Cow's Milk Research Articles

Draft genome sequencing of Pediococcus acidilactici strains isolated from cow's milk: unlocking insights into functional traits and applications.

Pediococcus acidilactici is a potential probiotic bacteria isolated from diverse sources. However, strains isolated from milk, especially from raw milk of healthy cows, have not been thoroughly studied. Here, we report the draft genome sequence of P. acidilactici strains MBBL5 and MBBL7, isolated from milk samples of healthy cows.

Q2-metnet: QIIME2 package to analyze 16S rRNA data via high-quality metabolic reconstructions of the human gut microbiota.

16S rRNA gene sequencing is the most frequent approach for the characterization of the human gut microbiota. Despite different efforts in the literature, the inference of functional and metabolic interpretations from 16S rRNA gene sequencing data is still a challenging task. High-quality metabolic reconstructions of the human gut microbiota, such as AGORA and AGREDA, constitute a curated resource to improve functional inference from 16S rRNA data, but they are not typically integrated into standard bioinformatics tools. Here, we present q2-metnet, a QIIME2 plugin that enables the contextualization of 16S rRNA gene sequencing data into AGORA and AGREDA. In particular, based on relative abundances of taxa, q2-metnet determines normalized activity scores for the reactions and subsystems involved in the selected metabolic reconstruction. Using these scores, q2-metnet allows the user to conduct differential activity analysis for reactions and subsystems, as well as exploratory analysis using PCA and hierarchical clustering. We apply q2-metnet to a dataset from our group that involves 16S rRNA data from stool samples from lean, allergic to cow's milk, obese and celiac children, and the Belgian Flemish Gut Flora Project cohort, which includes faecal 16S rRNA data from obese and normal-weight adult individuals. In the first case, q2-metnet outperforms existing algorithms in separating different clinical conditions based on predicted pathway abundances and subsystem scores. In the second case, q2-metnet complements competing approaches in predicting functional alterations in the gut microbiota of obese individuals. Overall, q2-metnet constitutes a powerful bioinformatics tool to provide metabolic context to 16S rRNA data from the human gut microbiota. Python code of q2-metnet is available in https://github.com/PlanesLab/q2-metnet and https://figshare.com/articles/dataset/q2-metnet_package/26180446. Supplementary data are available at Bioinformatics online.

Phytochemical characterization and quality evaluation of stirred goat milk yoghurt with added fruit and vegetable puree during storage

PurposeGoat milk yoghurt differs from cow milk yoghurt in that it has a different casein composition and content, which presents several technical challenges, including consistency with an appropriate flavor.Design/methodology/approachIn this study, the antioxidant potential and phytochemical profiling of the fruits (pineapple and papaya) and vegetable (carrot) extracts was evaluated and the effect of their purees on the quality and stability of stirred goat milk yoghurt (GMY) were investigated. The qualities of stirred GMY with carrot (CrY), pineapple (PaY) and papaya (PpY) purees were assessed against the product without puree (CY).FindingsThe carrot puree had the highest moisture, ash contents and pH value. The carrot extract had the highest DPPH radical scavenging activity, while the pineapple extract had the highest total phenolic value (1.59 µg GAE/g) and flavonoids content (0.203 µg CE/g). The scanning of all the puree extracts in GC-MS indicated that 5-hydroxymethylfurfural was a major component. The phytochemical quantification of the extracts through multiple reaction monitoring (MRM) against 16 compounds showed the presence of sinapic acid, cinnamic acid, pthalic acid, ferulic acid, 4-OH-benzoic acid, 3-OH-benzoic acid, p-coumaric acid, caffeic acid and vanillic acid in different quantities. The addition of purees and storage period had a significant (p < 0.05) effect on the moisture, pH, titratable acidity, syneresis, viscosity, color values and sensory properties of the products. In all the samples after 15 days of storage, Streptococcus thermophilus and Lactobacillus bulgaricus counts remained above the recommended level of 106CFU/g. Stirred GMY sample produced with pineapple puree showed a higher syneresis and viscosity, but the CrY sample demonstrated the highest antioxidant activity. The developed formulations remained stable with minimum changes in quality and sensory attribute during refrigerated storage for 10 days.Originality/valueThis study suggests that addition of fruit and vegetable improve the viscosity and sensory perception of the product with minimal use of synthetic flavor and preservatives.

