Raw milk kefir: microbiota, bioactive peptides, and immune modulation.

This study aims to characterize the microbiota and peptidomic composition of raw milk kefir, and to address the potential anti-allergic effects of raw milk kefir using validated research models for food allergy. Raw milk kefir was produced after incubation with a defined freeze-dried starter culture. Kefir was sampled during fermentation at seven time intervals. For comparison, kefir was also prepared from heat-treated milk. Peptide compositions were determined for the raw and heated milk, and kefir end products made from these milks. In a murine food allergy model, the two kefir end products were investigated for their allergy modulating effects. In both kefirs, we identified amplicon sequence variants identical to those in the starter culture, matching the bacteria Lactococcus lactis, Streptococcus thermophilus, Leuconostoc and the yeast Debaryomyces. In raw milk kefir, additional sequence variants of Lactococcus lactis and the yeasts Pichia and Galactomyces could be identified, which were absent in heated milk kefir. Analysis of peptide compositions in both kefirs indicated that the number and intensity of peptides drastically increased after fermentation. Heating of the milk negatively affected the diversity of the peptide composition in kefir. Only raw milk kefir suppressed the acute allergic skin response to the food allergen ovalbumin in sensitised mice. These effects coincided with differences in the T-cell compartment, with lower percentages of activated Th1 cells and IFNg production after treatment with kefir made from heated milk. The results of this study indicate specific properties of raw milk kefir that may contribute to its additional health benefits.

Early detection of mastitis in cows using the system based on 3D motions detectors.

Mastitis is one of the major health problems in dairy herds leading to a reduction in the leading to a reduction in the quality of milk and economic losses. The research aimed to present the system, which uses electronic 3D motion detectors to detect the early symptoms of mastitis. The system would allow more effective prevention of this illness. The experiment was carried out on 118 cows (64 Holstein Friesian and 54 Brown Swiss). The animals were kept in free-stall barn with access to pasture. The occurrence of mastitis cases was noticed in veterinary register. Microbiological culture was taken from milk in order to confirm the development of infection. Data from motion detectors were defined as time spent by animals on feed intake, ruminating, physical activity and rest, and were expanded by adding information about feeding group, breed type and lactation number. During analyses, two approaches were used to process the same dataset: artificial neural networks (ANN) and logistic regression. The obtained ANN and the logistic regression models proved to be satisfactory from the perspective of applied criteria of goodness of fit (area under curve-exceed 0.8). Quality parameters (accuracy, sensitivity and specifity) of logistic regression are relatively high (larger than 0.73), whereas the ranks of significance of the studied variables varied across datasets. These proposed models can be useful for automating the detection of mastitis once integrated into the farm's IT system.

Diagnosis and Rationale for Action against Cow's Milk Allergy (DRACMA) Guidelines update - III - Cow's milk allergens and mechanisms triggering immune activation.

Background: The immunopathogenesis of cow's milk protein allergy (CMPA) is based on different mechanisms related to immune recognition of protein epitopes, which are affected by industrial processing. Purpose: The purpose of this WAO DRACMA paper is to: (i) give a comprehensive overview of milk protein allergens, (ii) to review their immunogenicity and allergenicity in the context of industrial processing, and (iii) to review the milk-related immune mechanisms triggering IgE-mediated immediate type hypersensitivity reactions, mixed reactions and non-IgE mediated hypersensitivities. Results: The main cow's milk allergens - α-lactalbumin, ß-lactoglobulin, serum albumin, caseins, bovine serum albumins, and others - may determine allergic reactions through a range of mechanisms. All marketed milk and milk products have undergone industrial processing that involves heating, filtration, and defatting. Milk processing results in structural changes of immunomodulatory proteins, leads to a loss of lipophilic compounds in the matrix, and hence to a higher allergenicity of industrially processed milk products. Thereby, the tolerogenic capacity of raw farm milk, associated with the whey proteins α-lactalbumin and ß-lactoglobulin and their lipophilic ligands, is lost. Conclusion: The spectrum of immunopathogenic mechanisms underlying cow's milk allergy (CMA) is wide. Unprocessed, fresh cow's milk, like human breast milk, contains various tolerogenic factors that are impaired by industrial processing. Further studies focusing on the immunological consequences of milk processing are warranted to understand on a molecular basis to what extent processing procedures make single milk compounds into allergens.

