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.

Potentially toxic element bioaccumulation in consumed indoor shrimp farming associated with diet, water and sediment levels.

Shrimp production is an important industry for many countries and shrimp consumption is increasing worldwide. Shrimps are a highly nutritional food, but can pose a risk for human health if subject to high levels of environmental contaminants. This work studies the presence of As, Cd, Co, Cr, Cu, Hg, Ni, Pb and Zn in shrimps from Ecuador and compares them to such contents noted in other shrimp-production areas in the world to evaluate the possible risks associated with these elements for consumer health, and to relate them to potentially toxic element (PTE) contents in water, sediments and diets, and also to animal biometric parameters. The PTE levels (mg kg-1 DM) obtained are as follows: in the head-As (3.52-6.11), Cd (0.02-0.10), Co (0.14-0.49) Cr (0.23-4.89), Cu (99.9--233.0), Ni (0.52-1.86), Pb (0.24-1.09), Zn (51.8-100.5) and Hg (µg kg-1 DM) (10.00-66.81); in the tail-(0.91-3.21), Cd (0.01-0.02), Co (0.01-0.43) Cr (0.01-6.52), Cu (20.0-72.44), Ni (0.15-2.03), Pb (0.01-0.69), Zn (31.2-66.1) and Hg (µg kg-1 DM) (10.00-67.18). The concentration of all the PTEs is generally lower than the limits set for seafood by European regulations, except for As in the cephalothorax (4.63 mg kg-1). Different behaviours for PTE accumulation in shrimps were found, which preferentially tend to accumulate in the cephalothorax, except for Hg (40.13 µg kg-1 DM), which accumulates in muscle (body) and is associated with contents of proteins, lipids and total shrimp weight. Nonetheless, the target hazard quotient (THQ) values for PTEs indicate that the consumption of shrimp muscles from Ecuador does not pose a human health risk because the values of these indices are below 1 in all cases.

Evaluation of the interrelated effects of slaughtering, drying, and defatting methods on the composition and properties of black soldier fly (Hermetia illucens) larvae fat.

The interrelated effect of different slaughtering, drying and defatting methods of black soldier fly larvae (BSFL) on the lipid composition and properties of the fat was studied. Blanching and freezing were compared as slaughtering methods, oven or freeze-drying as drying methods, and mechanical pressing or supercritical fluid extraction (SFE) as defatting methods. The different modes of slaughtering, drying, and defatting, along with both binary and ternary interactions caused significant effects on processes yields, lipid composition, moisture content and thermal properties. Thus, considering the defatting degree and the yield in total valued products (defatted meal plus fat), the combination of blanching, freeze-drying plus mechanical pressing was the worst option (51.2% and 87.5%, respectively). In contrast, the other combinations demonstrated better and comparable efficiency, although SFE is preferable for defatting (83.2% and 96.9%, respectively). The content of major fatty acids (lauric, palmitic and myristic acids) was significantly affected by the BSFL treatments, although with unsignificant impact on the total saturated fatty acids content. To preserve the integrity of the fat, the combination of blanching and oven-drying was preferred, as non-thermal methods of slaughtering and drying caused intense lipolysis, releasing free fatty acids (FFA) in the range of 18.6-23.5%. To achieve the lowest moisture content in the fats (≤0.1%), oven-drying with mechanical pressing were desired, regardless of the slaughtering method; while values > 1% were reached for freezing, freeze-drying and SFE. Both differences in FFA and moisture contents caused different thermal behaviors in the samples. Specially, the melting temperature was lower for samples with higher FFA and moisture contents, with a notable difference when freezing, freeze-drying and SFE were combined (14.5 °C vs 30.6 °C, as the mean value for the rest of samples). The different modes of processing did not affect the minor lipid compounds. Therefore, the modes employed for slaughtering, drying, and defatting of BSFL determine, either individually or in combination, the process yields, composition, and properties of the fat.

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.

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.

Effects of chemical and handling exposure on fatty acids, oxidative stress and morphological welfare indicators in gilt-head sea bream (Sparus aurata).

This work investigates the changes in the morphological traits, body composition, body fat and oxidative stress of Sparus aurata under two stress conditions: a low weekly exposure to the pesticide Diuron and chronic handling stress. The fish exposed to handling stress showed lower values in the morphological traits and perivisceral fat, whereas chemical stress induced a decrease in the percentage of muscle and carcase and an increase in the percentage of spleen and hepatosomatic index. The two stress situations produced changes in the FRAP (ferric reducing antioxidant power) levels in all organs studied with a particularly marked response for Diuron in the gill. No significant differences in the ROS (reactive oxygen species) level were found for the digestive tract or the gill. In muscle, only Diuron produced a significant increase in ROS, while in liver, both treatments increased the ROS levels. This supports the oxidative stress sensitivity to chemical stress and shows that also could be an appropriate indicator of handling stress. The morphological indexes and body components examined could be practical and easy welfare indicators. Nevertheless, further works must be needed to use under the production conditions in fish farms.

Depletion of veterinary drugs used in aquaculture after administration in feed to gilthead seabream (Sparus aurata).

This study was conducted to evaluate the depletion of residues of the antibiotics flumequine, oxytetracycline, sulfadiazine, trimethoprim, and oxolinic acid after in-feed administration to gilthead seabream. Fish were treated with the target antibiotics at doses of 30 mg/kg of body weight per day for 10 days at two seawater temperatures. Fish in each of five tanks were fed with a different medicated feed. After in-feed administration, five fish were randomly selected at different times, and antibiotic presence was analyzed in a mixture of muscle and skin. Antibiotic concentrations were determined through a validated analytical method based on liquid chromatography separation and mass spectrometry detection. Two trials were carried out with fish at different temperatures (14.0 and 19.5°C). Depletion of antibiotics occurred more rapidly at the higher temperature. Elimination rates for all antibiotics assayed were high, which indicates that the withdrawal period for these antibiotics could be reduced. The results suggest that in gilthead seabream maintained at these two temperatures no detectable concentrations of the antibiotics used in this study will remain in edible tissues 35 days after treatment. For flumequine and oxolinic acid, the elimination time is shorter (4 and 20 days, respectively).