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Orthopteran insects have high reproductive rates leading to boom-bust population dynamics with high local densities that are ideal for short, episodic disease epidemics. Viruses are particularly well suited for such host population dynamics, due to their supreme ability to adapt to changing transmission criteria. However, very little is known about the viruses of Orthopteran insects. Since Orthopterans are increasingly reared commercially, for animal feed and human consumption, there is a risk that viruses naturally associated with these insects can adapt to commercial rearing conditions, and cause disease. We therefore explored the virome of the house cricket Acheta domesticus, which is both part of the natural Swedish landscape and reared commercially for the pet feed market. Only 1% of the faecal RNA and DNA from wild-caught A. domesticus consisted of viruses. These included both known and novel viruses associated with crickets/insects, their bacterial-fungal microbiome, or their plant food. Relatively abundant among these viral Operational Taxonomic Units (OTUs) was a novel Iflavirus, tentatively named Acheta domesticus Iflavirus (AdIV). Quantitative analyses showed that AdIV was also abundant in frass and insect samples from commercially reared crickets. Interestingly, the wild and commercial AdIV strains had short, extremely divergent variation hotspots throughout the genome, which may indicate specific adaptation to their hosts’ distinct rearing environments.

Commercial cricket production has been plagued by viral disease outbreaks that have decimated their potential outputs and profit margins. To date, no epidemiological studies have been performed to estimate the prevalence of major viruses affecting crickets raised in commercial settings. A cross-sectional study was performed to estimate the prevalence of three important viruses (Acheta domesticus densovirus [AdDV], Acheta domesticus volvovirus [AdVVV], and invertebrate iridovirus 6 [CrIV]). Samples were collected across age groups (2-, 4-, and 6-week-olds) and seasons (January, May, August, and October) to determine the effect that these variables had on the prevalence rates. Quantitative real-time PCR was performed and revealed the following high overall prevalence rates: 46.7, 100, and 100% for AdDV, AdVVV, and CrIV, respectively. Viral loads varied from 101–5 for AdDV, 102–7 for AdVVV, and 102–9 for CrIV. AdDV prevalence rates were statistically significant across age and season (age: χ2 = 8.3, p = 0.015; season: χ2 = 59.7, p < 0.001), with crickets more likely to be infected as they aged and during the colder winter months. CrIV followed similar trends when looking at changes in viral loads between ages and seasons. AdVVV experienced a spike in viral loads across all ages during the month of August. Understanding the epidemiology of these viruses is instrumental in determining best management practices for commercially raised crickets.

Insects generally have high reproductive rates leading to rapid population growth and high local densities; ideal conditions for disease epidemics. The parasites and diseases that naturally regulate wild insect populations can also impact when these insects are produced commercially, on farms. While insects produced for human or animal consumption are often reared under high density conditions, very little is known about the microbes associated with these insects, particularly those with pathogenic potential. In this study we used both target-free and targeted screening approaches to explore the virome of two cricket species commonly reared for feed and food, Acheta domesticus and Gryllus bimaculatus . The target-free screening of DNA and RNA from a single A. domesticus frass sample revealed that only 1% of the nucleic acid reads belonged to viruses, including known cricket, insect, bacterial and plant pathogens, as well as a diverse selection of novel viruses. The targeted screening revealed relatively high levels of Acheta domesticus densovirus, invertebrate iridovirus 6 and a novel iflavirus, as well as low levels of Acheta domesticus volvovirus, in insect and frass samples from several retailers. Our findings highlight the value of multiple screening approaches for a comprehensive and robust cricket disease monitoring and management strategy. This will become particularly relevant as-and-when cricket rearing facilities scale up and transform from producing insects for animal feed to producing insects for human consumption.

Oil portion of Tenebrio molitor (yellow mealworm) and Acheta domesticus ( house cricket) were examined and it was investigated how the physicochemical properties of the oils changed with High Hydrostatic Pressure Assisted Extraction (HHP-E) and conventional solvent extraction (CE) with hexane. The effect of HHP-E at 500 MPa and 30 and 40 °C for 15 min on the properties of oils was compared with the CE. Following the extraction of oil, fatty acid composition, peroxide value, crystallization and melting points, total phenolic content and antioxidant activities were determined. Oil yield was found in the range of 22.75–24.22% for mealworm and 16.17–18.09% for cricket with significant amount of Ω-3 and Ω-6 fatty acids. Fatty acid composition of insect oils was significantly affected from HHP-E and extraction temperature (p < 0.05). The difference between crystallization and melting point of mealworm were found to be higher than cricket (p < 0.05). HHP-E insect oil had desirable characteristics to be used as a food ingredient. Graphic Abstract

