Low-Field NMR Study of Shortcake Biscuits with Cricket Powder, and Their Nutritional and Physical Characteristics.

The growing human population renders challenges for the future supply of food products with high nutritional value. Here, we enhanced the functional and nutritional value of biscuits, a popular sweet snack, by replacing the wheat flour with 2%, 6%, or 10% (w/w) cricket powder. Consumer acceptance ratings for reference and 2% augmented cookies were comparable, whereas the higher levels of enhancement received inferior consumer scores. This relatively small change in biscuit recipe provided significant and nutritionally desirable enhancements in the biscuits, observed in a series of analyses. An increase in the protein content was observed, including essential amino acids, as well as minerals and fat. This conversion also affected the physical properties of the biscuits, including hardness, and water molecular dynamics measured by 1H NMR. Cricket powder-augmented biscuits join the line of enhanced, functionally superior food products. This and similar food augmentation provide a viable scenario to meet the human food demands in the future.

Sexual selection and population divergence III: Interspecific and intraspecific variation in mating signals.

A major challenge for studying the role of sexual selection in divergence and speciation is understanding the relative influence of different sexually selected signals on those processes in both intra- and interspecific contexts. Different signals may be more or less susceptible to co-option for species identification depending on the balance of sexual and ecological selection acting upon them. To examine this, we tested three predictions to explain geographic variation in long- versus short-range sexual signals across a 3,500 + km transect of two related Australian field cricket species (Teleogryllus spp.): (a) selection for species recognition, (b) environmental adaptation and (c) stochastic divergence. We measured male calling song and male and female cuticular hydrocarbons (CHCs) in offspring derived from wild populations, reared under common garden conditions. Song clearly differentiated the species, and no hybrids were observed suggesting that hybridization is rare or absent. Spatial variation in song was not predicted by geography, genetics or climatic factors in either species. In contrast, CHC divergence was strongly associated with an environmental gradient supporting the idea that the climatic environment selects more directly upon these chemical signals. In light of recently advocated models of diversification via ecological selection on secondary sexual traits, the different environmental associations we found for song and CHCs suggest that the impact of ecological selection on population divergence, and how that influences speciation, might be different for acoustic versus chemical signals.

Functionality of Cricket and Mealworm Hydrolysates Generated after Pretreatment of Meals with High Hydrostatic Pressures.

The low consumer acceptance to entomophagy in Western society remains the strongest barrier of this practice, despite these numerous advantages. More positively, it was demonstrated that the attractiveness of edible insects can be enhanced by the use of insect ingredients. Currently, insect ingredients are mainly used as filler agents due to their poor functional properties. Nevertheless, new research on insect ingredient functionalities is emerging to overcome these issues. Recently, high hydrostatic pressure processing has been used to improve the functional properties of proteins. The study described here evaluates the functional properties of two commercial insect meals (Gryllodes sigillatus and Tenebrio molitor) and their respective hydrolysates generated by Alcalase®, conventionally and after pressurization pretreatment of the insect meals. Regardless of the insect species and treatments, water binding capacity, foaming and gelation properties did not improve after enzymatic hydrolysis. The low emulsion properties after enzymatic hydrolysis were due to rapid instability of emulsion. The pretreatment of mealworm meal with pressurization probably induced protein denaturation and aggregation phenomena which lowered the degree of hydrolysis. As expected, enzymatic digestion (with and without pressurization) increased the solubility, reaching values close to 100%. The pretreatment of mealworm meal with pressure further improved its solubility compared to control hydrolysate, while pressurization pretreatment decreased the solubility of cricket meal. These results may be related to the impact of pressurization on protein structure and therefore to the generation of different peptide compositions and profiles. The oil binding capacity also improved after enzymatic hydrolysis, but further for pressure-treated mealworm hydrolysate. Despite the moderate effect of pretreatment by high hydrostatic pressures, insect protein hydrolysates demonstrated interesting functional properties which could potentially facilitate their use in the food industry.

Optimisation of a cheap and residential small-scale production of edible crickets with local by-products as an alternative protein-rich human food source in Ratanakiri Province, Cambodia.

