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The extensive investigation into the capacity of mealworms to digest diverse food by-products, as well as plastic wastes, has been a focal point in recent years. The transition from traditional diet sources like brans to food wastes has the potential to impact the physiological properties of mealworms. This study explored the utilization of various industrial food wastes such as okara, barley spent grain (BSG), sesame oil meal (SOM), and spent coffee grounds (SCG) as feed alternatives, and reports on their survival rate, biomass variations, and nutritional composition. In additional, the shift in their gut microbiota was also assessed. Among the range of industrial food wastes, mealworms exhibited the most robust growth performance when nourished with BSG. This particular group showed a survival rate of 98.33 % and a biomass increase of 23.06 %. In contrast, mealworms fed with SCG demonstrated the lowest survival rate and experienced a significant reduction in biomass. Although the groups fed with okara and SCG displayed moderate growth performance, both exhibited protein levels comparable to those observed in the oatmeal-fed group (used as the positive control). Notably, the inclusion of BSG in the mealworm diet exhibited the potential to enrich their omega-3 fatty acid content, suggesting potential benefits for applications as animal feed or even human consumption. Furthermore, an analysis of the gut microbiome was conducted to investigate the associations between specific diets and the composition of mealworm gut microbiota. In summary, food wastes such as BSG may be repurposed as feed substrates for mealworms before converting them into an alternative source of protein.

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Elderly people are susceptible to malnutrition due to many factors. An essential public health priority is ensuring that seniors have appropriate nutrition to prevent and treat malnutrition and dehydration as well as fulfilling the nutrition necessity. This study was conducted to utilize selected legumes and mushrooms to develop a high-protein instant soup (HPIS) product that is nutritious and suitable for the consumption of the elderly. Black bean, along with oyster mushroom and split-gill mushrooms, exhibited considerable amounts of protein (19.13 ± 1.13, 2.77 ± 0.09, and 4.65 ± 0.61 g/100 g, respectively), calcium (2308.65 ± 113.07, 640.19 ± 0.80, and 743.89 ± 0.66, respectively), iron (40.84 ± 2.42, 7.31 ± 0.05, and 40.10 ± 2.15, respectively), and zinc (18.06 ± 1.07, 3.87 ± 0.03, and 26.23 ± 0.78, respectively) content, and were incorporated into the HPIS formula. Drum dryer rotation speed significantly affected the HPIS properties. The study on the effect of different rotation speeds (3, 5, 7, and 9 RPM) used during the drying process revealed the use of 7 RPM resulting in the soup product with considerable quality. The optimized HPIS formula, which was supplemented with minerals, was moderately liked (7.1-7.5) by the elderly consumer. The majority of the consumers accepted the product (97 %) and were interested in purchasing the product (91 %) if it was available on the market. Incorporating selected legumes and mushrooms resulted in a soup product containing nutrition conforming to the Thai recommended daily intake (RDI), possessing adequate physicochemical and sensory properties for the consumption of the elderly.

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This study aimed to determine the compatibility of pre-fermented sugar beet pulp to support the growth of Pleurotus ostreatus mycelium in submerged fermentation. The goal was to create a meat alternative based on mycelial-fermented pulp. It was further explored whether pre-fermentation with lactic acid bacteria (LAB) on the pulp increased meat-like properties, such as aroma, springiness, and hardness, in the final product. Three strains were selected from a high throughput screening of 105 plant-derived LAB based on their acidification and metabolite production in the pulp. Two homofermentative strains (Lactococcus lactis) and one heterofermentative strain (Levilactobacillus brevis) were selected based on their low ethanol production, high lactic acid production, and overall acidification of the pulp. Mycelium of P. ostreatus was grown in submerged fermentations on the pre-fermented pulp, and the biomass was removed by centrifugation. The fungal strain consumed all available sugars and acids and released arabinose to the media. Volatiles were detected using GC-MS, and a large increase in concentrations of hexanal, 1-octen-3-ol, and 2-octenal was measured. Concentration of 1-octen-3-ol was lower in the pre-fermented samples vs. the non-pre-fermented. LC-MS amino acid analysis showed the presence of all essential amino acids on day 0 and 7 of fermentation. The highest concentration of amino acids was for glutamic acid/glutamine and aspartic acid/asparagine. A decrease in all amino acids after 7 days of fungal fermentation was measured for all fermentations. The decrease was more significant for pre-fermented samples. This was also confirmed through a total protein determination, except for samples pre-fermented with Lactococcus lactis strain NFICC142 which increased in total protein content after fungal fermentation. The protein digestibility increased after fungal fermentation, and the highest increase was seen for non-pre-fermented samples. The springiness of the fermented product indicated similarities to meat alternatives, while the hardness was much lower than other meat alternatives. The results indicate that dried sugar beet pulp can be used for submerged cultivation of P. ostreatus, but that pre-fermentation does not improve the physical or nutritional properties of the end product significantly, except for an increased protein content for NFICC142 pre-fermented media. This is the first known attempt to use LAB and P. ostreatus in mixed fermentation to produce fungal mycelium, as well as the first attempt at using SBP in a liquid fermentation for mycelial production of P. ostreatus.

