The aim of this study was characterization of some dairy drinks based on Milk Serum regarding major whey proteins (WP) and free amino acids (FAAs) using reversed phase high performance liquid chromatographic (RP-HPLC) methods. The studied WP, -lactalbumin (-La), bovine serum albumin (BSA), -lactoglobulin A (-Lg A) and -lactoglobulin B (-Lg B) were separated on Aeris XB-C18 column at 214 nm detection. The RP-HPLC method was validated by selectivity, linearity (R2 ≥0.99), sensitivity (LOQ, 1.35–10.08 µg mL−1), accuracy (recovery 96.79-103.07%) and precision (% RSD ≤ 4.13%). The total studied WP in studied dairy drinks varied between 1.42 and 3.047 g·L-1. The chromatographic profile of FAAs (aspartic acid, glutamic acid, serine, histidine, arginine, glycine, threonine, alanine, tyrosine, cysteine, tryptophan, methionine, valine, phenylalanine, isoleucine, leucine and lysine) was determined in lyophilized concentrate of Milk Serum by RP-HPLC using pre-column derivatization reaction with orthophthalaldehyde (OPA). The total studied FAAs in studied samples varied between 1.103 and 1.119 mg·g-1. Moreover, the Milk Serum showed bacteriostatic activity against two bacterial strains Escherichia coli and Staphylococcus aureus. The obtained results confirm that dairy drinks based on the Milk Serum constitutes a valuable sources of bioactive components with benefits for human healthy nutrition.

Sheep’s milk is a significant source of nucleotides monophosphate (NMPs) but can also contain undesirable residues from veterinary drugs, posing a potential human health risk. This study introduces a novel application of 2D-LC, in heart-cutting mode, for the simultaneous determination of nucleotides and veterinary drug residues in sheep’s milk. Two-dimensional liquid chromatography (2D-LC) allows for the separation of these compounds in a single chromatographic run despite their differing physicochemical properties. The proposed method separates six veterinary drug residues and five NMPs in a single injection. The compounds were separated using a C18 reversed-phase column in the first dimension and a Primesep SB analytical column in the second dimension. The method performance was evaluated in terms of linearity range, detection and quantification limits, matrix effects, precision, and accuracy. The results demonstrated good linearity and sensitivity, with quantification limits allowing for the quantification of veterinary drugs at the maximum residue level and nucleotides at typical levels found in milk samples. The method has been successfully applied to the analysis of sheep’s milk samples acquired from local supermarkets, with recoveries within a range of 70-119 % and 82-117 % for veterinary residues and NMPs, respectively.

The objective of this study was to determine the effects of supplementing L-isoleucine (L-Ile) on milk protein synthesis, using an immortalized bovine mammary epithelial (MAC-T) cell line. In this case, the cells were treated with 0, 0.3, 0.6, 0.9, 1.2 and 1.5 mM of supplemental Isoleucine (Ile), and the most efficient time for protein synthesis for each amino acid was determined by measuring the cell, medium and total protein at 0, 24, 48, 72 and 96 h. Confirmatory tests showed that 48h incubation time and 0.6 mM dosage of L-Ile are considered as the optimal time and dosage. The mechanism of milk protein synthesis was elucidated through proteomics analysis to clarify the metabolic pathway. When the L-Ile was supplemented, extracellular protein (medium protein) reached a peak at 48h, whereas in the case of the intracellular cell protein, it was shown to have reached to its peak at 24h in all L-Ile dosage treatments. In total, it is noted that there were 63 upregulated and 52 downregulated proteins. The results of the protein pathway analysis showed that the L-Ile group stimulated insulin/IGF pathway-mitogen activated protein kinase kinase/MAP kinase cascade, insulin/IGF pathway-protein kinase B signaling cascade, p53 pathway, de novo purine biosynthesis, Wnt signaling pathway, glycolysis, pentose phosphate pathway, and ATP synthesis which are pathways involved and related to protein and energy metabolism. Together, these results demonstrate that L-Ile supplementation was effective in stimulating β-casein synthesis by stimulating genes and pathways which are significantly related to protein and energy metabolism.

There are guidelines on lactation following maternal analgo-sedative exposure, but these do not consider the effect of maternal fasting, nor fluid abstention on human milk macronutrient composition. We therefore performed a structured search (PubMed) on ‘human milk composition’ and screened title, abstract and full paper on ‘fasting’ or ‘abstention’ and ‘macronutrient composition’ (lactose, protein, fat, solids, triglycerides, cholesterol). This resulted in 6 papers and one abstract related to religious fasting (n=129 women) and observational studies in lactating women (n=23, healthy volunteers, fasting). These data reflect two different ‘fasting’ patterns: an acute (18-25h) model in 71 (healthy volunteers, Yom Kippur/Ninth of Av) women and a chronic fasting (Ramadan) model in 81 women. Changes were most related to electrolytes and were moderate, with almost no changes in macronutrients during acute fasting. We therefor conclude that neither short term fasting nor fluid abstention (18-25h) affect human milk macronutrient composition, so that women can be reassured when this topic were raised during consulting. Besides the nutritional relevance, this also matters as clinical research samples – especially to estimate analgo-sedative exposure by lactation - are commonly collected after maternal procedural sedation, associated with maternal fasting and physiology-based pharmacokinetic (PBPK) models assume stable human milk composition.

The prevalence of food allergy has increased over the last decades and consequently the food labeling policies have improved over the time in different countries to regulate allergen presence in foods. In particular, Reg 1169 in EU mandates the labelling of 14 allergens whenever intentionally added to foods, but the inadvertent contamination by allergens still remains uncovered topic. In order to warn consumers on the risk of cross-contamination occurring in certain categories of foods, a precautionary allergen labelling system has been put in place by food industries on voluntary basis. In order to reduce the overuse of PAL, reference doses and action limits have been proposed by the VITAL project representing a guide in this jeopardize scenario. Development of sensitive and reliable mass spectrometry methods are therefore of paramount importance in this regard to check the contamination levels in foods. In this paper we describe the development of a managed time MRM method based on a triple quadrupole platform for milk and egg quantification in processed food. The method was in house validated and allowed to achieve levels of proteins lower than 0.2mg of total milk and egg proteins respectively in cookies, challenging the doses recommended by VITAL. The method was finally applied to cookies labeled as milk and egg-free. This method could represent in perspective a promising tool to be implemented along the food chain to detect even tiny amounts of allergens contaminating food commodities.

