Effect of ripening time on the content of bioactive peptides and fatty acids profile of Artisanal Coalho cheese.

The present study aimed to investigate the influence of ripening on the physicochemical, microbiological aspects, and fatty acid profile of Artisanal Coalho Cheeses and to detect if there are peptides with bioactive potential in their composition. Artisanal Coalho Cheese samples were kindly provided by a dairy farm located in Brazil in the Rio Grande do Norte state. A completely randomized design was adopted, with four maturation periods (0, 30, 45, and 60 days). Physicochemical traits (pH, total solids, moisture, non-fat solids, fat in total solids, protein, ash, fatty acid profile) and microbiological characterization (Salmonella sp, Listeria monocytogenes, total and thermotolerant coliforms, Staphylococcus aureus) were analyzed on cheese samples. Additionally, assays were performed for antioxidant and antihypertensive bioactivity through ACE and antimicrobial inhibition of the peptides extracted from the samples. There was a linear increase in total solids and ash content and a decrease in moisture content with increasing maturation time. The matured cheese samples had a lower pH than fresh Artisanal Coalho Cheese. Twenty-seven fatty acids were identified in the cheeses: 15 saturated, 07 monounsaturated, and 05 polyunsaturated, with a linear reduction of essential fatty acids (n6 and n3) during maturation. The microbiological quality of the cheeses was satisfactory, with an absence of undesirable bacteria in 92% of the cheese samples. Water-soluble peptide fractions from all periods tested showed antioxidant and antihypertensive potential with ACE control, and the maturation process potentiated these capacities, with a decline in these activities observed at 60 days. The antimicrobial activity against Gram-positive and Gram-negative bacteria increased with maturation, reaching better results until 60 days. The maturation process on wooden planks in the periods of 30, 45, and 60 days allows the production of Artisanal Coalho Cheese of an innovative character, safe to consumers from the microbiological point of view, with differentiated physicochemical and functional characteristics and good quality of lipid fraction compared to fresh cheese, enabling the addition of value to the dairy chain.

Effect of milk exposure on the redox profile of Caenorhabditis elegans.

The consumption of bovine milk and its derivatives is associated with inflammation, gastrointestinal disorders and the development of diseases in humans. Most studies related to milk effects are based on either clinal trials or experimental models such as mice and cell cultures. In this study we present the nematode Caenorhabditis elegans as an alternative model to evaluate the effects of milk on oxidative stress in other animal models. The toxicological effect of 20% milk exposure for 8 h on C. elegans was evaluated by progeny quantification, body size and pharyngeal pumping rate. Treating the worms with milk did not affect the worms brood size but interfered with their fecundity by delaying the average number of eggs in the first day of oviposition when compared to the control group. The size of worms treated with milk were significantly smaller compared to control. The pharyngeal pumping rate of milk-treated animals was not significantly different compared to untreated animals. Taking together, the results suggest that 20% milk treatment is not toxic for the worms but induces a minor delay achieving its adulthood and therefore its reproduction period. Milk exposure did not reduce the worms' survival under stress conditions and increase endogenous ROS levels. This study contributes to characterize the effects of milk exposure on the C. elegans nematode.

Characterization of Physicochemical and Sensory Properties of Cheeses Added with Bovine Colostrum.

The objective of this study was to develop fresh and matured cheeses with different bovine colostrum levels, aiming to promote the consumption of dairy products with the addition of colostrum. Four different cheese formulations were produced with a mixture of 0:100, 15:85, 20:80, and 25:75, bovine colostrum:milk (v:v), and aged for 0, 10, 20, and 40 days. Milk, colostrum, and fresh and matured cheeses were submitted to physicochemical characterization. Moreover, microbiological quality, yield, texture profile, color, and sensory acceptance of cheese samples were evaluated. Colostrum supplementation favored low acidity, high moisture, a pH range of 5.0-6.2, and water activity of 0.94-99. Sensory attributes and overall evaluation of all cheese formulations achieved an Acceptability Index above 70, indicating good acceptability. Since cheese with colostrum presented the potential to be used as human food, assessing the presence of colostrum bioactive components in those dairy products is a promising goal for further research.

An Overview of the Occurrence of Bioactive Peptides in Different Types of Cheeses.