Loss of allergy-protective capacity of raw cow's milk after heat treatment coincides with loss of immunologically active whey proteins.

The allergy-protective capacity of raw cow's milk was demonstrated to be abolished after heat treatment. The heat-sensitive whey protein fraction of raw milk is often implied to be the source of this allergy-protective effect, but a direct link between these proteins and the protection against allergic diseases is missing. This study therefore aimed at investigating the mechanistic relation between heat damage to whey proteins and allergy development. Raw cow's milk was heated for 30 min at 50, 60, 65, 70, 75, or 80 °C and the native whey protein profile of these differentially heated milk samples was determined using LC-MS/MS-based proteomics. Changes in the native protein profile were subsequently related to the capacity of these milk samples to prevent the development of ovalbumin-induced food allergy in a murine animal model. A substantial loss of native whey proteins, as well as extensive protein aggregation, was observed from 75 °C. However, whey proteins with immune-related functionalities already started to denature from 65 °C, which coincided with the temperature at which a loss of allergy protection was observed in the murine model. Complement C7, monocyte differentiation antigen CD14, and polymeric immunoglobulin receptor concentrations decreased significantly at this temperature, although several other immunologically active whey proteins also showed a decrease around 65 °C. The current study demonstrates that immunologically active whey proteins that denature around 65 °C are of importance for the allergy-protective capacity of raw cow's milk and thereby provides key knowledge for the development of microbiologically safe alternatives to raw cow's milk.

Development and Validation of the Immune Status Questionnaire (ISQ).

The self-assessment of perceived immune status is important, as this subjective observation leads individuals to decide whether or not to seek medical help or adapt their lifestyle. In addition, it can be used in clinical settings and research. The aim of this series of studies was to develop and validate a short questionnaire to assess perceived immune functioning. Five surveys were conducted among Dutch and International young healthy adults (18-30 years old), and two others among older age groups with various health complaints. For the first study, an existing immune functioning scale was modified and elaborated resulting in 23 immune-health-related items, of which the occurrence was rated on a 5-point Likert scale. A student sample was surveyed, and the results were used to shorten the 23-item listing into a 7-item scale with a predictive validity of 85%. Items include "sudden high fever", "diarrhea", "headache", "skin problems (e.g., acne and eczema)", "muscle and joint pain", "common cold" and "coughing". The scale is named Immune Status Questionnaire (ISQ), and it aims to assess perceived immune status over the preceding year. The second study revealed that the ISQ score correlated significantly with a 1-item perceived immune functioning (r = 0.383, p < 0.0001). In the third study, the final Likert scale descriptors were determined ("never", "sometimes", "regularly", "often" and "(almost) always)". The fourth study showed that the test-retest reliability of the ISQ is acceptable (r = 0.80). The fifth study demonstrated the association of ISQ scores with various neuropsychological and health correlates in an international sample, including perceived health and immune fitness, as well as levels of stress, fatigue, depression and anxiety. Study 6 demonstrated significant associations between ISQ scores and experiencing irritable bowel syndrome (IBS) symptoms in a sample of insomnia patients. Study 7 compared the effect of a dietary intervention in participants reporting "poor health" versus "normal health". It is shown that ISQ scores can differentiate between those with poor and normal health, and that an effective intervention is associated with a significant improvement in ISQ scores. Data from Study 7 were further used to determine an ISQ cut-off value for reduced immune functioning, and a direct comparison with 1-item perceived immune functioning scores enabled constructing the final scoring format of the ISQ. In conclusion, the ISQ has appropriate face, content, and construct validity and is a reliable, stable and valid method to assess the past 12 month's perceived immune status.

Effect of raw milk consumption on perceived health, mood and immune functioning among US adults with a poor and normal health: A retrospective questionnaire based study.

