Bioinformatic Surveillance Leads to Discovery of Two Novel Putative Bunyaviruses Associated with Black Soldier Fly.

The black soldier fly (Hermetia illucens, BSF) has emerged as an industrial insect of high promise because of its ability to convert organic waste into nutritious feedstock, making it an environmentally sustainable alternative protein source. As global interest rises, rearing efforts have also been upscaled, which is highly conducive to pathogen transmission. Viral epidemics have stifled mass-rearing efforts of other insects of economic importance, such as crickets, silkworms, and honeybees, but little is known about the viruses that associate with BSF. Although BSFs are thought to be unusually resistant to pathogens because of their expansive antimicrobial gene repertoire, surveillance techniques could be useful in identifying emerging pathogens and common BSF microbes. In this study, we used high-throughput sequencing data to survey BSF larvae and frass samples, and we identified two novel bunyavirus-like sequences. Our phylogenetic analysis grouped one in the family Nairoviridae and the other with two unclassified bunyaviruses. We describe these putative novel viruses as BSF Nairovirus-like 1 and BSF uncharacterized bunyavirus-like 1. We identified candidate segments for the full BSF Nairovirus-like 1 genome using a technique based on transcript co-occurrence and only a partial genome for BSF uncharacterized bunyavirus-like 1. These results emphasize the value of routine BSF colony surveillance and add to the number of viruses associated with BSF.

Large-scale production of house fly, Musca domestica (Diptera: Muscidae), larvae fed 3 manure types.

House flies, Musca domestica, L., (Diptera: Muscidae) are well-known pests at animal facilities; however, they can be used for manure biodegradation. Utilizing house flies to process animal manure offers a means to recycle nutrients and reduce contaminants (e.g., pathogens and heavy metals), while also producing multiple revenue streams (e.g., protein for feed, fat for biodiesel, frass as a soil amendment). This study determined house fly larval performance on a larger scale (kilogram of wastes; thousands of larvae; single feeding) as a follow-up to a previous experiment performed at a bench-top scale (g of wastes; hundreds of larvae; incremental feeding). Four thousand larvae were fed 1 kg of swine, dairy, or poultry manure, or a control (Gainesville diet: 50% wheat bran, 30% alfalfa meal, and 20% corn meal). Peak larval weight occurred 4 days after inoculation and no significant difference in development time to first pupariation occurred across diets. However, percent survivorship to pupariation varied, with the highest occurring in Gainesville (74%), swine (73%), and poultry (67%) manure, whereas 50% survived when fed dairy manure. The highest pupal weight was found for those fed Gainesville (27 mg), and similar weights were found for those fed swine (21 mg), dairy (24 mg), and poultry (25 mg) manure. Although using house flies to manage manure has received little consideration in Western countries, other regions have this practice in place. Results may provide insight on differences between small- and large-scale studies, which is valuable for industrialization of this species for waste management and creating a more circular economy.

Odorant binding proteins fromHermetia illucens: potential sensing elements for detecting volatile aldehydes involved in early stages of organic decomposition.

Organic decomposition processes, involving the breakdown of complex molecules such as carbohydrates, proteins and fats, release small chemicals known as volatile organic compounds (VOCs), smelly even at very low concentrations, but not all readily detectable by vertebrates. Many of these compounds are instead detected by insects, mostly by saprophytic species, for which long-range orientation towards organic decomposition matter is crucial. In the present work the detection of aldehydes, as an important measure of lipid oxidation, has been possible exploiting the molecular machinery underlying odour recognition inHermetia illucens(Diptera: Stratiomyidae). This voracious scavenger insect is of interest due to its outstanding capacity in bioconversion of organic waste, colonizing very diverse environments due to the ability of sensing a wide range of chemical compounds that influence the choice of substrates for ovideposition. A variety of soluble odorant binding proteins (OBPs) that may function as carriers of hydrophobic molecules from the air-water interface in the antenna of the insect to the receptors were identified, characterised and expressed. An OBP-based nanobiosensor prototype was realized using selected OBPs as sensing layers for the development of an array of quartz crystal microbalances (QCMs) for vapour phase detection of selected compounds at room temperature. QCMs coated with four recombinantH. illucensOBPs (HillOBPs) were exposed to a wide range of VOCs indicative of organic decomposition, showing a high sensitivity for the detection of three chemical compounds belonging to the class of aldehydes and one short-chain fatty acid. The possibility of using biomolecules capable of binding small ligands as reversible gas sensors has been confirmed, greatly expanding the state-of the-art in gas sensing technology.