BACKGROUND: The health status of the indigenous people of Ratanakiri Province, Cambodia, is significantly lower compared with that of the rest of the nation. The domestication and mass production of insects may represent a sustainable, cost effective and high quality alternative source of protein to traditional livestock. This study aimed to optimise a cheap and residential cricket breeding system based on unused wild resources. The development of crickets, Teleogryllus testaceus (Walker), under seven diets composed of taro aerial parts, young cassava leaves, young cashew leaves and brown rice flour (with or without banana slices), versus a traditionally used broiler feed diet was studied. RESULTS: Cricket mortality was low in all diets, except the two cashew-based diets. Total biomass was significantly higher under the broiler feed, in addition to the two diets containing a combination of cassava leaf powder and brown rice. Yet, crickets fed with the taro diet had the highest percentage of protein. Concerning the breeding system cost, units using cassava leaves were the cheapest. CONCLUSION: Diets based on cassava leaves seem to be the most promising. Nevertheless, to produce crickets with a high body mass and a high protein level, a new experiments must be devised in which cassava leaf maturity will be adapted to fit with the cricket growth stage. Moreover, to reduce the cost of the breeding units, handmade local products should be used instead of purchased components.

Gene expression profiling and inhibition of adipose tissue accumulation of G. bimaculatus extract in rats on high fat diet.

BACKGROUND: Molecular genetic mechanisms underlying the anti-inflammatory effects of ethanol extract (GB) from G. bimaculatus, a type of cricket, are not fully elucidated. G. bimaculatus was reported to be rich in unsaturated fatty acid and to decrease the omega-6/omega-3 fatty acid ratio when fed to chickens. GB may reduce the amount of fat or increase the unsaturated fatty acid ratio. METHODS: Male Wistar rats fed a high-fat diet (HFD) were orally administered with 5 groups: phosphate buffered saline (PBS, control), GB (100 mg/kg or 200 mg/kg), Pravastatin or Isaria sinclairii (IS) extract, which is reported to have fat-reducing effects, for either 1 or 2 months. GB's sero-biochemial, hematological and anti-oxidizing hepato-cellular biomarker levels were evaluated to dertermine their antilipidemic, anti-inflammatory, and anti-coagulant effect in rats after 1 or 2 month GB treatments on HFD (fat 60 %) Wistar rat. The abdominal and epididymidal fat weight were measured and the composition of fatty acid was analyzed by GC/MS. Microarray analyses were performed with a rat 28 K cDNA clone set array to identify the gene-expression profiles for the GB exposed high fat dieted Wistar rat. RESULTS: The weight and fatty acid composition of abdominal fat and epididymidal fat, total cholesterol, LDL-cholesterol, and triglyceride in GB treated rats were at lower levels than those of the control group. The anti-oxidant hepato-cellular biomarker levels, protein carbonyl content and malondialdehyde concentration in GB treated rats were significantly decreased. Compared to the control, the GB treated rat group (treated at a dose of 100 and 200 mg/kg), had 190 up-regulated genes including Gpm6a (glycoprotein m6a), Tmem14a (transmembrane protein 14A) and Fasin (fatty acid synthase), with down-regulated 235 genes including Cc121b (chemokine ligand 21b), Glycan1 (glycosylation dependent cell adhesion moleule, Serpinb1a (serine proteinase inhibitor) and Tcrb (T-cell receptor beta chain). CONCLUSION: The data suggest Fasin-related fatty acid synthesis and adipose differentiation related protein (Adfp), which is related to obesity, were upregulated by GB treatment, indicating their potential therapeutic markers for anti-atheriosclerosis or inflammation.

Barriers to gene exchange in hybridizing field crickets: the role of male courtship effort and cuticular hydrocarbons.

