Enteric methane emission reduction potential of natural feed supplements in ewe diets.

Research into the potential use of various dietary feed supplements to reduce methane (CH4) production from ruminants has proliferated in recent years. In this study, two 8-wk long experiments were conducted with mature ewes and incorporated the use of a variety of natural dietary feed supplements offered either independently or in combination. Both experiments followed a randomized complete block design. Ewes were offered a basal diet in the form of ad libitum access to grass silage supplemented with 0.5 kg concentrates/ewe/d. The entire daily dietary concentrate allocation, incorporating the respective feed supplement, was offered each morning, and this was followed by the daily silage allocation. In experiment 1, the experimental diets contained 1) no supplementation (CON), 2) Ascophyllum nodosum (SW), 3) A. nodosum extract (EX1), 4) a blend of garlic and citrus extracts (GAR), and 5) a blend of essential oils (EO). In experiment 2, the experimental diets contained 1) no supplementation (CON), 2) A. nodosum extract (EX2), 3) soya oil (SO), and 4) a combination of EX2 and SO (EXSO). Twenty ewes per treatment were individually housed during both experiments. Methane was measured using portable accumulation chambers. Rumen fluid was collected at the end of both experiments for subsequent volatile fatty acid (VFA) and ammonia analyses. Data were analyzed using mixed models ANOVA (PROC MIXED, SAS v9.4). Statistically significant differences between treatment means were considered when P < 0.05. Dry matter intake was not affected by diet in either experiment (P > 0.05). Ewes offered EO tended to have an increased feed:gain ratio relative to CON (P < 0.10) and SO tended to increase the average daily gain (P < 0.10) which resulted in animals having a higher final body weight (P < 0.05) than CON. Ewes offered EX1 and SO emitted 9% less CH4 g/d than CON. The only dietary treatment to have an effect on rumen fermentation variables relative to CON was SW, which enhanced total VFA production (P < 0.05). In conclusion, the A. nodosum extract had inconsistent results on CH4 emissions whereby EX1 reduced CH4 g/d while EX2 had no mitigating effect on CH4 production, likely due to the differences in PT content reported for EX1 and EX2. SO was the only dietary feed supplement assessed in the current study that enhanced animal performance whilst mitigating daily CH4 production.

Reducing methane emissions from agriculture is vital to minimize the effects of global warming and to meet greenhouse gas reduction targets set by EU policy. In this experiment, a range of natural feed supplements were offered to mature ewes through the concentrated portion of their diet. Soya oil and brown seaweed extract reduced daily methane emissions by 9% when offered independently of each other; however, no reduction in methane was observed when combined. Additionally, inclusion of soya oil improved animal weight gain. Results from the current experiment may contribute to the development of a targeted dietary strategy to reduce methane emissions from livestock.

Applications of Microalgae in Foods, Pharma and Feeds and Their Use as Fertilizers and Biostimulants: Legislation and Regulatory Aspects for Consideration.

Microalgae are a rich resource of lipids, proteins, carbohydrates and pigments with nutritional and health benefits. They increasingly find use as ingredients in functional foods and feeds as well as in cosmetics and agricultural products including biostimulants. One of their distinct advantages is their ability to grow on wastewaters and other waste streams, and they are considered an environmentally friendly and cheap method to recover nutrients and remove pollutants from the environment. However, there are limits concerning their applications if grown on certain waste streams. Within, we collate an overview of existing algal applications and current market scenarios for microalgal products as foods and feeds along with relevant legislative requirements concerning their use in Europe and the United States. Microalgal compounds of interest and their extraction and processing methodologies are summarized, and the benefits and caveats of microalgae cultivated in various waste streams and their applications are discussed.

Potential of the Red Macroalga Bonnemaisonia hamifera in Reducing Methane Emissions from Ruminants.