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This study aims to implement and increase values regarding the use of cricket powder in food. This is due to the need to increase the range of products enriched with cricket powder in order to increase the market and acceptance of insects in the diet. The work analyzed properties of yoghurt with cricket powder added (chemical composition, acidity, colour, consistency, degree of syneresis, texture and sensory analysis). Research has shown that the greater the addition of cricket powder, the higher the protein, fat and dry matter content and the lower the hardness of the yoghurt. As the content of cricket powder in yoghurt increased, the brightness (L*) and redness (a*) decreased, while the yellowness (b*) of the samples increased. A change in the colour of the yoghurt was observed with the increase in the cricket powder content; the yoghurt had a darker, browner colour. The best overall sensory rating compared to the control variant (6.4) was given to the yoghurt with 1.5% cricket powder added (5.7), while the worst rating was given to the yoghurt with 5% powder added (2 points out of 7).

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Mycoprotein is an alternative protein produced through fungal fermentation. However, it typically relies on refined glucose syrup derived from starch, which can be costly and unsustainable. This study investigates the potential of soybean processing by-products (okara and soy whey) as alternative substrates for producing mycoprotein using Aspergillus oryzae. A. oryzae was cultured for 7 days at 30 °C in diluted okara (1:50) and soy whey (1:1) with or without agitation (100 rpm). Soy whey produced higher biomass yields (369.2-408.8 mg dry biomass/g dry substrate), but had a lower biomass concentration (0.783-0.867 g dry weight/L). Conversely, okara produced a higher biomass concentration (2.02 g dry weight/L) with a yield of 114.7 mg dry biomass/g dry substrate. However, biomass formation in okara was only observed in static conditions, as agitation caused biomass to entangle with soy pulp, hampering its production. Additionally, okara tended to release protein into the media, while soy whey accumulated protein within the biomass, reaching up to 53% w/w protein content. The results of this study provide a promising approach to addressing both soybean processing waste reduction and food security concerns.

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Ribosome profiling and mass spectrometry have revealed thousands of previously unannotated small and alternative open reading frames (sm/alt-ORFs) that are translated into micro/alt-proteins in mammalian cells. However, their prevalence across human tissues and biological roles remains largely undefined. The placenta is an ideal model for identifying unannotated microproteins and alt-proteins due to its considerable protein diversity that is required to sustain fetal development during pregnancy. Here, we profiled unannotated microproteins and alt-proteins in human placental tissues from preeclampsia patients or healthy individuals by proteomics, identified 52 unannotated microproteins or alt-proteins, and demonstrated that five microproteins can be translated from overexpression constructs in a heterologous cell line, although several are unstable. We further demonstrated that one microprotein, XRCC6P1, associates with translation initiation factor eIF3 and negatively regulates translation when exogenously overexpressed. Thus, we revealed a hidden sm/alt-ORF-encoded proteome in the human placenta, which may advance the mechanism studies for placenta development as well as placental disorders such as preeclampsia.

