Alternative and New Protein Sources

The present study was conducted to examine the effect of different levels of essential and nonessential amino acid in NCSU-23 medium on the in vitro-produced porcine embryo as it develops from the zygote to the blastocyst stage. Four experiments were performed, each with a completely randomized design involving 5 to 8 replications of treatments. In order to know the effect of nonessential amino acids in NCSU-23 medium, 0, 5, 10 and 20 μl/ml MEM were supplemented there to, (Exp. 1) and the medium was supplemented with same level of essential amino acids (Exp. 2). The combined effect of nonessential (0, 5, 10 and 20 μl/ml MEM) and essential amino acids (0, 5, 10 and 10 μl/ml MEM) in NCSU-23 medium (Exp. 3), first 72 h with non-essential amino acids (at 0, 5, 10 and 20 μl/ml MEM), and last 4 d with essential amino acids with the same level as NEAA (Exp. 4) were examined. The embryo development was monitored and the quality of blastocysts was evaluated by counting the number of total cells and determining the ratio of inner cell mass (ICM) to trophoectoderm (TE) cells. When Eagle's nonessential amino acids (MEM) added to NCSU-23 medium, it significantly increased the likelihood of development to the 2- to 4-cell stage and subsequent blastocyst development. Supplementation of different levels of essential amino acids in the NCSU-23 medium decreased cleavage rate, rate of morula and blastocyst development and the number of ICMs. In the case of the combined effect of essential and nonessential amino acids, better and significant results were found for blastocysts, hatching blastocysts and for ICM numbers which were also dose dependent. With respect to the biphasic effect of nonessential and essential amino acids, nonessential amino acids increased cleavage whereas essential amino acids increased the total cell number. Neither the nonessential nor the essential group of amino acids, on their own, affected blastocyst cell number or the differentiation of cells in the blastocyst. In conclusion, this study determined the role of nonessential and essential amino acids in the culture of the porcine embryo and showed that the embryo requires different levels of amino acids as it develops from the zygote to the blastocyst stage.

The article presents a literature review on the risks of the global problem of fodder protein and protein amino acids in the feeding of agricultural animals and poultry. Alternative ways of solving it are due to the production and use of non-traditional feed raw materials in feed diets. A number of scientists consider atypical agricultural biotechnological products obtained from insects to be one of the alternative sources of fodder protein and amino acids and see prospects in the development of their production. The most common subjects for study in this regard are crickets, locusts, silkworm larvae, flies, worms, etc. Their protein value, amino acid, fatty acid and mineral composition, the content of vitamins, and other biologically active substances are studied. The development of this direction is considered one of the ways to solve a number of urgent environmental and fodder problems on a biological basis. Рurpose of the research was to study the protein value and complete amino acid composition of flour from insects, namely flour from the larvae of the BSF fly, crickets and snails, and the methods of its determining: crude protein by the Kjeldahl method, digestible protein with the help of 0,2% pepsin, and the digestibility coefficient, and the principles of conducting researches, as well as the characteristics of methods for determining the content of replaceable and essential amino acids, including sulfur-containing, using the Kapel-105M capillary electrophoresis system. As a result of the research, it was found that cricket flour is the least valuable in terms of protein value (digestible protein and digestibility coefficient), and it is also poor in the content of nonessential and essential amino acids, compared to other studied types of insect flour. The best indicators of protein value were noted in the flour from BSF fly larvae in terms of the content of crude and digestible proteins, as well as the digestibility coefficient, which is 75.6%, which indicates its good digestibility by the bodies of animals and birds. The highest content of amino acids is contained in the flour from BSF fly larvae. Taking into account the above results of crude protein content, digestibility, and digestibility in this sample, amino acid digestibility will also be higher compared to other flour samples. In addition, flour from BSF fly larvae is rich in sulfur-containing amino acids. The content of tryptophan and histidine, compared to other types of raw materials, is the highest. The studied samples of insect flour contain a complete set of essential and nonessential amino acids, but BSF fly larvae flour and snail flour are more valuable for essential amino acids.