The Use of Blondo (by Product from Lauric Acid Production) for Cheese Fruiters: The Study Ratio of Milk with Blondo and Citric Acid Concentration as a Coagulant

The processing of coconut milk into lauric acid produced Blondo as by product. Blondo was rich in protein, oil (fat), carbohydrates, vitamins and minerals. In addition, the fatty acids from coconut milk blondo were short and medium chain of saturated fatty acids which were very beneficial for health. From the nutritional content and physical properties, blondo had similarities with cow's milk. Thus, blondo had the potential to be processed into cheese. This study aimed to study the effect of the ratio of cow's milk with blondo and citric acid concentration as a coagulant on the quality of cheese. This research method used factorial RAK (Randomized Block Design). There were 2 factorial, namely the proportion of milk with coconut milk cream (25:75, 50:50, 75:25, 100:0) and citric acid concentration (2% and 4%). The results showed that the proportion of milk with blondo and citric acid concentration was significantly affected in the water content and protein content. However, it did not significantly affect with fat content, pH value and organoleptic taste, aroma, texture and color. Keywords: Cow's Milk, Coconut Milk, Blondo, Cheese, Citric Acid

Production and Quality Assessment of Plant-based Yoghurt from Coconut Milk Fortified with Date Syrup

Aim: To evaluate the physicochemical, microbial, sensory attributes and storage stability of coconut yoghurt fortified with date syrup. Methodology: Yoghurt was prepared from coconut milk and date syrup. Five samples were formulated as follows: T0 (100% cow milk), T1 (100% coconut milk), T2 (85% coconut milk, 15% date syrup), T3 (80% coconut milk, 20% date syrup), T4 (75% coconut milk, 25% date syrup) and T5 (70% coconut milk, 30% date syrup) The yoghurt samples were subjected to physicochemical and microbial analysis using standard methods. The sensory attributes assessed were appearance, aroma, taste, consistency and overall acceptability. The pH, titratable acidity and viscosity were evaluated during 4 weeks of storage at refrigerated condition (4- 6o c). Results: From the result, pH, titratable acidity, viscosity, and total solid ranged from 4.50 - 4.41%, 0.82 – 0.94%, 12.0 -72.40 cP and 11.64 – 18.81% respectively. These parameters increased significantly (p<0.05) with increase in the proportion of date syrup. The result showed a significant increase in ash, protein, carbohydrate, and fiber with a corresponding decrease in fat and moisture content as the proportion of date syrup increased. The total bacterial count ranged from 6.95 - 8.20cfu/ml, with Yeast, molds and coliforms undetected. The produced yoghurt samples all have quality attributes of animal milk yoghurt and better storage stability. Conclusion: Therefore, production of commercial yoghurt using plant-based milk (coconut milk) blended with nutritiously rich sweeteners like date fruit is encouraged.

The Impact of Seasonal Changes on the Composition of Cow Milk in Jalalabad City, Afghanistan

The study investigated the impact of seasonal variations on the components of cow milk in Jalalabad City, Afghanistan. A total of 80 milk samples were collected from six different local points within the city from January to August 2023. The research spanned two distinct periods: winter (January-February-March) and summer (June-July-August). During the winter, cows were fed hay, while in the summer; they grazed on green grass in pastures. Analysis revealed significant differences in fat content and titratableacidity between the winter and summer periods, with both factors being significant at the p<0.05 level according to paired t-test results. The study concluded that seasonal variations significantly affected the protein content, total solids, fat, and titratableacidity, with significance levels of p<0.05 and p<0.01, respectively.