Researchers have been exploring seaweeds to reduce methane (CH4) emissions from livestock. This study aimed to investigate the potential of a red macroalga, B. hamifera, as an alternative to mitigate CH4 emissions. B. hamifera, harvested from the west coast of Sweden, was used in an in vitro experiment using a fully automated gas production system. The experiment was a randomized complete block design consisting of a 48 h incubation that included a control (grass silage) and B. hamifera inclusions at 2.5%, 5.0%, and 7.5% of grass silage OM mixed with buffered rumen fluid. Predicted in vivo CH4 production and total gas production were estimated by applying a set of models to the gas production data and in vitro fermentation characteristics were evaluated. The results demonstrated that the inclusion of B. hamifera reduced (p = 0.01) predicted in vivo CH4 and total gas productions, and total gas production linearly decreased (p = 0.03) with inclusion of B. hamifera. The molar proportion of propionate increased (p = 0.03) while isovalerate decreased (p = 0.04) with inclusion of B. hamifera. Chemical analyses revealed that B. hamifera had moderate concentrations of polyphenols. The iodine content was low, and there was no detectable bromoform, suggesting quality advantages over Asparagopsis taxiformis. Additionally, B. hamifera exhibited antioxidant activity similar to Resveratrol. The findings of this study indicated that B. hamifera harvested from temperate waters of Sweden possesses capacity to mitigate CH4 in vitro.

Generation of Bioactive Peptides from Porphyridium sp. and Assessment of Their Potential for Use in the Prevention of Hypertension, Inflammation and Pain.

Inflammation, hypertension, and negative heart health outcomes including cardiovascular disease are closely linked but the mechanisms by which inflammation can cause high blood pressure are not yet fully elucidated. Cyclooxygenase (COX) enzymes play a role in pain, inflammation, and hypertension development, and inhibition of these enzymes is currently of great interest to researchers and pharmaceutical companies. Non-steroidal anti-inflammatory drugs are the drug of choice in terms of COX inhibition but can have negative side effects for consumers. Functional food ingredients containing cyclooxygenase inhibitors offer a strategy to inhibit cyclooxygenases without negative side effects. Several COX inhibitors have been discovered, to date, from marine and other resources. We describe here, for the first time, the generation and characterization of a bioactive hydrolysate generated using Viscozyme® and Alcalase from the red microalga Porphyridium sp. The hydrolysate demonstrates in vitro COX-1 inhibitory activity and antihypertensive activity in vivo, assessed using spontaneously hypertensive rats (SHRs). Peptides were identified and sequenced using MS and assessed using an in silico computational approach for potential bioactivities. The peptides predicted to be bioactive, including GVDYVRFF, AIPAAPAAPAGPKLY, and LIHADPPGVGL were chemically synthesized and cyclooxygenase inhibition was confirmed. Peptides AIPAAPAAPAGPKLY and LIHADPPGVGL had COX-1 IC50 values of 0.2349 mg/mL (0.16 µM) and 0.2193 mg/mL (0.2 µM), respectively. The hydrolysate was included in a food carrier (jelly candies) and an antihypertensive effect was observed in SHRs.

Maximizing Use of Pelagic Capture Fisheries for Global Protein Supply: Potential and Caveats Associated with Fish and Co-Product Conversion into Value-Add Ingredients.

Globally, capture fisheries contribute significantly to protein supply and the food security of a third of the world's population. Although capture fisheries production has not significantly increased in tonnes landed per annum during the last two decades (since 1990), it still produced a greater tonnage of protein than aquaculture in 2018. Policy in the European Union and other locations favors production of fish through aquaculture to preserve existing fish stocks and prevent extinction of species from overfishing. However, aquaculture production of fish in order to feed the growing global population would need to increase from 82 087 kT in 2018 to 129 000 kT by 2050. The Food and Agriculture Organization states that global production of aquatic animals was 178 million tonnes in 2020. Capture fisheries contributed 90 million tonnes (51%) of this. For capture fisheries to be a sustainable practice in alignment with UN sustainability goals, ocean conservation measures must be followed and processing of capture fisheries may need to adapt food-processing strategies already used extensively in the processing of dairy, meat, and soy. These are required to add value to reduced fish landings and sustain profitability.

Potential of Seaweeds to Mitigate Production of Greenhouse Gases during Production of Ruminant Proteins.

