Gryllus Bimaculatus Research Articles

Cricket Meal (Gryllus bimaculatus) as a Protein Supplement on In Vitro Fermentation Characteristics and Methane Mitigation.

Simple SummaryProtein sources of high quality and sustainability are found in insects. In many regions, insects are a primary food source, such as in Africa, South America, Asia, and Oceania. Insects are considered promising alternative feed sources, in particular as a source of protein. The use of edible insects as high-protein sources is widespread, and cricket has been proved to be a potential food and feed insect species. Cricket (Gryllus bimaculatus) also contain 54.10% crude protein, 6.90% crude fiber, 26.90% fat, and 78.90% total digestible nutrient, as well as a variety of essential amino acids, including methionine, lysine, histidine, valine, and leucine. In addition, insects have been investigated as a source of protein in diets of poultry, swine, and fish. However, there are currently little data on the utilization of insects as ruminant feed. The objective of this experiment was to conduct the effects of Cricket meal (Gryllus bimaculatus) (CM) as a protein replacement for soybean meal on in vitro fermentation end products, gas production, nutrient degradability, and methane mitigation.The aim of this work was to conduct the effects of cricket (Gryllus bimaculatus) meal (CM) as a protein supplement on in vitro gas production, rumen fermentation, and methane (CH4) mitigation. Dietary treatments were randomly assigned using a completely randomized design (CRD) with a 2 × 5 factorial arrangement. The first factor was two ratios of roughage to concentrate (R:C at 60:40 and 40:60), and the second factor was the level of CM to replace soybean meal (SBM) in a concentrate ratio at 100:0, 75:25, 50:50, 25:75, and 0:100, respectively. It was found that in vitro DM degradability and the concentration of propionic (C3) were significantly increased (p < 0.05), while the potential extent of gas production (a + b), acetate (C2), acetate and propionate (C2:C3) ratio, and protozoal population were reduced (p < 0.05) by lowering the R:C ratio and the replacement of SBM by CM. In addition, rumen CH4 production was mitigated (p < 0.05) with increasing levels of CM for SBM. In this study, CM has the potential to improve rumen fermentation by enhancing C3 concentration and DM degradability, reduced methane production, and C2:C3 ratio. The effects were more pronounced (p < 0.05) at low levels of roughage.

The Effects of Antioxidants and Packaging Methods on Inhibiting Lipid Oxidation in Deep Fried Crickets (Gryllus bimaculatus) during Storage.

This study aimed to investigate the effect of processing methods on inhibiting lipid oxidation of deep fried crickets (Gryllus bimaculatus) during storage. Four antioxidants and two packaging methods were used. The effects of different antioxidants and packaging methods on composition of fatty acids, contents of free fatty acids (FFA), peroxide value (PV), and thiobarbituric acid reactive substances (TBARSs) value of deep fried Gryllus bimaculatus were analyzed during 150 days of storage. The composition of fatty acids changed and the content of FFA, PV, and TBARs value also increased with the extension of storage time, indicating that the lipid oxidation dominated by oxidation of unsaturated fatty acids could occur in deep fried Gryllus bimaculatus during storage. In the same storage period, the total content of FFA, PV, and TBARs value of samples treated with antioxidants and vacuum-filling nitrogen packaging were lower than those of controls, suggesting that antioxidants and vacuum-filling nitrogen packaging have noticeable effects on inhibiting lipid oxidation and improving the quality of deep fried crickets, and dibutyl hydroxyl toluene (BHT) was found as the most effective antioxidant in this study. The results may provide a reliable reference for processing of deep fried edible insects.

Use of edible insects in thermal-induced protein gels containing porcine myofibrillar protein

Edible insects are considered an environmentally friendly source of protein; however, products containing only edible insects are associated with poor quality and processing properties. Therefore, we examined and compared the thermal stability and rheological properties among thermal-induced myofibrillar protein gels prepared using porcine myofibril protein and five different edible insect species: larvae: Tenebrio molitor L., Protaetia brevitarsis, and Allomyrina dichotoma and adult insects: Gryllus bimaculatus and Oxyachinensis sinuosa. The myofibrillar protein gels containing edible insects had a higher pH than that of the control. The water-holding capacity of the myofibrillar gel containing G. bimaculatus (T4) was higher than that of the other samples. The hardness, cohesiveness, gumminess, and chewiness of the thermal-induced myofibrillar gel containing T4 were the highest. The myofibrillar gel containing edible insects had a higher peak temperature than that of the control, except for T4, as well as a similar or lower enthalpy, which is required for denaturation. All myofibrillar gels with/without edible insects exhibited Hershel-Bulkley fluid properties, and rigid structures were developed during heating. The myofibrillar protein gels containing T4 revealed excellent processing properties. Therefore, myofibrillar protein gels containing T4 may be used for producing protein-rich foods.