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The traditional approach to formulating pig diets is based only on minimizing cost while meeting nutritional requirements and thus does not consider the environmental impacts associated with producing feed ingredients. To reduce the overall environmental impact of pork production, feed ingredients can be considered to formulate environmentally friendly diets. However, their potential effects on pig performance could decrease environmental benefits at the farm gate. The objective of this study was to quantify the effects of such eco-friendly pig diets on nitrogen (N) and energy (E) balances, the components of heat production (HP), and the performance of growing pigs. Digestibility coefficients of dry matter (84.5% vs. 88.2%, P < 0.01) and N (80.4% vs. 86.3%, P < 0.01) were significantly lower for the eco-friendly diet than the Control-diet (a commercial diet used in France). N excretion in feces was significantly higher for the group of pigs fed the eco-friendly diet than for the group fed the Control-diet (9.8 vs. 6.9 g/d, respectively, P = 0.01), while the N retention tended to be lower (27.8 vs. 30.3 g/d, respectively; P = 0.06). The metabolizable E:digestible E ratio did not differ between diets, but total HP was significantly lower for the eco-friendly diet group than for the Control-diet group (1,340 vs. 1,388 kJ/kg body weight0.60/d, respectively, P = 0.03). Using feed ingredients with lower environmental impacts, such as locally produced protein or co-products from wheat processing, is an effective way to decrease the environmental impacts of pig production. However, the nutritional composition of these eco-friendly ingredients could be overestimated, in particular the true digestibility of amino acids. This indicates the need to better estimate and consider the true digestibility of eco-friendly diets to decrease environmental impacts of livestock production without decreasing animal performance.

Livestock production is a significant contributor to global environmental change. In pig production, animal feed contributes to 55% to 75% of climate change impacts, 70% to 90% of nonrenewable energy use, and 85% to 100% of land occupation. To reduce the overall environmental impact of pork production, feed ingredients can be considered to formulate environmentally friendly diets. These feeding strategies that minimized environmental impacts contained lower proportions of cereals and oilseeds and a higher proportion of high-protein crops and co-products from wheat processing which have lower impacts. The use of these eco-friendly ingredients could influence performance due to their variability in energy, fiber, or protein contents, with an overestimation of their nutritional composition. Thus, decreasing environmental impacts of livestock production without decreasing animal performance requires better characterizing the ingredients that have lower environmental impacts.

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The demand for sustainable and ethically farmed animal products is on the rise as consumers become more environmentally and animal welfare conscious. The need to diminish the consumption of soybean meal is urgent, and companies are looking for ways to respond to this necessity by looking for alternatives to soybean meal. This study assessed the impact of introducing whole dehydrated and live black soldier fly larvae (BSFL) into the diet of an indigenous chicken breed as environmental enrichment. A total of 144 39-day-old male Bianca di Saluzzo chickens were distributed among 18 pens and assigned to three different experimental groups. The control group received a diet where soybean meal was entirely replaced by alternative ingredients. The two experimental groups were given the same diet supplemented with 5% of the expected daily feed intake of whole dehydrated BSFL or whole live BSFL. Throughout the trial period (from the bird age of 39-174 days of age), live weight was recorded every 21 days, and the average daily gain, daily feed intake, and feed conversion ratio were calculated. The time required for the birds to consume the larvae was recorded three times a week. At age 147 and 174 days, 12 birds per treatment were selected based on mean live weight and slaughtered. Measurements included hot and chilled carcass weights, organ weights (spleen, liver, heart, stomach), breast and thigh muscle weights, and the corresponding yields were calculated. Acid protease activity was measured in proventriculus extract, and chitinase and chitosanase activity was calculated based on the release of reducing sugars from chitin or chitosan. The results showed little improvement in final live weights and daily feed intakes of the animals fed the insect larvae compared with control birds. Larva supplementation had no negative impact on the overall well-being of the animals assessed by blood analysis and histopathological assessment of the intestinal tract, spleen, and liver. No differences were found between the dehydrated vs live insect larvae consumption times, with all larvae being eaten up very rapidly (< 3 min). The birds fed BSFL showed an increase in chitinase activity. These findings support the potential use of whole BSFL as a form of environmental enrichment, particularly in their dehydrated form, being more convenient to use and store, which would also encourage the uptake of this practice by farmers.