The potential of seaweed to mitigate methane is real and studies with red seaweeds have found reductions in methane produced from ruminants fed red seaweeds in the region of 60-90% where the active compound responsible for this is bromoform. Other studies with brown and green seaweeds have observed reductions in methane production of between 20 and 45% in vitro and 10% in vivo. Benefits of feeding seaweeds to ruminants are seaweed specific and animal species-dependent. In some instances, positive effects on milk production and performance are observed where selected seaweeds are fed to ruminants while other studies note reductions in performance traits. A balance between reducing methane and maintaining animal health and food quality is necessary. Seaweeds are a source of essential amino acids and minerals however, and offer huge potential for use as feeds for animal health maintenance once formulations and doses are correctly prepared and administered. A negative aspect of seaweed use for animal feed currently is the cost associated with wild harvest and indeed aquaculture production and improvements must be made here if seaweed ingredients are to be used as a solution to control methane production from ruminants for continued production of animal/ruminant sourced proteins in the future. This review collates information concerning different seaweeds and how they and their constituents can reduce methane from ruminants and ensure sustainable production of ruminant proteins in an environmentally beneficial manner.

Identification of Bioactive Peptides from a Laminaria digitata Protein Hydrolysate Using In Silico and In Vitro Methods to Identify Angiotensin-1-Converting Enzyme (ACE-1) Inhibitory Peptides.

Bioactive peptides range in size from 2-30 amino acids and may be derived from any protein-containing biomass using hydrolysis, fermentation or high-pressure processing. Pro-peptides or cryptides result in shorter peptide sequences following digestion and may have enhanced bioactivity. Previously, we identified a protein hydrolysate generated from Laminaria digitata that inhibited ACE-1 in vitro and had an ACE-1 IC50 value of 590 µg/mL compared to an ACE-1 IC50 value of 500 µg/mL (~2.3 µM) observed for the anti-hypertensive drug Captopril©. A number of peptide sequences (130 in total) were identified using mass spectrometry from a 3 kDa permeate of this hydrolysate. Predicted bioactivities for these peptides were determined using an in silico strategy previously published by this group utilizing available databases including Expasy peptide cutter, BIOPEP and Peptide Ranker. Peptide sequences YIGNNPAKGGLF and IGNNPAKGGLF had Peptide Ranker scores of 0.81 and 0.80, respectively, and were chemically synthesized. Synthesized peptides were evaluated for ACE-1 inhibitory activity in vitro and were found to inhibit ACE-1 by 80 ± 8% and 91 ± 16%, respectively. The observed ACE-1 IC50 values for IGNNPAKGGLF and YIGNNPAKGGLF were determined as 174.4 µg/mL and 133.1 µg/mL. Both peptides produced sequences following simulated digestion with the potential to inhibit Dipeptidyl peptidase IV (DPP-IV).

Identification of Bioactive Peptides from Nannochloropsis oculata Using a Combination of Enzymatic Treatment, in Silico Analysis and Chemical Synthesis.

In vitro ACE-1 inhibitory peptides were characterised previously from a number of microalgal species including Spirulina platensis (peptide IAPG), Chlorella vulgaris (peptides FDL, AFL, VVPPA), Isochrysis galbana (peptide YMGLDLK), Chlorella sorokiniana (peptides IW and LW) and indeed Nannochloropsis oculata (peptides GMNNLTP and LEQ). The isolation of protein from Nannochloropsis oculata using a combination of ammonium salt precipitation and xylanase treatment of resulting biomass combined with molecular weight cut off filtration to produce a permeate and characterisation of bioactive peptides is described. The Angiotensin-1-converting enzyme (ACE-1) IC50 value for the generated permeate fraction was 370 µg/mL. Ninety-five peptide sequences within the permeate fraction were determined using mass spectrometry and eight peptides were selected for chemical synthesis based on in silico analysis. Synthesized peptides were novel based on a search of the literature and relevant databases. In silico, simulated gastrointestinal digestion identified further peptides with bioactivities including ACE-1 inhibitory peptides and peptides with antithrombotic and calcium/calmodulin-dependent kinase II (CAMKII) inhibition. This work highlights the potential of Nannochloropsis oculata biomass as both a protein and bioactive peptide resource, which could be harnessed for use in the development of functional foods and feeds.

Angiotensin-I-Converting Enzyme Inhibitory Activity of Protein Hydrolysates Generated from the Macroalga Laminaria digitata (Hudson) JV Lamouroux 1813.