Hermetia illucens (L.) (Diptera: Stratiomyidae) Odorant Binding Proteins and Their Interactions with Selected Volatile Organic Compounds: An In Silico Approach.

The black soldier fly (BSF), Hermetia illucens (Diptera: Stratiomyidae), has considerable global interest due to its outstanding capacity in bioconverting organic waste to insect biomass, which can be used for livestock, poultry, and aquaculture feed. Mass production of this insect in colonies requires the development of methods concentrating oviposition in specific collection devices, while the mass production of larvae and disposing of waste may require substrates that are more palatable and more attractive to the insects. In insects, chemoreception plays an essential role throughout their life cycle, responding to an array of chemical, biological and environmental signals to locate and select food, mates, oviposition sites and avoid predators. To interpret these signals, insects use an arsenal of molecular components, including small proteins called odorant binding proteins (OBPs). Next generation sequencing was used to identify genes involved in chemoreception during the larval and adult stage of BSF, with particular attention to OBPs. The analysis of the de novo adult and larval transcriptome led to the identification of 27 and 31 OBPs for adults and larvae, respectively. Among these OBPs, 15 were common in larval and adult transcriptomes and the tertiary structures of 8 selected OBPs were modelled. In silico docking of ligands confirms the potential interaction with VOCs of interest. Starting from the information about the growth performance of H. illucens on different organic substrates from the agri-food sector, the present work demonstrates a possible correlation between a pool of selected VOCs, emitted by those substrates that are attractive for H. illucens females when searching for oviposition sites, as well as phagostimulants for larvae. The binding affinities between OBPs and selected ligands calculated by in silico modelling may indicate a correlation among OBPs, VOCs and behavioural preferences that will be the basis for further analysis.

Evaluation of Development Datasets for Hermetia illucens (L.) (Diptera: Stratiomyidae) for Estimating the Time of Placement of Human and Swine Remains in Texas, USA.

A basic tenet of forensic entomology is development data of an insect can be used to predict the time of colonization (TOC) by insect specimens collected from remains, and this prediction is related to the time of death and/or time of placement (TOP). However, few datasets have been evaluated to determine their accuracy or precision. The black soldier fly, Hermetia illucens (L.) (Diptera: Stratiomyidae) is recognized as an insect of forensic importance. This study examined the accuracy and precision of several development datasets for the black soldier fly by estimating the TOP of five sets of human and three sets of swine remains in San Marcos and College Station, TX, respectively. Data generated from this study indicate only one of these datasets consistently (time-to-prepupae 52%; time-to-eclosion 75%) produced TOP estimations that occurred within a day of the actual TOP of the remains. It is unknown if the precolonization interval (PreCI) of this species is long, but it has been observed that the species can colonize within 6 d after death. This assumption remains untested by validation studies. Accounting for this PreCI improved accuracy for the time-to-prepupae group, but reduced accuracy in the time-to-eclosion group. The findings presented here highlight a need for detailed, forensic-based development data for the black soldier fly that can reliably and accurately be used in casework. Finally, this study outlines the need for a basic understanding of the timing of resource utilization (i.e., duration of the PreCI) for forensically relevant taxa so that reasonable corrections may be made to TOC as related to minimum postmortem interval (mPMI) estimates.

Black soldier fly, Hermetia illucens (L.) (Diptera: Stratiomyidae), and house fly, Musca domestica L. (Diptera: Muscidae), larvae reduce livestock manure and possibly associated nutrients: An assessment at two scales.

The industrial production of insects for waste management or as a protein source is becoming vital to our society. Large volumes of manure are produced by concentrated animal facilities around the globe that must be managed, utilized, and disposed of properly. Flies offer a partial solution with their abilities to reduce these wastes and heavy metal pollutants. Meat and crop proteins are being supplemented by insect proteins for many feeds across the globe, yet science-based studies behind the mass-rearing of insects are still in their infancy. In the current study, the percent change in the composition of nutrients, heavy metals, and fiber, in dairy, poultry, and swine manure degraded by either black soldier fly (BSF) or house fly (HF) larvae was explored. Pre-digested and post-digested manure samples were collected from four independent studies that differed in production scale (number of larvae and feeding regimen): 1) BSF small-scale (100 larvae fed incrementally), 2) HF small-scale (100 larvae fed incrementally), 3) BSF large-scale (10,000 larvae fed a single time), and 4) HF large-scale (4,000 larvae fed a single time). Results indicate that nitrogen is a key nutrient impacted by larval digestion of manure by both species, regardless of scale. However, scale significantly impacted reductions of other nutrients, as did the type of manure in which the insects were reared. Ultimately, this study demonstrated that manure type and rearing scale impact the ability of BSF and HF larvae to reduce nutrients and heavy metals in manure, and thus insect management procedures need to be congruent with production emphases of the insects for waste management or protein products. Failure to take scale into consideration could lead to inaccurate assumptions related to industrialized efforts on this topic.