Encounter with heavier females changes courtship and fighting efforts of male field crickets Gryllus bimaculatus (Orthoptera: Gryllidae)

The effects of mating experience on male mating behavior are mediated by four factors: mating cost, such as resource depletion, perception of mating opportunities, self-perception of attractiveness, and female quality. For example, encountering females might increase male expectations of prospective mating opportunities, while copulation increases self-perception of attractiveness in males. To determine the relative importance of these factors, the effect of mating on the two components of reproductive effort (courtship and fighting effort) in Gryllus bimaculatus was examined. Calling activity before and after encountering females was measured, and copulation success was recorded. Subsequently, the intensity and outcome of male–male fighting behavior was recorded. Female encounter increased calling activity irrespective of copulation, thereby indicating that the perception of mating opportunities is important factor for the males. Changes in courtship effort of males were larger and fighting success was lower when they were previously paired with relatively heavier females. These results indicate that male reproductive effort is also affected by quality of previous mating partners.

The Central Organisation of Courtship and Rivalry Behaviour in Gryllus Bimaculatus (Degeer) as Revealed by Lesions of Abdominal Connectives

The Central Organisation of Courtship and Rivalry Behaviour in Gryllus Bimaculatus (Degeer) as Revealed by Lesions of Abdominal Connectives

Evaluation of Anti-Cancer Activity of Gryllus bimaculatus Water Extract on Non-Small Cancer Lung Cell via Apoptosis.

Gryllus bimaculatus, traditionally used in oriental medicine, demonstrates functional and pharmacological potential through demonstrating immunomodulatory, hepato-protective properties, anti-inflammatory, antioxidant, and neuroprotective effects. In this study, we examined the effect of G. bimaculatus on cell proliferation and apoptosis in lung cancer cells. This is the first study to examine the anti-cancer effects of G. bimaculatus extracts on non-small cell lung cancer. Frozen G. bimaculatus was obtained, homogenized, and dissolved in distilled water. Using a freeze dryer, samples were concentrated until almost all the water was removed, and extracts were diluted in solutions of phosphate buffered saline. Anti-cancer effects of extracts on human non-small cancer lung cells were estimated based on cell cytotoxicity, western blot, and flow cytometry, using lipopolysaccharides as a positive control. H460 and A549 human non-small cell cancer lung cells were treated with G. bimaculatus water extracts of various concentrations, with lipopolysaccharide used as a pos-itive control. The results showed that treatment with the extract for 24 or 48 h inhibited H460 proliferation, demonstrated by reduced cell numbers with morphological changes. Furthermore, flow cytometry analysis demonstrated that the extracts induced cell death on H460. However, extracts did not show cytotoxic effects on A549 cells. In conclusion, the extract induced apoptosis of lung cancer cells, possibly via caspase, Bcl-2 family signaling pathways. Therefore, G. bimaculatus water extracts are safe and efficient natural materials that may have great potential in the treatment of lung cancer.

Tolerant pattern recognition: evidence from phonotactic responses in the cricket Gryllus bimaculatus (de Geer).

When the amplitude modulation of species-specific acoustic signals is distorted in the transmission channel, signals become difficult to recognize by the receiver. Tolerant auditory pattern recognition systems, which after having perceived the correct species-specific signal transiently broaden their acceptance of signals, would be advantageous for animals as an adaptation to the constraints of the environment. Using a well-studied cricket species, Gryllus bimaculatus, we analysed tolerance in auditory steering responses to ‘Odd’ chirps, mimicking a signal distorted by the transmission channel, and control ‘Silent’ chirps by employing a fine-scale open-loop trackball system. Odd chirps on their own did not elicit a phonotactic response. However, when inserted into a calling song pattern with attractive Normal chirps, the females' phonotactic response toward these patterns was significantly larger than to patterns with Silent chirps. Moreover, females actively steered toward Odd chirps when these were presented within a sequence of attractive chirps. Our results suggest that crickets employ a tolerant pattern recognition system that, once activated, transiently allows responses to distorted sound patterns, as long as sufficient natural chirps are present. As pattern recognition modulates how crickets process non-attractive acoustic signals, the finding is also relevant for the interpretation of two-choice behavioural experiments.