Ameliorative effects of probiotic Limosilactobacillus fermentum and Enterococcus lactis isolated from cameroonian traditionally processed milk and palm wine against chronic constriction injury induced neuropathic pain in mice

Ethnopharmacological relevanceFermented milk and palm wine are regularly used by several ethnic groups in Cameroon in traditional treatment rituals for infections, inflammatory, cardiovascular disorders, and even metabolic diseases such as diabetes, hypercholesterolemia etc. Reports from many studies have demonstrated that fermented milk and palm wine are potential sources of probiotic bacteria. However, the capacity of probiotics isolated from these natural sources to alleviate neuropathic pain has not been experimentally tested. Aim of the studyThis study aimed at investigating the ameliorative potential of lactic acid bacteria isolated from palm wine and traditional fermented cow milk on the chronic constriction injury (CCI) induced neuropathic pain in mice. Materials and methodsPour plating technique on De Man Rogasa (MRS) agar was utilised for isolation of lactic acid bacteria from fermented cow milk and palm wine, and identified using the 16S r RNA gene sequencing. Neuropathic pain was induced by chronic constriction injury of the sciatic nerve. These bacteria were orally administered at different concentrations to Balb/c mice by gavage for 14 consecutive days. Cold allodynia, mechanical hyperalgesia and exploratory behaviour were evaluated on day 0, 7th and 14th respectively. The total level of calcium, oxidative stress markers and myeloperoxidase were also quantified in the sciatic nerve homogenate. Cyclooxygenase-2(COX-2) and cytokine profile were determined from serum. ResultsLactic acid bacteria were isolated from fermented cow milk and palm wine and two isolates were chosen according to their probiotic potentials and identified as strain of Limolactobacillus fermentum and Enterococcus lactis. Their 16 S rRNA gene sequences were deposited in NCBI genbank with accession number of OP896078 and OR619545, respectively. Pretreatment with Limosilactobacillus fermentum and Enterococcus lactis significantly alleviated mechanical hyperalgesia and cold allodynia with similar effect to the reference drug, morphine. These two isolates ameliorated CCI induced neuropathic pain by increasing antioxida776nts (GSH, CAT and SOD, P < 0.01) and decreasing pro-oxidants (MDA and NO, P < 0.01). Also, they inhibited the release of proinflammatory cytokines (IL-1β, TNF-α, IFN-γ, and IL-6; P < 0.01) and IL-10 level was significantly (P < 0.01) increased when compared to the negative control. Treatment with these bacteria significantly dropped the level of total calcium (P < 0.01), COX-2 (P < 0.01) and MPO (P < 0.01) when compared with the negative control. ConclusionThe neuroprotective potentials of these selected lactic acid bacteria against CCI induced neuropathic pain may be attributed to their anti-oxidant, anti-inflammatory properties and reduced calcium deposition in sciatic nerve.

Label-free-based proteomics analysis reveals differential proteins of sheep, goat and cow milk

Regarding the limited information on species protein differences between sheep, goat, and cow milk, the differentially expressed proteins in sheep, goat, and cow milk and their functional differences are analyzed using label-free proteomics technology to identify potential biomarkers. 770 proteins and 2914 peptide segments were identified. The statistical analysis showed significant differences in the relative abundances of the 74 proteins among the sheep, goat, and cow milk. CSN3 and LALBA can be used as potential biomarkers for goat milk, XDH can be used as potential biomarkers for cow milk, and CTSB and BPIFB1 can be used as potential biomarkers for sheep milk. The functional analysis using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes showed that these significantly different proteins were enriched by different pathways including thyroid hormone synthesis and glycerol phospholipid metabolism. The data revealed differences in the amounts and physiological functions of the milk proteins of different species, which may provide an important basis for research on the nutritional composition of dairy products and adulteration identification technology.

Bacteriocin characterization of Enterococcus faecium isolates and evaluation of their in situ anti-Listerial activity in Beyaz cheese