BACKGROUND: Pre-zygotic barriers often involve some form of sexual selection, usually interpreted as female choice, as females are typically the choosier sex. However, males typically show some mate preferences, which are increasingly reported. Here we document previously uncharacterized male courtship behavior (effort and song) and cuticular hydrocarbon (CHC) profiles in the hybridizing crickets Gryllus firmus and G. pennsylvanicus. These two species exhibit multiple barriers to gene exchange that act throughout their life history, including a behavioral barrier that results in increased time to mate in heterospecific pairs. RESULTS: We demonstrated that male mate choice (as courtship effort allocation) plays a more important role in the prezygotic behavioral barrier than previously recognized. In gryllids females ultimately decide whether or not to mate, yet we found males were selective by regulating courtship effort intensity toward the preferred (conspecific) females. Females were also selective by mating with more intensely courting males, which happened to be conspecifics. We report no differences in courtship song between the two species and suggest that the mechanism that allows males to act differentially towards conspecific and heterospecific females is the cuticular hydrocarbon (CHC) composition. CHC profiles differed between males and females of both species, and there were clear differences in CHC composition between female G. firmus and G. pennsylvanicus but not between the males of each species. CONCLUSION: Although many barriers to gene exchange are known in this system, the mechanism behind the mate recognition leading to reduced heterospecific mating remains unknown. The CHC profiles might be the phenotypic cue that allow males to identify conspecifics and thus to adjust their courtship intensity accordingly, leading to differential mating between species.

A comparison of convergently evolved insect silks that share ß-sheet molecular structure.

Raspy crickets produce silk webs that are used to build shelters. These webs have been found to consist of both fiber and film components. Raman spectra obtained from both components were found to be very similar for a given species. The protein structure of the fibers and films produced by both species was predominately ß-sheet with lesser amounts of ß-turns, unordered and α-helical protein also detected. The orientation of the ß-sheet backbone in the fiber was determined to be parallel to the fiber axis. Compared to cocoon and dragline silk the orientation distribution exhibits a significant randomly orientated protein component. Amino acid analysis confirmed the presence of glycine, serine, and alanine in both species, which are known to form antiparallel ß-sheet structures. Both species, although at significantly different concentrations, where found to contain proline. This amino acid is uncommon in insect silks, and likely involved in increasing fiber elasticity.

Anti-inflammatory effect of glycosaminoglycan derived from Gryllus bimaculatus (a type of cricket, insect) on adjuvant-treated chronic arthritis rat model.

Anti-inflammatory effects of glycosaminoglycan (GAG) derived from cricket (Gryllus bimaculatus, Gb) were investigated in a complete Freund's adjuvant (CFA)-treated chronic arthritic rat model. This GAG produced a significant anti-edema effect as evidenced by inhibition of C-reactive protein (CRP) and rheumatoid factor, and interfered with atherogenesis by reducing proinflammatory cytokine levels of (1) vascular endothelial growth factor (VEGF) production in human umbilical vein endothelial cells (HUVEC), (2) interleukin-6, (3) prostaglandin E2-stimulated lipopolysaccharide in RAW 264.7 cells, and (4) tumor necrosis factor (TNF)-α production in normal splenocytes, in a dose-dependent manner. This GAG was also found to induce nitric oxide (NO) production in HUVEC cells and elevated endothelial nitric oxide synthase (eNOS) activity levels. Histological findings demonstrated the fifth lumbar vertebrae (LV) dorsal root ganglion, which was linked to the paw treated with Gb GAG, was repaired against CFA-induced cartilage destruction. Further, combined indomethacin (5 mg/kg)-Gb GAG (10 mg/kg) inhibited more effectively CFA-induced paw edema at 3 h and 2 or 3 d after treatment to levels comparable to only the anti-inflammatory drug indomethacin. Ultraviolet (UV)-irritated skin inflammation also downregulated nuclear factor κB (NFκB) activity in transfected HaCaT cells. Data suggest that the anti-inflammatory effects of GAG obtained from cricket (Gb) may be useful for treatment of inflammatory diseases including chronic arthritis.

Purine derivate content and amino acid profile in larval stages of three edible insects.

BACKGROUND: Considering their high content of protein, insects are a valuable alternative protein source. However, no evaluation of their purine content has so far been done. High content of purine derivates may lead to the exclusion of such food from the diet of people with specific diseases. The aim of this study was to analyse the content of selected purine derivates and amino acid profile in the three insect species most often used for entomophagy in Europe and compare them with the purine content in egg white and chicken breast. RESULTS: The content of individual purine derivates and their total content were significantly dependent on insect species. The purine content in all three species was significantly higher (P < 0.05) than in egg white, but some values were significantly lower (P < 0.05) than in chicken breast. The total protein content was 548.9 g kg(-1) dry matter (DM) in mealworm (Tenebrio molitor), 551.6 g kg(-1) DM in superworm (Zophobas atratus) and 564.9 g kg(-1) DM in cricket (Gryllus assimilis). CONCLUSION: Larvae of mealworm and superworm are protein-rich and purine-low meat alternatives. In contrast, cricket nymphs are protein-rich and purine-rich and cannot be recommended for people with hyperuricemia or gout.