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A total of 23 studies were identified in a literature search performed in the Scopus, Science Direct and Google Scholar databases for meta-analysis. The criteria used include studies that were published from 2015 to 2023 and those reporting the effects of insect meal utilisation in poultry diets. Data on live weight (LW), carcass weight (CW), moisture, meat pH, lightness (L*), redness (a*), yellowness (b*), proximate composition (protein, fat and ash content) and shear force in broilers were subjected to OpenMEE software, and data were pooled using a random-effect model. Subgroup analysis and meta-regression were performed to ascertain the influence of dietary insect meals on the response of meat aspects and the source of heterogeneity, respectively, using the following moderators (insect species, dosage level, feeding duration and age at slaughter). The results indicated that dietary insect meal did not affect LW, CW, meat L*, pH, shear force, moisture, fat and ash content. In contrast, dietary insect meal increased the a* of the meat (standardised mean differences (SMDs) = 1.03; 95% confidence intervals (CIs) = 0.484-1.578; p ≤ 0.001), b* (SMD = 1.117; 95% CI = 0.334-1.90; p = 0.005), and meat protein content (SMD = 0.365; 95% CI = 0.031-0.7; p = 0.032). The subgroup analysis showed that insect meal dosage of ≤10% and age at slaughtered ≤35 days had improved the LW, CW and meat L*. In addition, the meat a*, protein and ash content were also influenced by insect species, dosage levels and age at slaughter. In conclusion, ≤10% of either Hermetia illucens or Tenebrio molitor can be included in broiler diets without compromising the LW, CW, meat pH, colour, shear force, moisture, fat and ash content in broilers. The study therefore indicated that insect meals have a bright future as an alternative protein source in poultry diets.

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Cell-cultured meat holds significant environmental value as an alternative protein source. Throughout the 21st century, cell-cultured meat has progressively penetrated commercial markets. However, a systematic review encompassing the entire field needs improvement. Employing Citespace, Vosviewer, and R-Bibliometrix software, a bibliometric analysis was used to present the research progress and general development trends of 484 articles on cell-cultured meat from 2000 to 2022 based on countries, authors, institutions, and keywords. This analysis provides ideas for the future development of cell-cultured meat in different countries or regions worldwide. Research on cell-cultured meat from 2000 to 2022 has undergone two phases: fluctuating growth (2000-2013) and rapid growth (2013-2022). Noteworthy contributions to cell-cultured meat studies emerge from author groups in the United States of America, the United Kingdom, and China, with influential institutions like the University of Bath significantly impacting pertinent research. Furthermore, over the past two decades, research has leaned towards exploring topics such as "biomaterials", "cultured", "land use", "public opinion", "animal welfare", and "food safety". Furthermore, this study reveals differences in nomenclature between regions and institutions. "Cultured meat" is more popular in some countries than in other forms. Institutions in Asia use "cultured meat" more frequently; however, institutions in the Americas adopt "cultivated meat" and rarely adopt "in vitro meat", and institutions in the European region have no particularly prominent tendency towards a specific nomenclature. Future research should emphasize aligning the labeling of cell-cultured meat with effective management strategies and referencing regulatory policies across various countries. For the first time, we use three different bibliometric methods to analyze temporal and spatial variation in research on cellular meat. The results of this study have a multiplier effect. We provide a theoretical basis and a practical reference for the identification of alternatives in the dual context of "food crisis and food security" and "climate crisis". At the same time, we also provide a reference for the sustainable development of the food system.

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Researchers are exploring solutions to meet the growing demand for protein due to the expected increase in global population by 2050. Interest in alternative protein sources like insects has risen, driven by concerns about environmental impact and the need for sustainable food production. This study aimed to develop and evaluate the physicochemical properties of soy-protein-based burgers enriched with insect protein from Alphitobius diaperinus. Three formulations were developed: a control (B0) and burgers with 5% (B5) and 10% (B10) insect protein-Whole Buffalo Powder (WBP). The results showed that adding insect protein decreased the burger analogue's pH. A clear trend was observed of increasing total lipids and saturated fatty acids (SFA) and decreasing monounsaturated (MUFA) and polyunsaturated fatty acids (PUFA) as the WBP concentration increased from 0% to 10%. No significant differences with increasing WBP concentration in the protein content of the burger analogue, as well as the cooking yield, were noted. The WBP addition had a notable effect on the color change, especially a decrease in brightness (L*). It was shown that as the WBP concentration increased, there were no significant differences in the texture profile of the burger analogues. The formulation with 5% WBP concentration was the most acceptable in sensory analysis.

