Dairy doi: 10.3390/dairy7040065

Authors: Fernando Mata Meirielly Jesus Joana Santos

European Protected Designation of Origin (PDO) and Protected Geographical Indication (PGI) cheeses represent a diverse sector in which geographical origin, production rules, species, territorial context, and market orientation shape product value. This study aimed to examine product, territorial, and market factors associated with variation in estimated retail price among European protected cheeses. A cross-sectional product-level dataset was constructed using 253 PDO and PGI cheeses registered in the EU geographical indication system. The dataset combined information on country, production region, species, production volume, territorial characteristics, registration status, market orientation, and estimated retail price. Retail price, expressed in €/kg, was analysed using a generalised linear model with a normal distribution and an identity link. The mean estimated retail price was 17.85 €/kg, with a range of 7.00 to 30.00 €/kg. Cow-milk cheeses represented 51.8% of the dataset, followed by mixed/other cheeses, sheep cheeses, and goat cheeses. The model was statistically significant overall (p < 0.001), indicating that the selected explanatory variables were jointly associated with estimated retail price. Higher estimated retail prices were associated with silage prohibition, mountain location, goat milk, and older registration age, whereas export dependency was negatively associated with price. Cow cheeses were significantly cheaper than mixed/other cheeses, while goat cheeses were significantly more expensive. These findings suggest that protected cheeses are economically heterogeneous and that estimated retail price is associated with more than protected status alone. Production constraints, territorial embeddedness, species identity, and market orientation were all associated with price differentiation. This study provides an exploratory quantitative basis for understanding value formation in the European protected cheese sector.

Dairy doi: 10.3390/dairy7040064

Authors: Karen Melo Borges Abias Santos Silva Jaciara Diavão Jean Marcelo Albuquerque Fernando César Ferraz Lopes Mariana Magalhães Campos Maria de Fátima Avila Pires Mirton José Frota Morenz

Heat stress is frequently associated with reduced dry matter intake (DMI), milk yield, and physiological alterations in Holstein dairy cows. The aim was to evaluate how prior exposure to heat stress affects feed intake, milk production, energy balance, blood metabolites, and infrared thermography, with special emphasis on the cows’ ability to recover and re-establish normal physiological and productive responses. Thirty multiparous lactating Holstein cows (days in milk = 87 ± 19.6 days) and previous milk yield (30.8 ± 3.95 kg/d) were monitored over 21 days, divided into three 7-day phases: thermoneutral (TN), cows kept at a constant temperature–humidity index (THI) of 69; heat stress (HS), cows at a THI of 82 for 9 h and 69 for 15 h, and recovery (REC), cows at a constant THI of 69. Milk yield decreased by 16.1% during HS and remained 17.3% lower during REC compared with TN. Dry matter intake was reduced by 5.8 and 4.2 kg/d for HS and REC compared to TN cows. Plasma insulin was higher in REC compared to HS. Under the conditions of the present study, heat stress was associated with impaired productive performance, altered metabolism, and changes in the physiological status of mid-lactation Holstein dairy cows, with some effects persisting after the cows returned to thermoneutral conditions.

Dairy doi: 10.3390/dairy7040063

Authors: Lilli Bittner-Schwerda Rüdiger D. Ollhoff Adriana Wöckel Anne Kaiser Sofija Džakula Anja Gretzschel Tilman Kühn Joachim Spilke Alexander Starke

Milk sample collection before attaching the milking cluster for bacteriological and cytological examination is a standard clinical diagnostic procedure in dairy production. Collecting samples premilking can be difficult under certain field conditions. Therefore, we aimed to determine the effect of the timing of milk sampling on the results of bacteriologic analysis and somatic cell count. The study was conducted on a dairy herd of 1520 German Holstein cows. From 110 cows selected because of previously high somatic cell count, paired milk samples (premilking and postmilking) were taken and submitted for bacteriological and cell count analysis. The somatic cell score (SCS) was significantly higher in postmilking samples than in premilking samples (p = 0.001). A total of 113 of 433 premilking and 69 of 433 postmilking samples had positive bacteriologic culture. Quarter samples with a positive bacteriologic culture had numerically higher SCS estimates than negative samples. Mammary quarters with positive premilking and negative postmilking cultures had a significantly lower SCS than quarters with positive premilking and postmilking cultures (p < 0.05). Our findings suggest that premilking and postmilking samples differ in bacteriological and cytological composition. Therefore, the sampling strategy has to be taken into account when making treatment and prevention strategy decisions based on milk samples.

Dairy doi: 10.3390/dairy7040062

Authors: Kathrin M. Weber Romina T. Albersdörfer Christine Schmidt Prisca V. Kremer-Rücker

Tail inflammation and necrosis in dairy cows are increasingly reported, yet the early stages of their development and dynamics remain unclear. We conducted a longitudinal study to characterize the macroscopic pathogenesis of tail lesions throughout the entire lactation period to identify the initial stages and vulnerable phases of lactation. Fifty-one cows were examined weekly. We scored the distal spineless tail for bloody lesions. Change point analysis and first-order Markov chains were used to describe the timing and transitions probabilities. Overall, 60.8% of the cows developed at least one bloody lesion, with 49.0% experiencing lacerations and 25.5% undergoing autoamputation. On average, autoamputation shortened tails by 4.0 cm. Three progression types were identified: laceration with healing and preservation of the tip, autoamputation after laceration and spontaneous autoamputation. Regarding the frequency of occurrence, 65.5% began as bloody lacerations and healed in 93.2% of cases. Of these, 4.0% progressed to autoamputation, 12.7% underwent spontaneous autoamputation from an intact tip. Significant changes in lesion severity occurred in lactation weeks 8 and 40. These patterns are consistent with an endogenous microcirculatory mechanism and support the hypothesis that tail inflammation and necrosis may represent a manifestation of Bovine Inflammation and Necrosis Syndrome.

Dairy doi: 10.3390/dairy7040061

Authors: Burim N. Ametaj

Lameness is a major welfare and economic disorder in dairy production and a clinical sign with multiple causes. This review focuses on claw horn disruption lesions (CHDL), a major cause of periparturient lameness. Although hoof-centered prevention and treatment remain essential, preclinical systemic alterations, recurrence, and clustering with other transition-period disorders suggest that CHDL may sometimes represent the local expression of broader pathophysiology. We propose the Lameness Triad, in which three mechanisms converge: sustained endotoxemic pressure and TLR4-mediated innate immune activation, endothelial glycocalyx degradation and vascular dysfunction, and metabolic triage that reallocates resources from production toward defense. Their combined pathogenic pressure is proposed to produce clinical disease when it exceeds a cow-specific Susceptibility–Resilience Gate comprising mucosal barrier integrity, hepatic endotoxin clearance, immune calibration, redox reserve, hoof structural reserve, and prior damage. Ruminal dysbiosis, mammary involution, and postpartum uterine contamination may contribute to inflammatory and endotoxemic pressure, whereas the confined, continuously loaded digital corium may become a principal site of clinical expression. Longitudinal multi-omics studies identified inflammatory, urinary, and milk-metabolite alterations before diagnosis, with within-cohort serum and urine models discriminating cows that later became lame as early as eight weeks prepartum. These findings require prospective validation and do not establish the complete causal sequence. The framework extends, rather than replaces, established hoof-health programs and provides testable hypotheses for earlier detection, prevention, and selection for resilience.

Dairy doi: 10.3390/dairy7040059

Authors: Milan Bogdanović Dušan Stevanović Ksenija Čobanović Sara Panseri Maria Nobile Radoslava Savić Radovanović Nataša Golić Nikola Popović

The chemical composition of donkey and sheep milk showed significant differences, with donkey milk containing less fat and protein and a higher level of lactose, while sheep milk had more dry matter and a higher proportion of casein and saturated, unsaturated, monounsaturated, and polyunsaturated fatty acids. This study investigated the biological effects of in vitro-digested donkey and sheep milk, and their blends at different ratios, on differentiated human intestinal Caco-2 cells. Gene expression analysis showed that treatment with digested donkey milk (100%) significantly altered the expression of the autophagy-related gene (SQSTM1), tight junction (CLDN4), and innate defense genes (DEFB1, MUC2, and MUC5), showing a non-cytotoxic response and a profile consistent with a barrier-related transcriptional response. In contrast, sheep milk (100%) and certain milk blends (70% and 50% sheep’s milk) down-regulated genes involved in the autophagy process (ULK1, AMBRA, BECN1, ATG5, GABARAP, and SQSTM1), tight junction genes (OCLN and CDH1), and innate defense genes (DEFB1 and MUC2), suggesting a distinct epithelial response that warrants further confirmation. Metabolomic profiling revealed clear compositional and functional distinctions among the digested milks, identifying 166 metabolites that were differentially regulated (136 compounds were down-regulated and 30 up-regulated). Donkey milk digestion yielded a metabolite profile enriched in polar peptides and compounds with reported antioxidant associations, whereas sheep milk digestion showed a different pattern dominated by hydrophobic peptides and lipid-related metabolites. These findings indicate that, under the tested in vitro conditions, donkey milk digestion is associated with gene expression and metabolomic patterns consistent with epithelial barrier support.

Dairy doi: 10.3390/dairy7040060

Authors: Mohammad Javed Ashar Paula A. Gonzalez-Rivas Frank R. Dunshea Christina D. Marth Surinder S. Chauhan

Injectable trace mineral supplementation (ITMS) has been proposed to support physiological adaptation during critical stages in dairy cattle; however, field-based evidence from pasture-based seasonal-calving systems remains limited. This study evaluated the effects of ITMS, administered at birth in calves and approximately 30 days before fixed-time artificial insemination in early-lactation cows, primarily on erythrocyte glutathione peroxidase activity and plasma trace mineral concentrations, while health, growth, production, and fertility were monitored as secondary outcomes. Thirty newborn calves receiving isotonic saline control (CNTRL; n = 15) or ITMS (n = 15) and 50 early-lactation cows (CNTRL, n = 24; ITMS, n = 26) were enrolled. In calves, erythrocyte glutathione peroxidase activity was higher in ITMS calves (322 vs. 359 U/g Hb; p < 0.01), and plasma selenium was higher on day 15 (0.57 vs. 0.62 µmol/L; p = 0.03). A treatment × week interaction was detected for body weight, but overall body weight and average daily gain did not differ. In cows, plasma selenium was transiently higher in multiparous ITMS cows on day 15 (1.04 vs. 1.09 µmol/L; p = 0.03), while antioxidant and inflammatory biomarkers, production, mastitis incidence, and fertility were unaffected. Overall, a single ITMS administration elicited modest biochemical and selenium responses without statistically detectable improvements in growth, health, production, or fertility.