Challenges with effective nutrient supplementation for amphibians: A review of cricket studies.

Over the last 25 years, numerous studies have investigated the impact of insect supplementation on insect nutrient content. In light of recent nutrition related challenges with regards to zoo amphibians fed an insect based diet, this review attempts to comprehensively compile both anecdotal and published data in the context of practical application on this topic. Insects, primarily crickets, used for amphibian diets historically demonstrate low concentrations of key nutrients including calcium and vitamin A. Commonly used practices for supplementation involving powder dusting or gut loading have been shown to improve delivery of calcium and vitamin A, though often not reaching desired nutrient concentrations. The large variety of factors influencing insect nutrient content are difficult to control, making study design, and results often inconsistent. Formulation and availability of more effective gut loading diets, combined with a standardized protocol for insect husbandry and dietary management may be the most effective way to supplement insects for use in amphibian feeding programs. Ideally, the nutritional improvement of feeder insects would begin at the breeder level; however, until this becomes a viable choice, we confirm that supplementation of crickets through both gut-loading and dusting appear necessary to support the nutritional health of amphibians and other insectivores in managed collections.

Effect of dietary carotenoids on vitamin A status and skin pigmentation in false tomato frogs (Dyscophus guineti).

Several species in captivity develop nutritional diseases including vitamin A deficiency; cases of this disease have been documented in amphibians, which may be linked to an insectivorous diet lacking in vitamin A or carotenoids. Adults and young of Dyscophus guineti were fed three diets over 9 weeks to evaluate effects on carotenoids and vitamin A status and skin pigmentation. Feeder crickets were either supplemented with soy oil (control, CON), soy oil enriched with ß-carotene (BC) or mixed carotenoids (MIX) by direct injection of known dosages. Vitamin A from feeder crickets (measured as retinol) was higher in insects supplemented with both BC and MIX; (P=0.0001) and plasma retinol concentrations were significantly higher in frogs fed MIX (P<0.02). Results suggest that both false tomato frogs and feeder crickets could receive some provitamin A activity through consumption of diets supplemented with ß-carotene, and xanthophylls like lutein and zeaxanthin. Pigmentation was evaluated weekly through the use of visual color charts, as well as quantitatively using a hand-held spectrophotometer. MIX diets had a significant effect on skin color values (P<0.0001), as well as on lightness (P=0.0005) and hue (P=0.0022). Results indicated that frogs fed with BC changed to yellower colors, and frogs fed with MIX changed to oranger colors. Visual color chart observations also scored significantly different between CON and MIX diets (P<0.05); the animals fed MIX also appeared oranger according to the qualitative observations. Dietary supplements with carotenoids resulted in color changes and higher circulating retinol concentrations in false tomato frogs. These pigments may provide provitamin A activity in diets, thus may support improved nutrition and health of captive-fed insectivorous amphibians.

A new species of Cardiodactylus (Orthoptera: Grylloidea: Eneopterinae) from Singapore.

Cardiodactylus is a speciose cricket genus belonging to the subfamily Eneopterinae. One new species of Cardiodactylus from Singapore is described: Cardiodactylus admirabilis Tan & Robillard n. sp. Acoustic analysis is also performed on the male calling song. A key to species of Eneopterinae from Singapore is provided.

Cuticular hydrocarbons as a basis for chemosensory self-referencing in crickets: a potentially universal mechanism facilitating polyandry in insects.

Females of many species obtain benefits by mating polyandrously, and often prefer novel males over previous mates. However, how do females recognise previous mates, particularly in the face of cognitive constraints? Female crickets appear to have evolved a simple but effective solution: females imbue males with their own cuticular hydrocarbons (CHCs) at mating and utilise chemosensory self-referencing to recognise recent mates. Female CHC profiles exhibited significant additive genetic variation, demonstrating that genetically unique chemical cues are available to support chemosensory self-referencing. CHC profiles of males became more similar to those of females after mating, indicating physical transfer of CHCs between individuals during copulation. Experimental perfuming of males with female CHCs resulted in a female aversion to males bearing chemical cues similar to their own. Chemosensory self-referencing, therefore, could be a widespread mechanism by which females increase the diversity of their mating partners.