Background: The house cricket, Acheta domesticus, is one of the most farmed insects worldwide and the foundation of an emerging industry using insects as a sustainable food source. Edible insects present a promising alternative for protein production amid a plethora of reports on climate change and biodiversity loss largely driven by agriculture. As with other crops, genetic resources are needed to improve crickets for food and other applications. Methods: We present the first high quality annotated genome assembly of A. domesticus from long read data and scaffolded to chromosome level, providing information needed for genetic manipulation. Results: Gene groups related to immunity were annotated and will be useful for improving value to insect farmers. Metagenome scaffolds in the A. domesticus assembly, including Invertebrate Iridescent Virus 6 (IIV6), were submitted as host-associated sequences. We demonstrate both CRISPR/Cas9-mediated knock-in and knock-out of A. domesticus and discuss implications for the food, pharmaceutical, and other industries. RNAi was demonstrated to disrupt the function of the vermilion eye-color gene producing a useful white-eye biomarker phenotype. Conclusions: We are utilizing these data to develop technologies for downstream commercial applications, including more nutritious and disease-resistant crickets, as well as lines producing valuable bioproducts, such as vaccines and antibiotics.

Current resource and processing constraints on conventional chitin production call for novel sources and more sustainable methods for its production. Herein, domestic cricket (Acheta domesticus L.) meal obtained from supercritical CO2 oil extraction was investigated as a viable source of chitin via a one-pot approach using acidic (choline chloride: glycerol, CCG) and alkaline (potassium carbonate: glycerol, KG) deep eutectic solvents (DESs). The chitin samples obtained were compared with those obtained using conventional acid-alkaline extraction (CE) and commercial crab shell chitin (CS chitin) by robust characterization of their composition and physicochemical properties employing color, FTIR, XRD, XPS, and SEM analysis. The results showed that KG DES and recovered KG DES exhibited high demineralization and deproteinization capacity, producing chitin with high purity, α-chitin form, high acetylation degree (>77%), crystallinity (crystallinity index > 81%), and micro-fibrous morphology closely similar to those of CE chitin and CS chitin. Whereas CCG DES demonstrated excellent demineralization, it was less effective at deproteinization, leading to chitin with lower purity and crystalline properties. Together, the results demonstrated that cricket meal could be an alternative source of chitin, while KG DES one-pot extraction holds strong potential as a sustainable and eco-friendly approach for obtaining commercial-grade chitin.

This study aimed to determine the influence and optimal conditions of incubation temperature and relative humidity (RH) on the egg hatchability patterns of two-spotted (Gryllus bimaculatus) and house (Acheta domesticus) crickets. Experiment I involved 100 cricket eggs per hatching box for each species, with six replications for each controlled incubation temperature of 23, 25, 27, 29, 30, 31, 32, and 33 °C at 70% RH. Experiment II used all the same procedures as Experiment I, except for incubation temperatures of 29, 30, 31, and 32 °C tested with varied RH levels of 65%, 70%, and 75%. In Experiment I, two-spotted crickets (9.47 ± 1.99 days) exhibited faster hatching than house crickets (13.70 ± 2.78 days). Additionally, the onset of hatching decreased with higher incubation temperatures for both types of crickets. In Experiment II, an incubation temperature of 31 °C and 70% RH resulted in a hatching rate of 79.75% for two-spotted crickets, with hatching beginning in 6 days. For house cricket eggs, the optimal conditions of 30 °C and 65–75% RH led to a peak daily hatching rate of 62.00–65.50% and hatching onset in 12 days. Thus, this study established the optimal incubation temperature and RH for egg hatching of two-spotted and house crickets.