This study aims to evaluate the effect of different drying methodologies (room, microwave, convective, oven and freeze-drying) on the chemical composition, the microbiological quality of extracts and the biological activities namely clotting milk and antioxidant activities for both spontaneous and cultivated Onopordum nervosum ssp. platylepis. The results showed that the drying methodology has significantly affected the phenolic composition. Freeze dried flowers showed the best amounts of total phenols, flavonoids and condensed tannins followed by the microwave dried flowers. However, the latest presented the lowest protein content. Finally, the biological activities were significantly dependent on the used drying process. The effective concentration EC50 values let to classify the samples according to their decreasing power to inhibit DPPH free radicals: dryer<oven<room <microwave<freeze-drying. The drying process significantly affected the clotting milk activity, freeze dried flowers showed the highest activity. All the obtained results do not reveal a significant difference between cultivated and spontaneous plants.

The lactic acid bacteria Streptococcus thermophilus and Lactobacillus helveticus are commonly used as starter cultures in dairy product production. This study aimed to investigate the characteristics of fermented milk using different ratios of these strains and analyze the changes in volatile compounds during fermentation and storage. The 10:1 ratio of Streptococcus thermophilus CICC 6063 to Lactobacillus helveticus CICC 6064 showed optimal fermentation time (4.2 h), viable cell count (9.64 log10 colony-forming units/mL), and sensory evaluation score (79.1 points). In total, 56 volatile compounds were identified and quantified by solid-phase microextraction and gas chromatography–mass spectrometry (SPME-GC-MS), including aldehydes, ketones, acids, alcohols, esters, and others. Among these, acetaldehyde, 2,3-butanedione, 2,3-pentanedione, 2-heptanone, acetoin, and 1-heptanol had the highest odor activity values and contributed to the characteristic flavor of fermented milk. The concentrations and types of key flavor compounds reached their peak at the end of fermentation. Throughout the fermentation and storage process, the levels of 2,3-pentanedione and acetoin exhibited synergistic dynamics. These findings enhance our understanding of the chemical and molecular characteristics of milk fermented with Streptococcus thermophilus and Lactobacillus helveticus, providing a basis for improving the flavor and odor of dairy products during fermentation and storage.

Human milk composition is dynamic and substitute formulae are intended to mimic its protein content. The purpose of this study was to investigate the potentiality of matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS) followed by multivariate data analyses as a tool to analyze peptide profiling of mammalian, human and formula milks. Breast milk samples from women at different lactation stages (2 (n = 5), 30 (n = 6), 60 (n = 5), and 90 (n = 4) days postpartum), and milk from donkeys (n = 4), cows (n = 4), buffaloes (n = 7), goats (n = 4), ewes (n = 5), and camels (n = 2) were collected. Different brands (n = 4) of infant formulae were also analyzed. Protein content (<30 kDa) was analyzed by MS and data were exported for statistical elaborations. Mass spectra for each milk closely clustered together, whereas different milk samples resulted well separated. Human samples formed a cluster in which colostrum constituted a well-defined subcluster. None of the milk formulae correlated with animal or human milk, although specifically characterized and well correlated each other. These findings propose MALDI-TOF MS milk profiling as an analytical tool to discriminate, in a blinded way, different milk types. As each formula has a distinct specificity, shifting a baby from one to another formula implies a specific proteomic exposure. These profiles may assist in milk proteomics for easiness of use and low cost consuming, suggesting that the MALDI-TOF MS pipelines may result useful for milk adulteration assessment but also for the characterization of banked milk specimens in paediatric clinical settings.

Zn is an essential element for children related to optimal growth, development and nutritional metabolism. We measured Zn bioaccesibility (Zn-BA) levels of foods by Inductively coupled plasma mass spectrometry (ICP-MS) after in vitro digestion-fermentation method from children with gluten related disorders (GRD-CH), obesity (OB-CH) and allergy/intolerance to cow’s milk proteins (AICM-CH). Zn-BA levels in sick children (34.7 ± 28.8, 29.6 ± 30.1 and 26.7 ± 30.4%, respectively) were significantly lower (p &lt; 0.05) than in healthy norm-weight children (HE-CH). For AICM-CH, Zn-BA in animal foods was significantly lower than in plant foods, and in both plant and animal foods from sick and healthy children (p &lt; 0.05). In foods of animal origin in sick and healthy children, Zn-BA in foods heated in liquid media (frying and boiling) was significantly higher than in foods heated by hot air (roasting and grilling). In conclusion, in children the studied diseases diminished the Zn-BA, probably by the alteration of the gut microbiota which could negatively affect their appropriate long-term development. In AICM-CH, the allergy to cow's milk proteins could induce intestinal dysbiosis and inflammatory processes, leading to allergies to other animal food proteins, and to a more pronounced decrease of Zn-BA.

Global warming has been increasing heat stress threat in animals which can be monitored by Temperature Humidity Index (THI). The present study describes the relationship of THI, calculated using 35 years period weather station data, and production performances of dairy cattle in a se-lected area of Bangladesh. The month January and June were identified as the coolest and hottest, respectively. Based on this outcome, every year in the month of January and June production performances of 10 crossbred cows with homogenous characteristics were monitored for a period of 5 years. The average THIMEAN value was found 17% higher in June as compared to January, and with this increment of THIMEAN average milk production was decreased 24.4% (p<0.05). The milk fat and protein content were also reduced (p<0.05) by 14.5 and 15.2%, respectively suggesting negative correlation as like as milk production. However, ash content was increased 15.3% that indicates a positive correlation. In addition, multiple regression analysis revealed each point increase in THIMEAN and rectal temperature, there was a decrease in milk yield 0.04 and 1.17 kg ECM, respectively. In contrary, each point increase in THIMEAN resulted 0.059 C increase of rectal temperature. Taken together, THIMEAN value calculated using metrological station data has distinct relationship with the production performances of lactating crossbred dairy cows.