Nowadays, as the interest in natural preservatives increases, research on bioprotective cultures that produce bacteriocins and studies on the use of these cultures in foods are also increasing day by day. Present study aimed to investigate bacteriocin characterization and safety of Enterococcus faecium H108, H206 isolates and their potential utilization in the reduction of Listeria monocytogenes ATCC7644 in Beyaz cheese. Enterocin producing E. faecium H108 and H206 strains were isolated from raw cow's milk and identified by 16S rRNA sequencing. Enterocin containing supernatants of E. faecium H108 and H206 were found to be stable to heat various chemicals used in food industry, wide pH range, lysozyme and catalase enzymes. On the other hand, activity of enterocins were inactivated by proteolytic enzymes. Maximum activity of E. faecium H108 was 6400 AU/mL, while that of H206 was 12800 AU/mL, and the activities remained stable for 24 h. Supernatant of both E. faecium H108 and H206 inhibited L. monocytogenes growth in vitro. E. faecium H108 and H206 did not show haemolytic and DN-ase activities and were susceptible to most of the antibiotics tested. The isolates were used in Beyaz cheese and were found to significantly, but not completely inhibit L. monocytogenes growth. The findings of this research suggest that E. faecium H108 and H206 isolates, after a detailed safety assessment, might be employed as bioprotective cultures in Beyaz cheese with starter culture or hurdle technologies.

Purification of α-lactalbumin and β-lactoglobulin from cow milk

Whey, a valuable byproduct of dairy processing, contains essential proteins like β-lactoglobulin (βLG) and α-lactalbumin (αLA), making it a focus of research for its nutritional benefits. Various techniques, including chromatography and membrane filtration, are employed for protein extraction, often requiring multiple purification steps. One approach that has gained prominence for the purification and concentration of proteins, including those present in whey, is the use of polyethylene glycol (PEG) in aqueous two-phase systems. Our study simplifies this process by using PEG alone for whey protein purification. This approach yielded impressive results, achieving 92% purity for βLG and 90% for αLA. These findings underscore the effectiveness of PEG-based purification in isolating whey proteins with high purity.

Analysis of milk adulteration by means of a potentiometric electronic tongue

The adulteration of milk presents significant challenges in the food industry, promoting the need for efficient detection methods. This study introduces a potentiometric electronic tongue for rapid and accurate milk adulteration detection. Utilizing polymeric membranes integrated with various additives, the electronic tongue distinguishes between different milk types and detects common adulterants. Experimental results demonstrate its effectiveness in discriminating raw, pasteurized, and medicated cow milk, as well as goat milk. Moreover, it successfully identifies adulterants like water and bovine milk in goat milk samples. Chemometric analyses, including Principal Component Analysis and Partial Least Squares regression, correlate sensor responses with traditional milk parameters such as fat, protein, and lactose content with up to a 0.97 R2 on the validation step. Strong correlations validate the electronic tongue's potential for rapid milk quality assessment. This innovative approach offers a cost-effective, reliable solution for detecting milk adulteration in contrast with current techniques that require numerous, time consuming experiments.

Allergen-specific B cell responses in oral immunotherapy-induced desensitization, remission, and natural outgrowth in cow's milk allergy.

Antigen-specific memory B cells play a key role in the induction of desensitization and remission to food allergens in oral immunotherapy and in the development of natural tolerance (NT). Here, we characterized milk allergen Bos d 9-specific B cells in oral allergen-specific immunotherapy (OIT) and in children spontaneously outgrowing cow's milk allergy (CMA) due to NT. Samples from children with CMA who received oral OIT (before, during, and after), children who naturally outgrew CMA (NT), and healthy individuals were received from Stanford biobank. Bos d 9-specific B cells were isolated by flow cytometry and RNA-sequencing was performed. Protein profile of Bos d 9-specific B cells was analyzed by proximity extension assay. Increased frequencies of circulating milk allergen Bos d 9-specific B cells were observed after OIT and NT. Milk-desensitized subjects showed the partial acquisition of phenotypic features of remission, suggesting that desensitization is an earlier stage of remission. Within these most significantly expressed genes, IL10RA and TGFB3 were highly expressed in desensitized OIT patients. In both the remission and desensitized groups, B cell activation-, Breg cells-, BCR-signaling-, and differentiation-related genes were upregulated. In NT, pathways associated with innate immunity characteristics, development of marginal zone B cells, and a more established suppressor function of B cells prevail that may play a role in long-term tolerance. The analyses of immunoglobulin heavy chain genes in specific B cells demonstrated that IgG2 in desensitization, IgG1, IgA1, IgA2, IgG4, and IgD in remission, and IgD in NT were predominating. Secreted proteins from allergen-specific B cells revealed higher levels of regulatory cytokines, IL-10, and TGF-β after OIT and NT. Allergen-specific B cells are essential elements in regulating food allergy towards remission in OIT-received and naturally resolved individuals.