Dairy doi: 10.3390/dairy7040058

Authors: Carlos Niño de Guzmán Pablo Pinedo Haipeng Yu Nikolay Bliznyuk Albert De Vries

Our first objective was to quantify the associations between health-related events (HRE) before insemination, the relative increase in estrus intensity (REI) at insemination, and the probability of cow-level pregnancy per artificial insemination (P/AI) in organic Holstein dairy cows. Quantifying these associations may aid on-farm decision-making, such as setting the voluntary waiting period, choice of type of semen, do-not-breed and culling decisions. A second objective was to develop predictive models to estimate P/AI based on readily available data, and present common goodness-of-fit results also used in the machine learning community. All data were collected from a certified organic dairy farm in the western USA from 2019 to 2021. Health-related and reproduction data were obtained through Dairy Records Management Systems (DRMS; Raleigh, NC, USA). Activity data were collected using pedometers (IceRobotics, Stirling, UK) mounted on the rear legs. The REI, defined as walking steps per hour before insemination divided by the cow’s baseline steps per hour, was available for 17,238 inseminations from 4759 cows. The REI was categorized as ≤200%, >200–400%, >400–600%, or >600%. The HRE were available for 65,684 inseminations from 13,365 cows. The HRE were categorized as mastitis, metabolic disease (i.e., hypocalcemia, ketosis, displaced abomasum, digestive problems), reproductive disease (i.e, metritis, endometritis, pyometra, retained fetal membranes), lameness, 2 different diseases, ≥3 different diseases, or as healthy (none of these diseases prior to insemination). Combinations (COMBO) between REI categories and 0, 1, or ≥2 HRE were also created. Data were split into training and test sets. The training data were used to fit three logistic regression models that included either HRE, or REI, or COMBO. Each of the three models also included the covariates of 3-mo herd-average P/AI prior to insemination, days in milk, and the fixed effects of parity, insemination season, days after the previous insemination or days to 1st insemination. A random effect accounted for repeated inseminations within cow. Parameter estimates, odds ratios, and the estimated marginal means of the estimated P/AI of the fixed effects were obtained from the logistic regression models. The models’ estimates were applied to the test datasets, and discrimination and calibration statistics were calculated to judge goodness-of-fit. Unadjusted mean P/AI were 0.31, 0.28 and 0.28 for the HRE, REI and COMBO training datasets. For the HRE model, estimated P/AI ranged from 0.20 (≥3 different HRE) to 0.30 (healthy). The estimated P/AI associated with four REI categories were not different from 0.27 in the REI model. The estimated P/AI associated with the combinations of HRE and REI in the COMBO model varied from 0.18 after ≥2 HRE and >200–400% REI, to 0.30 when inseminations were in healthy cows with REI >600%. Inseminations in older cows, in the spring, and outside 18–24 d after the previous insemination were also associated with lower estimated P/AI. The area underneath the Receiver Operating Characteristic curve ranged from 0.57 (COMBO) to 0.60 (HRE) for the test data, indicating fair discrimination ability of the models. Calibration plots showed that the prediction models produced unbiased predicted P/AI. In conclusion, the results showed no conclusive evidence of greater estimated P/AI related to greater REI as a measure of estrus activity. More HRE were associated with lower estimated P/AI. Combinations of low REI and more HRE were associated with notably decreased estimated P/AI. The logistic regression models produced unbiased predicted P/AI. We found no evidence that the strength of the relationship between REI and P/AI depended on the HRE category. The applications of the results are as follows. First, these predictive models may help inform insemination decisions in organic dairy cows, although further external validation is recommended, and the discriminatory performance is weak. Second, a variety of goodness-of-fit statistics were calculated to allow comparisons of the current logistic regression analyses with future analyses made by other machine learning techniques.

Dairy doi: 10.3390/dairy7040057

Authors: Mst. Ishrat Zerin Moni Asif Uzzaman Rezoanul Haque Md. Niamot Ali Amira A. Goma Md. Reazul Islam Mohammad Mahbubur Rahman Jashim Uddin

Dairy cattle are particularly susceptible to welfare challenges that can negatively impact their behaviour, physiological responses, milk yield, and overall health. These challenges include rough handling by unfamiliar people, extreme climatic conditions, inconsistent feeding, regrouping, and transportation. This pilot study investigated the interrelationships between behavioural and physiological responses and milk production of dairy cows reared in a smallholder stall housing system, restricting cow movement to standing and lying down in the Rajshahi district of Bangladesh. Fifty cows were observed, with all cows assessed over two days. Video recordings were made using a smartphone during the morning milking routine, focusing on specific behaviours: ear position (upright, forward, backward, downward), sniffing, stepping, and tail movement. Observations were conducted during the first two minutes after the start of the milking process manually. Physiological responses recorded included rectal temperature, vaginal temperature, and heart rate. Productivity outcomes include body condition score (BCS), milk yield per milking, daily milk yield, and milking frequency. Environmental data were collected to calculate the Temperature-Humidity Index (THI), which averaged 84.5 (±1.78 SD), exceeding the thermal comfort zone for dairy cattle (THI < 72). Statistical analyses comprised principal component analysis (PCA) to determine the most descriptive variables. Pearson correlation assesses the direction and strength of relationships between variables, and simple linear regression to evaluate milk production variables using all other variables as predictors. Results showed that cows displaying an upright ear position during milking had a positive relation with average daily milk yields (r = 0.456, p = 0.001). Milk yield was negatively correlated with THI (r = −0.331, p = 0.02), forward ear position (r = −0.356, p = 0.01), kicking (r = −0.531, p < 0.001), and sniffing behaviour during milking (r = −0.511, p < 0.001). The daily milk yields were positively correlated with body condition scores of milking cows (r = 0.661, p < 0.001). Rectal and vaginal temperatures were positively associated with each other (r = 0.777, p < 0.001). Similarly, rectal temperature and heart rate were positively associated with each other (r = 0.511, p < 0.001). Therefore, ear position, kicking, and sniffing behaviour during milking were used to assess emotional states, with forward ears associated with signs of stress. Healthy cows with higher body condition scores (BCS) produced more milk, requiring farmers to milk them more than twice a day. This pilot study highlights key behaviour and physiological responses relevant to stress and milk production, and thus overall welfare. For greater generalisability, future research should incorporate larger sample sizes and repeated environmental assessments over extended periods.

Dairy doi: 10.3390/dairy7040056

Authors: Flávio G. Silva Ana Geraldo Cátia Antas Alfredo Pereira Cristina Conceição

Teat-end hyperkeratosis (THK) is a welfare concern in dairy cows, associated with pain, discomfort, and increased mastitis risk. Identifying risk factors associated with THK may support targeted preventive management. This cross-sectional observational study aimed to determine the prevalence of THK and identify factors associated with its occurrence. A total of 492 Holstein-Friesian cows were evaluated at the milking parlor of a dairy farm located in Évora, Portugal. All four teats per cow (n = 1968) were scored on a 4-point scale: no lesion (1), smooth ring (2), rough (3), and very rough skin (4). Parity, days in milk, teat length, teat-end shape, udder health, and milking procedures were assessed as potential factors associated with THK. A mixed-effects ordinal logistic regression model was used to evaluate the associations between THK and the selected factors. Overall THK prevalence was 54.4%, with a median score of 2 (range: 1–4; IQR: 1–2). Score distribution was 45.6% (score 1), 43.0% (score 2), 10.3% (score 3), and 1.1% (score 4). Round teat ends were associated with greater odds of severe THK compared with flat teat ends (OR = 6.4; p = 0.016). Longer milking duration (OR = 2.7 per 100 s; p = 0.002) and a history of mastitis (OR = 80.0; p < 0.001) were also positively associated with THK severity. These findings highlight opportunities to mitigate THK through optimized milking management and breeding strategies. Notably, factors such as mastitis and prolonged milking time are also associated with culling, potentially masking the true burden of THK on animal welfare.

Dairy doi: 10.3390/dairy7040055

Authors: Elisa Rampacci Marcello Guadagnini Sebastiano Carra Cecilia Tolasi Fabrizio Passamonti

Effective colostrum management is essential to ensure adequate transfer of passive immunity (TPI) and reduce neonatal morbidity in calves. This study evaluated the impact of pasteurization on colostrum microbiological and immunological quality and its association with TPI and calf health on a dairy farm in Italy. A total of 79 colostrum samples were included: 39 pasteurized (PC) and 40 non-pasteurized (NPC). Immunoglobulin concentration was estimated by Brix refractometry, while bacterial contamination was assessed by total bacterial count (TBC) and total coliform count (TCC). In a second phase of the study, 80 newborn calves received either pasteurized (C-PC, n = 40) or non-pasteurized colostrum (C-NPC, n = 40). TPI was evaluated at 48 h and calves were clinically monitored for 21 days. Overall, colostrum showed not only high Brix values but also elevated bacterial contamination. Pasteurization significantly reduced the microbial load in colostrum (TBC, 5.7 vs. 2.9 log CFU/mL; TCC 4.6 vs. 1.5 log CFU/mL) (p < 0.0001). Brix values differed significantly between PC and NPC colostrum (p < 0.001), although the magnitude of the difference was minimal (27.8% in NPC vs. 27.6% in PC), suggesting limited biological relevance. Calves receiving pasteurized colostrum showed significantly lower odds of developing neonatal diarrhea than calves receiving unpasteurized colostrum (C-PC: OR 0.1, 95% CI 0.0–0.3; C-NPC: OR 9.9, 95% CI 3.4–32.6). These findings indicate that colostrum pasteurization reduces contamination effectively even in colostrum with high bacterial loads without substantial impact on its Brix values and was associated with improved calf diarrhea outcomes in the investigated herd.

Dairy doi: 10.3390/dairy7040054

Authors: Maria Belkacemi Naziha Fedala Teresa Gazzotti Elisa Zironi Giacomo Depau Giulia Rampazzo Carlo Boselli Valentina D’Onofrio Angela Costa Moussa Mokhtari Giampiero Pagliuca

Chemical contamination of milk represents a relevant food safety concern, particularly in countries where systematic monitoring programs are still limited. In Algeria, information on human exposure to aflatoxin M1 (AFM1) and per- and polyfluoroalkyl substance (PFAS) contamination remains fragmented or lacking. This study assessed the occurrence of AFM1 and 17 PFAS in cow and goat milk and explored farm-level management practices potentially influencing contamination. Bulk-tank milk samples were collected from 26 dairy cattle and 11 dairy goat farms. Farm characteristics, including feeding strategies, water sources, grazing practices, and herd size, were recorded using structured questionnaires to characterize farm-level practices and explore their potential association with contamination patterns. AFM1 was detected in all samples. In cow milk, 96.1% exceeded the EU limit (0.05 µg/kg), with concentrations ranging from 0.048 to 0.410 µg/kg (mean: 0.102 µg/kg), although none exceeded the Codex Alimentarius limit (0.50 µg/kg). Goat milk showed a lower prevalence of samples above the EU limit (36.4%), but higher concentrations were observed (range 0.030–0.578 µg/kg; mean 0.193 µg/kg), with two samples (18.2%) exceeding the Codex limit. PFAS contamination was generally low: 73.0% of cow and 64.0% of goat samples were below detection limits, with only PFBS (cow milk) and PFDA (goat milk) quantified above LOQ. Because of the limited number of farms and the single sampling period, no robust associations could be established between farm-level variables and contaminant concentrations. This preliminary study provides an exploratory overview of AFM1 and PFAS occurrence in bovine and goat bulk milk from the sampled Algerian farms. These findings reveal widespread AFM1 contamination within the investigated farms, consistent with possible feed-borne AFB1 exposure, whereas PFAS occurrence appeared sporadic and generally low.

Dairy doi: 10.3390/dairy7040053

Authors: Kelsey Pasch Felicitas Vignati William Brown

Postpartum dairy cows fed more rapidly fermentable starch sources have depressed dry matter intake (DMI), compounding the risk of negative energy balance and hepatic lipid accumulation. Rumen-protected choline (RPC) is supplemented to periparturient dairy cows to facilitate hepatic lipid export. Our objective was to evaluate the interaction of dietary starch fermentability (DF) and RPC supplementation on postpartum DMI and performance. Prepartum supplementation of a low dose of RPC (no RPC [C−] vs. RPC [C+; 30 g/d]) began 21 d before the expected calving date for Holstein cows with at least one parity. Postpartum, cows were assigned to 1 of 4 postpartum treatments for 21 d with a 2 × 2 factorial arrangement of starch fermentability rate (low [dry-rolled corn; LFERM] vs. high [dry-rolled wheat; HFERM]) and RPC (C− vs. C+). Prepartum, C+ decreased DMI by 2.3 kg compared with C−, but there was no evidence of treatment effect on DMI postpartum. Time and DF interacted on milk yield, with HFERM increasing milk yield after d 3 compared with LFERM. Compared with LFERM, HFERM decreased milk fat content but not fat yield. For blood metabolites, C+ decreased plasma beta-hydroxybutyrate by 0.3 mmol/L and tended to increase glucose concentration compared to C−. In conclusion, supplementation with RPC at a low rate of inclusion reduced DMI in prepartum cows and decreased postpartum plasma BHB concentrations. Further work is required to elucidate potential mechanisms of action for RPC-mediated reductions in DMI.