Protein malnutrition is one of the global nutritional problems encountered in the less developed countries of the world, prompting researchers to search for alternative dietary protein sources that would alleviate problems of protein shortage and could also provide, to a certain extent, some important medicinal purposes. The leaves of the plant were washed with distilled water to remove dust, authenticated at the forest research institute Jericho Ibadan and processed for leaf protein concentrates to evaluate for proximate composition, mineral content, phytochemical analysis and amino acid profile using the standard method of analysis. The results of proximate analysis of the sample in g/100g showed moisture content (12.58±0.053), Ash (2.14±0.061), Fat (15.67±0.031), Crude protein (43.78±0.031), Crude fat (17.23±0.071) and Carbohydrate (8.58±0.113). The mineral composition of the sample had 0.19±0.00mg/g of magnesium, Na; 41.382±0.35mg/g, Fe; 1.292±0.002, K; 9.098±0.02, Fe; 1.293±0.00mg/g, Zn; 0.013±0.00mg/g, Cr; 0.026±0.00mg/g phosphorus; 19.520±0.29 Ni; 0.001±0.00, Cd; 0.0143±0.00mg/g, while lead and selenium were not detected in the sample. The results of the phytochemical analysis showed the presence of Tannin, Saponin, Alkaloids, Flavonoids, and phenol in reasonable amounts, revealing that the sample would possess certain important medicinal characteristics. The amino acid content of the samples showed valuable essential and non-essential amino acids. It is evident from the study that the leaf protein concentrates from Mentha piperita plants' source of nutrients, phytochemicals, and essential and non-essential amino acids, indicating that the plant could still be classified as nutraceuticals apart from cosmeceuticals due to the concentration of phytochemicals and nutritive parameters of the sample.

Food consumption play a crucial role in human life, yet conventional food production and consumption patterns can be detrimental to the environment. Thus, research and development has been directed towards alternative proteins, with edible insects being promising sources. Edible insects have been recognised for their sustainable benefits providing protein, with less emission of greenhouse gas, land and water usage compared to sources, such as beef, chicken, and dairy products. Among the over 2000 known edible insect species, only four, namely yellow mealworm (Tenebrio molitor), migratory locust/grasshopper (Locusta migratoria), grain mould beetle, also known as lesser mealworm which is a larval form of Alphitobius diaperinus (from the family of Tenebrionidae of darkling beetles) and house cricket (Acheta domesticus), are currently authorised in specific products through specific producers in the EU. The expansion of such foods into Western diets face challenges such as consumer barriers, gaps in microbiological and chemical safety hazard data during production and processing, and the potential for fraudulent supply chain activity. The main aim of this study was to map the supply chain, through interviews with personnel along the supply chain, coupled with searches for relevant publications and governmental documents. Thus, the main potential points of food safety and fraud along the edible insect supply chain were identified. Feed substrate was identified as the main area of concern regarding microbiological and chemical food safety and novel processing techniques were forecast to be of most concern for future fraudulent activity. Despite the on-going authorisation of insect species in many countries there are substantial food safety and authenticity information gaps in this industry that need to be addressed before edible insects can be viewed as a safe and sustainable protein sources by Western consumers.

Apparent digestibility of raw materials by yellowtail kingfish (Seriola lalandi)

How cells utilize nutrients to produce the ATP needed for bioenergetic homeostasis has been well-characterized. What is less well-studied is how resting cells metabolically shift from an ATP-producing catabolic metabolism to a metabolism that supports anabolic growth. In metazoan organisms, the discovery of growth factors and the ability of their receptors to induce new transcription and translation led to the hypothesis that the bioenergetic and synthetic demands of cell growth were primarily met through the replacement of nutrients consumed during net macromolecular synthesis, a demand-based system of nutrient uptake. Recent data have challenged this hypothesis. Instead, there is increasing evidence that cellular nutrient uptake is a push system. Growth factor signaling has been linked to direct stimulation of nutrient uptake. The ability of growth factor signaling to increase the uptake of glucose, lipids, and amino acids to levels that exceed a cell's bioenergetic and synthetic needs has been documented in a wide variety of settings. In some tissues, this leads to the storage of the excess nutrients in the form of glycogen or fat. In others, the excess is secreted as lactate and certain nonessential amino acids. When growth factor signaling stimulates nutrient uptake to levels that exceed a cell's bioenergetic needs, adaptive changes in intermediate metabolism lead to the production of anabolic precursors that fuel the net synthesis of protein, lipids, and nucleic acids. Through the increased production of these precursors, growth factor signaling provides a supply-side stimulation of cell growth and proliferation.