English

In the recent past, cricket production has attracted a myriad of interests in the Global Food Sector. However, low production, limited input resources and rearing systems constrain the utilization of crickets. Scanty information exists on sundry input resources for upscaling of cricket production and how such inputs can be efficiently managed. This study sought to determine economic efficiency of improvised cricket rearing system using a generalized additive stochastic frontier approach (GAM-SFA) to assess the efficiency in cricket production under the new technology. Twenty-day old Acheta domesticus and Gryllus bimaculatus were separately reared in improvised cage system comprising bamboo hideouts, scrap blankets, cut bamboo stems and the plywood-based cages. GAM-SFA was used to estimate efficiency scores. Results revealed that the production was efficient. Feed, labor and water were positive and significant at 5% suggesting their importance and positive influence on cricket output. Similarly, the cost of feed, labor, water and scrap blanket were positive and significant suggesting that increase in these costs of inputs would increase the total cost. The mean TE, AE and EE were 85, 92 and 79%, respectively implying that there still exist potential to increase output using present technology and costs of production. Assessing key determinants of economic efficiency in cricket production under the system is necessary. Key words: Economic efficiency, crickets, GAM-SFA, improvised system.

A small, computationally flexible network produces the phenotypic diversity of song recognition in crickets.

How neural networks evolved to generate the diversity of species-specific communication signals is unknown. For receivers of the signals, one hypothesis is that novel recognition phenotypes arise from parameter variation in computationally flexible feature detection networks. We test this hypothesis in crickets, where males generate and females recognize the mating songs with a species-specific pulse pattern, by investigating whether the song recognition network in the cricket brain has the computational flexibility to recognize different temporal features. Using electrophysiological recordings from the network that recognizes crucial properties of the pulse pattern on the short timescale in the cricket Gryllus bimaculatus, we built a computational model that reproduces the neuronal and behavioral tuning of that species. An analysis of the model's parameter space reveals that the network can provide all recognition phenotypes for pulse duration and pause known in crickets and even other insects. Phenotypic diversity in the model is consistent with known preference types in crickets and other insects, and arises from computations that likely evolved to increase energy efficiency and robustness of pattern recognition. The model's parameter to phenotype mapping is degenerate - different network parameters can create similar changes in the phenotype - which likely supports evolutionary plasticity. Our study suggests that computationally flexible networks underlie the diverse pattern recognition phenotypes, and we reveal network properties that constrain and support behavioral diversity.

Toll signalling promotes blastema cell proliferation during cricket leg regeneration via insect macrophages.

Hemimetabolous insects, such as the two-spotted cricket Gryllus bimaculatus, can recover lost tissues, in contrast to the limited regenerative abilities of human tissues. Following cricket leg amputation, the wound surface is covered by the wound epidermis, and plasmatocytes, which are insect macrophages, accumulate in the wound region. Here, we studied the function of Toll-related molecules identified by comparative RNA sequencing during leg regeneration. Of the 11 Toll genes in the Gryllus genome, expression of Toll2-1, Toll2-2 and Toll2-5 was upregulated during regeneration. RNA interference (RNAi) of Toll, Toll2-1, Toll2-2, Toll2-3 or Toll2-4 produced regeneration defects in more than 50% of crickets. RNAi of Toll2-2 led to a decrease in the ratio of S- and M-phase cells, reduced expression of JAK/STAT signalling genes, and reduced accumulation of plasmatocytes in the blastema. Depletion of plasmatocytes in crickets using clodronate also produced regeneration defects, as well as fewer proliferating cells in the regenerating legs. Plasmatocyte depletion also downregulated the expression of Toll and JAK/STAT signalling genes in the regenerating legs. These results suggest that Spz-Toll-related signalling in plasmatocytes promotes leg regeneration through blastema cell proliferation by regulating the Upd-JAK/STAT signalling pathway.