Corn silage (Zea mays L.) is the most widely used energy resource in the diets of dairy cattle around the world; it stands out for its higher biomass yields, good palatability, homogeneous quality at harvest and ease of silage due to its higher soluble sugar content. It was carried out a search of studies related to dry matter yield (ton ha-1), population density (plant density ha-1), dry matter (DM), crude protein (CP), neutral detergent fiber (NDF), non-fibrous carbohydrates (NFC), organic matter (OM), DM digestibility (DMD) and neutral detergent fiber digestibility (NDFD) and milk production per hectare (kg of milk ha-1) that was determined using MILK2006®. It was carried out a cluster analysis (CL, PROC CLUSTER) obtaining six groups of corn silage: CL1: "Starch", which included DMD, DFDN, TDN1xDM, Mega calories per kg of DM and kg of milk/ton/DM; CL2: "Fats" which included TDN1xMS, Mcal/kg/DM and kg of milk/ton/DM; CL3: "Dry Matter", conformed only by DFDN; CL4: "Plant Density" including Yield of DM ha-1 and Milk Production ha-1; CL5: "Neutral Detergent Fiber" and CL6: "Raw Protein". It is concluded that CL1 was characterized by a higher DMD, DFND, CNF and starch that allow a higher TNDx1DM and an energy concentration (Mcal/kg/DM) that shows a higher milk production (kg of milk/ton/DM ha-1). The characteristics of CL2 with higher EE, allow a higher TNDx1DM and an energy concentration (Mcal/kg/DM) with a higher milk production (kg of milk/ton/DM ha-1).

The latest EU regulation on geographical indications (EU Regulation No. 1151/2012) has intro-duced a set of new tools for the protection and enhancement of food products in rural areas, under the group name of optional quality term (OQT). The Commission Delegated EU Regulation, No. 665/2014, regulated the conditions for the use of the optional quality term «mountain product» (MP), to support the implementation of a mountain value chain. This new tool is aimed at pro-moting local development, maintaining the economic activities in mountain areas and redistrib-uting wealth, whilst, at the same time, promoting the territory. Pecorino and goat cheeses are typ-ical Italian cheeses made usually with whole raw ewe's or raw goat's milk, without starter cul-ture addition. In an attempt to characterize these productions, the aim of this study was to inves-tigate the evolution of enterococci during the production and ripening of Pecorino cheese made in three different farms, located in Umbria, Italy in areas facing natural or other specific constraints as stipulated by Regulation 1305/2013 on support for rural development by the European Agri-cultural Fund for Rural Development (EAFRD). Enterococci are enteric organisms which are commonly isolated from ewe and goat's milk production in Umbria, Italy. Counts of enterococci in raw milk ranged from 1.75 for ovine milk to 3.62 for ewe milk and a marked reduction was observed after thermization especially in ovine milk. Out of 100 isolates, 69 were E. faecium, 23 E. durans, 8 E. faecalis and 2 E. casseliflavus and the distribution of species between farms and be-tween samples showed a prevalence of E. faecium in ovine farms and E. durans in ewes farms, with an equal distribution between samples. High percentages of susceptible isolates were found for amoxicillin/clavulanic acid, ampicillin, chloramphenicol, sulphamethoxazole, sulphameth-oxazole/trimethoprim, ticarcillin, vancomycin. A high prevalence of resistant strains (> 30%) was observed for amikacin, ciprofloxacin, ceftriaxone, kanamycin, tetracycline. A comparison of this results with those of previous works on similar dairy products revealed high levels of resistance to antimicrobials which needs to be addressed.

Recent advances in DNA sequencing technology have shown that the human milk microbiota of healthy women varies substantially. The gDNA extraction method may influence the observed variation, biasing the microbiological reconstruction after all. In this study, a genomic DNA extraction method for DNA isolation from human milk samples was standardized and compared with commercial and standard hose make methods. Spectrophotometric measurements, gel electrophoresis, and PCR amplifications were used as criteria for evaluating the quantity, quality, and functionality of the extracted DNA. Furthermore, the standardized method of extracting gDNA from human milk was evaluated for its ability to isolate functional DNA from gram-positive, and gram-negative bacteria and fungi, to improve the reconstruction of microbiological profiles. The novel DNA extraction method increased the quantity and quality of the gDNA extracted compared with commercial and standard house-make protocols. This method even allowed PCR amplification of the V3-V4 regions of the 16S ribosomal gene in all samples, and the ITS-1 region of the fungal 18S ribosomal gene in 95 % of the samples as well. It is concluded that the proposed method provides better performance for the extraction of gDNA from complex samples such as human milk.

The present study was conducted to estimate the exposure and risk characterization of the children population of Serbia to Aflatoxin M1 (AFM1) from milk and milk-based food. A total of 3404 samples comprising of milk and different milk-based food samples were collected from various regions of Serbia from 2017 to 2019. Evaluation of the AFM1 exposure was carried out using the deterministic method, whereas risk characterization was evaluated with the calculation of the Margin of Exposure (MOE) and the risk of Hepatocellular Carcinoma (HCC). Detection rates for AFM1 in milk and milk-based food samples ranged between 2% and 79%, with the highest incidence (79%) and mean level (22.34  0,018 ng kg-1) of AFM1 was detected in pasteurized and UHT milk. According to the three consumption estimates, the values of EDI were higher for toddlers as compared with children. Children aged 1–3 years had the highest risk of exposure to AFM1 in milk, with an estimated daily intake of 0.164 and 0.193 ng kg-1 bw day-1 using a lower bound (LB) and the upper bound (UB) exposure scenarios, respectively. Such difference could result from the higher consumption versus weight. Based on the EDI found in this study, the risk of AFM1 due to consumption of milk and milk-based food was low since MOE values obtained were 10000. In addition, the risk of HCC cases/year/105 individuals of different age groups showed that the value of HCC using potency estimates of 0.0017 (mean) was maximum (0.00034) in the age group 1–3 years which indicates no health risk for the evaluated groups. The present study revealed the importance of controlling and preventing AFM1 contamination in milk through continuous monitoring and regular inspection to reduce the risk of AFM1 exposure, especially in children.