Edible insects as an alternative protein source have gained consumers' attention, leading to new market possibilities. Several investigations have generated and characterized ingredients from insects and assessed their potential application in food, feed, and their biological potential. Insects are a rich source of protein, ranging from 30% to 65%. Insect derived ingredients show great potential to be added to food products. Protein isolates or concentrates, protein hydrolysates and peptides are obtained from edible insects using different methods. Insect protein techno-functional properties include water and oil-holding capacity, emulsifying, solubility and gelling properties. Depending on protein techno-functional properties, food applications can be designed to improve their development in the formulated food. Nowadays, several commercially available food and feed products are formulated including insect ingredients. However, research related to insect-derived peptides biological potential is limited. In-depth biological assays are needed to understand the potential health benefits of insects bioactive components. Potential future research could focus on the technological properties of insect proteins, oriented to the physicochemical interactions with food matrices, including sensory quality, texture and rheological properties of new food products.

Chinese (CH) lose approximately 2/3 as much nitrogen (N) as Caucasians (CA) on a high fat, high carbohydrate, zero protein intake. The justification theory of hereditary non-essential amino acid (NEA) deficiency states that deficiencies in NEA can occur on marginal protein intake by heterozygotes for any of the 32 enzymes required to synthesize all the NEA. N loss during starvation depends on the ability to resynthesize protein which is turning over. If an essential amino acid is lost then all of the NEA lost by that molecule of protein cannot be reutilized. Efficiency of reutilization depends upon the available constellation of NEA. The low protein intake of the CH, who have subsisted for 50 centuries on cereals, would make it difficult for a heterozygote for NEA synthesis to survive. The CA, a hunter and fisherman, would not lack for protein. Though total caloric nutrition might be poor there would be no selection against mutations in the NEA pathways. This interpretation is supported by data which show that the essential amino acids in the plasma of CH and CA after a 2-week fast are equal whereas the plasma of CH contains 50% more of the NEA. This is associated with a mean loss of 1.8 mgs. of nitrogen per basal calorie in the CA and 1.2 mgs. per basal calorie in the CH. The justification hypothesis may not only supply an explanation of differential response to the same diet, but might even provide dietary suggestions for prevention of many problems of protein synthesis.

The consumption of edible insects can be anadvantageous alternative to the conventional food supply chain, which involves global water waste, land deficit, undernutrition, and starvation. Besides the nutritional aspects, insect proteins have demonstrated a wide range of functional properties such as foamability, emulsifying and gelling abilities. The protein content and amino acid profile of some insects have revealed a good nutritional value and interesting functional properties. However, it is crucial to comprehend how the protein quality is affected by insect feeding, drying, and defatting. There is a knowledge gap about the impact of industrial treatment, such as pH, ionic strength, and heat treatment, on insect proteins' functional properties. In this review, we have aimed to highlight the potential application of insect proteins as a nutritional source and their promising technological applications. The study reported the principal insect protein characterization methodologies that have been investigated in the literature aiming to correlate the physicochemical parameters to possible protein functionalities. The research on the functional properties of insect proteins is at the exploratory level. Further detailed studies are needed to clarify the structure-function relation of insect proteins and how these functionalities and insect processing can increase consumer acceptance.

Use of peanut meal as an alternative protein source in diets for Channel Catfish Ictalurus punctatus was evaluated in a 9‐week study under controlled laboratory conditions. Five practical diets (28% crude protein and 6% crude lipid) were formulated to contain 0, 10, 15, 20, and 25% peanut meal as a replacement for cottonseed meal and soybean meal in the control diet. The control diet was a plant‐based diet containing principally soybean meal (25%), cottonseed meal (20%), corn (20%), and corn germ meal (20%). All diets met or exceeded all known nutrient requirements of Channel Catfish. Twenty Channel Catfish fingerlings (mean initial weight: 12.8 g/fish) were stocked into 25 water flow‐through glass aquaria (110 L). The fish were fed once daily to apparent satiation. Among dietary treatments, there were no significant differences in feed consumption, weight gain, feed conversion ratio, and survival, and no differences in muscle protein, fat, or moisture levels. The results demonstrate up to 25% peanut meal can be used as a protein source alternative to cottonseed meal or soybean meal in Channel Catfish diets without adversely affecting growth, feed efficiency, and body composition.