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Fish meal (FM) replacement is essential for the sustainable expansion of aquaculture. This study focussed on the feasibility of replacing FM with a single-cell protein (SCP) derived from methanotrophic bacteria (Methylococcus capsulatus, Bath) in barramundi fry (Lates calcarifer). Three isonitrogenous and isoenergetic diets were formulated with 0%, 6.4% and 12.9% inclusion of the SCP, replacing FM by 0%, 25% and 50%. Barramundi fry (initial body weight 2.5 ± 0.1 g) were fed experimental diets for 21 days to assess growth performance, gut microbiome composition and gut histopathology. Our findings revealed that both levels of SCP inclusion induced detrimental effects in barramundi fry, including impaired growth and reduced survival compared with the control group (66.7% and 71.7% survival in diets replacing FM with SCP by 25% and 50%, respectively; p < .05). Both dietary treatments presented mild necrotizing enteritis with subepithelial oedema and accumulation of PAS positive, diastase resistant droplets within hepatocytes (ceroid hepatopathy) and pancreatic atrophy. Microbiome analysis revealed a marked shift in the gut microbial community with the expansion of potential opportunistic bacteria in the genus Aeromonas. Reduced overall performance in the highest inclusion level (50% SCP) was primarily associated with reduced feed intake, likely related to palatability issues, albeit pathological changes observed in gut and liver may also play a role. Our study highlights the importance of meticulous optimization of SCP inclusion levels in aquafeed formulations, and the need for species and life-stage specific assessments to ensure the health and welfare of fish in sustainable aquaculture practices.

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Climate change and changing consumer demand are the main factors driving the protein transition. This shift toward more sustainable protein sources as alternatives to animal proteins is also reflected in the rapid upscaling of meat and dairy food analogues. Such changes could challenge food safety, as new food sources could result in new and unexpected food safety risks for consumers. This review analyzed the current knowledge on chemical and microbiological contamination of emerging alternative protein sources of plant origin, including soil-based (faba bean, mung bean, lentils, black gram, cowpea, quinoa, hemp, and leaf proteins) and aquatic-based (microalgae and duckweeds) proteins. Moreover, findings on commercial analogues from known alternative protein sources were included. Overall, the main focus of the investigations is on the European context. The review aimed to enable foresight approaches to food safety concerning the protein transition. The results indicated the occurrence of multiple chemical and microbiological hazards either in the raw materials that are the protein sources and eventually in the analogues. Moreover, current European legislation on maximum limits does not address most of the "contaminant-food" pairs identified, and no legislative framework has been developed for analogues. Results of this study provide stakeholders with a more comprehensive understanding of the chemical and microbiological safety of alternative protein sources and derived analogues to enable a holistic and safe approach to the protein transition.

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Introduction: Food neophobia (FN) is a psychological trait that inhibits one's willingness to eat unfamiliar foods. It is related to the acceptance of insect foods and cultured meat, which are major protein alternatives to conventional meat, and is an important personality trait for understanding the near-future food industry. However, the factor structure of Pliner and Hobden's FN scale (FNS) is unstable due to respondents' cultural backgrounds. Thus, we aimed to develop a Japanese version based on the alternative FNS (FNS-A), the most recent revised version, and to examine its validity. Methods: Four online surveys (preliminary 1: n = 202; preliminary 2: n = 207; main: n = 1,079; follow-up: n = 500) were conducted on the FNS-A. For the main survey, Japanese respondents (aged 20-69 years) answered the Japanese version of the FNS-A (J-FNS-A), their willingness to eat (WTE), and their familiarity with hamburgers containing regular protein foods (ground beef, tofu) and alternative protein foods (soy meat, cultured meat, cricket powder, algae powder, and mealworm powder). Results: Consistent with the FNS-A, confirmatory factor analysis assuming a two-dimensional structure (approach and avoidance) showed satisfactory model fit indices. The mean J-FNS-A score (Cronbach's α for 8 items = 0.83) was 4.15 [standard deviation (SD) = 0.93]. J-FNS-A scores were not associated with age and gender, whereas a greater than moderate association was found with WTE hamburgers containing alternative protein foods (rs = -0.42 to -0.33). The strength of these negative associations increased as food familiarity decreased (r = 0.94). The test-retest reliability at 1 month was also satisfactory (r = 0.79). Discussion: The validity of the J-FNS-A was confirmed. Higher J-FNS-A scores (mean = 41.51, SD = 9.25, converted to Pliner and Hobden's FNS score) of the respondents suggest that Japanese people prefer conservative foods. This scale could predict the negative attitudes toward foods with low familiarity, such as alternative proteins. The J-FNS-A appears to be a useful psychological tool for assessing Japanese food neophobia tendencies and predicting novel food choices of Japanese individuals.