Due to their nutritional value and sustainability, edible insect-based foods are gaining popularity in Europe. Their use is regulated by EU legislation, which defines authorised species and sets labelling requirements. Molecular tools are being developed to authenticate such products. In this study, yellow mealworm (Tenebrio molitor) larvae authorised for human consumption were fed wheat flour-based diets containing varying proportions of house cricket (Acheta domesticus) flour for 21 days. This was followed by a 48 h starvation period to assess the persistence of insect DNA in the digestive tract. Two novel, species-specific, single-copy markers were designed: ampd gene for the Acheta domesticus and MyD88 gene for the Tenebrio molitor. These were applied using qPCR and ddPCR. Both methods successfully detected cricket DNA in the guts of starved larvae. Linear regression analysis revealed a strong, statistically significant correlation between the proportion of Acheta domesticus flour in the diet and the normalised relative quantity of DNA. ddPCR proved to be more sensitive than qPCR, particularly in the detection of low DNA levels. These results suggest that the presence of DNA from undeclared insect species in edible insects may be indicative of their diet rather than contamination or adulteration. This highlights the importance of contextual interpretation in food authenticity testing.

The behavioural response of adult maize weevil, Sitophilus zeamais, to different types of semolina pasta enriched or not enriched with increasing proportions (5%, 10%, and 15%) of house cricket (Acheta domesticus) powder was investigated in olfactometer arena bioassays by using trap devices. In the five-choice behavioural bioassays, the number of S. zeamais adults attracted to 100% durum wheat semolina pasta was significantly higher than those attracted to the other pasta types enriched with A. domesticus powder. In the two-choice behavioural bioassays, the Response Index for each pasta type was positive and significant. However, although not significant, there was a progressive reduction in the Response Index as the cricket powder content increased. In similar experiments, there were no significant differences between cricket powder alone and the control in the number of attracted S. zeamais, indicating a neutral effect towards insects. These observations suggest that the lower attractiveness of pasta enriched with house cricket powder is mainly due to the masking of host food odours. Solid-phase microextraction coupled to gas chromatography–mass spectroscopy identified a total of 18 compounds in the head-space samples of the different types of pasta, highlighting differences in volatile composition. Some volatile compounds were only present in the pasta produced with cricket powder. In particular, 1-octen-3-ol and phenol were present in the samples containing 5%, 10%, or 15% cricket powder; pentanal, benzaldehyde, and dimethyl disulphide were present in samples containing 10% or 15% cricket powder; and 2,5-dimethyl-pyrazine was present in the sample containing 15% cricket powder. Further investigation with individual compounds and mixtures is needed to define the chemical basis of the differences in the insect olfactory preference observed in this study.

Food safety for cricket production is a crucial factor in producing edible crickets with safety for consumers and sustainability for two-spotted (Gryllus bimaculatus) as well as house (Acheta domesticus) cricket production. This study was conducted by simultaneously rearing two cricket species, comprising two-spotted crickets (G. bimaculatus) and house crickets (A. domesticus). A total of 16 rearing crates were used for the present study, which were allocated into 8 rearing crates for each studied cricket species, including paper egg cartons. Cricket eggs were incubated in the rearing crates. Once the crickets hatched, tap water and powdered feed were provided ad libitum throughout the experiment. At the end of this study (35 and 42 days for the two-spotted and house crickets, respectively), all crickets were harvested, rinsed in tap water, and boiled in water for 5 min. During the rearing and harvesting processes, samples were collected from various potential contamination points for bacteria, including E. coli and Salmonella spp. There were samples of the initial input (feed, drinking water, and staff hands), rearing environment (water pipe, crate wall, living cartons, frass, and cricket surface), and harvesting crickets (harvested, washed, and boiled crickets), with a 2-week sampling interval, except for the last round of sampling for the two-spotted crickets. Subsequently, all samples were submitted to isolate and identify contaminated bacteria. The samples from the last round of sampling for both kinds of crickets were submitted to quantify the level of contamination for E. coli and Salmonella spp., including antimicrobial resistance by the disk diffusion method for the positive isolate. The results showed that bacterial contamination was found in the rearing of both cricket species, primarily involving Klebsiella spp. and Enterobacter spp., mainly found in prepared drinking water and the water pipes of drinking water supply equipment, which are potential sources of contamination with cricket frass. E. coli was found in 4.8% and 4.3% of the two-spotted and house crickets, respectively, while no presence of Salmonella spp. was detected in any submitted samples. The quantification of E. coli and Salmonella spp. indicated E. coli contamination near the water pipe and the frass of two-spotted crickets, but Salmonella spp. was undetectable in both two-spotted and house crickets. The antimicrobial resistance of isolated E. coli mainly involved penicillin G, amoxicillin, ampicillin, erythromycin, lincomycin, and tiamulin. Thus, good farm management with proper sanitation practices (such as cleaning and keeping the environment dry), as well as boiling crickets during the harvesting process, may help ensure the safety of edible cricket production.