Larval Rearing Methods for Two Endangered Species of Diving Beetle, Cybister chinensis Motschulsky, 1854 and Cybister lewisianus Sharp, 1873 (Coleoptera: Dytiscidae), Using Laboratory-Bred Food Prey

To facilitate ex-situ conservation of predaceous endangered insect species large numbers of live prey organisms are typically collected from the natural environment, potentially exacerbating biodiversity decline. To address this issue, two endangered beetle species (Cybister chinensis Motschulsky, 1854 and Cybister lewisianus Sharp, 1873) were reared using only laboratory-bred food. The beetle larvae were provided with laboratory-bred two-spotted cricket (Gryllus bimaculatus DeGeer; Orthoptera: Gryllidae) larvae as prey. The survival rates of the beetle larvae were as follows: C. chinensis, first instar larvae, 100% (n = 25), second instar larvae, 100% (n = 25), and third instar larvae, 92% (n = 23); C. lewisianus, first instar larvae 96% (n = 24), second instar larvae 92% (n = 23), and third instar larvae 92% (n = 23). The body length of adult insects fed laboratory-bred crickets did not significantly differ compared to the body length of field-collected adults.

Different transcriptional levels of Corazonin, Elevenin, and PDF according to the body color of the two-spotted cricket, Gryllus bimaculatus.

Body color in insects changes according to the living environment and physiological stresses possibly involved in endocrine factors. To date, 3 predominant bioactive peptides, Corazonin, Elevenin, and pigment-dispersing factor (PDF), have been illuminated to be involved in the body color in insects and crustaceans. Here, we examined the possibilities that these 3 factors would contribute to body color changes via melanization in the two-spotted cricket, Gryllus bimaculatus, whose body color changes according to population density drastically. Quantitative analyses revealed the higher transcriptional levels of Corazonin and Elevenin in the crowded-conditioned crickets, whereas the transcriptional level of PDF was higher in the isolated-conditioned crickets. However, the body color was not changed by knockdown of Corazonin, Elevenin, and PDF by RNA interference. The present data indicated that coloration mechanisms in G. bimaculatus is differently controlled from the previous observation in Locusta migratoria, a closely related orthopteran species.

Transcriptional expression changes during compensatory plasticity in the\xa0terminal ganglion of the adult cricket Gryllus bimaculatus

BackgroundDamage to the adult central nervous system often leads to long-term disruptions in function due to the limited capacity for neurological recovery. The central nervous system of the Mediterranean field cricket, Gryllus bimaculatus, shows an unusual capacity for compensatory plasticity, most obviously in the auditory system and the cercal escape system. In both systems, unilateral sensory disruption leads the central circuitry to compensate by forming and/or strengthening connections with the contralateral sensory organ. While this compensatory plasticity in the auditory system relies on robust dendritic sprouting and novel synapse formation, the compensatory plasticity in the cercal escape circuitry shows little obvious dendritic sprouting and instead may rely on shifts in excitatory and inhibitory synaptic strength.ResultsIn order to better understand what types of molecular pathways might underlie this compensatory shift in the cercal system, we used a multiple k-mer approach to assemble a terminal ganglion transcriptome that included ganglia collected one, three, and 7 days after unilateral cercal ablation in adult, male animals. We performed differential expression analysis using EdgeR and DESeq2 and examined Gene Ontologies to identify candidates potentially involved in this plasticity. Enriched GO terms included those related to the ubiquitin-proteosome protein degradation system, chromatin-mediated transcriptional pathways, and the GTPase-related signaling system.ConclusionFurther exploration of these GO terms will provide a clearer picture of the processes involved in compensatory recovery of the cercal escape system in the cricket and can be compared and contrasted with the distinct pathways that have been identified upon deafferentation of the auditory system in this same animal.