Fucosylated carbohydrates and glycoproteins from human breast milk are essential for the development of the gut microbiota in early life because they are selectively metabolized by bifidobacteria. In this regard, α-L-fucosidases play a key role in this successful bifidobacterial colonization allowing the utilization of these substrates. Although a considerable number of α-L-fucosidases from bifidobacteria have been identified by computational analysis, only a few of them have been characterized. Hitherto, α-L-fucosidases are classified into 3 families, GH29, GH95 and GH151 based on their catalytic structure. However, bifidobacterial α-L-fucosidases belonging to a particular family show significant differences in their sequence. Because this fact could underlie distinct phylogenetic evolves, here extensive similarity searches and comparative analyses of the bifidobacterial α-L-fucosidases identified were carried out with the assistance of previous physicochemical studies available. This work reveals 4 and 2 paralogue bifidobacterial fucosidase groups within GH29 and GH95 families, respectively. Moreover, Bifidobacterium logum subsp. infantis species exhibited the greatest number of phylogenetic lineages in their fucosidases clustered in every family GH29, GH95 and GH151. Since α-L-fucosidases phylogenetically descended from other glycosyl hydrolase families, we hypothesized that could exhibit additional glycosidase activities other than fucosidase, raising the possibility about their application to transfucosylate other substrates than lactose in order to synthesis novel prebiotics.

The prevalence of type-2 diabetes mellitus (T2D) is increasing worldwide and there are no long-term preventive strategies to stop this growth. Emerging research shows that perturbations in the gut microbiome significantly contribute to the development of T2D, while microbiome modulators may be beneficial for T2D prevention. However, microbiome modulators that are effective, safe, affordable, and able to be integrated daily in the diet are not yet available. Based on our previous pro- and prebiotic studies, we developed a novel synbiotic yogurt comprised of human-origin probiotics and plant-based prebiotics and investigated its impact on diet- and streptozotocin-induced T2D in mice. We compared the effects of our synbiotic yogurt to those of a commercially-available yogurt (control yogurt). Interestingly, we found that feeding of this synbiotic yogurt significantly reduced the development of hyperglycemia (diabetes) in response to high-fat diet feeding and streptozotocin compared to milk-fed controls. Surprisingly, the control yogurt exacerbated diabetes progression. Synbiotic yogurt beneficially modulated the composition of gut microbiota compared to milk; conversely, the control yogurt negatively modulated the gut microbiota by significantly increasing the abundance of detrimental bacteria like Proteobacteria and Enterobacteriaceae. In addition, the synbiotic yogurt protected intact pancreatic islet morphology compared to the milk control, while the commercial yogurt demonstrated worse effects on pancreatic physiology. These results suggest that our newly developed synbiotic yogurt protects against diabetes in mice and can be used as a modality to prevent diabetes progression.

The objective of this study was to evaluate biomarkers of heat stress (HS) from automatic milking system (AMS), the relationships between measurements of temperature-humidity index (THI) and reticulorumen ph and temperature and some automatic milking systems parameters in dairy cows (rumination time (RT), milk traits, body weight (BW) and consumption of concentrate (CC)) during summer period. The experiment was carried out on a dairy farm at 54.9587408, 23.784146. Lithuanian Black and White dairy cows (n=365) were selected. The cows were milked with Lely Astronaut® A3 milking robots with free traffic. The rations were calculated to meet physiological requirements of the animals. Daily milk yield, rumination time, body weight, milk fat and protein ratio were collected from the Lely T4C management program for analysis. The pH and temperature of the contents of cow reticulorumen were measured using specific smaX-tec boluses manufactured for animal care. The daily humidity and air temperature in the farm zone were obtained from the adjacent weather station (2 km away). According this study during HS, the higher THI had positive correlation with ML, which show tendencies to increase risk of mastitis, and decrease CC, RT, BW, MY, reticulorumen ph and F/P. Some biomarkers of HS can be milk yield, milk lactose, somatic cell count, concentrate conception, rumination time, body weight, reticulorumen ph and milk fat – protein ratio. We can recommend to monitoring these parameters in the herd management program to identify possibility of heat stress.

Sustainable innovation in the agro-food system has become a strategy increasingly used by companies as a means to increase their competitiveness and position themselves in the market. In this context, the objective of this work is to identify the attitudes and sensory perceptions of consumers towards sustainable food technology through two scales (Food Technology Neophobia and Domain Specific Innovativeness) and hedonic tastings. For this, a new product was selected in the market: powder to prepare rice with milk. Most consumers have attitudes toward low neophobia to products with food technology, but there is also some caution, situation that is corroborated by the moderate predisposition towards innovations. Color and flavor attributes can make the difference in positive perceptions. It should not be forgotten that there is a segment of innovative sustainable consumers that represent a key market.

Microplastics are small plastic pieces with sizes less than 5 mm. They are becoming a global concern due to the potential risk to human health. The potential risks of microplastics may be greater for infants because they do not have sufficiently developed metabolizing enzymes, have a lower ability to remove them, high sensitivity of target organs. Infants should be breastfed for the first six months of life. Breast milk is considered to be the most complete and suitable source of nutrition. However, if breastfeeding during this period is not possible, it is necessary to use formulas designed for infant initial feeding. Infants may be exposed to higher levels of MPs through infant foods or plastics products. The aim of this study is to describe the possible sources of exposure to microplastics such as human placenta, plastic feeding bottles, toys as well as the presence of released microplastics in infants feces, breast milk and infant formulas. There is still not enough data available from the studied area. Therefore, it is necessary to pay increased attention to minimizing the negative effects of microplastics on human health.

Background: Functional beverages are ordinary foods with components or ingredients able to provide specific health effects other than purely nutritional effect. Aim: This study aims to formulate a new functional beverage obtained from the recovery of YoAlp® whey, a bovine fermented milk obtained using autochthonous starter cultures, added with Renetta Canada/Golden Delicious-Raspberry (YWL), Renetta Canada-Aronia (YWA) and Ravèntse (YWR) fruit juices to improve the functionality of the beverage. Methods: Microbiological analysis and chemical characterization, by a proteomic approach and spectrophotometric microassay, have been conducted on beverages to investigate the expected shelf-life and potential health promoting effects. Results: The microbiological assessment confirmed the safety of the beverages and the probiotic bacteria vitality over a shelf life of seven days. Biochemical analysis performed highlight the presence of different bioactive peptides (ACE inhibitory and antioxidant), and the betacasomorphin 9 (BCM-9), marker of A2 variant of β-casein typically present in Aosta Valley cattle breeds. Moreover, high amounts of total polyphenols, which concur to the antioxidant activity values observed, have been found. Furthermore, ACE inhibitory activity is well correlated to the bioactive peptides detected. Finally, a consumer test confirmed the YoAlp® whey-based beverages appreciation. Conclusions: The production of these beverages could represent a huge opportunity for Aosta Valley’s farms to increase their income and competitiveness, creating a circular economy by recovering a by-product like whey and following sustainability and healthy trends, which are driving the food sector.