Seaweeds have a long history of use as both food and medicine, especially in Asian cultures. Moreover, there is growing interest in the use of seaweed ingredients and bioactive compounds in pharmaceutical and nutraceutical products. One ailment that seaweed bioactive compounds may impact is hypertension caused by the enzyme Angiotensin Converting Enzyme 1 (ACE-1; EC 3.4.15.1), found within the Renin-Angiotensin Aldosterone System (RAAS), which causes vasoconstriction of blood vessels, including veins and arteries. The aim of this paper is to generate bioactive peptide containing protein hydrolysates from the brown seaweed Laminaria digitata (Hudson) JV Lamouroux 1813. Proteins were extracted from this seaweed by disrupting the seaweed cell wall using a combination of carbohydrases and proteolytic enzymes. Bioactive peptide containing permeates were generated from L. digitata protein hydrolysates, and both hydrolysates and permeates were screened for their ability to inhibit the enzyme ACE-1. The protein content of the permeate fractions was found to be 23.87% compared to the untreated seaweed, which contained 15.08% protein using LECO analysis. Hydrolysis and filtration resulted in a "white" protein powder, and the protein content of this powder increased by 9% compared to the whole seaweed. The total amino acid (TAA) content of the L. digitata protein permeate was 53.65 g/100 g of the sample, and contains over 32% essential amino acids (EAA). Furthermore, the L. digitata permeate was found to inhibit the ACE-1 enzyme by 75% when compared to the commercial drug Captopril© when assayed at a concentration of 1 mg/mL. The inhibition of ACE-1 (the IC50 value) of 590 µg/mL for the L. digitata permeate compares well with Captopril©, which had 100% inhibition of ACE-1, with an IC50 value of 500 µg/mL. This study indicates that there is potential to develop protein powders with ACE-1 inhibitory bioactivities from the brown seaweed L. digitata using enzymatic hydrolysis as a cell disruption and protein extraction/hydrolysate generation procedure.

The Prebiotic Effect of Australian Seaweeds on Commensal Bacteria and Short Chain Fatty Acid Production in a Simulated Gut Model.

Diet is known to affect the composition and metabolite production of the human gut microbial community, which in turn is linked with the health and immune status of the host. Whole seaweeds (WH) and their extracts contain prebiotic components such as polysaccharides (PS) and polyphenols (PP). In this study, the Australian seaweeds, Phyllospora comosa, Ecklonia radiata, Ulva ohnoi, and their PS and PP extracts were assessed for potential prebiotic activities using an in vitro gut model that included fresh human faecal inoculum. 16S rRNA sequencing post gut simulation treatment revealed that the abundance of several taxa of commensal bacteria within the phylum Firmicutes linked with short chain fatty acid (SCFA) production, and gut and immune function, including the lactic acid producing order Lactobacillales and the chief butyrate-producing genera Faecalibacteria, Roseburia, Blautia, and Butyricicoccus were significantly enhanced by the inclusion of WH, PS and PP extracts. After 24 h fermentation, the abundance of total Firmicutes ranged from 57.35−81.55% in the WH, PS and PP samples, which was significantly greater (p ≤ 0.01) than the inulin (INU) polysaccharide control (32.50%) and the epigallocatechingallate (EGCG) polyphenol control (67.13%); with the exception of P. comosa PP (57.35%), which was significantly greater than INU only. However, all WH, PS and PP samples also increased the abundance of the phylum Proteobacteria; while the abundance of the phylum Actinobacteria was decreased by WH and PS samples. After 24 h incubation, the total and individual SCFAs present, including butyric, acetic and propionic acids produced by bacteria fermented with E. radiata and U. ohnoi, were significantly greater than the SCFAs identified in the INU and EGCG controls. Most notably, total SCFAs in the E. radiata PS and U. ohnoi WH samples were 227.53 and 208.68 µmol/mL, respectively, compared to only 71.05 µmol/mL in INU and 7.76 µmol/mL in the EGCG samples. This study demonstrates that whole seaweeds and their extracts have potential as functional food ingredients to support normal gut and immune function.

In Vitro Protein Digestibility of Selected Seaweeds.