Nutritional Value and Anti-inflammation Activity of Misutkaru with Added Gryllus bimaculatus Powder

Purpose: This study investigated the nutritional value of Misutkaru with added &lt;i&gt;Gryllus bimaculatus&lt;/i&gt; powder (GBM) and its applicability as a healthy functional food.Methods: Chemical analysis of the moisture, crude fat, protein, and mineral contents was performed in accordance with the Association of Official Analytical Chemists (AOAC) guidelines. The amino acid and fatty acid compositions were analyzed using an automatic amino acid analyzer and gas chromatography, respectively. The levels of inflammatory cytokines tumor necrosis factor‑alpha (TNF-α), interleukin 1 beta (IL-1β), and interleukin 6 (IL‑6) induced by lipopolysaccharides in RAW 264.7 cells were measured.Results: The general composition per 100 g of GBM was 41.87 g protein, 19.75 g fat, and 28.52 g carbohydrates. The mineral content per 100 g of GBM was 889.66 mg calcium, 1189.73 mg potassium, 220.36 mg magnesium, 207.51 mg sodium, 694.81 mg phosphorus, and 15.50 mg zinc. In particular, valine (21.361 mg/kg), leucine (29.180 mg/kg), and isoleucine (15.562 mg/kg) were abundant in GBM. GBM also effectively downregulated the production of the inflammatory cytokines TNF-α, IL-1β, and IL-6 in RAW 264.7 macrophages.Conclusion: Misutkaru with added &lt;i&gt;Gryllus bimaculatus&lt;/i&gt; powder may have potential for application in the development of food materials or foods to prevent muscle loss in elderly individuals and sarcopenia patients, build muscle, and prevent increase in blood lipid concentrations in middle aged people. In particular, as &lt;i&gt;Gryllus bimaculatus&lt;/i&gt; is low in fat and carbohydrates, it can be used as a diet material.

Individual Scores for Associative Learning in a Differential Appetitive Olfactory Paradigm Using Binary Logistic Regression Analysis.

Numerous invertebrates have contributed to our understanding of the biology of learning and memory. In most cases, learning performance is documented for groups of individuals, and nearly always based on a single, typically binary, behavioural metric for a conditioned response. This is unfortunate for several reasons. Foremost, it has become increasingly apparent that invertebrates exhibit inter-individual differences in many aspects of their behaviour, and also that the conditioned response probability for an animal group does not adequately represent the behaviour of individuals in classical conditioning. Furthermore, a binary response character cannot yield a graded score for each individual. We also hypothesise that due to the complexity of a conditioned response, a single metric need not reveal an individual's full learning potential. In this paper, we report individual learning scores for freely moving adult male crickets (Gryllus bimaculatus) based on a multi-factorial analysis of a conditioned response. First, in an absolute conditioning paradigm, we video-tracked the odour responses of animals that, in previous training, received either odour plus reward (sugar water), reward alone, or odour alone to identify behavioural predictors of a conditioned response. Measures of these predictors were then analysed using binary regression analysis to construct a variety of mathematical models that give a probability for each individual that it exhibited a conditioned response (Presp). Using standard procedures to compare model accuracy, we identified the strongest model which could reliably discriminate between the different odour responses. Finally, in a differential appetitive olfactory paradigm, we employed the model after training to calculate the Presp of animals to a conditioned, and to an unconditioned odour, and from the difference a learning index for each animal. Comparing the results from our multi-factor model with a single metric analysis (head bobbing in response to a conditioned odour), revealed advantageous aspects of the model. A broad distribution of model-learning scores, with modes at low and high values, support the notion of a high degree of variation in learning capacity, which we discuss.

Gryllus bimaculatus Extract Protects against Lipopolysaccharide-Derived Inflammatory Response in Human Colon Epithelial Caco-2 Cells.

