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Efforts to recycle organic wastes using black soldier fly (BSF) Hermetia illucens into high-nutrient biomass that constitutes a sustainable fat (biodiesel) and high-quality protein ingredient in animal feeds have recently gained momentum worldwide. However, there is little information on the most suitable rearing conditions for growth, development and survivorship of these flies, which is a prerequisite for mass production technologies. We evaluated the physiological requirements for growth and reproduction of H. illucens on two diets [spent grains supplemented with brewers' yeast (D1) and un-supplemented (D2)]. Development rates at nine constant temperatures (10-42°C) were fitted to temperature-dependent linear and non-linear day-degree models. Thereafter, life history table parameters were determined within a range of favourable temperatures. The thermal maximum (TM) estimates for larval, pre-pupal and pupal development using non-linear model ranged between 37.2 ± 0.3 and 44.0 ± 2.3°C. The non-linear and linear day-degree model estimations of lower developmental temperature threshold for larvae were 11.7 ± 0.9 and 12.3 ± 1.4°C for D1, and 10.4 ± 1.7 and 11.7 ± 3.0°C for D2, respectively. The estimated thermal constant of immature life stages development of BSF was higher for the larval stage (250±25 DD for D1 and 333±51 for D2) than the other stages evaluated. Final larval wet weight was higher on D1 compared to D2. The population growth rate was most favourable at 30-degree celsius (°C) with higher intrinsic rate of natural increase (rm = 0.127 for D1 and 0.122 for D2) and shorter doubling time (5.5 days for D1 and 5.7 days for D2) compared to the other temperatures. These results are valuable for the optimization of commercial mass rearing procedures of BSF under various environmental conditions and prediction of population dynamics patterns using computer simulation models.

Generally, the DNA barcode relying on a short fragment of the cytochrome c oxidase I (COI) gene is a powerful tool for facilitating species discovery and taxonomic resolution in Diptera, including black flies. However, the COI barcode lacks sufficient resolution to identify several species or infer phylogenetic relationships of black flies in the Simulium striatum species-group, whereas the fast-evolving nuclear big zinc finger (BZF) gene has been suggested as a key marker for identifying the species. In this study, a new species of black fly in the S. striatum species-group from Kamphaeng Phet province, central Thailand, was discovered and characterized through an integrated method combining morphological analysis and molecular data based on the BZF gene. The new species, Simulium (Simulium) concitatum sp. nov., was morphologically described for all life stages, excluding the egg. It shares many morphological similarities with other species of the S. striatum species-group, particularly S. thilorsuense Takaoka, Srisuka & Saeung, 2022 described from Tak province, western Thailand. Sequence analysis and phylogeny inferred from the BZF gene further confirmed that S. concitatum sp. nov. is a distinct species of the S. striatum species-group and revealed its close genetic relationship to S. wangkwaienseTakaoka, Srisuka & Saeung, 2020. The morphological differences between the new species and all known species of the S. striatum species-group documented inThailand and other countries are provided to assist in species identification. Furthermore, this study underscores the BZF gene as an effective genetic marker to differentiate the species.

Black soldier fly (BSF) is a generalist species able to reduce large quantities of organic substrates and is thus considered as an interesting solution for waste management. Moreover, as BSF larvae accumulate high quantities of nutrients during their growth, they are valued because of their potential to produce products such as protein meal or fat for livestock feeds. Abiotic factors can influence larva growth, and a more detailed knowledge and control of these parameters can lead to the development of mass BSF breeding for the production of innovative products for animal feeds. As little information is available on the effects of the pH of substrates and feeding systems, the aim of this study was to evaluate the impact of these two factors on the activities of BSF larvae, prepupae, and adults. An experiment was performed with two fixed factors: i) pH (4.0; 6.1; 7.5; 9.5) and ii) feeding system (batch feeding system (TFS) or daily feeding system (DFS)). The pH treatments impacted larval weight on the first, third, and fifth day, but not at the end of the trial. Larval activity increased pH values from the fourth day onward, with final values of around 8.9-9.4 in all the treatments. The weight of the prepupae ranged from between 0.094 and 0.100 g. The final weight of the larvae and pupae, sex ratio, ingested food, larval mortality, percentage of emergence, and time to reach the pupa stadium were all affected by the feeding system. DFS showed the heaviest final larval weight (0.149 g), but required a longer time (11.3 d) than TFS to reach the prepupa stadium. The findings of this research could be useful for the mass production of BSF. Evaluation of an appropriate feeding system and initial pH value of the substrate are important parameters to reduce the time and to increase the weight in the production of larvae.