Given the broad and intense use of plastic, society is being increasingly affected by its degradation and by-products, particularly by microplastics (MPs) -fragments smaller than 5 mm in size-, and nanoplastics (NPs) -with sizes less than 1 µm. MPs and NPs may enter the body primarily through inhalation, consumption, and skin contact. Once ingested, MPs can penetrate tissues, deviating to other parts of the body, and hence potentially affecting important cellular pathways such as nonconforming chemokine receptors that control the communication between the fetus and the mother. Consequently, the potential health harm induced by MP internalization is a major issue, evidenced by multiple studies demonstrating harmful consequences in diverse animal models and human cells. Here, an overview of the various modes of exposure to MPs and NPs is presented, including inhalation, placental transfer, ingestion, breastmilk consumption, and skin absorption, as well as placental and fetal toxicity due to plastic particles based on animal and in vitro studies. Though MPs in our environment are becoming more recognized, their developmental toxicity is still scarcely known. Besides negatively affecting pregnancy, MPs and NPs have been shown to potentially harm the developing fetus, given their ability to cross the placental barrier. Still, considerable gaps remain in our understanding of the dispersion and toxicity of these particles in the environment, as well as the precise types of NPs and MPs bearing the greatest dangers. As a result, we advocate for larger-scale epidemiological investigations, the development of novel approaches for measuring NP and MP exposures, and the necessity of understanding the toxicity of various kinds of NPs to guide future research efforts.

The quality and safety of fermented milk products, which make up a sizeable portion of human nutrition, are greatly influenced by both the milk itself and the starting cultures employed to ferment it. Lactic acid bacteria (LAB) create a variety of metabolites throughout the fermentation process, altering the organoleptic properties of the substrates. The final product’s nutritional value and digestibility are enhanced by fermentation of the raw materials, which also adds vitamins, vital amino acids, and fatty acids. The quality and safety of fermented milk products, which make up a sizeable portion of human nutrition, are greatly influenced by both the milk itself and the starting cultures employed to ferment it. The inhibition of the growth of pathogenic microbes, which lowers the likelihood of the emergence of foodborne diseases, is another crucial function of LAB. A variety of pathogenic bacteria are frequently present in raw (unpasteurized) milk, and other raw materials; these pathogenic bacteria should be removed during the fermentation process. As a result, a variety of LAB metabolites, such as hydrogen peroxide, organic acids, and bacteriocins, function as bio preservative agents, enhancing food safety and lengthening the shelf life of the finished products.

Food allergy has been recognised by the World Health Organisation as a chronic disease that could lead to fatality. The symptoms can mediate from mild to severe consequences. Some cases have reported that food allergy sufferers are bullied and are discriminated at workplace, thus increasing their stress level, apart from facing difficulty in consuming certain food due to allergy. This has an adverse effect on the quality of life amongst food allergy sufferers. Medical bills are increased when food allergy sufferers become unknowingly exposed to food allergen. Hence, this study investigated food allergens among food allergy sufferers in order to make their lives better. A total of 250 respondents participated in an online survey. Surprisingly, apart from chicken, soybean and other food allergens that have been listed under the Big Eight Allergens, the respondents mentioned some fruits that could trigger allergic reactions, such as rambutans and watermelons. Food allergy sufferers can be allergic to seafood and seafood products as well, such as shrimp paste or locally known as belacan, which refers to fermented shrimp paste cured under the sun. It is a common food ingredient used among Malaysians. The study findings offer practical insights, such as avoidance of food allergens amidst food allergy sufferers and help the government to carefully manage for menu planning. The knowledge contribution of this study reveals several specific food and fruits, as aforementioned, which appear to be common and nutritious food, yet harmful to food allergy sufferers.

The first 1000 days of life is a critical period that contributes significantly to the programming of an individual's future health. Among the many changes that occur during this period, there is growing evidence that the establishment of a healthy gut microbiota early in life plays an important role in the prevention of both short- and long-term health problems. Numerous publications suggest that the quality of gut microbiota colonization depends on several dietary factors, including breastfeeding. In this respect, a relationship between breastfeeding and the risk of inflammatory bowel disease (IBD) has been suggested. IBD are chronic intestinal diseases in which perinatal factors may be partly responsible for its onset. We propose to review the existence of links between breastfeeding and IBD, based on experimental and clinical studies. Overall, despite encouraging experimental data in rodents, the association between breastfeeding and the development of IBD remains controversial in humans, partly due to considerable heterogeneity between clinical studies. The duration of exclusive breastfeeding is probably decisive for its lasting effect on IBD. Thus, specific improvements in our knowledge could support dietary interventions targeting the gut microbiome, such as the early use of prebiotics, probiotics or postbiotics in order to prevent the disease.

A nanostructured electrochemical bi-sensor system for analysis of milks has been developed using the Layer by Layer technique. The non-enzymatic sensor [CHI+IL/CuPcS]2, is a layered material containing a negative film of the anionic sulfonated copper phthalocyanine (CuPcS) acting as electrocatalytic material, and a cationic layer containing a mixture of an ionic liquid (IL) (1-butyl-3-methylimidazolium tetrafluoroborate) that enhances the conductivity and chitosan (CHI) that facilitates the enzyme immobilization. The biosensor ([CHI+IL/CuPcS]2-GAO) results from the immobilization of galactose oxidase on the top of the LbL layers. FTIR, UV-vis and AFM have confirmed the proposed structure and cyclic voltammetry has demonstrated the amplification caused by the combination of materials in the film. Sensors have been combined to form an electronic tongue for milk analysis. Principal Component Analysis has revealed the ability of the sensor system to discriminate between milk samples with different lactose content. Using PLS-1 calibration models, correlations have been found between the voltammetric signals and chemical parameters measured by classical methods. PLS-1 models provide excellent correlations with lactose content. Additional information about other components such as fats, proteins and acidity can also be obtained. The method developed is simple and the short response time permits its use in assaying milk samples on-line.