Healthy Effects of Milk and Dairy Product Consumption in the Mediterranean Area and Japan.

Milk is a food enriched in essential components for human health. Especially, in the Mediterranean area, besides cow's milk, milk from goats, sheep, and donkeys, is largely used. The consumption of animal milk is an important component of the Mediterranean (MED) diet, even if in moderate amounts. Milk is a complete food since it contains proteins, carbohydrates, and fats, as well as micronutrients (minerals and vitamins). Milk-fermented products are largely consumed in the MED diet, such as cheese and yogurt, which are rich in essential metabolites, bioactive compounds, vitamins, minerals, and exopolysaccharides. A large body of evidence suggests that consumption of milk and dairy products does not increase the risk of all-cause mortality, type 2 diabetes, and cardiovascular disease, even if some earlier studies have reported harmful effects associated with their higher consumption. Also, in Japan, despite the lower consumption of milk than in Western countries, intake of bovine milk is associated with healthy effects. The present review describes the effects of the various constituents of animal milk on human health, with special reference to the Mediterranean area and Japan. Experimental data and clinical trials support the ability of milk and dairy products to lower the risk of chronic diseases.

МИГРАЦИИОННАЯ АКТИВНОСТЬ 137CS В АГРОПРОДУКЦИЮ БИОГЕОЦЕНОЗОВ КРАСНОЯРСКОГО КРАЯ

The purpose of the study is to determine the intensity of 137Cs movement along the trophic chain “soil – diet – livestock products” in the agricultural landscapes of the Krasnoyarsk Region with additional radiation load. A literature review of scientific data characterizing the features of the migration activity of 137Cs in the links “soil – feed – livestock products” in agrobiocenoses with radioactive contamination above established standards is given. Based on the results of a radioecological survey of agricultural landscapes in the central regions of the Krasnoyarsk Region and calculation of absorbed doses, the author identified agricultural landscapes with additional technogenic pollution and determined the specific activity of 137Cs in feed and cow's milk in agricultural landscapes with additional technogenic load. The specific activity values of 137Cs in agricultural landscape feeds with doses of 1.33 and 1.55 mGy/year are higher than the background va¬lue for the Krasnoyarsk Region. The content of 137Cs in livestock products produced in farm biogeocenoses with additional technogenic load is higher than the average for the region, but does not exceed the criterion of regulatory documents; accordingly, radiation-safe agricultural products are produced on the territory of the Krasnoyarsk Region. For agricultural landscapes with absorbed doses of 1.33 and 1.55 mGy/year, a KP for the link “feed – livestock products” has been established. When the radioecological situation changes, the established Kp values can be used to predict the migration of 137Cs in the chain “soil – feed – livestock products”. A direct linear moderate Spearman rank correlation was identified between the specific activity of 137Cs in feed and the concentration of 137Cs in cow's milk. Linear regression equations have been determined to determine the specific activity of cow's milk based on data on the specific activity of 137Cs in feed. The revealed direct linear relationship indicates that even a slight increase in the 137Cs activity of the diet leads to an equivalent increase in the specific activity of technogenic cesium in cow's milk.

Exploring the potential of omega-3 fatty acids in acne patients: A prospective intervention study.