The black soldier fly ( Hermetia illucens ) is a saprophagous insect known for bioconverting organic waste, potentially offering environmental benefits, such as contributing to waste reduction and nutrient cycling. The performance of larvae varies significantly with factors substrate moisture, larval density, and scale of production. Three experiments were conducted using a mix of spent mushroom substrate (SMS) and chicken feed (CF). In the first experiment, 250 larvae were reared on 100 g dry matter (DM) feed at moisture levels of 65–75%. Results showed that the average individual larval weight, total biomass, and feed conversion ratio (FCR) improved with increased moisture. In the second experiment, 300 and 350 larvae/box were tested at 70% and 75% moisture. The highest average individual larval fresh weight (158.6 mg) was observed at 70% moisture with 250 larvae, while the highest biomass was achieved at 75% moisture with 300 larvae. Finally, different scales (10–2,500 g feed with 25–6,500 larvae) were tested with a similar feeding rate. The highest individual larval weight was recorded at the 100 g scale, with no clear correlation between weight and scale. However, the 50 g scale achieved the highest substrate reduction (33.2%). Overall, this study underscores the need to adjust moisture, density, and scale to nutrient conversion efficiency when using SMS, CF or other diets. The optimal results for the SMS feed mix were observed at 75% substrate moisture, 250 larvae per 100 g DM, and at approximately 2 larvae per cm 2 .

Cytoforms (cytospecies and cytotypes) of the Simulium damnosum complex are the major onchocerciasis vectors in Africa; but they differ in vectorial efficiency.Transmission elimination of onchocerciasis requires understanding the vector dynamics and delineation of transmission zones, for monitoring and evaluation, and for stop-treatment decisions.This study sought to understand the vector population dynamics and to delineate OperationalTransmission Zones in the 3 administrative regions (Far North, North, and Adamawa) of northern Cameroon. Using precontrol and relief maps, larvae were sampled in both rainy and dry seasons (July 2021 to February 2023) and fixed in Carnoy's solution. Chromosomal inversions were identified by cytotaxonomy. Karyotypes and inversion frequencies were compared between larval populations. Preimaginal stages were strictly seasonal in Far North, mostly seasonal in North, and mostly perennial in Adamawa. Five cytoforms (4 cytospecies and 1 cytotype) were recorded with 2 of them being reported for the first time (including S. yahenseVajime & Dunbar, an efficient forest zone vector), while 2 previously reported cytoforms were absent in all samples.These cytoform changes may impact the duration to transmission elimination. Also, there was redistribution of inversions in some populations and occurrence of new inversions at very high frequencies, suggesting genetic shifts probably caused by climate change.The 3 regions were distinct based on their vector phenology, cytoform composition, inversions and inversion frequencies, thereby enabling their delineation as separate OperationalTransmission Zones. Graphical Abstract

The growing global population increases the demand for protein, while organic waste management has become more challenging. Alternative protein sources are essential to mitigate the environmental impact of food production. The black soldier fly (BSF; Hermetia illucens ) has emerged as an alternative to traditional protein sources (e.g., soybean meal, fishmeal) due to its ability to convert diverse organic waste, addressing both issues simultaneously. This makes the BSF a promising candidate for industrial rearing, with its successful development closely tied to microbial influences on growth and behaviour, particularly bacterial influences on oviposition. In this study, we focus on the microbiota throughout insect development with a special focus on egg surface microbiota and their origin. We analysed the microbiota in the haemolymph and gut of larvae raised on sterilized and non-sterilized feed, pupal cell pulp, the wash of the ovipositor, eggs directly collected after oviposition, ovarian eggs, the empty female abdomen, eggs exposed to adult BSF, and surface-sterilized eggs. Our analysis revealed distinct bacterial community profiles across life stages, indicating a transition from larval dominance of Enterobacteriaceae to Burkholderiaceae on all analysed eggs. At the genus level, larval stages were characterized by Morganella , Escherichia , and Proteus , transitioning to less diverse communities in egg samples predominated by Burkholderia-Caballeronia-Paraburkholderia . Our study reveals that while predominant microbiota persist throughout all life stages, microbial community composition transforms progressively during maturation, particularly before oviposition. Understanding egg surface microbiota and the cues guiding oviposition has the potential to boost egg production and simplify mass harvesting of BSF larvae.

The widespread nominal black fly Simulium (Simulium) rufibasis Brunetti was reexamined morphologically, chromosomally, and molecularly to determine the status of populations in Japan and Korea with respect to S. rufibasis from the type locality in India and to all other known species in the S. (S.) tuberosum species-group. Morphological comparisons established that the species previously known as S. rufibasis in Japan and Korea is distinct from all other species. Consequently, it was described and illustrated as a new species, Simulium (S.) yamatoense. Simulium yokotense Shiraki, formerly a synonym of S. rufibasis, was morphologically reevaluated and considered a species unplaced to species-group in the subgenus Simulium. Chromosomal analyses of S. yamatoense sp. nov. demonstrated that it is unique among all cytologically known species of the S. tuberosum group and is the sister species of the Taiwanese species tentatively known as S. (S.) arisanum Shiraki. Populations of S. yamatoense sp. nov. included two cytoforms, based on the sex chromosomes. Cytoform A, including topotypical representatives, was found in Kyushu, Japan, whereas cytoform B was found in Korea and Honshu, Japan. Molecular analysis based on the COI mitochondrial gene generally corroborated morphological and chromosomal data that S. yamatoense sp. nov. is a distinct species and, like the chromosomal data, indicate that it is most closely related to S. arisanum, with interspecific genetic distance of 2.92–4.63%.