Seaweed biomass is considered a valuable and potential, alternative protein source but it is currently under-exploited. Seaweed or Macroalgae do not require arable land and freshwater for their cultivation, they are fast growing and contain several health ingredients and beneficial macronutrients. In this study, we determined the in vitro k-Protein Digestibility-Corrected Amino Acid Score (k-PDCAAS) values of six different, Irish seaweeds using the rapid k-PDCAAS method. Based on the amino acid profile and protein content of each seaweed, the in vitro protein digestibility and k-PDCAAS scores were calculated. In addition, the limiting amino acid(s) for each of the six seaweeds was/were determined. Results suggest that although the in vitro digestibility was quite similar for all analyzed seaweeds, their k-PDCAAS scores varied significantly. The red seaweed Palmaria palmata had a k-PDCAAS score of 0.69 ± 0.014, while Fucus serratus had a value of 0.63 ± 0.084 and Alaria esculenta a value of 0.59 ± 0.021. The seaweeds were found to be rich in essential amino acids and taurine. Overall, the amino acid composition of the seaweeds studied suggests that they are suitable alternative protein sources for use in human nutrition providing both essential and non-essential amino acids to the consumer.

Macroalgal Proteins: A Review.

Population growth is the driving change in the search for new, alternative sources of protein. Macroalgae (otherwise known as seaweeds) do not compete with other food sources for space and resources as they can be sustainably cultivated without the need for arable land. Macroalgae are significantly rich in protein and amino acid content compared to other plant-derived proteins. Herein, physical and chemical protein extraction methods as well as novel techniques including enzyme hydrolysis, microwave-assisted extraction and ultrasound sonication are discussed as strategies for protein extraction with this resource. The generation of high-value, economically important ingredients such as bioactive peptides is explored as well as the application of macroalgal proteins in human foods and animal feed. These bioactive peptides that have been shown to inhibit enzymes such as renin, angiotensin-I-converting enzyme (ACE-1), cyclooxygenases (COX), α-amylase and α-glucosidase associated with hypertensive, diabetic, and inflammation-related activities are explored. This paper discusses the significant uses of seaweeds, which range from utilising their anthelmintic and anti-methane properties in feed additives, to food techno-functional ingredients in the formulation of human foods such as ice creams, to utilising their health beneficial ingredients to reduce high blood pressure and prevent inflammation. This information was collated following a review of 206 publications on the use of seaweeds as foods and feeds and processing methods to extract seaweed proteins.

Nutritional Profiling and Preliminary Bioactivity Screening of Five Micro-Algae Strains Cultivated in Northwest Europe.

This study aimed to map the nutritional profile and bioactivities of five microalgae that can be grown in Northwest Europe or areas with similar cultivation conditions. Next to the biochemical composition, the in vitro digestibility of carbohydrates, proteins, and lipids was studied for Chlamydomonas nivalis, Porphyridium purpureum, Chlorella vulgaris, Nannochloropsis gaditana, and Scenedesmus species biomass. These microalgae were also assessed for their ability to inhibit the angiotensin-1-converting enzyme (ACE-1, EC 3.4.15.1), which is known to play a role in the control of blood pressure in mammals. Large differences in organic matter solubility after digestion suggested that a cell disruption step is needed to unlock the majority of the nutrients from N. gaditana and Scenedesmus species biomass. Significant amounts of free glucose (16.4-25.5 g glucose/100 g dry algae) were detected after the digestion of C. nivalis, P. purpureum, and disrupted Scenedesmus. The fatty acid profiles showed major variations, with particularly high Ω-3 fatty acid levels found in N. gaditana (5.5 ± 0.5 g/100 g dry algae), while lipid digestibility ranged from 33.3 ± 6.5% (disrupted N. gaditana) to 67.1 ± 11.2% (P. purpureum). C. vulgaris and disrupted N. gaditana had the highest protein content (45-46% of dry matter), a nitrogen solubility after digestion of 65-71%, and the degree of protein hydrolysis was determined as 31% and 26%, respectively. Microalgae inhibited ACE-1 by 73.4-87.1% at physiologically relevant concentrations compared to a commercial control. These data can assist algae growers and processors in selecting the most suitable algae species for food or feed applications.