Short-term phenotypic plasticity in long-chain cuticular hydrocarbons.

Cuticular hydrocarbons provide arthropods with the chemical equivalent of the visually extravagant plumage of birds. Their long chain length, together with the number and variety of positions in which methyl branches and double bonds occur, provide cuticular hydrocarbons with an extraordinary level of information content. Here, we demonstrate phenotypic plasticity in an individual's cuticular hydrocarbon profile. Using solid-phase microextraction, a chemical technique that enables multiple sampling of the same individual, we monitor short-term changes in cuticular hydrocarbon profiles of individual crickets, Teleogryllus oceanicus, in response to a social challenge. We experimentally manipulate the dominance status of males and find that dominant males, on losing fights with other dominant males, change their hydrocarbon profile to more closely resemble that of a subordinate. This result demonstrates that cuticular hydrocarbons can be far more responsive to changes in social dominance than previously realized.

Pseudovitamin B12 and factor S are the predominant corrinoid compounds in edible cricket products.

In this study, we determined the vitamin B12 content of commercially-available edible insect products using a bioassay based on Lactobacillus delbrueckii ATCC 7830. Although the vitamin content of giant water bug, bee larva, grasshopper, and weaver ant products was low, we found that diving beetle and cricket products contained relatively high amounts of vitamin B12 (approximately 89.5 and 65.8 µg/100 g dry weight, respectively). In the cricket products most widely circulated as foods, specific corrinoid (vitamin B12) compounds were extracted and identified using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Despite the bioassay detecting high vitamin B12 content (approximately 50-75 µg/100 g dry weight) in these cricket products, UPLC-MS/MS analysis indicated that pseudovitamin B12 and 2-methylmercaptoadenyl cobamide (also known as factor S) were actually the predominant corrinoid compounds (~74% and ~21%, respectively), with authentic vitamin B12 making up only 5% of total corrinoids.

Allergenicity assessment of the edible cricket Acheta domesticus in terms of thermal and gastrointestinal processing and IgE cross-reactivity with shrimp.

The allergenic potency of the cricket Acheta domesticus, a promising edible insect, has never been assessed. This work aims to study the immunoreactivity of Acheta domesticus, and its cross-reactivity with the shrimp Litopenaeus vannamei, assessing the effect of cooking and gastrointestinal digestion on their allergenic properties. Different cricket proteins were detected by immunoblotting with shrimp-allergic patients' sera. Tropomyosin was identified as the most relevant IgE-binding protein, and its cross-reactivity with shrimp tropomyosin was demonstrated by ELISA. While shrimp tropomyosin showed scarce stability to gastric digestion, cricket tropomyosin withstood the whole digestion process. The sarcoplasmic calcium-binding protein, specifically detected in shrimp, showed exceptional stability to gastrointestinal digestion. IgE-binding proteins in a model of enriched baked products were partially protected from proteolysis. In conclusion, the ingestion of A. domesticus proteins poses serious concerns to the Crustacean-allergic population. The high stability of tropomyosin may represent a risk of primary sensitization and clinical cross-reactivity.

Development of ultra-high performance liquid chromatographic and fluorescent method for the analysis of insect chitin.

A precise quantification of insect chitin is needed in order to avoid overestimation of crude protein due to chitin-bound nitrogen. An UPLC/FLR method was optimized and validated for the determination of glucosamine (GlcN) hydrolyzed from chitin in insect materials. The method was applied for quantifying the chitin content in mealworms (Tenebrio molitor) and crickets (Acheta domesticus). A baseline separation was obtained using an Acquity HSS T3 C18 column, with an external calibration curve of excellent linearity, and a low limit of detection and quantification of GlcN. Even though the recovery of GlcN from spiked cricket material was slightly lower compared to that using spectrophotometric method, the UPLC/FLR method proved a sensitive and specific method of quantification of insect chitin. Chitin contents in T. molitor and A. domesticus were 4.6 ± 0.1% and 4.5 ± 0.0% on dry matter basis, respectively. Less than 0.01% of chitin was present in insect protein-enriched fractions extracted with 0.1 N NaCl at pH 10.