Density dependence in helminth establishment and heterogeneity in exposure to infection are known to drive resilience to interventions based on mass drug administration (MDA). However, the interaction between these processes is poorly understood. We developed a novel individual-based model for onchocerciasis transmission, EPIONCHO-IBM, which accounts for both processes. We fit the model to pre-intervention epidemiological data and explore parasite dynamics during MDA with ivermectin. Density dependence and heterogeneity in exposure to blackfly (vector) bites were estimated by fitting the model to matched pre-intervention microfilarial prevalence, microfilarial intensity and vector biting rate data from savannah areas of Cameroon and Côte d'Ivoire/Burkina Faso using Latin hypercube sampling. Transmission dynamics during 25 years of annual and biannual ivermectin MDA were investigated. Density dependence in parasite establishment within humans was estimated for different levels of (fixed) exposure heterogeneity to understand how parametric uncertainty may influence treatment dynamics. Stronger overdispersion in exposure to blackfly bites results in the estimation of stronger density-dependent parasite establishment within humans, consequently increasing resilience to MDA. For all levels of exposure heterogeneity tested, the model predicts a departure from the functional forms for density dependence assumed in the deterministic version of the model. This is the first, stochastic model of onchocerciasis, that accounts for and estimates density-dependent parasite establishment in humans alongside exposure heterogeneity. Capturing the interaction between these processes is fundamental to our understanding of resilience to MDA interventions. Given that uncertainty in these processes results in very different treatment dynamics, collecting data on exposure heterogeneity would be essential for improving model predictions during MDA. We discuss possible ways in which such data may be collected as well as the importance of better understanding the effects of immunological responses on establishing parasites prior to and during ivermectin treatment.

Black flies (Diptera: Simuliidae) are ecologically important components of lotic ecosystems and a public health concern due to their aggressive biting behavior, pathogen transmission, and induction of allergic and inflammatory reactions in humans and livestock. Despite their relevance, experimental research on Simuliidae remains limited compared with other dipteran vectors, largely because their immature stages require specific aquatic conditions that are difficult to reproduce in the laboratory. Larval development depends on continuous water flow, high dissolved oxygen, and suspended particulate food, factors that have hindered the establishment of reproducible rearing protocols. Here, we assess the feasibility and efficiency of rearing Simuliidae through their aquatic stages under controlled laboratory conditions approximating natural lotic environments. Egg masses were collected from submerged substrates in the São Pedro River (Macaé, Rio de Janeiro, Brazil) and transported to the Institute of Biodiversity and Sustainability (NUPEM/UFRJ) aquatic bioterium. Eggs were incubated in aerated containers until hatching, and larvae were transferred to a circular rearing system generating continuous unidirectional flow using submersible pumps. Physicochemical parameters were monitored daily (20-25 °C; pH 7.0; ammonia and nitrite ≤ 0.25 ppm). Larvae were fed a standardized suspension of yeast, spirulina, and chlorella. Eleven independent experiments monitored development from hatching to pupation. Additional assays tested larval densities (50, 100, 200 individuals) and survival following flow interruption. Pupal viability was assessed through adult emergence. Mean egg hatching success was 74%. Under continuous flow, larval survival averaged 92%, with pupation occurring within 13-15 days. Adult emergence reached 92.5%, indicating high pupal viability. Increased density caused only modest survival reduction, demonstrating system robustness. In contrast, flow interruption caused rapid mortality after 24 h, confirming strict dependence on oxygenated flow. Morphological identification revealed five species- , , , , and -indicating frequent multispecies oviposition. Simuliidae can be reliably reared through aquatic stages under laboratory conditions when hydrodynamic and physicochemical parameters are controlled. Although long-term colonization was not achieved, the protocol provides a reproducible framework for experimental studies on simuliid development, physiology, and biting-associated allergic responses.

Simulium quinquestriatum Shiraki (Diptera: Simuliidae), a human-biting fly that is distributed widely across Asia, is a vector for multiple pathogens. However, the larval development of this species is poorly understood. In this study, we determined the number of instars in this pest using three batches of field-collected larvae from Guiyang, Guizhou, China. The postgenal length, head capsule width, mandibular phragma length, and body length of 773 individuals were measured, and k-means clustering was used for instar grouping. Four distance measures—Manhattan, Euclidean, Chebyshev, and Canberra—were determined. The reported instar numbers, ranging from 4 to 11, were set as initial cluster centers for k-means clustering. The Canberra distance yielded reliable instar grouping, which was consistent with the first instar, as characterized by egg bursters and prepupae with dark histoblasts. Females and males of the last cluster of larvae were identified using Feulgen-stained gonads. Morphometric differences between the two sexes were not significant. Validation was performed using the Brooks–Dyar and Crosby rules, revealing that the larval stage of S. quinquestriatum is composed of eight instars.