Seaweed Components as Potential Modulators of the Gut Microbiota.

Macroalgae, or seaweeds, are a rich source of components which may exert beneficial effects on the mammalian gut microbiota through the enhancement of bacterial diversity and abundance. An imbalance of gut bacteria has been linked to the development of disorders such as inflammatory bowel disease, immunodeficiency, hypertension, type-2-diabetes, obesity, and cancer. This review outlines current knowledge from in vitro and in vivo studies concerning the potential therapeutic application of seaweed-derived polysaccharides, polyphenols and peptides to modulate the gut microbiota through diet. Polysaccharides such as fucoidan, laminarin, alginate, ulvan and porphyran are unique to seaweeds. Several studies have shown their potential to act as prebiotics and to positively modulate the gut microbiota. Prebiotics enhance bacterial populations and often their production of short chain fatty acids, which are the energy source for gastrointestinal epithelial cells, provide protection against pathogens, influence immunomodulation, and induce apoptosis of colon cancer cells. The oral bioaccessibility and bioavailability of seaweed components is also discussed, including the advantages and limitations of static and dynamic in vitro gastrointestinal models versus ex vivo and in vivo methods. Seaweed bioactives show potential for use in prevention and, in some instances, treatment of human disease. However, it is also necessary to confirm these potential, therapeutic effects in large-scale clinical trials. Where possible, we have cited information concerning these trials.

A National Survey Identifying the Factors Associated With Cardiovascular Care Nurses' Perceived Knowledge of International Practice Guidelines: The First Step in the Development of an Implementation Strategy.

BACKGROUND: The implementation of international guidelines within everyday practice remains problematic, which can have a detrimental impact on quality of care delivered. This study aimed to ascertain the factors associated with clinical nurses' perceived knowledge of international guidelines. METHODS: In this cross-sectional survey, nurses from 45 hospitals across Ireland were recruited. A previously validated anonymous questionnaire that assessed guideline knowledge, use, and barriers to implementation was used. Data were analyzed using SPSS 23 and logistic regression. RESULTS: Of the 542 responses, 54% had used international guidelines relevant to their practice and 50% had consulted within the last year. Most nurses perceived that poor patient follow-up, lack of time and resources, poor clinical leadership, workload, long guidelines, and not understanding guideline detail were barriers to guideline use and implementation. Forty-five percent rated their perceived knowledge of guidelines as "low." Logistic regression identified that "high" knowledge levels were significantly associated with having read guidelines in the last year and their use with practice. In contrast, low knowledge of the guidelines was associated with perceptions that they were lengthy and not easy to use, lack of confidence to challenge colleagues when guidelines are not implemented, or not being able to influence current practice. CONCLUSIONS: This study identified the specific knowledge needs in this cohort of mainly basic grade registered nurses, with low perceived guideline knowledge. A whole unit or team approach led by nurse champions is needed to develop and establish practice and educational strategies that would increase the availability, application, and knowledge of guidelines within everyday practice.

Aquaculture Production of the Brown Seaweeds Laminaria digitata and Macrocystis pyrifera: Applications in Food and Pharmaceuticals.

Seaweeds have a long history of use as food, as flavouring agents, and find use in traditional folk medicine. Seaweed products range from food, feed, and dietary supplements to pharmaceuticals, and from bioenergy intermediates to materials. At present, 98% of the seaweed required by the seaweed industry is provided by five genera and only ten species. The two brown kelp seaweeds Laminaria digitata, a native Irish species, and Macrocystis pyrifera, a native New Zealand species, are not included in these eleven species, although they have been used as dietary supplements and as animal and fish feed. The properties associated with the polysaccharides and proteins from these two species have resulted in increased interest in them, enabling their use as functional foods. Improvements and optimisations in aquaculture methods and bioproduct extractions are essential to realise the commercial potential of these seaweeds. Recent advances in optimising these processes are outlined in this review, as well as potential future applications of L. digitata and, to a greater extent, M. pyrifera which, to date, has been predominately only wild-harvested. These include bio-refinery processing to produce ingredients for nutricosmetics, functional foods, cosmeceuticals, and bioplastics. Areas that currently limit the commercial potential of these two species are highlighted.