The search of DNA polymorphisms useful for the genetic improvement of dairy farm animals lasts more than 40 years with relevant findings in cattle for milk traits, where the best combination of alleles for dairy processing have been found in casein genes and in the DGAT1. Nowadays, similar results are not reached yet in river buffaloes despite advanced genomic technologies and accurate phenotype records are available. The aim of the present study was to investigate and validate in a larger buffalo population the effect on six milk traits of four single nucleotide polymorphisms (SNP) in the CSN1S1, CSN3, SCD and LPL genes, previously reported to be associated with or affect dairy traits in smaller populations often belonging to one farm. A total of 800 buffaloes were genotyped. Daily milk yield (dMY, kg), protein yield (dPY, kg) and fat yields (dFY, kg), fat and protein contents (dFP, % and dPP, %), somatic cell count (SCC, 103cell/ml) and urea (mg/dl) were individually recorded in each month for the whole lactation from 2010 to 2021. A total of 15,742 individual milk test day records (2,496 lactations) were available on 680 buffalo cows with 3.6±1.7 parity (from 1 to 13) and 6.1±1.2 test day records per lactation on average. Three out four SNP in CSN1S1, CSN3 and LPL were associated with at least one of analyzed traits. In particular, the CSN1S1 (AJ005430:c.578C&gt;T) gave favorable associations with all yield traits (dMY, p=0.022; dPY, p=0.014 ; dFY, p=0.029) and SCS (p=0.032), whereas the CSN3 (HQ677596: c.536C&gt;T) positively associated with SCS (p=0.005) and milk urea (p=0.04). Favorable effect on dMY (p=0.028), dFP (p=0.027) and dPP (p=0.050) were observed for the LPL. Conversely, SCD did not show any association with milk traits. This is the first example of confirmation study carried out in the Mediterranean river buffalo for genes of economic interest in dairy field and it represents a very important indication for the preselection of young bulls destined to breeding programs in the view of a more sustainable dairy production.

With the increasing interest in CM as a potential source of nutrients and bioactive compounds, there arises a necessity to delve into methods for its processing and drying. Over recent decades, a significant amount of research has been conducted on the chemical composition of CM and its potential health benefits. Consequently, this study is directed towards examining CM drying technology, including an analysis of the influence of different drying methods on its chemical composition and nutri-tional properties. The objective of this review is to systematically summarize the available literature on CM drying processes and their impact on its chemical composition. Special attention is devoted to comparing various drying methods such as freeze-drying, spray drying, and freeze-drying, as well as investigating factors affecting the quality of dried CM. To achieve this goal, we scrutinize a series of studies published between 2003 and 2022 to identify optimal drying methods that promote the maximum preservation of CM's nutritional components and bioactive compounds. Additionally, we explore possibilities for reducing drying time to enhance production efficiency. This research is grounded in the analysis of data from diverse scientific research databases, including Scopus and Google Scholar, encompassing a systematic review of scientific literature and a comparative analysis of the obtained results.

Several animal food allergens assembly into amyloids under gastric-like environments. These aggregated structures provide Gad m 1 with an enhanced IgE interaction due to the amyloid assembly of the epitope regions. However, whether these properties are unique of Gad m 1 or common to other food allergens has not yet been addressed. Using Bos d 5, Bos d 12 and Gal d 2 as food allergen models and Gad m 1 as control, aggregation reactions and the sera of milk, egg and fish allergic patients we have analyzed the IgE interaction of the distinct amyloids. We found that amyloids formed by Bos d 12 and Gal d 2 full-length and truncated chains are recognized by the IgEs of milk and egg allergic patient sera. As with Gad m 1, in most cases amyloid recognition is higher than that of the precursor structure. Bos d 5 was not recognized under any fold by the IgE of the sera studied. These results support that formation of IgE-binding amyloids might be a common feature to animal food allergen.

Heat-stressed lactating dairy cattle exhibit unique metabolic symptoms; many of which are undoubtedly involved in heat-induced subfertility. Because of its known systemic effects, we hypothesized that γ-aminobutyric acid (GABA) participates in the regulation of insulin and progesterone during heat stress. Multiparous lactating Holstein cows (n=6) were studied during four experimental periods: 1) thermoneutral (TN; d 1-5), 2) TN + hyperinsulinemic-hypoglycemic clamp (d 6-10), 3) heat stress (HS; d 16-20), and 4) HS + euglycemic clamp (d 21-25). Blood samples were collected once daily via coccygeal venipuncture into heparinized evacuated tubes. Analysis of GABA concentrations from all four treatment periods yielded no differences. In direct comparison to TN concentrations, plasma GABA tended to be decreased during HS (16.57±2.64 vs 13.87±2.28 ng/ml, respectively, P = 0.06). Both milk production and plasma insulin were moderately correlated with plasma GABA (r=0.35, P<0.01; r = -0.32, P<0.01). Plasma progesterone was correlated with plasma GABA concentrations during TN but not HS. These results are the first to indicate that peripheral GABA could be involved in regulating aspects of production and reproduction during heat stress. More research is needed to determine its precise role(s).

Management of calcium and phosphate in biofluids is key to maintaining physiological mineral homeostasis (i.e., appropriate mineralization of hard tissues and an absence of mineral deposition in soft tissues). This review describes and contrasts the ways vertebrates manage calcium phosphate in two biological fluids (breast milk and serum) and illustrates the benefits of mineral sequestration by proteins. In milk, phosphoprotein-sequestered calcium magnesium phosphates provide nutritional support, whereas in serum, protein-sequestered calcium phosphates control transport and delivery of calcium and phosphate to tissues for biological function or excretion. In addition, subsets of sequestered phosphates in serum have been identified as culprits underlying ectopic deposition of calcium phosphates and toxicity.