Investigating edible insects as a sustainable food source: nutritional value and techno-functional and physiological properties.

This work is aimed to evaluate the nutritional composition, and the techno-functional and in vitro physiological properties of flours made using six different insect species and the sensorial feasibility of including them in bakery products. The insect flours exhibited high protein and fat contents as their main components, highlighting the presence of chitin in ant samples. The techno-functional properties showed high oil holding, swelling, and emulsifying capacities in all the analysed insect flours, whereas their bulk density, hydration properties, and foaming capacity showed average values and no gelation capacity. Moreover, these edible insect flours exhibited effective hyperglycaemia and hyperlipidaemia properties, which together with their high antioxidant capacity are associated with beneficial in vitro physiological effects. The beetle and caterpillar flours stand out in these properties, and thus were selected to make a cupcake. The sensory evaluation confirmed that the edible beetle powder can be successfully included in baked goods to provide excellent sensory properties and very high acceptance. Thus, these insect flours may be of great interest to the food industry as a healthy source of protein, exerting a positive impact on functional and sensory food properties, and with a potential role in the prevention of diseases associated with hyperglycaemia and hyperlipidaemia.

Gryllus bimaculatus extract protects against lipopolysaccharide and palmitate-induced production of proinflammatory cytokines and inflammasome formation.

Inflammation and the inflammasome complex formation are associated with numerous diseases, and palmitates or lipopolysaccharides (LPS) have been identified as potential links between these disorders. Recently, edible insects such as the Gryllus bimaculatus (GB) and the larva of Tenebrio molitor have emerged as alternative food sources. In the present study, the effect of GB on LPS­ or palmitate­induced production of inflammatory cytokines, the formation of the inflammasome complex, reactive oxygen species (ROS) generation, endoplasmic reticulum (ER) stress and cell death was investigated in RAW264.7 cells. The results revealed that GB extract downregulated the production of inflammatory cytokines (such as TNF­α, IL­1ß and IL­6). Since the role of the MAP kinase and NF­κB signalling pathways in the production of inflammatory cytokines is well established, the translocation of p65 into the nucleus and the phosphorylation of IκB and MAP kinases were further examined. Both these processes were upregulated following LPS and palmitate treatment, but they were inhibited by the GB extract. Moreover, GB extract decreased LPS/palmitate­induced inflammasome complex formation (assessed via analysing the levels of the apoptosis­associated speck­like protein containing a caspase­recruitment domain, NOD­like receptor family pyrin domain containing 3, cleaved caspase­1 and IL­1ß), the generation of ROS, ER stress and cell death. Treatment with SB203580 (a p38 inhibitor), SP600125 (a JNK inhibitor) and pyrrolidinedithiocarbamate ammonium (an NF­κB inhibitor) decreased the production of inflammatory cytokines, as well as helped in the recovery of LPS/palmitate­induced cell death. Overall, GB extract served an inhibitory role in LPS/palmitate­induced inflammation via inhibiting the MAP kinase and NF­κB signalling pathways, inflammasome complex formation, ROS generation, ER stress and cell death.

Extrafloral nectar content alters foraging preferences of a predatory ant.

We tested whether the carbohydrate and amino acid content of extrafloral nectar affected prey choice by a predatory ant. Fire ants, Solenopsis invicta, were provided with artificial nectar that varied in the presence of carbohydrates and amino acids and were then provided with two prey items that differed in nutritional content, female and male crickets. Colonies of fire ants provided with carbohydrate supplements consumed less of the female crickets and frequently did not consume the high-lipid ovaries of female crickets. Colonies of fire ants provided with amino acid supplements consumed less of the male crickets. While a number of studies have shown that the presence of extrafloral nectar or honeydew can affect ant foraging activity, these results suggest that the nutritional composition of extrafloral nectar is also important and can affect subsequent prey choice by predatory ants. Our results suggest that, by altering the composition of extrafloral nectar, plants could manipulate the prey preferences of ants foraging on them.