Omega-3 fatty acids (ω-3 FA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA), are essential nutrients known for their anti-inflammatory properties, which involve reducing pro-inflammatory cytokines, eicosanoids, and insulin-like growth factor-1. This suggests their potential to alleviate acne severity, especially when deficits are present. To elevate EPA/DHA levels in acne patients through dietary intervention and supplementation, observing subsequent clinical effects. Over 16 weeks, 60 patients without prescription medication (n = 23 acne comedonica [AC], n = 37 acne papulopustulosa [AP]) adhered to a Mediterranean diet, incorporating oral algae-derived ω-3 FA supplementation (600 mg DHA/300 mg EPA week 1-8, 800 mg DHA/400 mg EPA week 8-16). At four visits (V1-V4), blood EPA/DHA levels were tracked using the HS-omega 3 index® (EPA/DHA (%) of total identified fatty acids in erythrocytes; target 8%-11%, deficit <8%, severe deficit <4%), alongside clinical assessments and standardized questionnaires. At baseline, 98.3% of patients had an EPA/DHA deficit, with the mean HS-omega 3 index® rising from 4.9% at V1 to 8.3% at V4 (p < 0.001). AC showed significantly higher indices than AP at V4 (p = 0.035). Objective improvements in both inflammatory and non-inflammatory lesions were observed (p < 0.001). While self-reported appearance worsened in four patients, overall quality of life improved (p < 0.001), particularly in AP. Dietary triggers were more clearly defined than beneficial foods. Intake of cow's milk and dairy products reduced (p < 0.001). Compliance was good; no adverse events were reported. Many acne patients have a ω-3 FA deficit. The HS-omega 3 index® can be increased by a Mediterranean diet and oral supplementation with algae-derived ω-3 FA. Acne severity improved significantly in patients with target ω-3 FA levels.

Prevalence and Phage-Based Biocontrol of Methicillin-Resistant Staphylococcus aureus Isolated from Raw Milk of Cows with Subclinical Mastitis in Vietnam

S. aureus, particularly methicillin-resistant S. aureus, has been recognized as a main cause of bovine mastitis and food poisoning. This study investigated the prevalence, antibiotic resistance, and phage-based biocontrol of S. aureus and methicillin-resistant S. aureus isolated from raw milk of cows with subclinical mastitis. The results showed that the prevalence of S. aureus and methicillin-resistant S. aureus was 12% (48/400) and 1.5% (6/400), respectively. The S. aureus isolates were highly resistant to penicillin (72.92%), erythromycin (43.75%), and tetracycline (39.58%). Out of 48 S. aureus isolates, 6 were identified as methicillin-resistant strains. Among them, one isolate was found to harbor the sea gene. A total of 5 phages were recovered from 50 pork and 50 chicken meat samples, 1 from pork and 4 from chicken meat samples. Phage PSA2 capable of lysing all 6 methicillin-resistant isolates was selected for characterization. The use of phage PSA2 completely inactivated methicillin-resistant S. aureus SA33 in raw milk at both 24 °C and 4 °C, indicating its potential as a promising antibacterial agent in controlling methicillin-resistant S. aureus in raw milk and treating bovine mastitis.

Prevalence and Phage-Based Biocontrol of Methicillin-Resistant Staphylococcus aureus Isolated from Raw Milk of Cows with Subclinical Mastitis in Vietnam

Prevalence and Phage-Based Biocontrol of Methicillin-Resistant Staphylococcus aureus Isolated from Raw Milk of Cows with Subclinical Mastitis in Vietnam

A Comparative Study of Dairy and Non-Dairy Milk Types: Development and Characterization of Customized Plant-Based Milk Options

Plant-based milk has gained considerable attention; however, its high nutritional variation highlights the need for improved formulation designs to enhance its quality. This study aimed to nutritionally compare cow milk with plant-based milk produced from hazelnuts (H), Brazil nuts (BN), cashew nuts (CN), soybeans (S), and sunflower seeds (SS), and to perform physicochemical and technological characterization. The plant-based milk produced with isolated grains showed a nutritional composition inferior to that of cow milk in almost all evaluated parameters, protein content (up to 1.1 g 100 g−1), lipids (up to 2.7 g 100 g−1), color parameters, minerals, and especially calcium (up to 62.4 mg L−1), which were originally high in cow milk (up to 1030 mg L−1). However, the plant-based milk designed using a blend composition was able to promote nutritional enhancement in terms of minerals, especially iron (Fe) and magnesium (Mg), high-quality lipids (up to 3.6 g 100 g−1), and carbohydrates (3.4 g 100 g−1 using CN, BN, and S). The protein content was 1.3% compared to 5.7 in cow milk, and the caloric value of plant-based milk remained 32.8 at 52.1 kcal, similar to cow milk. Satisfactory aspects were observed regarding the shelf life, especially related to microbiological stability during the 11 d of storage at 4 °C. For the designed plant-based milk to be equivalent to cow milk, further exploration for optimizing the blends used to achieve better combinations is required. Furthermore, analyzing possible fortification and preservation methods to increase shelf life and meet the nutritional and sensory needs of the public would be interesting.