The search for improvements in quality of life has increasingly involved changes in the diet, especially the consumption of foods which, in addition to having good nutritional value, are characterized by offering health benefits. Among the molecules that trigger several beneficial responses are peptides, which are specific fragments of proteins known to produce positive effects on the human body. This review aimed to discuss the bioactive potential of peptides from cheeses. Studies show that the protein composition of some cheese varieties exhibits a potential for the release of bioactive peptides. The production of these peptides can be promoted by some technological procedures that affect the milk structure and constituents. The cheese maturation process stands out for producing bioactive peptides due to the action of enzymes produced by lactic acid bacteria. Thus, in addition to being proteins with high biological value due to their excellent amino acid profile, peptides from some types of cheeses are endowed with functional properties such as anti-hypertensive, antimicrobial, antioxidant, anticarcinogenic, opioid, and zinc-binding activities.

Nutritional and physicochemical quality of formulations based on colostrum and bovine whey.

The objective of this study was to investigate the nutritional quality of bovine colostrum and whey mixtures. Five whey with bovine colostrum formulations were prepared (90:10; 80:20; 70:30; 60:40 and 50:50 whey:colostrum v:v) to be subjected to low-temperature pasteurization (63°C to 65°C for 30 minutes) and freeze-drying. The samples underwent chemical composition characterization, fatty acid profile analysis, determination of contamination by Enterobacteriaceae, pH, and Dornic acidity measurements before and after vat pasteurization. The amount of protein, fat, total solids, defatted dry extract, Brix and density increased as the bovine colostrum concentration increased. The level of saturated fatty acids and the thrombogenicity and atherogenicity indices reduced, while unsaturated fatty acids increased as the level of added bovine colostrum increased. The low-temperature pasteurization of the formulations was possible and effective, eliminating contamination by Enterobacteriaceae in the samples. Mixing bovine colostrum and whey reduced the colostrum viscosity, allowing a successful pasteurization procedure. Due to colostrum composition, the formulations yielded a higher nutritional value when compared to whey alone. The parameters applied in the formulation of mixtures of bovine colostrum and whey resulted in valuable ingredients for preparing novel dairy products.

Lactoperoxidase potential in diagnosing subclinical mastitis in cows via image processing.

This report describes how image processing harnessed to multivariate analysis techniques can be used as a bio-analytical tool for mastitis screening in cows using milk samples collected from 48 animals (32 from Jersey, 7 from Gir, and 9 from Guzerat cow breeds), totalizing a dataset of 144 sequential images was collected and analyzed. In this context, this methodology was developed based on the lactoperoxidase activity to assess mastitis using recorded images of a cuvette during a simple experiment and subsequent image treatments with an R statistics platform. The color of the sample changed from white to brown upon its exposure to reagents, which is a consequence of lactoperoxidase enzymatic reaction. Data analysis was performed to extract the channels from the RGB (Red-Green-Blue) color system, where the resulting dataset was evaluated with Principal Component Analysis (PCA), Multiple Linear Regression (MLR), and Second-Order Regression (SO). Interesting results in terms of enzymatic activity correlation (R2 = 0.96 and R2 = 0.98 by MLR and SO, respectively) and of somatic cell count (R2 = 0.97 and R2 = 0.99 by MLR and SO, respectively), important mastitis indicators, were obtained using this simple method. Additionally, potential advantages can be accessed such as quality control of the dairy chain, easier bovine mastitis prognosis, lower cost, analytical frequency, and could serve as an evaluative parameter to verify the health of the mammary gland.

Identification of bioactive peptides released from in vitro gastrointestinal digestion of yam proteins (Dioscorea cayennensis).

Bioactive peptides have been broadly studied for their contribution to human health. This study aimed to identify bioactive peptides generated by in vitro gastrointestinal digestion of yam proteins. Yam protein concentrate (YPC) was submitted to simulated digestion. Gastric phase hydrolysate (GPH) and total gastrointestinal phase hydrolysate (GIPH) had their peptides identified by nanoLC-ESI-MS/MS. Peptide sequences were subjected to a database-driven (BIOPEP) bioactivity search. In vitro tests included: Antioxidant activity, DNA damage protection, ACE-inhibitory activity and antibacterial activity against the bacteria Escherichia coli, Salmonella sp. and Lysteria monocytogenes. Simulated digestion generated small peptides (mostly MW < 3500 Da), several of them with potential bioactive sequences predicted in silico. In both GPH and GIPH biological activities were detected, although GIPH displayed stronger DNA damage protection and antibacterial activity against Escherichia coli. The digestion of yam proteins releases promising biologically active peptides which can contribute to the prevention of bacterial infection and chronic degenerative diseases, with beneficial effects to human health.