Bioactive Peptides in Preventative Healthcare: An Overview of Bioactivities and Suggested Methods to Assess Potential Applications.

Food derived bioactive peptides can be generated from various protein sources and usually consist of between 2-30 amino acids with bulky, side-chain aromatic amino acids preferred in the ultimate and penultimate positions at the C-terminal end of the amino acid chain. They are reported to impart a myriad of preventative health beneficial effects to the consumer once ingested and these include heart health benefits through inhibition of enzymes including renin (EC 3.4.23.15) and angiotensin- I-converting enzyme (ACE-1; EC 3.4.15.1) within the renin angiotensin aldosterone system (RAAS) anti-inflammatory (due to inhibition of ACE-I and other enzymes) and anti-cancer benefits, prevention of type-2 diabetes through inhibition of dipeptidyl peptidase IV (DPP-IV), bone and dental strength, antimicrobial and immunomodulatory effects and several others. Peptides have also reported health benefits in the treatment of asthma, neuropathic pain, HIV and wound healing. However, the structure, amino acid composition and length of these peptides, along with the quantity of peptide that can pass through the gastrointestinal tract and often the blood-brain barrier (BBB), intact and reach the target organ, are important for the realisation of these health effects in an in vivo setting. This paper aims to collate recent important research concerning the generation and detection of peptides in the laboratory. It discusses products currently available as preventative healthcare peptide options and relevant legislation barriers to place a food peptide product on the market. The review also highlights useful in silico computer- based methods and analysis that may be used to generate specific peptide sequences from proteins whose amino acid sequences are known and also to determine if the peptides generated are unique and bioactive. The topic of food-derived bioactive peptides for health is of great interest to scientific research and industry due to evolving drivers in food product innovation, including health and wellness for the elderly, infant nutrition and optimum nutrition for sports athletes and the humanisation of pets. This paper provides an overview of what is required to generate bioactive peptide containing hydrolysates, what methods should be used in order to characterise the beneficial health effects of these hydrolysates and the active peptide sequences, potential applications of bioactive peptides and legislative requirements in Europe and the United States. It also highlights success stories and barriers to the development of peptide-containing food products that currently exist.

Alternative Proteins as a Source of Bioactive Peptides: The Edible Snail and Generation of Hydrolysates Containing Peptides with Bioactive Potential for Use as Functional Foods.

Members of the Phylum Mollusca include shellfish such as oysters and squid but also the edible garden snail known as Helix aspersa. This snail species is consumed as a delicacy in countries including France (where they are known as petit-gris), southern Spain (where they are known as Bobe), Nigeria, Greece, Portugal and Italy but is not a traditional food in many other countries. However, it is considered an excellent protein source with a balanced amino acid profile and an environmentally friendly, sustainable protein source. The aim of this work was to develop a different dietary form of snail protein by generating protein hydrolysate ingredients from the edible snail using enzyme technology. A second aim was to assess the bioactive peptide content and potential health benefits of these hydrolysates. H. aspersa hydrolysates were made using the enzyme Alcalase® and the nutritional profile of these hydrolysates was determined. In addition, the bioactive peptide content of developed hydrolysates was identified using mass spectrometry. The potential heart health benefits of developed snail hydrolysates were measured in vitro using the Angiotensin-I-converting Enzyme (ACE-1; EC 3.4.15.1) inhibition assay, and the ACE-1 inhibitory drug Captopril© was used as a positive control. The generated H. aspersa hydrolysates were found to inhibit ACE-1 by 95.60% (±0.011) when assayed at a concentration of 1 mg/mL (n = 9) compared to the positive control Captopril© which inhibited ACE-1 by 96.53% (±0.0156) when assayed at a concentration of 0.005 mg/mL (n = 3). A total of 113 unique peptide sequences were identified following MS analysis with peptides identified ranging from 628.35 Da (peptide GGGLVGGI-protein accession number sp|P54334|XKDO_BACSU) to 2343.14 Da (peptide GPAGVPGLPGAKGDHGFPGSSGRRGD-protein accession number sp|Q7SIB2|CO4A1_BOVIN) in size using the BIOPEP-UWM database.