Are alternative proteins increasing food allergies? Trends, drivers and future perspectives

Simple SummaryThis study aimed to evaluate diets with complete substitution of soybean meal (SBM) by an algae meal from Spirulina platensis (SM) or partly defatted larvae meal from Hermetia illucens (HM) as feed for piglets and growing pigs. Main feed ingredients of the experimental diets were wheat, barley, and SM or HM. The final diets contained 21% (piglets) and 13% (growing pigs) of SM or HM and nitrogen (N) balance studies were applied to measure parameters of apparent N digestibility, complex dietary protein quality, and individual amino acid (AA) efficiency according to the ‘Goettingen approach’. Diets were well accepted by the animals and an extended level of AA supplementation yielded improved dietary protein quality with both of the alternative feed proteins. However, HM based diets provided superior apparent N digestibility.Two age-dependent nitrogen (N) balance studies (average body mass 25 and 60 kg) utilized 16 male castrated piglets and 16 barrows to measure N utilization parameters of diets with complete substitution of SBM by alternative protein sources (SM, HM), but different AA fortifications. Lysine supplementation up to 80% of the recommended lysine (Lys) supply in diets HM (A) and SM (A) yielded similar protein quality data (63.6 ± 2.1 and 63.7 ± 3.4). Surprisingly, only in piglet diet HM (AA) did the extended AA supplementation (Lys, methionine (Met), threonine (Thr)) enhance protein quality (72.8 ± 6.7) significantly (p = 0.004). Similar trends were observed in growing pigs. However, when the level of histidine (His) in diet SM (AA) was increased, feed protein quality (71.8 ± 1.3) was significantly (p < 0.001) improved indicating the importance of adequate His supply in diets with a complete substitution of SBM by the algae meal (SM) under study. AA efficiency data extend the possibilities to explain the observed responses on protein quality. When an adequate AA balancing in the diet is guaranteed, from nutritional point of view both of the alternative proteins may replace SBM in pig diets.

The results of study of effect of freezing temperature on the change of amino acid content and the ratio of essential and nonessential amino acids in veal proteins are given. The meat of calves at the age of 3 months, bred in the Leningrad region of the RF, was the object of research. The amino acid composition of veal proteins was analyzed using the Shimadzu 20AD Chromatograph with precolumn phenylisothiocyanate derivatization and spectrophotometric detection at 254 nm by reversedphase gradient highperformance liquid chromatography (HPLC). The Kjeldahl method was used to define the protein content on the basis of the sample mineralization. It was shown that the change of the amount and ratio of essential and nonessential amino acids of veal proteins depended on the freezing temperature as well as on structural features of Rside radicals. Also it was established that the total amount of essential and nonessential amino acids in veal proteins during freezing at -24 and -35 °С temperatures decreased by 13,5 and 8,2%, respectively; the total of essential amino acids by 11 an 7%, nonessential ones by 22 and 11%, respectively. It was proved that significant changes were characteristic of amino acids with polar anionic and cationic radicals. So, the amount of amino acids with polar negatively charged radicals during veal freezing at temperatures -24 and -35 ° С decreased by 22 and 16,5%; with polar positively charged radicals - by 17 and 13,5%, respectively. At the same time, the freezing temperature affects slightly the change in the content of amino acids with nonpolar (hydrophobic) Rgroups and polar nonionogenic radicals. It was shown that the total amount of free amino acids increased independently of the freezing temperature. However, more essential amino acids were accumulated in veal frozen at a temperature of -24 ° С (2,33 times) than at -35 ° С (1,80 times). The amount of free nonessential amino acids changed to a lesser extent