Simple SummaryInflammatory bowel disease (IBD), a potentially life-threatening disease, is characterized by increased tight junction permeability and overproduction of proinflammatory cytokines. The long-term administration of recognized chemotherapeutic agents can cause serious potential side effects. As such, increasing attention has been paid to natural, low-toxicity products with anti-inflammatory properties for treating IBD. We assessed the potential utility of the edible cricket species Gryllus bimaculatus for anti-inflammatory and cytoprotective effects in the human epithelial cell line Caco-2, following treatment with an inflammatory lipopolysaccharide stimulus. We found that aqueous ethanolic G. bimaculatus extract (AE-GBE) treatment increased cell viability and significantly reduced inflammatory mediators. Moreover, AE-GBE significantly reduced inflammatory cytokine expression levels, intestinal epithelial permeability, and related tight junction protein expression levels. In conclusion, AE-GBE can protect epithelial cells from lipopolysaccharide-induced impaired barrier integrity by increasing tight junction proteins and preventing various inflammatory mediators. These results may be used to pursue further use of natural insect extracts in treating IBD.Increased tight junction permeability and overproduction of proinflammatory cytokines are crucial pathophysiological mechanisms in inflammatory bowel disease (IBD). This study evaluated anti-inflammatory effects of aqueous ethanolic Gryllus bimaculatus extract (AE-GBE) against intestinal permeability on lipopolysaccharide (LPS)-treated Caco-2 cells. Treatment with AE-GBE increased cell viability and significantly reduced inflammatory mediators such as nitric oxide and LPS-induced reactive oxidative stress. LPS increased the expression levels of iNOS, Cox-2, and 4-hydroxylnonenal; however, these levels were attenuated by AE-GBE treatment. Moreover, the mRNA and protein expression levels of the inflammatory cytokines TNFα, IL-6, IL-1β, and IFNγ were increased by LPS, but were significantly reduced by AE-GBE treatment. Intestinal epithelial permeability and the related expression of the proteins Zoula ocludence-1, occludin, and claudin-1 was increased by LPS treatment, and this effect was significantly reduced by AE-GBE treatment. The reduction in AMPK phosphorylation in LPS-treated Caco-2 cells was reversed in activation by co-treatment with AE-GBE. In conclusion, AE-GBE can protect epithelial cells from LPS-induced impaired barrier integrity by increasing tight junction proteins and preventing various inflammatory mediators. Thus, AE-GBE has the potential to improve inflammation-related diseases, including IBD, by inhibiting excessive production of inflammation-inducing mediators.

Lifelong exposure to artificial light at night impacts stridulation and locomotion activity patterns in the cricket Gryllus bimaculatus.

Living organisms experience a worldwide continuous increase in artificial light at night (ALAN), negatively affecting their behaviour. The field cricket, an established model in physiology and behaviour, can provide insights into the effect of ALAN on insect behaviour. The stridulation and locomotion patterns of adult male crickets reared under different lifelong ALAN intensities were monitored simultaneously for five consecutive days in custom-made anechoic chambers. Daily activity periods and acrophases were compared between the experimental groups. Control crickets exhibited a robust rhythm, stridulating at night and demonstrating locomotor activity during the day. By contrast, ALAN affected both the relative level and timing of the crickets' nocturnal and diurnal activity. ALAN induced free-running patterns, manifested in significant changes in the median and variance of the activity periods, and even arrhythmic behaviour. The magnitude of disruption was light intensity dependent, revealing an increase in the difference between the activity periods calculated for stridulation and locomotion in the same individual. This finding may indicate the existence of two peripheral clocks. Our results demonstrate that ecologically relevant ALAN intensities affect crickets’ behavioural patterns, and may lead to decoupling of locomotion and stridulation behaviours at the individual level, and to loss of synchronization at the population level.

Insects as Novel Ruminant Feed and a Potential Mitigation Strategy for Methane Emissions.

Simple SummaryIn the last decade, there has been a growing interest in using edible insects as animal feed due to their high nutritive value and environmental advantages over the conventional livestock feeds. Insects have been used in the diets of some animals (poultry, fish, and swine) however, their evaluation in ruminants is still limited. The current in vitro study evaluated the usage of four different kinds of edible insects to partially substitute soybean meal as an example to the conventional high-quality expensive protein sources in ruminants’ diets. This study showed that the evaluated insects had high protein and fat contents. Substitution of 25% of soybean meal with the tested insects in a ruminant diet had no adverse effect on rumen fermentation profile or nutrient digestibility. Moreover, the inclusion of some species in the diet led to a reduction of the methane production up to 16–18% which is an additional environmental benefit. The findings of this study are encouraging for further work in this promising area to improve the sustainability of livestock industry.This study is the first to evaluate the chemical composition and impacts of four different edible insects, Acheta domesticus (A.d), Brachytrupes portentosus (B.p), Gryllus bimaculatus (G.b), and Bombyx mori (B.m), on the digestibility, rumen fermentation, and methane production when used as a substitute for 25% of the soybean meal (SBM) in a ruminant diet through in vitro incubation. The dietary treatments were 100% grass hay, 60% grass hay + 40% SBM, 60% grass hay + 30% SBM + 10% A.d, 60% grass hay + 30% SBM + 10% B.p, 60% grass hay + 30% SBM + 10% G.b, and 60% grass hay + 30% SBM + 10% B.m. The experiment was conducted as a short-term batch culture for 24 h at 39 °C, and the incubation was repeated in 3 consecutive runs. Chemical analysis of the insects showed that they were rich in fat (14–26%) with a high proportion of unsaturated fatty acids (60–70%). Additionally, the insects were rich in protein (48–61%) containing all essential amino acids and the amino acid profiles of the insects were almost the same as that of SBM. The inclusion of insects did not affect nutrient digestibility or the production of volatile fatty acids but did increase the production of ammonia-nitrogen. The addition of G.b and B.m led to decrease in methane production by up to 18% and 16%, respectively. These results reveal that substitution of 25% SBM in the diet with the tested insects had no negative impacts, and their potential to reduce methane production is an environmental benefit.