Dairy doi: 10.3390/dairy7040052

Authors: Arhemy Pineda-Montes Javier R. Reyna-Granados Rosa I. Luna-Ramirez Mario R. Mondaca-Duarte Ulises Macías-Cruz Leonel Avendaño-Reyes Carolina García-Benitez José C. Leyva-Corona Juan F. Hernández-Chávez Pablo Luna-Nevárez

Heat stress (HS) is a major issue affecting productivity and physiological responses in dairy cattle. Advances in gene function research have made it possible to identify favorable genotypes for HS tolerance. This study evaluates the combined effects of temperature–humidity index (THI) and genomic load of favorable SNPs in the genes GRM8, SMAD3, and TLR4 on daily milk yield (DMY), rectal temperature (RT), and gene expression in Holstein cows (n = 160). Environmental conditions ranged from thermoneutral to severe HS, allowing the assessment of genotype by environment interactions. A subset of cows (n = 40) was selected based on genomic load to evaluate gene expression under opposite thermal conditions. Results showed that increasing THI was associated with reduced DMY and increased RT, with more pronounced effects in cows carrying fewer favorable genotypes. Slope analyses revealed that cows with higher genomic load exhibited a moderate decline in DMY and lower increase in RT, indicating enhanced resilience to HS. Significant THI × genomic load interactions were detected above a threshold of 79 units (p < 0.05). Gene expression analyses supported these findings, showing differential expressions of GRM8, SMAD3, and TLR4 under HS conditions. The functional and predictive value of these genes as markers of thermotolerance could help producers by supporting their application in genetic selection programs, thus providing another tool to combat HS and improve resilience and productivity in dairy systems.

Dairy doi: 10.3390/dairy7040051

Authors: Yasunori Shinozuka Takuya Kanda Keiichi Hisaeda Akira Goto Yoichi Inoue Naoki Yamamoto

This study investigated the impact of moderate teat-end hyperkeratosis on milk somatic cell count (SCC) and bacterial count (BC) in Holstein cows. Comparing teat-end scores (TS) 2 and 3, we found that TS3 quarters exhibited significantly higher SCC (p = 0.017) despite having shorter days in milk than TS2 quarters. While BC in foremilk (BC1) remained stable, post-milking samples (BC2) decreased during sequential sampling; however, a significant Group × Time interaction (p = 0.020) revealed a less pronounced bacterial reduction in TS3. These findings tentatively suggest that moderate hyperkeratosis (TS3) may be associated with subclinical mammary irritation and altered bacterial clearance dynamics, independent of the absolute bacterial load. Managing teat-end morphology may help optimize milk quality, even when bacterial contamination is minimal.

Dairy doi: 10.3390/dairy7040050

Authors: Vittorio Lo Presti

The dairy industry is increasingly seeking natural alternatives to synthetic additives to meet the growing demand for clean-label and functional foods. Bee-derived products (BDPs), including propolis, honey, bee pollen, bee bread, and royal jelly, represent a promising class of bioactive ingredients due to their antimicrobial, antioxidant, and immunomodulatory properties. This review critically examines their integration across the dairy value chain, adopting a farm-to-product perspective. At the farm level, BDPs can modulate rumen fermentation, influence microbial populations, and contribute to improved feed efficiency and reduced enteric methane emissions. These effects may translate into modifications in milk composition and functional properties. At the processing and product levels, the incorporation of BDPs into dairy matrices such as yogurt, cheese, and fermented milk enables the development of functional foods enriched with bioactive compounds and supports probiotic viability in synbiotic systems. However, their application is associated with technological and sensory challenges, including variability in chemical composition, dose-dependent antimicrobial effects, and potential impacts on texture and flavour. By bridging animal and food science, this review highlights the multifunctional role of BDPs in enhancing sustainability, safety, and nutritional value in dairy systems, while identifying current limitations and future research directions for their effective industrial implementation.

Dairy doi: 10.3390/dairy7040049

Authors: Giovanni Bittante Giorgia Stocco Alessio Cecchinato Luigi Gallo Stefano Schiavon

Breed and herd are major determinants of milk productivity and efficiency. This study evaluated 1508 lactating cows from three dairy (Holstein, Brown Swiss, Jersey) and three dual-purpose (Simmental, Rendena, Alpine Grey) breeds across 41 multi-breed mountain herds. Data on body size, production, and milk samples were collected. Daily milk yield, milk energy, and cheese yield were quantified, and herds were classified under high (>75 MJ/d) or low (<75 MJ/d) average daily milk energy production. Cheese-making traits were predicted from 508 individual model cheese-making trials. Nine productivity ratios and three efficiency indicators (energy efficiency, economic efficiency, and income over feed costs) were calculated. Data was analyzed using mixed models including herd productivity class, herd within class (random), breed, parity, and days in milk. Differences among breeds were smaller within herds than across herds and were more pronounced for quality and cheese-making traits. Dairy and dual-purpose breeds showed similar overall performance. Jerseys were the least productive in absolute terms but, when adjusted for body size, were as or more efficient than other dairy breeds. Holsteins had the highest milk yield, whereas Brown Swiss showed superior milk quality and cheese yield. Simmental outperformed local dual-purpose breeds in size and production but not in efficiency. Accurate assessment of energy and economic efficiency is essential for breeding and crossbreeding strategies.

Dairy doi: 10.3390/dairy7040048

Authors: Sana Ullah Sher Ali Usama Nasir Ana Raquel Pinardi Roice Eliana Rosim Muhib Ullah Carlos Augusto Fernandes de Oliveira

Mycotoxins are toxic fungal secondary metabolites that can contaminate the feed of dairy animals and can be transferred into milk and dairy products. This study aims to assess the occurrence of major mycotoxins in 190 samples of milk-based infant foods commercially available in São Paulo, Brazil, and characterize the associated risks. Aflatoxins (AFs) M1 (AFM1), B1 (AFB1), B2 (AFB2), G1 (AFG1) and G2 (AFG2), fumonisins (FBs) B1 (FB1) and B2 (FB2), ochratoxin A (OTA), zearalenone (ZEN), deoxynivalenol (DON), and T-2 toxin were analyzed using ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC–MS/MS). Ninety-eight samples (51.6%) were contaminated with at least one mycotoxin, while the co-occurrence of two to four mycotoxins was observed in 32 samples (16.8%). ZEN was the most prevalent mycotoxin (60 samples; 32%), followed by FBs (FB1 and FB2) (42 samples; 22%), OTA (11 samples; 5.8%), DON (8 samples; 4.2%), and T-2 toxin (2 samples; 1.1%). AFs were quantified in only two samples (1.1%), one milk-based product containing 0.630 μg/kg of AFM1 and one flavored milk powder with AFB1 at 5.850 μg/kg. The analyzed flavored milk powder samples showed the highest maximum concentrations of DON, FBs, and OTA (165.5, 93.79, and 86.77 μg/kg, respectively), while unflavored milk powder contained the highest concentration of ZEN (17.40 μg/kg). Infant formula provided the highest estimated daily intake (EDI) value of DON (1.226 μg/kg bw/day) for infants aged ≤one year. Hazard quotient (HQ) values > 1 revealed a potential health concern for DON in only one sample of infant formula intended for infants up to six months of age. Margin of exposure (MoE) values for AFs and OTA were <10,000, emphasizing the need for continuous monitoring due to their potential toxic effects. These findings emphasize the need for stringent preventive measures and regulatory compliance to prevent mycotoxin contamination in milk-based infant foods in Brazil.

Dairy doi: 10.3390/dairy7040047

Authors: Mauro Decandia Valeria Giovanetti Andrea Frongia Maria Gabriella Serra Andrea Cabiddu Maria Rosaria Carboni Maria Sitzia Marco Acciaro

This study evaluated the effects of replacing alfalfa hay with sulla [Sulla coronaria (L.) B.H. Choi & H. Ohashi] hay or sulla haylage in pasture-based diets for lactating Sarda ewes on milk production, milk composition, and nutrient utilization efficiency. Seventy-two mid-lactation ewes were assigned to three dietary treatments for 56 days—alfalfa hay (AH), sulla hay (SH), or sulla haylage (SHL)—all combined with 3 h/day grazing and a fixed amount of supplemental concentrate. The diets were formulated to be theoretically isoenergetic and isoproteic. Pasture nutritive value was generally comparable among the groups throughout the experimental period. Milk yield did not differ among treatments; however, fat- and protein-corrected milk (FPCM) and milk fat and protein concentrations were higher in AH compared with the sulla-based diets. Actual nutrient intake differed among treatments, with nitrogen intake (NI) being greatest in AH, consistent with the higher milk urea concentration observed in this group. Estimated apparent energy utilization efficiency (FPCM/UFL intake) showed treatment-associated differences in exploratory analyses, with lower values observed in SH compared with AH and SHL. Similarly, exploratory estimates of apparent nitrogen utilization efficiency (Milk N/NI) were highest in SHL, intermediate in SH, and lowest in AH (p < 0.001). Condensed tannins were not detected in the conserved sulla forages under the analytical conditions adopted in this study. Therefore, the observed responses were unlikely to be directly associated with detectable condensed tannin activity. Overall, the results suggest that forage conservation method may influence milk composition and estimated apparent nutrient utilization indices, with sulla haylage showing higher exploratory estimates of apparent nitrogen utilization efficiency without negative effects on milk yield under the conditions of the present study.

Dairy doi: 10.3390/dairy7030046

Authors: Dimitra V. Liagka George C. Fthenakis Vasia S. Mavrogianni Dafni T. Lianou Vassiliki Spyrou Natalia G. C. Vasileiou

The intensification and continuous evolution of dairy sheep and goat farming have played an essential role in the development and implementation of milking equipment. The increasing demand for time-efficient milking procedures, reduced labour costs, sustained milk production, and optimal mammary health have driven the widespread adoption and optimisation of machine milking technologies. The objectives of this article are (i) the review of milking systems and relevant technological developments in milking equipment and (ii) the evaluation and description of their impact on udder health, as applied on dairy small ruminant farms. Milking systems used on farms depend on the available space and number of animals on the farms. Appropriate settings in milking systems are important for ensuring good milk quality; among them, vacuum level, pulsation rate and ratio are important characteristics that must be monitored regularly. Further, use of appropriate teatcups specific to the animal species to be milked is significant. An important aspect of proper maintenance of the milking system is the cleaning procedure after completion of milking. Points for consideration are quality and temperature of the water used for cleaning, use of detergents and disinfectants, and maintenance schedule and teatcup replacement. Some technological features that are part of milking systems include automatic vacuum shut off, electronic milk recording, electronic identification of animals, automatic flushing of milking clusters and automatic pre-stimulators. Farms will benefit from applying precision technologies, which will use data from tools related to animal genetic background, animal behavioural indicators, environmental conditions and disease-related functions for more holistic and cost-effective farm management. In this context, integration of sensor-based technologies in milking systems will be able to provide real-time information regarding quality of milk produced at individual and farm levels. Moreover, the introduction of automatic system flushing in-between animals during the milking procedure can contribute to breaking chains of potential bacterial transfer and reducing animal infections during milking. Overall, although machine milking has certainly contributed to improved efficiency, milk quality and labour conditions, flaws in system function may adversely affect mammary health.