Rearing substrate impacts growth and macronutrient composition of Hermetia illucens (L.) (Diptera: Stratiomyidae) larvae produced at an industrial scale.

Organic waste is a rapidly increasing problem due to the growth of the agricultural production needed to meet global food demands. Development of sustainable waste management solutions is essential. Black soldier fly, Hermetia illucens (L.) (Diptera: Stratiomyidae) (BSF), larvae are voracious consumers of a wide range of organic materials ranging from fruits and vegetables to animal remains, and manure. Thanks to this ability and considering the larval high protein and lipid content, BSF larvae are a useful additive in animal feeds and biodiesel production. Unfortunately, the feasibility of using the black soldier fly as a tool for waste valorization and feed production has primarily been investigated at the benchtop scale. Thus, mobilization of current practices to an industrial scale is challenging because scaling up from small laboratory studies to large industrial studies is not necessarily linear. The goal of this study was to demonstrate the ability of the BSF to recycle organic waste at an industrial scale. To accomplish this goal, three organic waste streams were used (e.g., apples, bananas, and spent grain from a brewery) to test six diet treatments (1) apple, (2) banana, (3) spent grain, (4) apple and banana, (5) apple and spent grain, and (6) banana and spent grain. Working at scale of 10,000 BSF larvae life history traits, waste valorization, protein and lipid profiles were measured for each diet treatment. Differences were recorded across all variables, except substrate conversion, for larvae fed on fruit and spent grain (alone or with fruit). Growth rate significantly differed across treatments; larvae reared on spent grain grew twice as fast as those fed apples alone, but those reared on the apple and spent grain mixture produced twice as much insect biomass. However, it should be noted that larvae resulting from the apple diet contained 50% more fat than larvae fed the fruit and spent grain mixtures. Commonly-available organic wastes were successfully used at an industrial scale to produce BSF larvae that have the potential to substitute other sources of protein and lipids in different industrial applications. Industrialization efforts are encouraged to assess these impacts when integrating diverse ingredients into larval diets as a means to more precisely predict output, such as larval development time and final larval biomass.

Hermetia illucens and Hermetia fenestrata (Diptera: Stratiomyidae) Colonization of "Spoiled" Stingless Bee Geniotrigona thoracica (Hymenoptera: Apidae) Hives in Malaysia.

Meliponiculture, the keeping of domesticated stingless bees such as Geniotrigona thoracica (Smith, 1857) (Hymenoptera: Apidae), is an increasingly popular agricultural industry in Malaysia. This study reports the soldier fly (Diptera: Stratiomyidae) species of the genus Hermetia colonizing stingless bee colonies in Malaysia. The larvae were reared in the laboratory to the adult stage and identified through molecular and morphological approaches. Hermetia illucens (Linnaeus, 1758) and Hermetia fenestrata de Meijere, 1904 (Diptera: Stratiomyidae) were identified from the sample provided. Earlier records of stratiomyids in stingless bee nests were misidentified as H. illucens. This paper represents the first identified record of H. fenestrata colonizing a "spoiled" stingless bee colony. In addition, adult and larval morphological differences between both species and the roles of both species in bee nest decomposition are discussed.

Rethinking organic wastes bioconversion: Evaluating the potential of the black soldier fly (Hermetia illucens (L.)) (Diptera: Stratiomyidae) (BSF).

