed animal products is on the rise as consumers become more environmentally and animal welfare conscious. The need to diminish the consumption of soybean meal is urgent, and companies are looking for ways to respond to this necessity by looking for alternatives to soybean meal. This study assessed the impact of introducing whole dehydrated and live black soldier fly larvae (BSFL) into the diet of an indigenous chicken breed as environmental enrichment. A total of 144 39-day-old male Bianca di Saluzzo chickens were distributed among 18 pens and assigned to three different experimental groups. The control group received a diet where soybean meal was entirely replaced by alternative ingredients. The two experimental groups were given the same diet supplemented with 5% of the expected daily feed intake of whole dehydrated BSFL or whole live BSFL. Throughout the trial period (from the bird age of 39-174 days of age), live weight was recorded every 21 days, and the average daily gain, daily feed intake, and feed conversion ratio were calculated. The time required for the birds to consume the larvae was recorded three times a week. At age 147 and 174 days, 12 birds per treatment were selected based on mean live weight and slaughtered. Measurements included hot and chilled carcass weights, organ weights (spleen, liver, heart, stomach), breast and thigh muscle weights, and the corresponding yields were calculated. Acid protease activity was measured in proventriculus extract, and chitinase and chitosanase activity was calculated based on the release of reducing sugars from chitin or chitosan. The results showed little improvement in final live weights and daily feed intakes of the animals fed the insect larvae compared with control birds. Larva supplementation had no negative impact on the overall well-being of the animals assessed by blood analysis and histopathological assessment of the intestinal tract, spleen, and liver. No differences were found between the dehydrated vs live insect larvae consumption times, with all larvae being eaten up very rapidly (< 3 min). The birds fed BSFL showed an increase in chitinase activity. These findings support the potential use of whole BSFL as a form of environmental enrichment, particularly in their dehydrated form, being more convenient to use and store, which would also encourage the uptake of this practice by farmers.

Wheat starch processing by-products as rearing substrate for black soldier fly: does the rearing scale matter?

Rearing scale may influence black soldier fly (BSF) larvae traits when they are fed on a single diet, but different feeding substrates have not been tested yet. This study evaluated the effects of wheat starch processing by-products-based diets on growth performance, bioconversion efficiency, and nutritional profile of BSF larvae reared in different scales. Four diets (D1 and D2 [isonitrogenous, isolipidic and isoenergetic]; D3 and D4 [displaying 1:1 and 1:2 as protein to carbohydrate ratios, respectively]) were tested at 3 rearing scales (4 replicate boxes/diet, with a constant volume [0.84 cm3]/larva and feed [0.7 g]/larva): 1) small (S; 12 × 12 cm, substrate height: 4 cm, 686 6-day-old larvae (6-DOL)/box), 2) medium (M, 32 × 21 cm, substrate height: 7 cm, 5 600 6-DOL/box), and 3) large (L, 60 × 40 cm, substrate height: 7 cm, 20 000 6-DOL/box). Larval weight was recorded at the beginning of trial and every 4 days, and growth rate (GR), specific growth rate (SGR), feed conversion ratio (FCR), survival, bioconversion efficiency corrected for residue (BER), reduction rate (RR), and waste reduction index (WRI) calculated at the end of larval growth (frass DM ≥ 55%). Substrate pH, T and height were measured at the beginning, every 4 days, and end of trial. Larval proximate composition was analysed at the end of trial. Data were analysed by generalised linear mixed model (SPSS software, P < 0.05). The D1 larvae showed higher weight, GR, SGR and WRI (along with higher substrate T) than D2 at M scale, while increased SGR and FCR - as well as decreased survival, RR and WRI - were observed in D2 larvae at S scale (P < 0.05). Larval CP and ether extract (EE) contents were influenced by M and L scales only, being higher in D2 group than in D1 (P < 0.001). Differently, decreased ash was recorded in D2 larvae when reared at S and M scales, while L scale revealed higher ash in D2 group than D1 (P < 0.001). The D3 larvae displayed greater weight, SGR, survival, RR and WRI (along with greater substrate T) than D4 at M scale, with increased survival and substrate T being also highlighted in L scale (P < 0.05). The D3 larvae also showed lower DM and EE - as well as higher CP - than D4 at all the rearing scales (P < 0.001). In conclusion, D1 and D3 led to better BSF larval growth performance, bioconversion efficiency and nutritional profile mainly at M and L scales, as a consequence of their ability to facilitate larval aggregation and, in turn, allow achieving a higher substrate T.