Dairy doi: 10.3390/dairy7030045

Authors: Marcos Busanello Mariani Schmalz Lindorfer Alexandre Gallucci Toloi Maity Zopollatto

Automatic milking systems (AMSs) are increasingly adopted in dairy farms, and barn design, particularly regarding cow traffic design (CowTD), plays a key role in system performance. This study evaluated the association between different CowTDs and operational and production indicators of AMS Brazilian dairy farms. The data were obtained from 149 commercial dairy farms equipped with AMS (average of 1.6 AMS per farm) encompassing approximately 14,642 lactating cows recorded between June and December 2025. Cow traffic designs were classified as free or smart-gated systems, including milk-first (MF) and feed-first (FF) configurations. Mixed models were used to evaluate the effects of regions, housing system, and CowTD on the number of lactating cows per AMS (NCowsAMS), milking frequency (MFreq), milk yield per milking (MYMilking), daily milk yield per cow (MYcow), daily milkings per AMS (MilkingsAMS), and daily milk yield per AMS (MYAMS). On average, farms milked 58 cows per AMS with a mean MFreq of 2.69 milkings/cow per day and produced 2227 kg of milk per AMS per day. Smart-gated CowTD supported a greater number of cows per AMS than free CowTD systems (FF = 57 and MF = 58 vs. Free = 53 cows/AMS). Although free CowTD increased MFreq to approximately three milkings/cow/day, this advantage did not translate into greater or equivalent MYAMS, despite MF and free CowTD exhibiting similar MYcow (37.0 and 37.2 kg/day, respectively). Even though free CowTD achieved the highest MilkingsAMS (Free = 156 vs. MF = 151 and FF = 143 milkings/day), it milked fewer cows per robot, resulting in lower MYAMS. Consequently, FF and MF systems produced 86 and 180 kg/day more MYAMS, respectively, than free CowTD. These results suggest that AMS performance is primarily driven by the NCowsAMS rather than MFreq alone. Under Brazilian commercial conditions, smart-gated CowTD systems appear to be more efficient, as evidenced by higher MYAMS, while allowing higher stocking densities, potentially without increasing labor requirements.

Dairy doi: 10.3390/dairy7030044

Authors: Vasily Isakov Alexei Goncharov Vladimir Pilipenko Armida Sasunova Alla Kochetkova Vladimir Bessonov

Metabolic dysfunction-associated steatotic liver disease (MASLD) affects 25–30% of adults globally. Dietary sugar reduction is one of the key therapeutic targets, but elimination of sugar-sweetened foods may compromise adherence to calorie-restricted diets. Brazzein, a natural sweet protein that is 500–2000 times sweeter than sucrose, offers a promising substitute, yet clinical data in patients with MASLD are lacking. In a double-blind, randomized, two-period crossover trial, 103 adults with MASLD tasted iso-sweet vanilla ice cream sweetened with either brazzein or sucrose on two consecutive days. Overall impression and sensory attributes (appearance, color, aroma, taste, and texture) were rated on 5-point hedonic scales, and the percentage of the 100 g portion consumed was recorded. Brazzein-sweetened ice cream met the prespecified criteria for both non-inferiority and equivalence versus sucrose for overall impression. Top-2 box acceptance (ratings ≥ 4) was extremely high and nearly identical (96.1% for brazzein and 98.1% for sucrose). Mean consumption exceeded 98% of the portion for both products, with no significant difference between sweeteners. Secondary sensory ratings were closely similar, and multivariate analyses indicated highly overlapping sensory profiles. Exploratory subgroup analyses suggested consistent findings across most demographic and clinical characteristics, although participants with advanced liver fibrosis (LSM ≥ 9.6 kPa) showed numerically higher ratings for sucrose. In exploratory analyses, liver stiffness was associated with slightly lower intake at higher stiffness values. This study provides the first evidence that brazzein-sweetened ice cream maintains short-term sensory acceptability comparable to a conventional sucrose-sweetened product in adults with MASLD. These findings support further development and evaluation of brazzein-containing sugar-reduced foods, including repeated-exposure sensory studies and separate metabolic investigations.

Dairy doi: 10.3390/dairy7030043

Authors: Juthamas Nabthonglang Siriyakorn Niyomprapasakun Pakpoom Navanukraw Narinthip Laosuwan Thanapol Nongbua Jutarop Phetcharaburanin Chainarong Navanukraw

The objectives of this study were to evaluate the effects of different dietary omega-6 to omega-3 (n-6:n-3) fatty acid ratios during timed artificial insemination (TAI) on reproductive responses, luteal development, and offspring characteristics in lactating dairy cows. Holstein cows (n = 60) averaging 109 ± 10 days in milk (DIM) were randomly assigned to receive either a greater n-6:n-3 diet or a lower n-6:n-3 diet for 42 days. All cows were synchronized using a modified Ovsynch protocol. Although synchronized ovulation rates and pregnancy outcomes were not significantly affected by treatment, cows fed the lower n-6:n-3 ratio exhibited significantly greater corpus luteum (CL) volume and cross-sectional area on days 11 and 14 post-TAI, indicating enhanced luteal development. Pregnancy rates following the first AI and cumulative AI, as well as the number of services per conception, did not differ between treatments. However, cows fed the lower n-6:n-3 ratio tended to produce a greater proportion of female offspring (66.67% vs. 42.90%; p = 0.09). In addition, calves born to cows receiving the lower n-6:n-3 ratio had greater birth weights than calves born to cows fed the greater n-6:n-3 ratio diet. Additionally, the lower n-6:n-3 diet tended to increase milk yield and significantly increased lactose and solids-not-fat yields. In conclusion, a lower dietary n-6:n-3 ratio during a TAI program enhanced luteal development, tended to increase the proportion of female offspring and increased calf birth weight. These findings suggest that reducing the dietary n-6:n-3 ratio during the breeding period may enhance luteal development and may influence offspring sex ratio and calf birth weight in lactating dairy cows.

Dairy doi: 10.3390/dairy7030042

Authors: Carlo Boselli Antonella Chiariotti Valentina D’Onofrio Maria Concetta Campagna Giuliano Palocci Vittoria Lucia Barile Antonio Borghese

Milk flow dynamics during mechanical milking are strongly influenced by species-specific mammary anatomy, milk partitioning between cisternal and alveolar compartments, and milking management. The present study aimed to compare milkability traits across the main dairy species reared in central Italy using a large dataset collected over 20 years. A total of 7315 animals were included: dairy cows (1103), buffaloes (2870), goats (2399), sheep (754), and donkeys (189). Milk flow curves were recorded using a portable Lactocorder® device. The following traits were analyzed: milk yield (MY), lag time (LT), milk ejection time (MET), total milking time (TMT), peak flow rate (PFR), average flow rate (AFR), plateau phase (PL), bimodal phase (BM), and bimodality incidence (Bimo). Marked interspecific differences emerged. Dairy cows showed the highest MY and PFR, with bimodality occurring in 23.7% of curves. Buffaloes exhibited lower flow rates, prolonged LT, and extended TMT, reflecting their strong dependence on oxytocin-mediated alveolar milk ejection. Sheep demonstrated short milking times and low bimodality (13.5%), consistent with their large cisternal milk fraction. Goats displayed breed-dependent variability, with specialized dairy breeds showing higher PFR and longer TMT. Donkeys produced low milk volumes but exhibited rapid and efficient milk flow, with the lowest incidence of bimodality (7.4%). Overall, milk flow patterns reflected species-specific udder morphology and physiological mechanisms of milk ejection. Although this field-based study faces inherent limitations in environmental and protocol standardization across farms, the resulting long-term dataset remains highly representative. These findings highlight the importance of tailoring milking machine settings and prestimulation protocols to species and breed characteristics to optimize milking efficiency, labor management, and animal welfare.

Dairy doi: 10.3390/dairy7030041

Authors: Khue Minh Tran Oanh Thi Hoang Lan Thi Nguyen

The increasing demand for sustainable and health-oriented dairy and alternative products has enhanced interest in reducing or modifying fat in cheese systems. However, such modifications often lead to undesirable changes in texture and flavor, highlighting the multifunctional roles of milk fat within the cheese matrix. Rather than serving solely as a compositional component, milk fat contributes fundamentally to structure organization and flavor development through its physicochemical properties, interactions with the protein network, and lipid-derived pathways. This review examines these roles from a mechanistic perspective and evaluates emerging lipid structuring approaches for texture modulation, while also discussing complementary approaches with potential to address flavor attributes. Collectively, it provides insights for rational formulation and guides future research toward the design of improved dairy and alternative cheese products.

Dairy doi: 10.3390/dairy7030040

Authors: Sébastien Czaplicki Jean-Luc Hornick Noémie Besserve Bertille Dumont Saint Priest Françoise Lessire Hélène Bartholomé Isabelle Dufrasne

Heat stress (HS) alters rumen function and may compromise fermentation efficiency in dairy cows. This in vitro study evaluated the effects of six additives—betaine, sodium bicarbonate, bentonite clay, protected fat, Saccharomyces cerevisiae yeast, and Melissa officinalis (lemon balm)—on ruminal fermentation under increasing incubation temperatures (39.0 °C, 40.5 °C, 41.5 °C). This study was designed as an initial in vitro screening approach aimed at evaluating additive resilience under a conservative, worst-case thermal challenge. The highest incubation temperature (41.5 °C) was not used to reproduce the exact physiological rumen environment but rather served to identify additives capable of maintaining fermentation when exposed to extreme conditions that may occur during severe heat load. These results therefore constitute a preliminary step before in vivo validation under realistic HS scenarios. Fermentation was assessed after 24 h by measuring gas production, pH, redox potential, volatile fatty acids (VFAs), and ammonia (NH3) and performing protozoa counts. Temperature alone produced limited effects in the control, with numerical increases in gas, acetate, total VFAs, and NH3 at 41.5 °C. Betaine and yeast maintained overall stable fermentation across all temperatures, preserving gas production, pH, redox potential, and VFA profiles. Sodium bicarbonate increased the pH but reduced gas production. Lemon balm enhanced VFAs and propionate at 39.0 °C but showed reduced activity at high temperature. Protected fat and bentonite clay resulted in lower gas production with minimal influence on other parameters. The protozoa counts were not affected by temperature and showed statistically detectable differences among additives. Overall, betaine and yeast exhibited the highest thermal stability and appear to be suitable candidates for inclusion in feeding strategies aimed at supporting rumen fermentation during periods of HS.