Population growth and unprecedented economic growth and urbanization, especially in low- and middle-income countries, coupled with extreme weather patterns, the high-environmental footprint of agricultural practices, and disposal-oriented waste management practices, require significant changes in the ways we produce food, feed and fuel, and manage enormous amounts of organic wastes. Farming insects such as the black soldier fly (BSF) (Hermetia illucens) on diverse organic wastes provides an opportunity for producing nutrient-rich animal feed, fuel, organic fertilizer, and biobased products with concurrent valorization of wastes. Inclusion of BSF larvae/pupae in the diets of poultry, fish, and swine has shown promise as a potential substitute of conventional feed ingredients such as soybean meal and fish meal. Moreover, the bioactive compounds such as antimicrobial peptides, medium chain fatty acids, and chitin and its derivatives present in BSF larvae/pupae, could also add values to the animal diets. However, to realize the full potential of BSF-based biorefining, more research and development efforts are necessary for scaling up the production and processing of BSF biomass using more mechanized and automated systems. More studies are also needed to ensure the safety of the BSF biomass grown on various organic wastes for animal feed (also food) and legalizing the feed application of BSF biomass to wider categories of animals. This critical review presents the current status of the BSF technology, identifies the research gaps, highlights the challenges towards industrial scale production, and provides future perspectives.

Mass Production of the Black Soldier Fly, Hermetia illucens (L.), (Diptera: Stratiomyidae) Reared on Three Manure Types.

Recent interest in the mass production of black soldier fly (BSF) larvae has resulted in many studies being generated. However, a majority of the studies are benchtop, or small-scale, experiments. Results generated from such studies may not translate to large-scale/industrial production. The current study was conducted at a conventional large-scale (10,000 larvae/treatment fed seven kg) to determine the impact on selected life-history traits when BSF were fed seven kg of manure (swine, dairy, or poultry) or a control diet (Gainesville diet: 50% wheat bran, 30% alfalfa meal, and 20% corn). Results showed larvae fed dairy manure took one to two days longer to develop to prepupation, with lower survivorship (45%) compared to those fed poultry or swine manure (>70%). Furthermore, the maximum larval weight was reached on day six for those fed swine manure, while other treatments achieved the maximum weight on day seven. However, larvae fed swine manure averaged 150 mg, while those fed the other diets ranged between 175 and 200 mg. Data from this study may be valuable for the industrialization of BSF. Companies using a scale varying from previously published work, including this study, should conduct pilot studies to optimize their system prior to implementation.

Interspecific Competition between the House Fly, Musca domestica L. (Diptera: Muscidae) and Black Soldier Fly, Hermetia illucens (L.) (Diptera: Stratiomyidae) When Reared on Poultry Manure.

Few studies have examined the competitive interaction between the house fly (HF) and the black soldier fly (BSF). The fact that the BSF deters HF oviposition is widely cited in BSF literature, but this interaction has not been assessed in over three decades. In this study, the competitive interaction of BSF and HF larvae was observed on fresh (day 0) and aged poultry manure (manure aged for two, four, six, or eight days). Specifically, a priority effect study was conducted to determine if colonization sequence influences time to first pupariation (HF) or pre-pupation (BSF), survivorship, and weight. Results show >70% of HFs reached pupariation in all treatments except when placed on manure eight days after the initial inoculation with BSF. However, age of the resource negatively impacted time to first pupariation and puparium weight when HFs were reared alone or introduced two to eight days after BSF. No BSF pre-pupae resulted from treatments in which HFs were the pioneering species. BSFs reached the highest percent pre-pupation when reared alone on fresh manure, but BSFs may be more susceptible to the negative impacts of an aging resource, as no pre-pupae were observed when provided with six- or eight-day-old manure. Similar to HFs, age of the resource may have impacted development and survivorship; other factors such as moisture content, chemical composition, and amount of resource provided may have also impacted our results. These data may be useful in implementing BSFs as biological control agents of the HF, as well provide valuable information for facilities mass-producing HFs or BSFs for food or feed.

Life-History Traits of the Black Soldier Fly, Hermetia illucens (L.) (Diptera: Stratiomyidae), Reared on Three Manure Types.

Structural changes and growth of animal production systems have resulted in greater volumes of manure. Current manure storage methods pose a potential environmental threat. Lessening these issues is a key concern for the animal production industry. The primary aim of this research was to evaluate black soldier fly (BSF) performance when fed poultry, swine, or dairy manure at different rates (18 or 27 g/2 d until 40% prepupation). The results indicated that larvae fed with the control diet (Gainesville diet) were the heaviest (+31-70%); however, for other life-history traits, those fed the higher feed rate of poultry manure produced comparable results to the control. Larvae fed more resource, regardless of manure type, weighed more as larvae (+3-9%), pupae (+22-48%), and adults (+18-42%), developed faster (up to 3-4 d), had a higher percentage reach the prepupal stage (+2-16%), lived longer as adults (+1 d), and converted more resource to biomass (up to 1% more) than those fed at the lower rate. Yet, no difference was detected in dry matter (DM) reduction across feed rate for a given manure type. Based on these results, all three manure types can be digested by black soldier fly larvae, thus demonstrating their potential for waste management.

