The Effects of Slaughter Methods and Drying Temperatures on the Protein Hydrolysis of Black Soldier Fly Larvae Meal.

In recent years, the potential of insects as a sustainable protein alternative to feed the growing world population has been explored. Differences in the ways insects are processed can affect their proximate composition and digestibility. This work studied the effects of the combination of different types of slaughter methods and drying temperatures on the proximate composition, organic matter digestibility (OMd), hydrolysis degree (DH/NH2 and DH/100 g DM), total hydrolysis (TH), and hygienic and sanitary characteristics of BSFL (black soldier fly larvae) meal. Four types of slaughter methods were used including freezing (F), blanching + freezing (B), Melacide® + freezing (M), and liquid nitrogen slaughter (N). Each of these was used with three drying temperatures (50, 70, and 90 °C). A negative correlation between the acid detergent fiber (ADF) and protein digestibility parameters was obtained. The most suitable drying temperature was 70 °C, as it produced higher values of protein digestibility (DH and TH), resulting in hygienic and sanitary conditions suitable for food use. Slaughtering with liquid nitrogen and blanching was more conducive to achieving high protein digestibility results than traditional freezing or the use of Melacide®.

Antimicrobial and antioxidant activity of encapsulated tea polyphenols in chitosan/alginate-coated zein nanoparticles: a possible supplement against fish pathogens in aquaculture.

Regulation of antibiotic use in aquaculture calls for the emergence of more sustainable alternative treatments. Tea polyphenols (GTE), particularly epigallocatechin gallate (EGCG), have various biological activities. However, tea polyphenols are susceptible to degradation. In this work, EGCG and GTE were encapsulated in zein nanoparticles (ZNP) stabilized with alginate (ALG) and chitosan (CS) to reduce the degradation effect. ALG-coated ZNP and ALG/CS-coated ZNP encapsulating EGCG or GTE were obtained with a hydrodynamic size of less than 300 nm, an absolute ζ-potential value >30 mV, and an encapsulation efficiency greater than 75%. The antioxidant capacity of the encapsulated substances, although lower than that of the free ones, maintained high levels. On the other hand, the evaluation of antimicrobial activity showed greater efficiency in terms of growth inhibition for ALG/CS-ZNP formulations, with average overall values of around 60%, reaching an inhibition of more than 90% for Photobacterium damselae. These results support encapsulation as a good strategy for tea polyphenols, as it allows maintaining significant levels of antioxidant activity and increasing the potential for antimicrobial activity, in addition to increasing protection against sources of degradation.

Evaluation of In Vitro Protein Hydrolysis in Seven Insects Approved by the EU for Use as a Protein Alternative in Aquaculture.

Rapid population growth is leading to an increase in the demand for high-quality protein such as fish, which has led to a large increase in aquaculture. However, fish feed is dependent on fishmeal. It is necessary to explore more sustainable protein alternatives that can meet the needs of fish. Insects, due to their high protein content and good amino acid profiles, could be a successful alternative to fishmeal and soybean meal traditionally used in sectors such as aquaculture. In this work, seven species of insects (Hermetia illucens, Tenebrio molitor, Acheta domestica, Alphitobius diaperinus, Gryllodes sigillatus, Gryllus assimilis, and Musca domestica) approved by the European Union (UE) for use as feed for farmed animals (aquaculture, poultry, and pigs) were studied. Their proximate composition, hydrolysis of organic matter (OMd), hydrolysis of crude protein (CPd), degree of hydrolysis (DH/NH2 and DH/100 g DM), and total hydrolysis (TH) were analyzed. The results showed that Tenebrio molitor had digestibility similar to that of fishmeal, while Acheta domestica and Hermetia illucens provided similar digestibility to that of soybean meal. The acid detergent fiber (ADF) data were negatively correlated with all protein digestibility variables. The differences in the degree of hydrolysis (DH) results and the similarity in total hydrolysis (TH) results could indicate the slowing effects of ADF on protein digestibility. Further in vivo studies are needed.

In Vitro Crude Protein Digestibility of Insects: A Review.