Background: Little is known about the natural history of immunoglobulin (Ig) E-mediated diseases in preterm infants, further hampered by the lack of noninvasive investigations. We aimed at developing a non-invasive tool for the study of IgE and eosinophil-derived neurotoxin (EDN) in neonatal fecal samples and evaluating its predictive value for the development of IgE-mediated diseases (cow’s milk allergy, asthma, or atopic dermatitis) during the first year of life. Methods: We developed a stool extraction protocol, followed by freeze-drying and solubilization. sIgE responses were investigated in neonatal fecal samples from 21 preterm infants with a 300-allergen multiplex containing whole and molecular allergens and confirmed by capillary Western blot with nano-immunoassay. The local eosinophilic component was investigated by measuring the concentration of EDN. Results: The multiplexed allergen assay detected sIgE in all samples. Confirmation was obtained with Western blot. Frequency and levels of sIgE in neonatal fecal samples differed between infants who developed IgE-mediated diseases and controls. Neonatal fecal sIgE directed to milk proteins predicted later development of cow’s milk allergy (specificity 88%, sensitivity 78%). Allergen specificity of neonatal fecal sIgE was associated with later development of cow’s milk allergy and asthma. Neonatal fecal EDN levels predicted the development of IgE-mediated diseases (sensitivity 100%, specificity 75%). Conclusion: Non-invasive investigation of neonatal fecal sIgE is a promising tool for the prediction of subsequent development of IgE-mediated diseases.

The Australian dairy sheep industry is small and mostly based on a natural grass grazing system which can limit productivity. The current study tested different plant oil-infused and rumen protected polyunsaturated fats and their interactions with sire breeds to improve lactation traits and body condition score (BCS) of ewes grazing low quality pastures. It was hypothesised that supplementing lactating ewe diets plant-derived polyunsaturated oils will improve milk production and composition without compromising BCS. Sixty ewes (n=10/treatment) in mid-lactation, balanced by sire breed, parity, milk yield, body condition score, and liveweight were supplemented with: 1) control: wheat-based pellets without oil inclusion; wheat-based pellets including 2) canola oil (CO); 3) rice bran oil (RBO); 4) flaxseed oil (FSO), 5); safflower oil (SFO) and 6) rumen protected fat containing eicosapentaenoic acid and docosahexaenoic acid (RPO). Except for the control group, all supplementary diets included the same level of 50 ml/kg DM of oil and all diets were isocaloric and isonitrogenous. Experimental animals were grazed in the same paddock with ad libitum access to pasture, hay and water during the 10-week study. RPO was the most effective diet that enhanced milk, fat and protein yields by approximately 30, 13, and 31% respectively (P<0.0001). Significant increase in milk production was also observed in CO, RBO, and SFO (P<0.0001). Breed significantly influenced animal performance with higher milk yield recorded for crossbred Awassi x East Friesian (AW x EF) (578 g/day) vs purebred Awassi (452 g/day) (P<0.0001). This study provides empirical evidence for the use of rumen-protected and plant-derived oil-infused pellets as supplements under low quality pasture grazing conditions, to improve production performance of purebred Awassi and crossbred AW x EF ewes.

This study investigated the presence, distribution, and antimicrobial resistance profiles of extend-ed-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae in a dairy herd located in Northern Italy. A questionnaire was administered, and the feces of clinically healthy pre-weaned dairy calves, their mothers, and the cows treated for mastitis, waste milk, water, and environmental samples, were collected and inoculated on Chromagar ESBL™ medium. The questionnaire identi-fied several risk factors including administration of waste milk to male calves, lack of routine cleaning of calf feeding and housing equipment, and blanket dry cow therapy. Escherichia coli was identified by MALDI-TOF MS in the feces of 28 of 37 (75.67%) calves, 2 of 3 (66.67%) treated cows, 8 of 14 (57.15%) environmental samples, and the waste milk. Six of 14 (42.85%) environmental samples were also positive for ESBL Acinetobacter baumannii, while all fecal samples were negative. ESBL production was confirmed for all E. coli isolates based on the double disk susceptibility test (DDST), while none were carbapenemase producers. Upon minimal inhibitory concentration (MIC) analysis, all of them showed multiple resistances and were categorized as multi-drug resistant (MDR). This study confirms the widespread presence of MDR, ESBL-E. coli in dairy calves and highlights their diffusion in the farm environment.

Background and Objectives: Acute lung injury (ALI) is an inflammatory response in the lung caused by bacteria, viruses, trauma and other factors that results in lack of alveolar surfactant and subsequently respiratory distress. Endothelial-to-mesenchymal transition (EndoMT) is closely related to ALI-induced pulmonary fibrosis development and progression. This study examined the effect of milk fat globulin factor 8 (MFGE8) knockdown and overexpression on lipopolysaccharide (LPS)-induced EndoMT. Methods: Total MFGE8 activity in patients with acute respiratory distress syndrome (ARDS) and healthy volunteers was assessed using a colorimetric kit. In vitro, cell morphology was observed by microscopy, and invasion and migration were tested by Transwell and scratch assays. Specific siRNAs were transfected into HLMECs to downregulate MFGE8, and CD31 and α-SMA protein expression was detected by fluorescence microscopy and FCM. MFGE8, BMP, Smad 1, Smad4, Smad5, Snail, CD31 and α-SMA protein and gene expression was assessed by western blotting (WB) and qRT‒PCR. In vivo, the changes in cell morphology and alveolar septum in the lung of mice (C57BL/6, aged 7-8 weeks, male) were observed by H&E staining. In addition, ECM deposition in lung tissue was detected by Masson staining. CD31 and α-SMA protein expression in lung tissue was measured by WB. Results: The serum level of MFGE8 was lower in the ARDS group than in the control group. MFGE8 was identified as a protective factor associated with hospital survival. In vitro, the treatment of human lung microvascular endothelial cells (HLMECs) with LPS for 96 h clearly changed the cells from the typical cobblestone shape of ECs to spindle-like fibroblasts. The scratch and Transwell assay results indicated that endothelial cell migration and invasion were enhanced in the LPS group compared with the control group. Fluorescence microscopy, FCM, WB and PCR showed that LPS greatly inhibited CD31 (endothelial marker) expression and increased α-SMA (mesenchymal marker) expression. These data showed that LPS could induce EndoMT in HLMECs. Notably, siRNA-treated HLMECs yielded the same results. The administration of rhMFGE8 to HLMECs in the LPS or LPS+siRNA group ameliorated the changes in cell morphology and decreased cell migration and invasion. rhMFGE8 attenuated the effect of LPS or LPS+siRNA on EndoMT induction by increasing CD31 and decreasing α-SMA protein and gene expression. Moreover, activation of BMP/Smad1/5-Smad4 signalling in response to LPS and Snail (related to EndoMT transcription factors) expression were increased by MFGE8 knockdown but inhibited by rhMFGE8. In vivo, H&E staining revealed a thickened alveolar septum in the LPS group, and the thickness increased over time; in contrast, rhMFGE8 reversed this effect. ECM deposition occurred early in ALI induced by LPS and increased over time, and the administration of rhMFGE8 reversed this effect. WB showed that LPS inhibited CD31 protein expression and increased α-SMA protein expression in lung tissue. Conclusions: rhMFGE8 exerts a protective effect early in LPS-induced EndoMT through BMP/Smad1/5/Smad4 signalling and could be a therapeutic target in ALI.