INTRODUCTION: Raw milk consumption is controversially discussed, and people are looking for raw milk due to expected positive health impacts. The purpose of this consumer survey was to evaluate health conditions prior to and after consuming of raw milk (RM). METHODS: An on-line survey was distributed in Spring 2018 among existing consumers of raw milk. One-Item health score, 1-item immunity score, immune status (ISQ), mood, bowel and skin conditions were rated retrospectively based on validated questionnaires. Data from 327 participants (age 54 years) were included, of which 156 (48%) were allocated to the poor health group after they reported being immune depressed or suffering from a chronic disease. Others were allocated to the normal health group. Milk consumption pattern before and after changing of the milk diet were recorded. All health outcomes were evaluated according a linear mixed model in SPSS. RESULTS: Health, perceived immunity, bowel and mood scores increased post RM consumption with around 35% in the poor health group (P < 0.001), and around 9% in the normal health group (P < 0.001). Bowel and mood scores were overall lower in women than in men. Outcomes were independent of the origin of raw farm milk. CONCLUSIONS: This consumer survey suggests that positive health and mood changes are associated with the consumption of raw milk. Effects were strongest in people with a self-reported poor health status as well as in women.

Milk processing increases the allergenicity of cow's milk-Preclinical evidence supported by a human proof-of-concept provocation pilot.

BACKGROUND: Several studies demonstrated the adverse effect of milk processing on the allergy-protective capacity of raw cow's milk. Whether milk processing also affects the allergenicity of raw milk is hardly investigated. OBJECTIVE: To assess the allergenicity of raw (unprocessed) and processed cow's milk in a murine model for food allergy as well as in cow's milk allergic children. METHODS: C3H/HeOuJ mice were either sensitized to whole milk (raw cow's milk, heated raw cow's milk or shop milk [store-bought milk]) and challenged with cow's milk protein or they were sensitized and challenged to whey proteins (native or heated). Acute allergic symptoms, mast cell degranulation, allergen-specific IgE levels and cytokine concentrations were determined upon challenge. Cow's milk allergic children were tested in an oral provocation pilot with organic raw and conventional shop milk. RESULTS: Mice sensitized to raw milk showed fewer acute allergic symptoms upon intradermal challenge than mice sensitized to processed milk. The acute allergic skin response was low (103 ± 8.5 µm vs 195 ± 17.7 µm for heated raw milk, P < 0.0001 and vs 149 ± 13.6 µm for shop milk, P = 0.0316), and there were no anaphylactic shock symptoms and no anaphylactic shock-induced drop in body temperature. Moreover, allergen-specific IgE levels and Th2 cytokines were significantly lower in raw milk sensitized mice. Interestingly, the reduced sensitizing capacity was preserved in the isolated native whey protein fraction of raw milk. Besides, native whey protein challenge diminished allergic symptoms in mice sensitized to heated whey proteins. In an oral provocation pilot, cow's milk allergic children tolerated raw milk up to 50 mL, whereas they only tolerated 8.6 ± 5.3 mL shop milk (P = 0.0078). CONCLUSION AND CLINICAL RELEVANCE: This study demonstrates that raw (unprocessed) cow's milk and native whey proteins have a lower allergenicity than their processed counterparts. The preclinical evidence in combination with the human proof-of-concept provocation pilot provides evidence that milk processing negatively influences the allergenicity of milk.

Patterns of Biodynamic Milk Fatty Acid Composition Explained by A Climate-Geographical Approach.

BACKGROUND: Biodynamic dairy production is based on a land-related animal production without the additional input of N-fertilizers. The concentrate level per cow is low. This affects the yield level of animals and product quality outcomes. METHODS: We examined the milk fatty acid (FA) composition of European biodynamic farms in relation to the ecological region of production and the farm's climate conditions. Climate data were derived from existing maps describing ecological vegetation zones within Europe. Additionally, biodynamic shop milk was compared to conventional shop milk, based on a regional comparison. RESULTS: The largest differences in the FA composition were between biodynamic summer and winter milk. We found increased proportions of conjugated linoleic acid (CLA), alpha-linolenic acid (ALA-n3), monounsaturated FA (MUFA), and polyunsaturated FA (PUFA) in the summer milk. A principal component analysis expressed the structure that was present in the biodynamic farm milk samples, based on clusters of a single FA within four components. The components could be correlated with the season of production, the amount of precipitation, the elevation of the farm above sea level, and the length of the grazing season. Biodynamic shop milk in the summer had a lower n6/n3 PUFA ratio compared to the conventional shop milk in all regions of production. Mean values were 1.37 and 1.89, respectively. CONCLUSIONS: The differentiation of biodynamic milk FA composition is consistent with the existing knowledge about the effects of fresh grass, fodder, and ratio composition on the milk's FA composition. Based on the n6/n3 PUFA ratio, the average biodynamic dairy cow had a high intake (>82%) of fresh grass and conserved roughage (hay and grass silage), especially in the summer.