Differentiation between left and right wing stridulatory files in the field cricket Gryllus bimaculatus (Orthoptera: Gryllidae)

Male crickets produce acoustic signals by wing stridulation, attracting females for mating. A plectrum on the left forewing's (or tegmen) anal margin rapidly strikes along a serrated vein (stridulatory file, SF) on the opposite tegmen as they close, producing vibrations, ending in a tonal sound. The tooth strike rate of the plectrum across file teeth is equal to the sound frequency produced by the cricket (i.e., ∼5k teeth/s for ∼5 kHz in field crickets) and is specific to the forewing's resonant frequency. Sound is subsequently amplified using specialised wing cells. Anatomically, the forewings appear to mirror each other: both tegmina bear a SF and plectrum; however, most cricket species stridulate using right-over-left wing overlap making the stridulatory mechanism asymmetrical by default, rendering the left tegmen's SF unused. Therefore, we hypothesised structural differences between functional and unfunctional SFs. Three-dimensional mapping was used to accurately measure SF structures in Gryllus bimaculatus wings. We found that the left SF shows significantly greater variation in inter-tooth distance than the right, but less variation within the first sixty teeth (the functional part) than the right file. The left SF's slow evolutionary change over millions of years is discussed considering modern molecular phylogenies and fossil records.

Identification and Characterization of 24-Dehydrocholesterol Reductase (DHCR24) in the Two-Spotted Cricket, Gryllus bimaculatus.

Simple SummaryDHCR24 (24-dehydrocholesterol reductase) is a key enzyme for producing cholesterol from desmosterol and that is also involved in the conversion of plant sterols to cholesterol in most plant-feeding insects. This study extensively examined the possibility of DHCR24 involved in the sterol conversion in omnivorous insects, which feed on multiple food origins. Homologs of DHCR24 (GbDHCR24-1 and -2) were identified and characterized by using the two-spotted cricket, Gryllus bimaculatus, as an experimental model. The quantitative expression analyses and RNA interference experiments revealed that GbDHCR24-1 rather than GbDHCR24-2 facilitates the desmosterol-to-cholesterol conversion in crickets. Our data suggested that the omnivorous species produced cholesterol from desmosterol in the same manner as the plant-feeding species do.Arthropods, including insects, convert sterols into cholesterol due to the inability to synthesise cholesterol de novo. 24-dehydrocholesterol reductase (DHCR24) plays an important role in the conversion. Not only involving the cholesterol biosynthesis in vertebrates, DHCR24 is required for the conversion of desmosterol into cholesterol in phytophagous insects. The current study extensively examined DHCR24 in omnivorous insects, which feed on both plants and animals, using Gryllus bimaculatus as the experimental model. We identified cDNAs encoding two homologues of DHCR24 from G. bimaculatus, which were designated as GbDHCR24-1 and GbDHCR24-2. Both homologues contained the flavin adenine dinucleotide binding domain, which is a feature of DHCR24. Quantitative polymerase chain reaction revealed that among tissues of adult crickets, fat body and anterior midgut expressed high levels of GbDHCR24s. Both fat body and anterior midgut demonstrated DHCR24 activities in which one of the functions is the conversion of desmosterol into cholesterol in vitro. Knockdown of GbDHCR24-1 significantly reduced the conversion activity in the anterior midgut while knockdown of the GbDHCR24-2 did not. Additionally, the accumulation of desmosterol was detected in a feeding experiment with a specific DHCR24 inhibitor, azacosterol. We finally concluded that GbDHCR24-1 is the major enzyme that facilitates the desmosterol-to-cholesterol-conversion in crickets.