Lactoperoxidase (LP), which is a high-value minor whey protein, has recently drawn extensive attention from research scientists and industry due to its multi-function and potential therapeutic applications. In this study, the separation and optimization of two similar-sized proteins, LP and lactoferrin (LF) were investigated using strong cation exchange column chromatography. Optimization was started with central composite design based experiments to characterize the importance of different decision variables. The three variables used in the optimization were flow rate, length of gradient and final salt concentration in the linear elution gradient step. The obtained empiric functional model represented the effect of the significant factors on the yield as the objective function. Afterwards, the calibrated mechanistic model was employed to predict accurate optimal set of variables. The optimal operating points were found and the results were compared with validation experiments. Predictions respecting yield confirmed a very good agreement with experimental results while keeping purity, a product quality characteristic, equal or above to a predefined value.

To evaluate the behaviour of the relevant microbial populations during stretched cheese production, the quantitative microbiological analysis was performed during the critical steps of the preparation. The obtained data distributions proved statistically significant increases in all indicators, on average by 4.55 ± 0.64 log CFU/g of presumptive lactococci counts, 4.06 ±0.61 of lactobacilli, 1.53 ± 0.57 log CFU/g of coliforms, 2.42 ± 0.67 log CFU/g of Escherichia coli, 1.53 ± 0.75 log CFU/g of yeasts and moulds, and 0.99 ± 0.27 log CFU/g of presumptive Staphylococcus aureus, from the early stage of milk coagulation until curd ripening (0–24 h). The following steaming/stretching process caused reductions in viable counts with the most significant inactivation effect on coliform bacteria, including E. coli (-4.0 ± 1.0 log CFU/g). Total viable counts and yeasts and moulds showed 2 and almost 3 log reduction (-2.2 ± 1.1 log CFU/g and -2.6 ± 0.9 log CFU/g), respectively. The lowest decreases in presumptive S. aureus counts were estimated at the level of -1.50 ± 0.64 log CFU/g. The counts of yeasts and moulds showed the best indicatory function during the entire storage period of vacuum-packaged cheeses at 6 °C.

Background: Glucose 6-phosphate dehydrogenase (G6PD) deficiency increases the risk of neonatal hyperbilirubinemia. The aim of the study is to evaluate the clinical findings of glucose 6 phosphate dehydrogenase (G6PD) and pyruvate kinase (PK) deficiency, to prevent further examinations for prolonged jaundice, and to contribute by regulating its treatment. At the same time, it is to determine whether systemic-immune-inflammation index (SII), neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) can be used in addition to conventional complete blood count (CBC) parameters in the diagnosis of neonatal prolonged jaundice. Methods: In full term neonates with hyperbilirubinemia who were admitted to Medicine Hospital between January 2019 and January 2024 with the complaint of jaundice, 167 infants with a serum bilirubin level above 5 mg/dL, whose jaundice persisted after the 10th day, were included in the study. Results: 63.5% of the patients were male and the mean age was 31±6 days. G6PD was negatively perfectly correlated with NLR (r: -1; p

Automatic milking systems (AMSs) are among the earliest Precision Livestock Farming developments that have transformed dairy farming worldwide. This review aims to gather, evaluate, and summarize papers that focus on the use of modeling approaches in the context of AMS. We provided a review of 60 articles with a specific focus on cows’ health, production, and behavior/management. The most used modeling approach was Machine Learning (ML, present in 63% of the studies), followed by statistical analysis (14%), fuzzy algorithms (9%), deterministic mod-els (7%), and detection algorithms (7%). Most of the reviewed studies (82%) focused on the detection of cows' health, specifically mastitis, while only 11% were concerned with milk production. Accurate forecasting of dairy cow milk yield and knowledge on the deviation between expected and observed milk yields of individual cows would be beneficial in dairy cow management. Likewise, the study of cows’ behavior and the herd management in AMSs is under-explored (7%). Despite the increasing use of ML techniques in this field there is still a lack of a robust methodology for their application. In particular, we identified a significant gap in the systematic balancing of positive and negative classes for health prediction models.

Background: allergic diseases and, in particular food allergy (FA), have been identified as one of the major diseases in the world. The first 1000 days of life can be a “window of opportunity” or a “window of susceptibility” during which several factors can predispose children to FA development. Changes in gut microbiota composition, from pregnancy to infancy, could have a pivotal role in this regard: some bacterial genus such as Lactobacillus or Bifidobacterium seem to be protective from FA development, on the contrary Clostridium and Staphylococcus appear to be unprotective. Methods: we conducted a research through the most recent literature (2013-2023), using Pubmed and Scopus as databases. We included English original papers, clinical trials, metanalysis and reviews. Case reports, series and letters have been excluded. Results: during pregnancy, maternal diet can play a fundamental role in influencing gut microbiota composition of newborns. After birth, human milk can promote the development of protective microbial species by human milk oligosaccharides (HMOs), that play a prebiotic role. Also complementary feeding can modify gut microbiota composition. Conclusions: the first two years of life are a critical period where several factors, in genetic predisposed children, can increase the risk of FA development.