Comparison of different applications of automatic herd control systems on dairy farms - a review.

Recent years have seen the rapid development of different devices which can be helpful in the daily work of livestock farmers. The growing size of livestock herds has led farmers to lose individual contact with their animals, while behavioral studies show that breeders can effectively and precisely monitor a herd of up to 100 cows. This was the main motivation for this study, which aims to identify and test various electronic devices which provide useful herd management data, including estrus detection, individual activity and body temperature measurement, monitoring rumen pH levels, milk quality and content as well as milk temperature and somatic cell count measurements. Some devices can detect the metabolic status of animals with a reasonable level of precision. Contemporary animal farms are offered a large number of systems for monitoring the behavior of the animals in the herd and helping to identify those that are intended for insemination or are too active or excessively apathetic. Monitoring devices support herd management and help to reduce costs through the early detection of animal diseases and nutritional problems. This review aims to compile and summarize the information currently available on the use of automatic herd control systems on dairy farms, as well as to discuss the interpretation of the results, providing a useful diagnostic tool in nutritional evaluations of dairy herds. © 2018 Society of Chemical Industry.

Raw Cow's Milk Prevents the Development of Airway Inflammation in a Murine House Dust Mite-Induced Asthma Model.

Epidemiological studies show an inverse relation between raw cow's milk consumption and the development of asthma. This protective effect seems to be abolished by milk processing. However, evidence for a causal relationship is lacking, and direct comparisons between raw and processed milk are hardly studied. Therefore, this study investigated the preventive capacity of raw and heated raw milk on the development of house dust mite (HDM)-induced allergic asthma in mice. Six- to seven-week-old male BALB/c mice were intranasally (i.n.) sensitized with 1 µg HDM or PBS on day 0, followed by an i.n. challenge with 10 µg HDM or PBS on days 7-11. In addition, mice were fed 0.5 mL raw cow's milk, heated raw cow's milk, or PBS three times a week throughout the study, starting 1 day before sensitization. On day 14, airway hyperresponsiveness (AHR) in response to increasing doses of methacholine was measured to assess lung function. Bronchoalveolar lavage fluid (BALF) and lungs were furthermore collected to study the extent of airway inflammation. Raw milk prevented both HDM-induced AHR and pulmonary eosinophilic inflammation, whereas heated raw milk did not. Both milk types suppressed the Th2-polarizing chemokine CCL17 in lung homogenates and reduced lung Th2 and Th17 cell frequency. IL-4 and IL-13 production after ex vivo restimulation of lung T cells with HDM was also reduced by both milk types. However, local IL-5 and IL-13 concentrations were only suppressed by raw milk. These findings support the asthma-protective capacity of raw cow's milk and show the importance of reduced local type 2 cytokine levels. Heated raw milk did not show an asthma-protective effect, which indicates the involvement of heat-sensitive components. Besides causal evidence, this study provides the basis for further mechanistic studies.

Fatty acid profiles and antioxidants of organic and conventional milk from low- and high-input systems during outdoor period.