Dairy doi: 10.3390/dairy7030039

Authors: Guanshi Zhang David S. Wishart Burim N. Ametaj

Ketosis is one of the most economically significant metabolic disorders affecting periparturient dairy cows, causing production losses and predisposing animals to secondary complications. Current blood-based diagnostics are invasive and provide limited insight into the underlying metabolic perturbations. This study employed an integrated three-platform metabolomics approach to characterize milk metabolite alterations in ketotic Holstein dairy cows and to evaluate milk-based biomarker panels for early ketosis detection. Milk samples from 20 healthy control (CON) cows and 6 ketotic cows were collected at 2 weeks postpartum and analyzed by direct injection/liquid chromatography–tandem mass spectrometry (DI/LC-MS/MS), proton nuclear magnetic resonance (1H-NMR) spectroscopy, and inductively coupled plasma mass spectrometry (ICP-MS). Ketosis was confirmed by serum β-hydroxybutyrate concentrations ≥ 1400 μmol/L. Principal component analysis, partial least squares-discriminant analysis, and receiver operating characteristic (ROC) curve analyses were applied. All three platforms discriminated ketotic cows from healthy cows, with clear cluster separation validated by 2000 permutation tests (p < 0.05). DI/LC-MS/MS identified 16 significantly altered metabolites (p < 0.05), with butyrylcarnitine (C4), phosphatidylcholine 30:0 (PC 30:0), ether-linked phosphatidylcholine O-38:3 (PC O-38:3), and citrulline identified as the top discriminatory biomarkers (AUC = 0.920; 95% CI: 0.85–0.98; sensitivity = 91.7%; specificity = 93.3%). ICP-MS revealed significantly reduced selenium (Se, p = 0.017), manganese (Mn, p = 0.045), and chromium (Cr, p = 0.037), as well as elevated cobalt (Co, p = 0.014) in ketotic milk (AUC = 0.870). 1H-NMR detected no individually significant metabolites; however, multivariate analysis distinguished groups (AUC = 0.890), with succinate (numerical fold change: +5.77×; p = 0.059), methanol (−1.94×; not significant), and acetate (+2.88×; not significant) as top VIP contributors. The combined multi-platform biomarker panel (joint classification using top VIP features from all three platforms, without formal data fusion) achieved superior diagnostic performance (AUC = 0.970; 95% CI: 0.93–1.00; sensitivity = 95.0%; specificity = 96.7%). These findings identify coordinated perturbations in glycerophospholipid metabolism, acylcarnitine profiles, amino acid homeostasis, antioxidant mineral status, and energy metabolism during early ketosis, and suggest that milk metabolomics is a promising non-invasive approach for precision dairy health monitoring, pending validation in independent cohorts. We acknowledge the small ketotic group size (n = 6) as a limitation; therefore, these findings should be considered discovery cohort observations requiring prospective validation before clinical translation.

Dairy doi: 10.3390/dairy7030038

Authors: Xin Li Md. Yousuf Ali Khan Xiling Hou Pin Chen Dandan Lang Yi Zhang

Genetic screening of monogenic traits is important for improving genetic health and increasing favorable milk protein in dairy cattle. This study developed and applied an amplification refractory mutation system PCR (ARMS-PCR) assay to simultaneously screen 21 causal variants underlying 18 monogenic traits in Holstein cattle, including 13 recessive genetic defects, 2 milk protein loci, and 3 morphological loci. The assay was designed as a unified multiplex PCR-capillary electrophoresis workflow, enabling clear detection of allele-specific products distinguished by fragment size and fluorescent color. Sanger sequencing validation of newly incorporated loci supported the accuracy of the assay. A total of 1656 cows from 12 commercial farms were genotyped using the multiplex ARMS-PCR panel, and amplicons were analyzed by capillary electrophoresis. Carriers were detected for all genetic defects except DUMPS, with carrier frequencies ranging from 0.12% to 6.64%. The highest frequencies were observed for HH5 (6.64%) and MWS (6.58%), whereas HH3, HH1, and HCD showed intermediate frequencies of 1.81% to 3.08%; all the remaining defects were below 1%. Overall, 22.10% of sampled cows carried at least 1 defect allele, including 20.47% carrying 1 defect, 1.51% carrying 2, and 0.12% carrying 3. For milk protein loci, the desirable β-casein A2A2 and κ-casein BB genotypes occurred at frequencies of 45.83% and 14.13%, respectively, and the favorable A2A2/BB combination was present in 6.22% of sampled cows. Dominant red, recessive red, and polled alleles were rare. These results indicate that multiplex fluorescent ARMS-PCR can serve as a practical targeted screening tool for the simultaneous management of known deleterious alleles and selection of favorable monogenic variants in dairy cattle breeding.

Dairy doi: 10.3390/dairy7030037

Authors: Ioannis Sakaridis Maria Ioannidou Martha Maggira Georgios Samouris

Milk adulteration is a common form of food fraud, particularly in high-value dairy products from small ruminants. A frequent practice involves dilution with water, often combined with the addition of sugars to mask physicochemical changes and avoid detection during routine quality control. This study aimed to develop an analytical approach for detecting combined adulteration in goat and sheep milk involving both water dilution and sucrose addition. Controlled experiments were conducted by diluting milk samples with water (1–15%) followed by the addition of sucrose solutions. Changes in physicochemical parameters, including fat, protein, total solids, lactose, density, freezing point depression, mineral content, and pH, were evaluated using an automated milk analyzer. In parallel, a suspected adulterant powder was characterized using conventional chemical analysis, ICP-AES, and HPLC-RI, revealing a composition predominantly of sucrose (91.4% w/w) with elevated sodium levels. Sucrose in milk samples was subsequently quantified using an enzymatic spectrophotometric method. Water dilution reduced protein, total solids, and density, while sucrose addition partially restored these parameters, masking adulteration effects. However, sucrose was reliably detected at concentrations above 0.1%. The proposed workflow may provide a practical and cost-effective complementary tool for routine dairy authenticity surveillance and fraud prevention.

Dairy doi: 10.3390/dairy7030036

Authors: Gorka Santamarina-García Min Yap Fiona Crispie Gustavo Amores Cathy Lordan Mailo Virto Paul D. Cotter

Artisanal raw milk cheeses harbour complex microbial communities that drive cheese making and shape sensory quality. Previous work on Idiazabal cheese identified rennet as a major microbial source, although all reservoirs contributed to varying degrees. However, their impact in terms of enzyme-encoding genes related to technological quality of cheese remained unexplored. Building on that, this study draws on metagenome-assembled genomes (MAGs) from cheeses and dairy environments to comprehensively identify enzyme-encoding genes involved in key biochemical processes. In cheese MAGs (Lacticaseibacillus paracasei), protease-encoding genes were dominated by ATP-dependent metalloproteases (M41), carbohydrate-active enzyme-encoding genes (CAZymes) by glycoside hydrolases (GH) and glycosyltransferases (GT), while esterase, lipase, and related-enzyme-encoding genes were restricted to sparse ‘GDXG’, type-B and esterase D families. Dairy environments emerged as major reservoirs of enzyme-encoding genes, with notable differences among sample types (p ≤ 0.001). The richest sources of protease-encoding genes were grass (610 genes), linked primarily to Pantoea agglomerans, and rennet (318), mainly related to Basfia sp. and Moraxella sp., dominated by metalloproteases (M23, M38) and serine proteases (S15). The largest reservoirs of CAZyme-encoding genes were food contact surfaces (1550), associated mainly with Salinisphaera sp. and Dietzia sp., and rennet (1505), related to, e.g., Bacteroides pyogenes, Alloprevotella sp., and Lentilactobacillus buchneri. Food contact surfaces were also the richest source of esterase, lipase and related-enzyme-encoding genes (1209), mainly linked to Dietzia sp., Corynebacterium sp., and Brevibacterium aurantiacum. Similarly, aroma-related enzyme-encoding genes (e.g., oppA, pepA, GH13, esterase D) were consistently detected in environmental matrices. These results provide novel insights into dairy microbiomes as functional reservoirs of aroma precursors, revealing their relevance for artisanal PDO cheese production and future biotechnological applications.

Dairy doi: 10.3390/dairy7030035

Authors: Sayoko Oishi Yui Kitagawa Karen Koshimizu Shiki Nakatsuji Shigeki Morikawa Yujiro Hagita Hisataka Iwata Ryotaro Miura Seizo Hamano Koumei Shirasuna

Repeat breeder (RB) cows fail to conceive after three or more breeding attempts, yet the uterine factors contributing to this condition remain unclear. Here, we compared uterine fluid composition, endometrial cell responses to uterine fluid, and uterine microbiota between normal pregnant (Normal) and RB cows. Uterine fluid was collected from multiparous Holstein cows classified as Normal (n = 5; conception within three breeding attempts) or RB (no conception after four attempts); RB cows were classified as RB-Rec (n = 4; conceived after uterine lavage) or RB-NoRec (n = 3; did not conceive after uterine lavage). Uterine fluids were applied to bovine endometrial cells, and microbiota composition was analyzed. Concentrations of lipopolysaccharide in the uterine fluid did not differ between the three groups, and uterine fluid from both RB groups significantly increased C-X-C motif chemokine ligand 8 secretion relative to the Normal group. RNA-seq revealed distinct endometrial cell responses to RB-NoRec uterine fluid, with enrichment of immune-related pathways including phagosome, ferroptosis, and cellular senescence. Uterine microbiota profiles differed among groups, with short-chain fatty acid-producing genera (e.g., Fusicatenibacter and Dorea) predominant in Normal cows and inflammation-associated genera (e.g., Trueperella and Helicobacter) predominant in RB-NoRec cows. These data indicate that uterine fluid properties and microbiota composition vary with reproductive status and may modulate endometrial immune activation associated with fertility outcomes.

Dairy doi: 10.3390/dairy7030034

Authors: Cindy Bande-De León David Delgado Domingo Fernández Massimo Mozzon Adela Abellán Lucia Aquilanti Luis Tejada

Plant-derived coagulants are increasingly explored as alternatives to animal rennet. This study provides the first evaluation of the coagulation kinetics and technological properties of Onopordum platylepis Murb. in ewe’s milk. Response surface methodology was applied to optimize temperature (30, 33, and 36 °C), pH (5.5, 6.0, and 6.5), and calcium chloride concentration (2, 4, and 6 mM). The optimal conditions for minimizing milk-clotting time were 36 °C, pH 5.5, and 6 mM CaCl2. Under standardized activity (50 IMCU/L), the Rheological properties of gels produced by Onopordum platylepis were compared with Cynara cardunculus, Cynara humilis, animal rennet, a commercial plant coagulant, and fermentation-produced chymosin. Onopordum platylepis showed slower curd-firming rates than animal rennet and Cynara cardunculus, but similar behavior to Cynara humilis. Gels produced with Onopordum platylepis exhibited firmness comparable to commercial plant coagulants. The water-holding capacity was similar to other coagulants, though protein losses were higher for Onopordum platylepis, Cynara humilis, and Cynara cardunculus than animal rennet. Overall, Onopordum platylepis demonstrates potential for ewe’s milk cheese production, in which highly proteolytic coagulants are used.

Dairy doi: 10.3390/dairy7030033

Authors: André L. A. Neves Moritz A. Pfeiffer Rajan Dhakal Svenja Woudstra Volker Krömker Hanne H. Hansen Ricardo Augusto Mendonça Vieira

The authors would like to make the following corrections to the published paper [...]

Dairy doi: 10.3390/dairy7030032

Authors: Atalay Ergül Celile Aylin Oluk Çiğdem Takma Serap Göncü Mervan Bayraktar

Milk color reflects the optical output of a complex colloidal system governed by protein micelles, fat globules, and serum phase interactions. In this study, we evaluated whether CIE Lab* color parameters can explain variation in milk composition and somatic cell count (SCC) using Lasso-based multiple linear regression and Extreme Gradient Boosting (XGBoost). A total of 119 Holstein milk samples were analyzed for fat, protein, lactose, dry matter, electrical conductivity, freezing point, and SCC, and five color indices (L*, a*, b*, Hue, and Chroma) were used as predictors. Model robustness was evaluated using 10-fold cross-validation and an independent 80/20 train–test split. In regression analyses, Lasso explained 32.7% of protein variation (R2 = 0.327), 26.3% of dry matter (R2 = 0.263), 22.8% of lactose (R2 = 0.228), and 19.1% of fat (R2 = 0.191). Spectral tone parameters (a*, Hue, and Chroma) were consistently retained as key predictors, whereas L* showed a limited contribution. SCC exhibited weak direct associations with color traits but was significantly related to electrical conductivity (p < 0.05), indicating inflammation-driven ionic changes rather than pigment effects. In classification analysis (SCC ≥ 200,000 cells/mL), the XGBoost model achieved 74% accuracy and an AUC of 0.69 in the independent test set, with Chroma and electrical conductivity identified as the most influential features. These findings suggest that, among the evaluated color variables, Chroma provided the most relevant information for discriminating SCC status, whereas the overall contribution of milk color traits to compositional prediction remained moderate. Therefore, color-derived measurements should be interpreted as instrument-based optical indicators that may complement, but not replace, conventional milk quality assessments.