Simple SummaryFish meal (FM) is the major protein source in aquafeed to achieve sustainable aquaculture production. However, the supply of FM is low due to high cost and low availability. There is ongoing conceited research to identify alternative viable protein sources to replace the finite FM. Novel camelina meal (CM) is a plant protein with high amino acids levels and has been tested as an alternative protein source in livestock feeding. However, there is limited information on supplementation of the CM in aquaculture diets. In this study, four diets were formulated to contain 0% plant protein, 205 g/kg soybean meal, and two levels of CM (205 g/kg and 330 g/kg) in diets fed to red seabream. Results indicated that adding CM 205 g/kg in diets-maintained growth performance, digestive enzyme activities, and nutrient digestibility regulated the immunity and stress resistance and modulated the growth-related genes in red seabream. These findings provide the first step in using novel CM and are essential for future practical formulations of feed for red seabream and other marine fish species.A 60-day experiment was designed to assess the effect of different ratios of fish meal (FM): camelina meal plant protein (CM) on growth response and relative gene expression of growth-promoting factors, feed utilization potency, digestive enzymes activities, apparent digestibility (ADC), stress response, non-specific immunity of Pagrus major. Four isonitrogenous (490.7 g/kg of crude protein) and isolipidic (91.5 g/kg total lipid) experimental diets were formulated and designated as camelina meal (CM0), soyabean meal (SBM20.5), CM20.5, and CM33 based on protein contents. At the end of the feed trial, significantly higher (p < 0.05) weight gain, specific growth rate, and feed intake but lower feed conversion ratio were recorded in fish fed CM0, SBM20.5, and CM20.5 than fish fed CM33. The lowest growth, feed utilization, enzyme activity, and digestibility were recorded in fish fed CM33. Significantly higher pepsin, amylase, and protease activities were observed in fish fed CM0, SBM20.5, and CM20.5 diets than fish fed CM33. The highest ADC of protein was recorded in fish fed CM0, SBM20.5, and CM20.5 diets. Hematocrit levels were depressed CM33 while total serum protein, total cholesterol, triglyceride, blood urea nitrogen, total bilirubin, aspartate aminotransferase, and alanine aminotransferase were not significantly changed by the inclusion of CM. Non-specific immune variables (lysozyme activity, peroxidase activity in serum and nitro blue tetrazolium) in fish fed CM0, SBM20.5, and CM20.5 were significantly higher than in fish fed CM33 diet. The superoxide dismutase of fish fed CM20.5 was not significantly different from CM0 and SBM20.5 (p > 0.05). Catalase and low salinity stress test show that CM0, SBM20.5, and CM20.5 were not significantly (p > 0.05) different, while CM33 was significantly lower than the rest of the diets. TBARs show that CM20.5 and CM33 diets were significantly different (p < 0.05), but CM20.5 was not significantly different from SBM20.5. Significantly higher hepatic IGF-1 and IGF-2 mRNA expression was found in fish-fed diet groups CM0, SBM20.5, and CM20.5 than fish fed CM33. The present study indicated that the addition of CM up 205 kg/kg to diet maintains growth, digestive enzymes, nutrient digestibility, immunity, stress resistance, and feed utilization efficiency of red sea bream.

Locusts have high nutritional value and are considered alternative protein sources for humans and animals. Diet can strongly affect the nutritional composition of insects. In this study, the influences of diets – wheat (Triticum aestivum) seedlings and rape (Brassica napus) leaves – on the chemical composition and amino acids of the migratory locust (Locusta migratoria) were assessed. The crude protein, soluble carbohydrate and triglyceride were analyzed using the Bradford method, anthrone colorimetry, and glycerol kinase chromogenic method, respectively. Additionally, amino acids were determined using cation exchange chromatography. The study revealed that wheat seedlings had lower water content than rape leaves. Although the crude protein content was comparable, the soluble carbohydrates of wheat seedlings were approximately 2.5 times higher than those of rape leaves. The body weight (fresh weight and dry weight), crude protein and triglyceride of locusts significantly increased when fed wheat seedlings compared with when fed rape leaves, with values of 48.08%, 31.58%, 43.83% and 25.00%, respectively. The high water content of rape leaves resulted in the high water content of rape leaf-fed locusts. Plants also affect the amino acid composition of locusts. The eight amino acids (four essential amino acids and four non-essential amino acids) and the total amino acids of rape leaf-fed locusts were significantly enhanced compared with those of wheat seedlings. The essential amino acid index and biological value indicate that the protein quality of locusts fed rape leaves has been significantly improved. These results demonstrate that the nutritional composition of L. migratoria can be altered by manipulating the diet. Insects with controllable nutrients can be provided by insect farms and further studies will improve the development of insects as food or feed.