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Beyond the key bitter compound kaempferol 3-O-(2‴-O-sinapoyl-ß-d-sophoroside) previously described in the literature (1), eight further bitter and astringent-tasting kaempferol glucosides (2-9) have been identified in rapeseed protein isolates (Brassica napus L.). The bitterness and astringency of these taste-active substances have been described with taste threshold concentrations ranging from 3.3 to 531.7 and 0.3 to 66.4 µmol/L, respectively, as determined by human sensory experiments. In this study, the impact of 1 and kaempferol 3-O-ß-d-glucopyranoside (8) on TAS2R-linked proton secretion by HGT-1 cells was analyzed by quantification of the intracellular proton index. mRNA levels of bitter receptors TAS2R3, 4, 5, 13, 30, 31, 39, 40, 43, 45, 46, 50 and TAS2R8 were increased after treatment with compounds 1 and 8. Using quantitative UHPLC-MS/MSMRM measurements, the concentrations of 1-9 were determined in rapeseed/canola seeds and their corresponding protein isolates. Depending on the sample material, compounds 1, 3, and 5-9 exceeded dose over threshold (DoT) factors above one for both bitterness and astringency in selected protein isolates. In addition, an increase in the key bitter compound 1 during industrial protein production (apart from enrichment) was observed, allowing the identification of the potential precursor of 1 to be kaempferol 3-O-(2‴-O-sinapoyl-ß-d-sophoroside)-7-O-ß-d-glucopyranoside (3). These results may contribute to the production of less bitter and astringent rapeseed protein isolates through the optimization of breeding and postharvest downstream processing.

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This study aimed to upcycle a byproduct of the edible oil industry, cold-pressed nettle seed meal (CPNSM), into a plant-based emulsifier, thereby increasing the sustainability of the food system. The protein content of the nettle seed protein (NSP) powder was 48.3% with glutamic acid (16.6%), asparagine (10.7%), and arginine (9.7%) being the major amino acids. NSPs had a denaturation temperature of 66.6 °C and an isoelectric point of pH 4.3. They could be used as emulsifiers to form highly viscous coarse corn oil-in-water emulsions (10% oil, 4% NSP). Nevertheless, 10-fold diluted emulsions exhibited rapid creaming under different pH (2-9), salt (0-500 mM NaCl) and temperature (>40 °C) conditions, but they were relatively stable to aggregation. Our findings suggest that NSPs could be used as emulsifiers in highly viscous or gelled foods, like dressings, sauces, egg, cheese, or meat analogs.

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Lentil (Lens culinaris) is a protein-rich legume consumed worldwide and it also has the potential to become an alternative source of protein ingredient for human nutrition. The aim of this study was to determine the best processing parameters for the whole grain protein wet extraction, as well as to analyze the techno-functional properties, and physical characteristics of the protein concentrate and its flour. It was also evaluated the application of the concentrate into a fish-like croquette. The processing route was carried out by alkaline extraction and acid precipitation of the proteins where the pH, stirring time and solute:solvent ratio were evaluated. The final dried protein concentrate presented 85% protein on dry basis and a mass yield of 14%. The results were reproducible when tested on a first scaling up test. For the techno-functional properties, solubility, water and oil retention capacities, emulsification and foaming capacities and stability, and gelling capacity were tested. As for the food application into fish-like croquettes, the lentil protein showed similar scores for sensory acceptance, flavor and texture when compared to a commercial clean-taste concentrate. The results observed in this study were compatible to other alternative pulse-protein ingredients on the market, positioning lentil protein as a promising alternative protein source to produce ingredients for the plant-based market.