Dairy doi: 10.3390/dairy7030031

Authors: Aurelio Guevara-Escobar Vicente Lemus-Ramírez José Guadalupe García-Muñiz Adolfo Kunio Yabuta-Osorio Claudia Andrea Vidales-Basurto Benjamín Valdés-Aguirre

Modeling milk yield in dairy cows is essential for improving management decisions, but traditional lactation curve models often fail to capture individual variability. Machine learning approaches offer greater flexibility; however, their performance in small, within-herd datasets and their reliance on explicit cow identification remain unclear, particularly in grazing systems. This study aimed to evaluate whether routinely measured biological traits can substitute for cow identification in machine learning models for predicting daily milk yield within a herd under limited data conditions. The dataset comprised 62 lactations from 48 Holstein–Friesian cows in a grazing system. Two machine learning models were developed: one including cow identification (With ID) and another excluding cow identification but incorporating milk quality traits, body weight, and body condition score (Without ID). Both models were compared with the Wood lactation model fitted to individual cows. The With ID and Without ID models achieved R2 values of 0.97 and 0.93 and RMSE values of 1.2 and 1.6 kg d−1, respectively. Both machine learning models outperformed the Wood model fitted individually to each cow (R2 < 0.90; RMSE > 2.03 kg d−1), which represents an implicitly cow-specific approach. The model including cow identification therefore served as a machine learning analogue to this benchmark. Importantly, the trait-based model closely matched the performance of the cow-specific model. These results demonstrate that machine learning models based on routinely measured traits provide a practical approach for predicting within-herd milk yield from small datasets, while retaining much of the accuracy of cow-specific models.

Dairy doi: 10.3390/dairy7030030

Authors: Giorgos Notis Maria Perroti Chrystalla Demosthenous Manos C. Vlasiou

Milk is a complex biochemical mixture in which proteins significantly influence the behaviour of xenobiotics and bioactive compounds. Interactions between milk proteins and substances such as veterinary drugs or natural bioactives can modify molecular stability, binding dynamics, and exposure pathways, affecting food safety and the One Health concept. This study presents a comparative, matrix-focused investigation on how three chemically distinct ligand classes, sulfanilamide antibiotics, naturally occurring phenolic compounds and zinc–polyphenol complexes, interact with major milk proteins, β-lactoglobulin and casein. Protein–ligand interactions were examined using steady-state fluorescence spectroscopy to assess quenching behaviour and comparative interaction trends. Molecular docking was employed as a qualitative tool to provide structural context. Distinct interaction patterns were observed across ligand classes, reflecting differences in molecular structure, hydrophobicity, and coordination chemistry. Importantly, zinc coordination modified interaction profiles relative to the corresponding free ligands, indicating that metal coordination can affect ligand–protein interactions within the milk matrix. These findings support the concept that milk proteins may function as matrix-dependent modulators of ligand behaviour. The study is positioned as a hypothesis-generating framework highlighting the importance of food matrices as active biochemical environments. Herein, we provide a foundation for hypothesising how the milk matrix affects residue behaviour and bioactive interactions, with relevance to veterinary pharmacology and food safety risk assessment.

Dairy doi: 10.3390/dairy7020029

Authors: Thomas Paravicini Magdalena Nüesch-Inderbinen Markus Mader Karin Darpel Roger Stephan Claudia Bachofen

Highly pathogenic avian influenza virus (HPAIV) H5N1 has been detected in dairy cattle in the United States, with high viral loads observed in milk from infected animals. This raises public health concerns regarding potential transmission through exposure to raw milk. The sale of raw milk via vending machines represents a well-established distribution model in many European countries, including Switzerland. Although a notice must be posted on these milk vending machines stating that it is raw milk, together with appropriate processing instructions (heating to over 70 °C required, storage below 5 °C, consumption within 3 days), these notices are sometimes missing, and consumers often do not follow these guidelines. Over a four-month period, spanning from June 2025 to September 2025, 124 raw milk samples were collected from vending machines across Switzerland. Samples were screened for influenza A using reverse-transcription quantitative PCR (RT-qPCR). No samples tested positive for influenza A virus. The data from this study demonstrate the feasibility of implementing a sampling and detection system for HPAIV H5N1 in direct-to consumer raw milk samples and highlight the currently very low risk of HPAIV in raw milk samples sold via vending machines in Switzerland.

Dairy doi: 10.3390/dairy7020028

Authors: Severino Guilherme Caetano Gonçalves dos Santos Carla Aparecida Soares Saraiva Severino Gonzaga Neto Vinícius de França Carvalho Fonsêca Aline Cristina Sant’Anna Maria Isabelly Leite Maia Luiz Arthur dos Anjos Lima Tarsys Noan Silva Veríssimo Larissa Kellen da Cunha Morais Wylke Alves de Azevedo Soares Pavlos Vinícius do Nascimento Delfino Isac Belarmino Afo Edilson Paes Saraiva

A cross-sectional study was carried out on small-scale dairy farms in Paraíba State, Brazil, to evaluate the prevalence and anatomical distribution of integument injuries in grazing dairy cows. Twelve herds (n = 12) were assessed, recording herd size, milk yield, and daily hours of pasture access. The integument of five body regions (back/shoulder/neck, carpus, flank/side/udder, tarsus, and hindquarters) was scored for hairless spots, lesions, and swellings (0 = absent; 1 = present). Among 335 cows observed, 267 (81.5%) presented at least one injury. The most frequent conditions were hairless patches on the hock (65.4 ± 5.03%), lesions on the flank/side/udder (34.5 ± 1.46%), and swelling in the back/shoulder/neck region (52.9 ± 5.86%). Cluster analysis revealed distinct patterns of integument injuries among cows. The high prevalence of integument injuries indicates that even in grazing systems, welfare concerns persist. Neck swelling and hairless spots on the hock were the most common problems, emphasizing the need for improved management and environmental practices to enhance welfare outcomes. These findings have practical implications for welfare assessment and management strategies in tropical grazing dairy systems.

Dairy doi: 10.3390/dairy7020027

Authors: Joel Bueso-Ródenas Gema Romero Alfonso Navarro Elena Pérez Pilar Gascó José Ramón Díaz

Infrared thermography has been proposed as a non-invasive tool for mastitis detection in dairy ruminants; however, the extent of environmental confounding and diagnostic performance in small ruminants remain poorly characterized. This study evaluated udder thermography in dairy goats through correlation analysis under winter and summer conditions, and an experimental intramammary inflammation challenge using Staphylococcus aureus lipoteichoic acid, with receiver operating characteristic analysis using somatic cell count >1500 × 103 cells/mL as the reference standard. Strong positive correlations between ambient temperature and udder surface temperatures intensified substantially from winter to summer, while surface temperatures showed weak or absent correlations with rectal temperature. Experimental inflammation induced a 12-fold increase in somatic cell count (305 vs. 3658 × 103 cells/mL, p < 0.001); however, thermographic responses remained minimal and spatially inconsistent, with area under the curve values approximating 0.5 and weak correlations between thermographic measurements and somatic cell count. Environmental temperature variation and physiological thermoregulatory adjustments substantially exceeded the minimal thermal signal generated by intramammary inflammation, limiting diagnostic utility. Infrared thermography showed poor diagnostic utility for detecting experimentally induced intramammary inflammation in dairy goats under the tested conditions.

Dairy doi: 10.3390/dairy7020026

Authors: Adriana Wöckel Wolf Wippermann Benno Waurich Erik Bannert Julia Wittich Christina Felgentreu Franz Fröhlich Fanny Rachidi Peter Hufe Detlef May Sven Dänicke Hermann H. Swalve Alexander Starke Melanie Schären-Bannert

Production diseases in dairy cattle impose economic and welfare burdens, yet few studies quantify costs using on-farm cases. This study aimed to estimate costs and lost revenues at the individual-animal level in 10 German dairy farms (average of 592 cows; 9694 kg marketed milk/cow/year; 32.9% culling rate). Each farm was visited for three weeks; diseased cows and calves were examined by a trained veterinarian. Diagnoses, treatments, labour times, and outcomes were recorded, and costs calculated for labour, products, veterinary and orthopaedic services, discarded milk, decreased milk yield, culling, book loss, and reduced carcass value. In total, 1272 single-animal cases were included: 68% were stand-alone diseases, 11% involved multiple diagnoses within one organ system, and 21% affected several organ systems. When several diseases occurred in the same animal, total costs and lost revenues were greater than the sum of stand-alone cases, indicating compounding effects. High-impact conditions included mastitis, claw disorders, left displaced abomasum, and multimorbidity; per-case losses ranged from €43 (digital dermatitis) to >€1200 (left displaced abomasum with complications). Labour and culling-related costs were higher than reported, and productivity losses exceeded treatment costs in many cases. Findings support farm-level decision-making, prevention, and parameterization of future dynamic models.

Dairy doi: 10.3390/dairy7020025

Authors: Ana Maria da Costa Goncalves Noronha Eslam Ahmed Ahmed O. Matti-Alapafuja Belgutei Batbekh Masaaki Hanada Naoki Fukuma Takehiro Nishida

The need for new feed ingredients that could reduce methane (CH4) emissions from dairy cattle while maintaining rumen function is essential for sustainable milk production. This study aimed to evaluate the CH4 mitigation potential of selected microalgae and macroalgae, along with an agro-industrial by-product, using two feeding strategies, and hypothesized that lipid- and polyphenol-rich materials would reduce CH4 production in an inclusion-dependent manner. An in vitro batch culture study (24 h) was conducted to evaluate microalgae (Euglena gracilis and Aurantiochytrium spp.), macroalgae (Undaria pinnatifida), and an agro-industrial by-product (grape marc) either as feed additives (5%) or as a partial replacement of the concentrate mixture (30%, 50%, and 70%) in a basal diet consisting of 50% Klein grass hay and 50% concentrate mixture. As a feed additive, grape marc stands out for its potential to reduce CH4 yield by about 43.3% without adversely affecting digestibility, pH, or total volatile fatty acid concentrations. When used as feed replacements, Euglena-, Aurantiochytrium-, and grape marc-based feeds reduced CH4 yield at the highest replacement levels (50 and 70%); however, these effects were accompanied by decreased total gas production and volatile fatty acid concentrations, indicating reduced fermentation activity. Meanwhile, at a 30% replacement level, they showed promising efficiency as alternative feeds. Overall, CH4 mitigation depends more strongly on inclusion strategy rather than feed type. Lipid-rich microalgae showed potential as concentrate replacements up to 30%, whereas grape marc was most effective as a feed additive for reducing CH4 emissions.