A Comparative Study of Dairy and Non-Dairy Milk Types: Development and Characterization of Customized Plant-Based Milk Options

Plant-based milk has gained considerable attention; however, its high nutritional variation highlights the need for improved formulation designs to enhance its quality. This study aimed to nutritionally compare cow milk with plant-based milk produced from hazelnuts (H), Brazil nuts (BN), cashew nuts (CN), soybeans (S), and sunflower seeds (SS), and to perform physicochemical and technological characterization. The plant-based milk produced with isolated grains showed a nutritional composition inferior to that of cow milk in almost all evaluated parameters, protein content (up to 1.1 g 100 g−1), lipids (up to 2.7 g 100 g−1), color parameters, minerals, and especially calcium (up to 62.4 mg L−1), which were originally high in cow milk (up to 1030 mg L−1). However, the plant-based milk designed using a blend composition was able to promote nutritional enhancement in terms of minerals, especially iron (Fe) and magnesium (Mg), high-quality lipids (up to 3.6 g 100 g−1), and carbohydrates (3.4 g 100 g−1 using CN, BN, and S). The protein content was 1.3% compared to 5.7 in cow milk, and the caloric value of plant-based milk remained 32.8 at 52.1 kcal, similar to cow milk. Satisfactory aspects were observed regarding the shelf life, especially related to microbiological stability during the 11 d of storage at 4 °C. For the designed plant-based milk to be equivalent to cow milk, further exploration for optimizing the blends used to achieve better combinations is required. Furthermore, analyzing possible fortification and preservation methods to increase shelf life and meet the nutritional and sensory needs of the public would be interesting.

A new colorimetric lactate biosensor based on CUPRAC reagent using binary enzyme (lactate-pyruvate oxidases)-immobilized silanized magnetite nanoparticles.

Anovel optical lactate biosensoris presented that utilizes a colorimetric interaction between H2O2 liberated by a binary enzymatic reaction and bis(neocuproine)copper(II) complex ([Cu(Nc)2]2+) known as CUPRAC (cupric reducing antioxidant capacity) reagent. In the first step, lactate oxidase (LOx) and pyruvate oxidase (POx) were separately immobilized on silanized magnetite nanoparticles (SiO2@Fe3O4 NPs), and thus, 2mol of H2O2 was released per 1mol of the substrate due to a sequential enzymatic reaction of the mixture of LOx-SiO2@Fe3O4 and POx-SiO2@Fe3O4 NPs with lactate and pyruvate, respectively. In the second step, the absorbance at 450nm of the yellow-orange [Cu(Nc)2]+ complex formed through the color reaction of enzymatically produced H2O2 with [Cu(Nc)2]2+ was recorded. The results indicate that the developed colorimetric binary enzymatic biosensor exhibits a broad linear range of response between 0.5 and 50.0µM for lactate under optimal conditions with a detection limit of 0.17µM. The fabricated biosensor did not respond to other saccharides, while the positive interferences of certain reducing compounds such as dopamine, ascorbic acid, and uric acid were minimized through their oxidative removal with a pre-oxidant (NaBiO3) before enzymatic and colorimetric reactions. The fabricated optical biosensor was applied to various samples such as artificial blood, artificial/real sweat, and cow milk. The high recovery values (close to 100%) achieved for lactate-spiked samples indicate anacceptableaccuracy of this colorimetric biosensor in the determination of lactate in real samples. Due to the increase in H2O2 production with the bienzymatic lactate sensor, the proposed method displays double-fold sensitivity relative to monoenzymatic biosensors and involves a neat color reaction with cupric-neocuproine having a clear stoichiometry as opposed to the rather indefinite stoichiometry of analogous redox dye methods.