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This study investigated the effects of fish protein hydrolysate derived from barramundi on growth performance, muscle composition, immune response, disease resistance, histology and gene expression in white shrimp (Penaeus vannamei). In vitro studies demonstrated FPH enhanced mRNA expressions of key immune-related genes and stimulated reactive oxygen species (ROS) production and phagocytic activity in shrimp hemocytes. To evaluate the effects of substituting fish meal with FPH in vivo, four isoproteic (43 %), isolipidic (6 %), and isoenergetic diets (489 kcal/100 g) were formulated with fish meal substitution levels of 0 % (control), 30 % (FPH30), 65 % (FPH65), and 100 % (FPH100). After 8-week feeding, the growth performance of FPH65 and FPH100 were significantly lower than that of control and FPH30 (p < 0.05). Similarly, the midgut histological examination revealed the wall thickness and villi height of FPH100 were significantly lower than those of control (p < 0.05). The shrimps were received the challenge of AHPND + Vibrio parahaemolyticus at week 4 and 8. All FPH-fed groups significantly enhanced resistance against Vibrio parahaemolyticus at week 4 (p < 0.05). However, this protective effect diminished after long-period feeding. No significant difference of survival rate was observed among all groups at week 8 (p > 0.05). The expressions of immune-related genes were analyzed at week 4 before and after challenge. In control group, V. parahaemolyticus significantly elevated SOD in hepatopancreas and Muc 19, trypsin, Midline-fas, and GPx in foregut (p < 0.05). Moreover, hepatopancreatic SOD of FPH65 and FPH100 were significantly higher than that of control before challenge (p < 0.05). Immune parameters were measured at week 8. Compared with control, the phagocytic index of FPH 30 was significantly higher (p < 0.05). However, dietary FPH did not alter ROS production, phenoloxidase activity, phagocytic rate, and total hemocyte count (p > 0.05). These findings suggest that FPH30 holds promise as a feed without adverse impacts on growth performance while enhancing the immunological response of white shrimp.

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This study focused on optimizing the production of fermented Spirulina (FS) products using a bioactivity-guided strategy with Lactobacillus helveticus B-4526 and Kluyveromyces marxianus Y-329 in a 3-L bioreactor. Various operating conditions, including aeration rates and pH modes, were tested. While both microorganisms thrived under all conditions, the "cascade" mode, controlling dissolved oxygen, enhanced protein hydrolysis and antioxidant activity, as confirmed by SDS-PAGE and DPPH/TEAC assays, respectively. Screening revealed that "cascade" FS significantly decreased viability of colon cancer cells (HT-29) in a dose-dependent manner, with up to a 72 % reduction. Doses ≤ 500 µg mL-1 of "cascade" FS proved safe and effective in suppressing NO release without compromising cellular viability. Additionally, "cascade" FS exhibited diverse volatile organic compounds and reducing the characteristic "seaweed" aroma. These findings highlight "cascade" FS as a promising alternative food source with improved bioactive properties, urging further exploration of its bioactive compounds, particularly bioactive peptides.

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Yeast protein can be a nutritionally suitable auxiliary protein source in livestock food. The breakdown of proteins and thereby generating high-quality peptide, typically provides nutritional benefits. Enzyme hydrolysis has been effectively uesed to generate peptides; however, studies on the potential applications of different types of enzymes to produce yeast protein hydrolysates remain limited. This study investigated the effects of endo- (alcalase and neutrase) and exotype (flavourzyme and prozyme 2000P) enzyme treatments on yeast protein. Endotype enzymes facilitate a higher hydrolysis efficiency in yeast proteins than exotype enzymes. The highest degree of hydrolysis was observed for the protein treated with neutrase, which was followed by alcalase, prozyme 2000P, and flavourzyme. Furthermore, endotype enzyme treated proteins exhibited higher solubility than their exotype counterparts. Notably, the more uniform particle size distribution was observed in endotype treated yeast protein. Moreover, compared with the original yeast protein, the enzymatic protein hydrolysates possessed a higher content of ß-sheets structures, indicating their higher structural stability. Regardless of enzyme type, enzyme treated protein possessed a higher total free amino acid content including essential amino acids. Therefore, this study provides significant insights into the production of protein hydrolysates as an alternative protein material.