BACKGROUND: Intensification of organic dairy production leads to the question of whether the implementation of intensive feeding incorporating maize silage and concentrates is altering milk quality. Therefore the fatty acid (FA) and antioxidant (AO) profiles of milk on 24 farms divided into four system groups in three replications (n = 71) during the outdoor period were analyzed. In this system comparison, a differentiation of the system groups and the effects of the main system factors 'intensification level' (high-input versus low-input) and 'origin' (organic versus conventional) were evaluated in a multivariate statistical approach. RESULTS: Consistent differentiation of milk from the system groups due to feeding-related impacts was possible in general and on the basis of 15 markers. The prediction of the main system factors was based on four or five markers. The prediction of 'intensification level' was based mainly on CLA c9,t11 and C18:1 t11, whereas that of 'origin' was based on n-3 PUFA. CONCLUSION: It was possible to demonstrate consistent differences in the FA and AO profiles of organic and standard conventional milk samples. Highest concentrations of nutritionally beneficial compounds were found in the low-input organic system. Adapted grass-based feeding strategies including pasture offer the potential to produce a distinguishable organic milk product quality.

The relationship between high-fat dairy consumption and obesity, cardiovascular, and metabolic disease.

PURPOSE: To comprehensively review the data on the relationship between the consumption of dairy fat and high-fat dairy foods, obesity, and cardiometabolic disease. METHODS: We have conducted a systematic literature review of observational studies on the relationship between dairy fat and high-fat dairy foods, obesity, and cardiometabolic disease. We have integrated these findings with data from controlled studies showing effects of several minor dairy fatty acids on adiposity and cardiometabolic risk factors, and data on how bovine feeding practices influence the composition of dairy fat. RESULTS: In 11 of 16 studies, high-fat dairy intake was inversely associated with measures of adiposity. Studies examining the relationship between high-fat dairy consumption and metabolic health reported either an inverse or no association. Studies investigating the connection between high-fat dairy intake and diabetes or cardiovascular disease incidence were inconsistent. We discuss factors that may have contributed to the variability between studies, including differences in (1) the potential for residual confounding; (2) the types of high-fat dairy foods consumed; and (3) bovine feeding practices (pasture- vs. grain-based) known to influence the composition of dairy fat. CONCLUSIONS: The observational evidence does not support the hypothesis that dairy fat or high-fat dairy foods contribute to obesity or cardiometabolic risk, and suggests that high-fat dairy consumption within typical dietary patterns is inversely associated with obesity risk. Although not conclusive, these findings may provide a rationale for future research into the bioactive properties of dairy fat and the impact of bovine feeding practices on the health effects of dairy fat.

Organic food quality: a framework for concept, definition and evaluation from the European perspective.

Consumers buy organic food because they believe in the high quality of the product. Furthermore, the EU legal regulatory framework for organic food and farming defines high quality of the products as an important goal of production. A major challenge is the need to define food quality concepts and methods for determination. A background is described which allows embedding of the quality definitions as well as evaluation methods into a conceptual framework connected to the vision and mission of organic agriculture and food production. Organic food quality is defined through specific aspects and criteria. For evaluation each criterion has to be described by indicators. The determination of indicators should be through parameters, where parameters are described by methods. Conversely, the conceptual framework is described according to underlying principles and starting definitions are given, but further work has do be done on the detailed scientific description of the indicators. Furthermore, parameters have to be defined for the evaluation of suitability of these indicators for organic food production.

Experimental improvement of cow milk fatty acid composition in organic winter diets.

BACKGROUND: Organic milk is seen as more healthy in terms of its fatty acid (FA) profile. In three on-farm crossover trials with 10-12 cows in each group, different forages were compared for their potential to improve the FA composition. Different hay qualities (hay of pasture vs. hay of leys), additional energy sources (fodder beets vs. wheat) and roughage qualities (hay of pasture vs. grass silage) were compared for their effect on the FA composition of the milk. RESULTS: Rumenic acid (CLA cis-9, trans-11) and α-linolenic acid (ALA) were selected as principal markers to evaluate effects. The overall CLA cis-9, trans-11 was low (3.6-6.3 g kg⁻¹ fat), whereas ALA levels were intermediate (6.8-9.4 g kg⁻¹ fat). Differences between the forages were explained by the fat metabolism of the ruminants. CONCLUSION: Organic winter milk is low in several desirable FAs. Diets rich in mature fodder and sugar were a poor choice for an improved FA composition.