An innovative optimized protocol for high-quality genomic DNA extraction from recalcitrant Shea tree (Vitellaria paradoxa, C.F. Gaertn) plant and its suitability for downstream applications.

BACKGROUND: It is not always easy to find a universal protocol for the extraction of genomic DNA (gDNA) from plants. Extraction of gDNA from plants such as shea with a lot of polysaccharides in their leaves is done in two steps: a first step to remove the polysaccharides and a second step for the extraction of the gDNA. In this work, we designed a protocol for extracting high-quality gDNA from shea tree and demonstrate its suitability for downstream molecular applications. METHODS: Fifty milligrams of leaf and root tissues were used to test the efficiency of our protocol. The quantity of gDNA was measured with the NanoDrop spectrometer and the quality was checked on agarose gel. Its suitability for use in downstream applications was tested with restriction enzymes, SSRs and RAPD polymerase chain reactions and Sanger sequencing. RESULTS: The average yield of gDNA was 5.17; 3.96; 2.71 and 2.41 µg for dry leaves, dry roots, fresh leaves and fresh roots respectively per 100 mg of tissue. Variance analysis of the yield showed significant difference between all tissue types. Leaf gDNA quality was better compared to root gDNA at the absorbance ratio A260/280 and A260/230. The minimum amplifiable concentration of leaf gDNA was 1 pg/µl while root gDNA remained amplifiable at 10 pg/µl. Genomic DNA obtained was also suitable for sequencing. CONCLUSION: This protocol provides an efficient, convenient and cost effective DNA extraction method suitable for use in various vitellaria paradoxa genomic studies.

Effect of household cooking techniques on the microbiological load and the nutritional quality of mealworms (Tenebrio molitor L. 1758).

Mealworms are new food products in Europe, but consumers do not know how to cook them. Although cooking could increase the safety, acceptability, palatability, and digestibility of insects, the heating process could have deleterious effects on protein and lipid quality. Therefore, this study characterized the effects of different household cooking methods (boiling, pan-frying, vacuum cooking, and oven cooking) on the microbial load and nutritive value of mealworms, with a focus on protein digestibility and fatty acid composition. Boiling and cooking under vacuum were the most efficient techniques to reduce microbial load while maintaining the high levels of protein and polyunsaturated fatty acids of mealworms. Cooking method-related changes were very low on macronutrients content except for pan-fried mealworms which exhibited the highest lipid content. Cooking slightly changed fatty acid composition of mealworms by principally decreasing their level of saturated fatty acids but also increased the in vitro crude protein digestibility of mealworms.

Microbiological Load of Edible Insects Found in Belgium.

Edible insects are gaining more and more attention as a sustainable source of animal protein for food and feed in the future. In Belgium, some insect products can be found on the market, and consumers are sourcing fresh insects from fishing stores or towards traditional markets to find exotic insects that are illegal and not sanitarily controlled. From this perspective, this study aims to characterize the microbial load of edible insects found in Belgium (i.e., fresh mealworms and house crickets from European farms and smoked termites and caterpillars from a traditional Congolese market) and to evaluate the efficiency of different processing methods (blanching for all species and freeze-drying and sterilization for European species) in reducing microorganism counts. All untreated insect samples had a total aerobic count higher than the limit for fresh minced meat (6.7 log cfu/g). Nevertheless, a species-dependent blanching step has led to a reduction of the total aerobic count under this limit, except for one caterpillar species. Freeze-drying and sterilization treatments on European species were also effective in reducing the total aerobic count. Yeast and mold counts for untreated insects were above the Good Manufacturing Practice limits for raw meat, but all treatments attained a reduction of these microorganisms under this limit. These results confirmed that fresh insects, but also smoked insects from non-European trades, need a cooking step (at least composed of a first blanching step) before consumption. Therefore, blanching timing for each studied insect species is proposed and discussed.

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.