Dairy doi: 10.3390/dairy7020024

Authors: Crina-Raluca Andrei Nicolae Tiberiu Constantin Remus Ioan Chiorean Adrian Bota Florin Petrișor Posastiuc Mariana Ioniță Ioan Liviu Mitrea

This study evaluated key reproductive and productive parameters of the Romanian buffalo reared under temperate-continental conditions. Data collected over a 30-year period (1990–2020) from 310 female buffaloes raised at the Şercaia Buffalo Research and Development Station (Brașov County, Romania) were analyzed. Only animals that completed their first gestation and calving within the herd and had complete lifetime records were included. Buffaloes were monitored longitudinally from first calving until the end of their productive life, regardless of the reason for exit. The evaluated traits included age at first calving, number of lactations, milk yield, milk fat content, and the effect of calf sex on milk production. Data were analyzed using descriptive statistics, group comparisons, and linear regression. Mean age at first calving was 51.6 ± 14.0 months, and females completed an average of 7.8 ± 3.0 lactations, indicating good reproductive longevity. Buffaloes calving male offspring produced significantly more milk per lactation (1272 ± 267 L) than those calving female offspring (1099 ± 244 L; p < 0.01). Milk fat content averaged approximately 7% and did not differ significantly between females giving birth to female offspring and those giving birth to male offspring (p > 0.05). Overall, the results indicate good productive stability and reproductive durability of the Romanian buffalo under semi-intensive management and confirm its adaptability to temperate environments.

Dairy doi: 10.3390/dairy7020023

Authors: Thierno Abdoul Rahim Sow Moncef Hammami Alpha Oumar Sily Diallo Romdhane Karoui

Acid-induced coagulation of milk plays an important role in the production of dairy products with high quality. The gel texture is significantly affected by processing conditions such as heat treatment and mineral composition. This study examines the effect of moderate heat treatment applied at 65 °C for 5 min and ionic calcium supplementation (10 mM CaCl2) on coagulation at 30 °C for 180 min of cow’s milk induced by glucono-δ-lactone (GDL). A slow and gradual protonation was induced, reaching pH values of 4.3–4.5. Rheological analysis revealed an increase in G′ from 59.28 Pa for raw milk gel to 224.1 Pa after the addition of CaCl2. An inverse trend was observed for gel produced with heated milk after the addition of CaCl2. However, the gel produced from calcium-fortified heated milk showed G′ values of 136.7 Pa. Turbiscan analysis showed the highest TSI for gels made from heated milk. Scanning electron microscopy analysis indicates that raw milk gels supplemented with CaCl2 exhibit dense and homogeneous networks, while heat-treated GDL gels show more porous networks. Mid-infrared (3000–2800 cm−1, 1700–1500 cm−1, and 1500–900 cm−1) and fluorescence spectra revealed changes in protein–protein, protein–water, and protein–protein–lipid interactions throughout coagulation.

Dairy doi: 10.3390/dairy7020022

Authors: Elizangela Domenis Marino Gercino Ferreira Virgínio Júnior Ariany Faria de Toledo Izally Carvalho Gervásio Simara Larissa Fanalli Carlos Eduardo Cardoso de Aguiar Freire Samuel Ferreira Gonçalves Carolina de Souza Moreira Matthew Flavel Luiz Carlos Roma Júnior Weber Vilas Bôas Soares Aline Silva Mello Cesar Carla Maris Machado Bittar

The present study evaluated the effects of sugarcane-derived polyphenol nutritional supplement on milk production, milk composition, and somatic cell count (SCC) in lactating Holstein cows. A second objective was to evaluate the quality and stability of yogurt produced with milk from supplemented cows. Sixteen cows (20 ± 2 L/d milk, 120 ± 21 days in milk, 550 ± 82 kg body weight) were assigned to either a control group or a polyphenol-supplemented group in a randomized block design for a six-week evaluation period. Milk yield and individual milk samples for composition analysis (fat, protein, lactose, non-fat solids, casein, free fatty acids, and milk urea nitrogen) were recorded weekly, and only SCC was monitored three times per week. Yogurt was produced from pooled milk of each treatment and evaluated for fermentation parameters (pH, titratable acidity), shelf life, and sensory attributes were evaluated using a triangle test (discriminative sensory analysis). Polyphenol supplementation did not significantly affect milk yield or composition (p > 0.05), but reduced SCC considerably (238 × 103 vs. 593 × 103 cells/mL; p < 0.01). Yogurt fermentation and storage parameters were not significantly affected by supplementation, although pH and titratable acidity changed considerably over time (p < 0.01). Sensory analysis indicated that only 37.3% of panelists correctly identified differences between yogurts, which did not differ from chance expectation in the triangle test. These results demonstrate that polyphenol supplementation can improve udder health, evidenced by reduced SCC, without compromising milk yield, composition, or yogurt quality. Polyphenols thus represent a promising nutritional supplement for enhancing dairy cow health and maintaining product quality.

Dairy doi: 10.3390/dairy7020021

Authors: Nikoleta Lugonja Milena Milojević Petar Ristivojević Maja Krstić Ristivojević Dalibor Stanković

Milk is a highly nutritious food and a cornerstone of the human diet, supplying not only essential macronutrients but also a wide range of bioactive compounds with important functional and health-promoting properties. This study presents the first comparative analysis of ruminant (cow, goat, sheep) and equine (mare, jenny) milk samples collected in Serbia, with emphasis on their physicochemical properties, protein profile, redox characteristics, and nutritional potential. Ruminant milk had significantly higher protein concentrations, with cow and sheep milk containing the highest levels of protein. Two equine milks demonstrated a distinctive whey-to-casein protein ratio to ruminants, and a higher content of active sulfhydryl groups, correlating with improved digestibility and functional properties. Antioxidant potential was determined using spectrophotometric and electrochemical methods, confirming superior redox potential in mare’s milk, followed by jenny’s and sheep’s milk. Nutritional properties of milk separated by Principal Component Analysis highlighted species-specific profiles of equine milks as a promising alternative for individuals with an allergy to cow’s milk protein, offering enhanced antioxidant protection, bioactive compounds, and digestibility. These results support the potential of equine milk as a functional food with added value in human nutrition.

Dairy doi: 10.3390/dairy7010020

Authors: Anri Timonen Rohish Kaura Annely Aleksejev Lea Tummeleht Kerli Mõtus Arvo Viltrop Toomas Orro

This cross-sectional study investigated the associations between the acute-phase proteins (APP) serum amyloid A (SAA) and haptoglobin (Hp), herd-level factors, and the seroprevalence of respiratory pathogens in Estonian dairy herds. Serum samples were analysed from 938 cows (95 herds) and 921 heifers (94 herds). Seroprevalence was tested for bovine herpesvirus 1 (BHV-1), bovine respiratory syncytial virus (BRSV), bovine parainfluenza virus 3, bovine viral diarrhoea virus, bovine coronavirus, bovine adenovirus, and Mycoplasma bovis (M. bovis). Farm visits included questionnaires on herd management practices. Linear random-intercept regression models showed higher serum SAA concentrations in cows from farms with BHV-1 seroprevalence of >50% and on BRSV-positive farms (p < 0.05), while farms employing a veterinarian had lower serum SAA concentrations. Cows had higher serum Hp concentrations in M. bovis-positive herds (p = 0.030). In heifers, serum SAA concentrations increased with low to moderate BHV-1 seroprevalence, decreased with higher M. bovis seroprevalence, and were higher in free-stall or mixed housing compared to tie-stall housing. Heifers’ serum Hp concentrations were lower in BHV-1-positive herds, but higher in herds with breeding bulls and larger herd sizes. To conclude, APP may reflect the herd health status and management-related effects on animals, supporting their use in herd-level monitoring.

Dairy doi: 10.3390/dairy7010019

Authors: Burim N. Ametaj

The transition from energy sufficiency to deficiency triggers complex metabolic and immune adaptations that have traditionally been viewed through a reductionist pathological lens. During early lactation, coordinated mobilization of adipose tissue, muscle protein, and bone minerals supports milk synthesis, with ketogenesis specifically arising from hepatic oxidation of non–esterified fatty acids. This review introduces the Keto–Inflammatory Network (KIN), a novel framework positioning ketonemia as an evolutionarily conserved adaptive response rather than inherent metabolic dysfunction. The KIN integrates β–hydroxybutyrate (BHB) signaling with immune modulation, epigenetic regulation, circadian rhythms, and microbiota interactions. Through mechanisms including NLRP3 inflammasome inhibition, HDAC–mediated epigenetic modifications, and HCAR2 receptor activation, ketone bodies orchestrate anti–inflammatory responses while maintaining metabolic flexibility. Building upon important precedent work recognizing beneficial roles of ketones in ruminant metabolism, this review synthesizes recent advances in immunometabolism and systems biology into an integrated framework. The KIN encompasses calcium–ketone integration through the Calci–Keto–Inflammatory Code (CKIC), temporal regulation via the Ketoinflammatory Clock, and trans–kingdom signaling through microbiota interactions. In dairy cattle, this perspective reframes periparturient ketonemia as existing on a continuum from adaptive to pathological, with biological meaning determined by integrated metabolic–inflammatory patterns rather than absolute ketone concentrations. The CKIC paradigm, while requiring prospective validation, suggests novel therapeutic approaches leveraging ketone signaling for inflammatory diseases, autoimmune conditions, and metabolic disorders while challenging traditional threshold–based ketosis management strategies. This systems–level understanding opens new avenues for precision interventions that work with, rather than against, evolved adaptive mechanisms refined through millions of years of mammalian evolution. By distinguishing ketonemia (measurable ketone elevation) from pathological ketosis (dysregulated ketone accumulation), and by integrating evidence from both ruminant and monogastric models, this review provides a comprehensive framework for next–generation metabolic medicine.

Dairy doi: 10.3390/dairy7010018

Authors: Tijana Ledina Matija Munjić Vladimir S. Kurćubić Ivana Branković Lazić Jasna Lončina

Enterococci are ubiquitous lactic acid bacteria frequently detected in dairy environments, where they represent an important component of the non-starter lactic acid bacteria community, particularly in artisanal cheeses produced from raw milk. Due to their metabolic versatility, enterococci may contribute to cheese ripening and the development of characteristic sensory attributes; however, their technological relevance is accompanied by growing concern regarding their role as opportunistic pathogens and reservoirs of antimicrobial resistance. This review critically summarizes current knowledge on antimicrobial resistance in enterococci isolated from milk and dairy products, with emphasis on both intrinsic and acquired resistance traits and their reported prevalence across different dairy matrices and geographical regions. Particular attention is given to artisanal cheeses, in which heterogeneous and region-specific resistance patterns have been described. Advances in whole-genome sequencing have substantially improved understanding of the genetic basis of antimicrobial resistance in dairy enterococci and have largely corroborated earlier findings obtained through phenotypic antimicrobial susceptibility testing combined with targeted resistance gene detection. Nevertheless, available data remain fragmented due to variability in study design, analytical approaches, and reporting practices. Overall, the evidence highlights the need for harmonized surveillance strategies integrating phenotypic and genomic data within a One Health framework to improve risk assessment and to better understand the role of dairy enterococci in the dissemination of antimicrobial resistance along the food chain.

Dairy doi: 10.3390/dairy7010017

Authors: Antônio Paulo de Oliveira Neto Tássia Barrera de Paula e Silva Kellen Ribeiro Oliveira Luis Henrique Rodrigues Silva Poliana Teixeira Rocha Salgado Simone Eliza Facioni Guimarães Alex Lopes Silva Polyana Pizzi Rotta

This study examined the effects of average daily gain (ADG) during gestation on growth, nutrient digestibility, metabolic response, and subsequent milk yield and composition in dairy heifers. Twenty pregnant Holstein × Gyr heifers (450 ± 5.0 kg; 18 ± 1.1 months) were randomly assigned to moderate (MOD; target 0.35 kg/day) or high (HIG; target 0.70 kg/day) ADG groups, and received a total mixed ration from day 70 of gestation until calving. Body growth, blood metabolites, and lactation performance after birth were measured. At calving, HIG heifers had greater body weight (p < 0.01) and thoracic perimeter (p = 0.02). Nutrient digestibility and most blood metabolites were not affected by ADG (p > 0.05), except for triiodothyronine concentrations, which differed between treatments over time (p < 0.01). Milk yield and energy-corrected milk were not affected by gestational ADG (p > 0.10), while milk fat and total solids showed numerical treatment × week interactions (p ≤ 0.10). These results indicate that higher ADG during gestation increases body reserves at calving but does not affect milk yield. The moderate ADG for Holstein × Gyr heifers during gestation may improve milk quality through higher fat and solids content, emphasizing the importance of tailoring growth strategies for heifers during gestation.