Growth performance, proximate composition and fatty acid profile of black soldier fly larvae reared on two grape pomace varieties.

The black soldier fly (Hermetia illucens) is attracting increasing interest for its ability to convert low-value substrates into highly nutritious feed. This study aimed at evaluating grape pomace from two varieties (Becuet - B; Moscato - M) as rearing substrates for black soldier fly larvae (BSFL), focusing on the related effects on larval growth performance, proximate composition, and fatty acid profile. A total of six replicates per treatment, and 1 000 BSFL per replica, were used. Larval development was assessed by larvae weight, which was recorded eight times during the trial: the day after the beginning of the trial, and then on days 5, 8, 13, 15, 20, 22, and 27 (day in which the 30% of BSFL reached the prepupal stage). Production and waste reduction efficiency parameters, namely the growth rate, substrate reduction and substrate reduction index, were calculated. The two grape pomace varieties were analysed for their proximate composition and fatty acid profile; the same analyses were conducted on BSFL (30 larvae per replica) that were collected at the end of the trial (day 27). The growth rate of BSFL showed a higher value when the larvae were reared on B substrate (4.4 and 3.2 mg/day for B and M, respectively; P < 0.01). The rearing substrate did not significantly affect the proximate composition of BSFL. The percentage of total lipids (TL) in M-fed BSFL was significantly higher than in B ones. Total saturated (P < 0.001) and monounsaturated fatty acids (P < 0.05) were significantly higher in M-fed BSFL, while an opposite trend was observed for total branched-chain (P < 0.001) and total polyunsaturated fatty acids (P < 0.001). Interestingly, some conjugated linoleic acid (CLA) isomers [i.e., C18:2 c9t11(+t7c9+t8c10) and t9t11] were detected in low amounts in both rearing substrates (total CLA equal to 0.085 and 0.16 g/100 g TL in B and M substrate, respectively). Some CLA isomers (i.e., C18:2 c9t11, t7c9, and t10c12) were also found in BSFL, reaching a total CLA concentration equal to 2.95 and 0.052 g/100 g of TL in B-fed and M-fed BSFL, respectively. This study demonstrates that winery by-products from different grape varieties can significantly affect the development and lipid composition of BSFL. The CLA biosynthesis potential of BSFL opens newsworthy perspectives for a new valorisation of winery by-products to produce full-fat black soldier fly meal and black soldier fly oil enriched in specific fatty acids of potential health-promoting interest.

Effect of feeding black soldier fly larvae meal based diet on canine skin barrier function, organic antioxidant defence and blood biochemistry.

Black soldier fly meal in pet diets is gaining acceptance. This study aimed to assess the use of black soldier fly larvae defatted meal (BSFL) and its impact on blood parameters, biochemical markers, organic antioxidant capacity, skin barrier function and skin and coat quality. A cross-over study involved eight beagle dogs with two periods of 50 days each and a washout period of seven days in between. Two approximately iso-nutritive extruded diets were evaluated, the first containing 29.5% BSFL meal and a control diet containing 26% poultry by-product meal (PBP) as protein source. Skin and coat evaluations and blood collections were conducted before and after each period. Skin barrier function was assessed by measurement of trans epidermal water loss (TEWL) and stratum corneum hydration (SCH) in belly and pinna of the dogs on days 0, 15, 30, and 45 of each period. A trend for higher antioxidant effect significant reduction in serum scavenging capacity was found with PBP for BSFL diet trough malondialdehyde and Vitamin E measurement in dog's serum 2,2-diphenyl-1-picryl-hydrazyl (DPPH) assay. When fed PBP diet dogs exhibited reduction in serum cholesterol triglycerides and decreased LDL levels after 50 days, while dogs fed BSFL presented significant reduction in ALT. TEWL was significantly reduced in belly and pinna over time when dogs were fed BSFL, and TEWL in belly was significantly lower in dogs fed BSFL in comparison to PBP. while Increased SCH was also higher for the BSFL group observed in the same along the feeding period in comparison to PBP, indicating improved ability of the dogs to retain water and keep skin moisture. Improvement skin barrier function could be related to fatty acids from BSFL and increased sebaceous lipids in skin. These are responsible for to avoid water loss and improve skin protection against microbial insults. Inclusion of BSFL as protein source did not promote negative changes in blood biochemistry and had minor antioxidant effect in healthy dogs. However, it proved effective in improving skin barrier function, making BSFL a valuable alternative protein source for dogs, particularly those with sensitive skin or allergies manifesting on the skin.