The high protein content of insects has been widely studied. They can be a good food alternative, and therefore it is important to study the effect of digestion on their protein. This review examines the different in vitro protein digestibility methodologies used in the study of different edible insects in articles published up to 2021. The most important variables to be taken into account in in vitro hydrolysis are the following: phases (oral, gastric and intestinal), enzymes, incubation time and temperature, method of quantification of protein hydrolysis and sample preprocessing. Insects have high digestibility data, which can increase or decrease depending on the processing of the insect prior to digestion, so it is important to investigate which processing methods improve digestibility. The most commonly used methods are gut extraction, different methods of slaughtering (freezing or blanching), obtaining protein isolates, defatting, thermal processing (drying or cooking) and extrusion. Some limitations have been encountered in discussing the results due to the diversity of methodologies used for digestion and digestibility calculation. In addition, articles evaluating the effect of insect processing are very limited. It is concluded that there is a need for the standardisation of in vitro hydrolysis protocols and their quantification to facilitate comparisons in future research.

Pre-Treatment of Fish By-Products to Optimize Feeding of Tenebrio molitor L. Larvae.

Fish discards are organic waste with high and good-quality protein levels, as well as a fatty acid profile rich in n-3 LCPUFAs, mainly eicosapentaenoic acid and docosahexaenoic acid. These discards can be used as food for Tenebrio molitor (Linnaeus, 1758) larvae, thus increasing the nutritional value of this insect. This study focused on increasing larval acceptance of fish through different pre-treatments of the diets provided, as well as increasing the accumulation of EPA and DHA in fish-fed larvae. Four different diets were prepared: control (broiler feed), DGF50: 50% dried ground fish (Pagellus bogaraveo, Brünnich, 1768) + 50% broiler feed, for different periods, FGF100: 100% fresh ground P. bogaraveo and DUF100: 100% dried whole unground P. bogaraveo. Growth, mortality, proximate composition, fatty acid profile and lipid nutritional indices were determined. Larvae fed with FGF100 displayed better results among treatments, doubling the initial weight, as well as increasing their protein level and decreasing fat levels. Regarding fatty acids, eicosapentaenoic acid and docosahexaenoic acid were only detected in larvae fed with a fish-based diet for a period longer than 5 days. These results show that pre-treatment of fish-based diets causes changes in the growth and compositional parameters of T. molitor larvae.

Fishmeal Dietary Replacement Up to 50%: A Comparative Study of Two Insect Meals for Rainbow Trout (Oncorhynchus mykiss).

The demand of optimal protein for human consumption is growing. The Food and Agriculture Organization (FAO) has highlighted aquaculture as one of the most promising alternatives for this protein supply gap due to the high efficiency of fish growth. However, aquaculture has been facing its own sustainability problem, because its high demand for protein has been traditionally satisfied with the use of fishmeal (FM) as the main source. Some of the most promising and sustainable protein substitutes for FM come from insects. The present manuscript provides insight into an experiment carried out on rainbow trout (Oncorhynchus mykiss) with a 50% replacement of FM with different larvae insect meals: Hermetia illucens (HI), and Tenebrio molitor (TM). TM showed better results for growth, protein utilization and more active digestive function, supported by intestinal histological changes. Liver histology and intermediary metabolism did not show relevant changes between insect meals, while other parameters such as antioxidant enzyme activities and tissue damage indicators showed the potential of insect meals as functional ingredients.

Effect on Intermediary Metabolism and Digestive Parameters of the High Substitution of Fishmeal with Insect Meal in Sparus aurata Feed.

Hermetia illucens and Tenebrio molitor were tested on account of their potential to replace fish protein in feed. Two levels of replacement for H. illucens, 30% and 50% (H30 and H50), and one for T. molitor, 50% (T50), as well as an additional diet with a modified fatty acid fraction (H50M), were investigated in relation to juvenile Sparus aurata growth indices, enzyme activities and gut microbiome. A T50 diet showed similar results to a control (C) diet, with no significant differences regarding morphological indices and minor differences for nutritional indices. Regarding the gut microbiome, H50M was the diet which showed the more similar prokaryotic community to C, which suggests that fatty acid fractions might influence the composition of the gut microbiome. Nevertheless, differences appeared to be related to a redistribution of dominant species, while changes in species affiliation were limited to minoritary species. The positive correlation between some of these minoritary species (Peptostreptococcus russellii, Streptococcus dysgalactiae and Weisella confusa) and several fish growth parameters might explain differences between control and insect diets. Deciphering such uncertainty and revealing the potential role these unusual species may play on fish performance should be addressed in future investigations.