Larval digestion of different manure types by the black soldier fly (Diptera: Stratiomyidae) impacts associated volatile emissions.

Volatile emissions from decomposing animal waste are known environmental pollutants. The black soldier fly, Hermetia illucens (L.), is being evaluated for industrialization as a means to recycle wastes and produce protein for use as food and feed. We examined the ability of black soldier fly larvae to reduce odorous compounds associated with animal wastes. Black soldier fly larvae were reared under laboratory conditions on poultry, swine, and dairy manure at feed rates of 18.0 and 27.0 g every other day until 40% reached the prepupal stage. Volatile emissions were collected and analyzed from freshly thawed as well as the digested waste when 90% of the black soldier fly larvae reached the prepupal stage. Volatiles were also collected simultaneously from manure not inoculated with black soldier fly larvae (non-digested) and held under similar conditions. Manure samples were analyzed for relative amounts of nine select odorous volatile organic compounds: phenol, 4-methylphenol, indole, 3-methylindole, propanoic acid, 2-methylpropanoic acid, butanoic acid, 3-methylbutanoic acid and pentanoic acid. Black soldier fly larvae reduced emissions of all volatile organic compounds by 87% or greater. Complete reductions were observed for 2-methly propanoic acid in digested poultry manure, phenol, 4-methylphenol, indole and all five fatty acids in digested swine manure, and 4-methylphenol, indole, 3-methylindole and all five acids in digested dairy manure. This study is the first to identify volatile emissions from manure digested by black soldier fly larvae and compare to those found in non-digested manure. These data demonstrate additional benefits of using black soldier fly larvae as a cost-effective and environmentally friendly means of livestock manure management in comparison to current methods. CAPSULE: Black soldier fly larvae are capable of altering the overall profile of volatile organic compounds and reducing levels of targeted odorous compounds in livestock manure.

The Impact of Diet Protein and Carbohydrate on Select Life-History Traits of The Black Soldier Fly Hermetia illucens (L.) (Diptera: Stratiomyidae).

This study examined the impact of diet protein and carbohydrate percentages as well as moisture on the immature development, survivorship, and resulting adult longevity and egg production of the black soldier fly, Hermetiaillucens (L.) (Diptera: Stratiomyidae). Moisture impacted development and corresponding life-history traits more than protein:carbohydrate content; larvae were unable to develop on diets at 40% moisture. Larvae fed diets at 70% moisture developed faster, grew larger, and required less food than those reared on diets at 55% moisture. Larvae reared on the balanced diet (21% protein:21% carbohydrate) at 70% moisture developed the fastest on the least amount of food and had the greatest survivorship to the prepupal stage. Adult emergence and longevity were similar across treatments, indicating immature life-history traits were impacted the most. The control (Gainesville house fly) diet was superior to the artificial diets for all parameters tested. These differences could indicate that other constituents (e.g., associated microbes) serve a role in black soldier fly development. These data are valuable for industrialization of this insect as a "green" technology for recycling organic waste, which can be highly variable, to produce protein for use as feed in the livestock, poultry, and aquaculture industries, as well as for bioenergy production.

Cool-weather activity of the forensically important hairy maggot blow fly Chrysomya rufifacies (Macquart) (Diptera: Calliphoridae) on carrion in Upstate South Carolina, United States.

The hairy maggot blow fly Chrysomya rufifacies (Macquart) (Diptera: Calliphoridae) has expanded its range in the United States since its introduction into Texas (ca. 1980) and has been collected in 15 states. We investigated the bionomics of immature and adult C. rufifacies collected from carcasses of a raccoon Procyon lotor (Linnaeus) and white-tailed deer Odocoileus virginianus Zimmerman in Upstate South Carolina during November 2007, and used these insects to estimate the minimum period of insect activity. Puparia of C. rufifacies were collected from deer carrion; 28% were parasitized by Nasonia vitripennis (Walker) (Hymenoptera: Pteromalidae). The mean daily ambient temperature during this study was 11.4+/-1.02 degrees C, representing the lowest recorded mean temperature for adult activity of C. rufifacies; adults of C. rufifacies were observed flying among the carcasses at 9.0 degrees C. Although C. rufifacies is considered a warm-weather blow fly, researchers should be aware of its activity at suboptimal conditions, behavior that might aid its expansion into more northern areas.