The Coconut Water Antimicrobial Peptide CnAMP1 Is Taken up into Intestinal Cells but Does Not Alter P-Glycoprotein Expression and Activity.

Coconut antimicrobial peptide-1 (CnAMP1) is a naturally occurring bioactive peptide from green coconut water (Cocos nucifera L.). Although biological activities have been reported, the physiological relevance of these reports remains elusive as it is unknown if CnAMP1 is taken up into intestinal cells. To address this open question, we investigated the cytotoxicity of CnAMP1 in intestinal cells and its cellular uptake into human intestinal cells. Considering the importance of the P-glycoprotein (P-gp) to the intestinal metabolism of xenobiotics, we also investigated the influence of CnAMP1 on P-gp activity and expression. Both cell lines showed intracellular fluorescence after incubation with fluorescein labelled CnAMP1, indicating cellular uptake of the intact or fragmented peptide. CnAMP1 (12.5-400 µmol/L) showed no signs of cytotoxicity in LS180 and differentiated Caco-2 cells and did not affect P-gp expression and activity. Further research is required to investigate the identity of CnAMP1 hydrolysis fragments and their potential biological activities.

Development and validation of a rapid reversed-phase liquid chromatography method for CnAMP1 peptide quantification in human intestinal cell lines.

Plant foods are rich sources of biologically active peptides that may have a role in the prevention of diseases. Coconut water is a valuable beverage due to its nutrient composition and the presence of bioactive compounds, such as the peptide CnAMP1. It is unknown if CnAMP1 can be absorbed into intestinal cells. We, therefore, aimed to develop and validate a simple reversed-phase liquid chromatographic method to quantify the peptide in Caco-2 and LS180 cell lysates. CnAMP1 standards (1-200 µmol/L) and spiked cell lysates were injected onto a Reprosil-Pur 120 C18-AQ column (4.6 × 250 mm) using acetonitrile:water:trifluoroacetic acid (14.0:85.9:0.1, by volume) as mobile phase in isocratic mode at flow rate of 1 mL/min. The method achieved rapid separation (total run time of 6 min), with linear response, good sensitivity (limit of detection, 8.2 ng; lower limit of quantification, 30.6 ng) and no interfering peaks. Best recoveries (84-96%), accuracies (7.6-14.8%) and precision (1.5-8%) were found for LS180 cell lysates spiked with medium (50 µmol/L) and high (100 µmol/L) amounts of the peptide. Uptake assays detected no peptides in the cell lysates; however, after the first 15-min incubation CnAMP1 underwent partial hydrolysis upon incubation with LS180 cells (29%) and extensive hydrolysis with Caco-2 cells (93%).

Growth Impairment Caused by Raw Linseed Consumption: Can Trypsin Inhibitors Be Harmful for Health?

Linseed (Linun usitatissimum L.) is an important oilseed whose nutritional value can be impaired due to presence of antinutritional factors and low protein digestibility. Protein fractions from raw linseed meal were extracted, isolated and analyzed in vitro and in vivo. Globulins, the major protein fraction of linseed, showed low in vitro susceptibility to trypsin and chymotrypsin, but its in vivo digestibility was 93.2 %. Albumin fraction had high trypsin inhibition activity (5250 Inhibition Units g(-1)) and presented low molecular mass protein bands, similar to known trypsin inhibitors. Raw linseed consumption caused negative effects on rat growth and reduction of intestinal villi. Results indicate that raw linseed meal must not be used as an exclusive source of protein regardless of the major proteins have high digestibility; digestive enzymes inhibitors in raw linseed probably reduces the protein utilization.

Marine sponge lectins: actual status on properties and biological activities.

Marine sponges are primitive metazoans that produce a wide variety of molecules that protect them against predators. In studies that search for bioactive molecules, these marine invertebrates stand out as promising sources of new biologically-active molecules, many of which are still unknown or little studied; thus being an unexplored biotechnological resource of high added value. Among these molecules, lectins are proteins that reversibly bind to carbohydrates without modifying them. In this review, various structural features and biological activities of lectins derived from marine sponges so far described in the scientific literature are discussed. From the results found in the literature, it could be concluded that lectins derived from marine sponges are structurally diverse proteins with great potential for application in the production of biopharmaceuticals, especially as antibacterial and antitumor agents.