γ-Linolenic and Stearidonic Acids from Boraginaceae of Diverse Mediterranean Origin.

Thirty Boraginaceae species from different tribes were evaluated in a search of γ-linolenic (GLA, 18:3n-6) and stearidonic acid (SDA, 18:4n-3)-rich oils. The high GLA percentages were found in the seed oils of Symphytum bulbosum and S. tuberosum subsp. tuberosum (27.6 and 27.2 % of total fatty acids (FA)), which are unusually high values for GLA-oils in the current literature. On the whole seed, noticeable GLA percentages were found in S. grandiflorum, S. tuberosum subsp. tuberosum and Borago officinalis (7.43, 4.90, and 4.51 g/100 g, respectively). The main SDA-taxa detected in this study were Buglossoides arvensis, B. incrassata and Glandora oleifolia (21.3, 18.9, and 16.3 % of total fatty acids). On total seed weight, Glandora rosmarinifolia showed the highest SDA content (3.57 g/100 g). Finally, the higher FA contents were found in S. grandiflorum and Paramoltkia doerfleri seeds (35.2 and 37.0 g/100 g, respectively). Principal component analysis showed that similarities in FA profiles allow grouping species as botanical criteria for Boraginaceae tribes do, while the FA groupings confirm the metabolic activities of desaturase and elongase enzymes. Data on the FA composition of the seed oils analyzed here suggest their potential use as functional foods and can be considered as novel sources of SDA and GLA.

SWATH Differential Abundance Proteomics and Cellular Assays Show In Vitro Anticancer Activity of Arachidonic Acid- and Docosahexaenoic Acid-Based Monoacylglycerols in HT-29 Colorectal Cancer Cells.

Colorectal cancer (CRC) is one of the most common and mortal types of cancer. There is increasing evidence that some polyunsaturated fatty acids (PUFAs) exercise specific inhibitory actions on cancer cells through different mechanisms, as a previous study on CRC cells demonstrated for two very long-chain PUFA. These were docosahexaenoic acid (DHA, 22:6n3) and arachidonic acid (ARA, 20:4n6) in the free fatty acid (FFA) form. In this work, similar design and technology have been used to investigate the actions of both DHA and ARA as monoacylglycerol (MAG) molecules, and results have been compared with those obtained using the corresponding FFA. Cell assays revealed that ARA- and DHA-MAG exercised dose- and time-dependent antiproliferative actions, with DHA-MAG acting on cancer cells more efficiently than ARA-MAG. Sequential window acquisition of all theoretical mass spectra (SWATH) - mass spectrometry massive quantitative proteomics, validated by parallel reaction monitoring and followed by pathway analysis, revealed that DHA-MAG had a massive effect in the proteasome complex, while the ARA-MAG main effect was related to DNA replication. Prostaglandin synthesis also resulted as inhibited by DHA-MAG. Results clearly demonstrated the ability of both ARA- and DHA-MAG to induce cell death in colon cancer cells, which suggests a direct relationship between chemical structure and antitumoral actions.

Life after Harvest: Circadian Regulation in Photosynthetic Pigments of Rocket Leaves during Supermarket Storage Affects the Nutritional Quality.

Vegetables, once harvested and stored on supermarket shelves, continue to perform biochemical adjustments due to their modular nature and their ability to retain physiological autonomy. They can live after being harvested. In particular, the content of some essential nutraceuticals, such as carotenoids, can be altered in response to environmental or internal stimuli. Therefore, in the present study, we wondered whether endogenous rhythms continue to operate in commercial vegetables and if so, whether vegetable nutritional quality could be altered by such cycles. Our experimental model consisted of rocket leaves entrained under light/darkness cycles of 12/12 h over 3 days, and then we examined free-run oscillations for 2 days under continuous light or continuous darkness, which led to chlorophyll and carotenoid oscillations in both constant conditions. Given the importance of preserving food quality, the existence of such internal rhythms during continuous conditions may open new research perspective in nutrition science. However, while chromatographic techniques employed to determine pigment composition are accurate, they are also time-consuming and expensive. Here we propose for the first time an alternative method to estimate pigment content and the nutritional quality by the use of non-destructive and in situ optical techniques. These results are promising for nutritional quality assessments.