Dairy doi: 10.3390/dairy7010016

Authors: Carmen Burner Hunter G. Perez Anderson A. C. Alves Valerie E. Ryman

Historically, methods to detect subclinical mastitis have been used to aid treatment decisions. The limitations of these tests have led to the exploration of the use of differential somatic cell count (DSCC). The objective of this study was to evaluate the relationship between DSCC and antibiotic success in subclinically infected cows. Cows were pre-screened to confirm an SCC ≥ 200,000 cells/mL and a positive bacterial culture. At enrollment, aseptic quarter milk samples were collected for culturing and evaluated using SCC, DSCC, the California Mastitis Test (CMT), and electrical conductivity. Enrolled quarters were treated with an intramammary antibiotic and resampled 7-, 14-, 28-, and 35-days post-treatment and retrospectively classified as “Cured” or failed to cure (Failed) based on a bacteriological cure. The overall cure rate was 51.5%. Pre-treatment SCC, total leukocyte count (TLC), neutrophils, and CMT were lower in Cured quarters compared to Failed. Meanwhile, macrophage percentage was greater in Cured quarters compared to Failed. No difference was found between lymphocyte percentage and conductivity. When evaluating the use of multiple threshold optimizations to predict cure outcome, TLC tended to have the highest ranking for the metric of interest; however, SCC tended to numerically perform as well in sensitivity and specificity outside of the optimized parameter.

Dairy doi: 10.3390/dairy7010015

Authors: Mohammad Javed Ashar Paula A. Gonzalez-Rivas Frank R. Dunshea Christina D. Marth Surinder S. Chauhan

The transition period, spanning approximately three weeks before and after parturition, represents one of the most critical physiological windows in dairy production. Profound metabolic, endocrine, and immune adaptations occur as the cow shifts from gestation to lactation, predisposing high-yielding cows to oxidative stress, inflammation, and metabolic imbalance. Negative energy and metabolisable protein balances stimulate extensive lipolysis and the accumulation of non-esterified fatty acids (NEFAs) and β-hydroxybutyrate (BHBA), which can impair hepatic function and postpartum performance. This review integrates recent advances in the understanding of metabolic and immunometabolic regulation during transition, emphasising the interplay among energy metabolism, oxidative stress, and immune dysregulation. It critically re-evaluates current nutritional interventions including controlled-energy and negative DCAD diets, rumen-protected polyunsaturated fatty acids, and methyl donor amino acids while highlighting the emerging potential of injectable trace minerals as adjunct strategies to improve metabolic resilience. The review concludes by outlining remaining knowledge gaps and proposing a framework linking physiological mechanisms with targeted nutritional management.

Dairy doi: 10.3390/dairy7010014

Authors: Aline Luiza do Nascimento Cristina B. da Silva Ana Luiza de Freitas dos Santos Beatriz Danieli Bruna Klein Lucas Henrique Bavaresco Aline Zampar Creciana Maria Endres Andréia Maria Faion Nathália Coelho Andrade Jocinei Dognini Ana Luiza Bachmann Schogor

Colonial cheese production represents a valuable cultural and economic activity in southern Brazil. This study evaluated the effect of oral supplementation of dairy cows with an essential oil blend (EOB)—a combination of eucalyptus oil, peppermint oil, and menthol crystals—on the chemical composition and quality parameters of Colonial cheese during 21 days of ripening. Nine dairy cows were randomly assigned to three groups: control, EOB3.6 (3.6 g/cow/day), and EOB7.2 (7.2 g/cow/day). Milk from each treatment was used to produce Colonial cheeses, which were analyzed for physicochemical composition, texture, color, lipid profile, thiobarbituric acid reactive substances (TBARS), and microbiological quality at different ripening stages. Data were analyzed by analysis of variance (ANOVA) using SAS® software, following verification of normality and homogeneity of variances. When assumptions were met, repeated-measures ANOVA was applied, and means were compared using Tukey’s test (p < 0.05). Sensory data were evaluated by ANOVA using XLSTAT® (Addinsoft, Paris, France). EOB supplementation maintained the physicochemical integrity of the cheeses, with a gradual increase in fat content during maturation (40 g/100 g at 21 days, p < 0.05). At seven days, the EOB7.2 treatment showed lower lipid oxidation (TBARS = 0.063, p < 0.05), indicating antioxidant potential. Significant interactions between treatment and maturation were observed for color parameters and polyunsaturated fatty acids (PUFA) (p < 0.05). Cheeses from EOB7.2 presented higher saturated fatty acids (SFA) and lower unsaturated fatty acids (UFA) compared with the control (p < 0.05). No Salmonella spp. or Staphylococcal enterotoxins were detected. Counts of coagulase-positive Staphylococcus, molds, and yeasts remained stable, while Escherichia coli counts were lower in EOB-supplemented cheeses throughout ripening. Overall, EOB supplementation improved oxidative stability and microbiological safety without compromising the technological or compositional quality of Colonial cheese.

Dairy doi: 10.3390/dairy7010013

Authors: Aleksandra Rizojeva-Silava Sandija Zeverte-Rivza

The study analyzes the long-term development prospects of the Latvian dairy sector until 2032, using an integrated modeling approach that combines the AGMEMOD partial equilibrium model with the TOPSIS multi-criteria evaluation method. The study addresses the main challenge facing the sector—how to maintain productivity in the context of structural consolidation and increasing environmental requirements. The AGMEMOD model was recalibrated using updated data for Latvia for 2015–2023. Two scenarios were developed: A1 “Targeted and intensive farm modernization” and A2 “Limited farm modernization”. Scenario A1 is characterized by gradual technological adoption, leading to higher productivity while keeping total milk production almost unchanged relative to the Baseline scenario, whereas scenario A2 reflects slower modernization and reduced productivity growth. The TOPSIS evaluation identified scenario A1 as the most attractive alternative, as it combines productivity gains and greater adaptability to policy and environmental requirements. The results confirm that technological modernization and flexible policy mechanisms are essential to maintain the competitiveness and productivity performance of Latvia’s dairy sector. The integrated AGMEMOD–TOPSIS approach provides a methodological tool for evidence-based policy analysis and strategic planning in agricultural market management.

Dairy doi: 10.3390/dairy7010012

Authors: Lilla Sándorová Péter Árpád Fehér Ana Clarissa Ambagan Katalin Nagy Miklós Gábor Szabari Szilvia Áprily Szilárd Bodó Ákos Bodnár Péter Póti Ferenc Pajor Gabriella Holló Viktor Stéger

The polymorphism of the bovine β-casein gene (CSN2) is of increasing interest due to its relevance for A2 milk production. This study genotyped 2773 Holstein-Friesian cows for five CSN2 alleles (A1, A2, A3, B, I) using both conventional DNA-based PCR and a newly evaluated direct PCR protocol. Eleven genotypes were detected, with A2/A2 (33.9%) and A1/A2 (30.3%) being the most common, resulting in an A2 allele frequency of 59.0%. Genetic diversity indices indicated moderate polymorphism and a significant deviation from Hardy–Weinberg equilibrium, consistent with ongoing selection for the A2 allele. Associations between CSN2 genotype and milk traits (305-day milk, fat, and protein yield; fat% and protein%) were evaluated using linear mixed-effects models including lactation number, age at calving, and calving year as covariates, and cow ID as a random intercept. Several genotype effects reached statistical significance (p < 0.05); however, all effect sizes were very small (partial η2 < 0.01), indicating that any influence of CSN2 on production traits is negligible within this population and management context. These findings suggest that A2-oriented selection is unlikely to compromise productivity. The direct PCR genotyping method achieved 96–100% success and enabled substantially faster and more cost-efficient processing (approximately 80–90% reduction in reagent costs), providing a rapid and scalable approach for large herds.

Dairy doi: 10.3390/dairy7010011

Authors: Didem Şahingil Hacer Gürkan Ali Adnan Hayaloğlu

Kefir grains, originating from the Caucasus, are irregularly shaped, semi-solid granules resembling popcorn or cauliflower (0.3 to 3.5 cm), composed of bacteria and fungi embedded in extracellular polysaccharides such as kefiran. In this study, kefir samples were produced using different ratios of goat’s and cow’s milk (100:0, 75:25, 50:50, 25:75, and 0:100) to examine microbial populations, physicochemical and volatile properties, rheological behaviour, antioxidant capacity, and organic acid content. The type of milk used significantly affected the chemical composition and pH (p < 0.05), although titratable acidity remained stable during storage. Increasing the proportion of goat’s milk decreased viscosity but enhanced the total free amino acid (FAA) content. Goat milk kefir exhibited stronger antioxidant activity than cow milk kefir due to the formation of bioactive peptides and FAAs through proteolysis, with the highest values observed in samples with a higher proportion of goat’s milk and the lowest in 100% cow’s milk kefir. Organic acid levels increased during fermentation and, in 100% goat milk kefir, lactic, acetic, propionic, hippuric, butanoic, and citric acids reached their highest concentrations. The diversity and intensity of volatile compounds also increased proportionally with the goat milk ratio. In conclusion, considering antioxidant activity, volatile aroma components, organic acid content, sensory properties, and viscosity values, sample D, which is 75% goat milk kefir, is recommended for consumers.

Dairy doi: 10.3390/dairy7010010

Authors: Victoria Luño Karen Hammand Felisa Martínez

Colostrum is crucial for the survival, development, and the future productivity of newborns. In this study, we evaluated the immunological and microbiological quality of colostrum in 28 Rasa Aragonesa ewes and its relationship with offspring growth during the first 48 h postpartum. Colostrum samples were collected by hand milking immediately after parturition. Immunoglobulin concentration was assessed using Brix refractometry and the samples were categorised according to their immunoglobulin content: high (>24 Brix value), medium (19–23 Brix value), and low (<19 Brix value). Bacterial counts of aerobes and coliforms were determined with the 3M Petrifilm™ system and the weight of the lambs was recorded using a digital suspension scale. The mean aerobic count (AC) was 3.63 ± 0.69 log10 CFU/mL after 24 h of incubation and the mean coliform count (CC) was 1.59 ± 0.82 log10 CFU/mL after 24 h of incubation. Colostrum with a high immunoglobulin concentration had lower aerobic count after 48 h of incubation than that with poor immunological quality. In relation to coliform counts, similar values were found in all groups. No significant differences were observed in terms of lamb weight gain according to colostrum quality. In conclusion, the immunological quality of colostrum affected the AC determined, but it did not affect CC or early postnatal lamb weight. These findings offer preliminary insights into the usefulness of the Petrifilm™ system in microbiological quality determination of colostrum and its relationship with immunological quality determined in vitro.

Dairy doi: 10.3390/dairy7010009

Authors: Roman Konečný Michaela Horčičková Martin Kváč Lucie Hasoňová Eva Samková Hana Nejeschlebová Oto Hanuš Klára Bartáková

Lactoferrin (LF) is an iron-binding immunoprotein of the mammary gland whose levels increase during mastitis and may be influenced by the metabolic status of the cow. During early lactation, dairy cows are exposed to a negative energy balance (NEB) and the associated increase in susceptibility to mastitis. However, the extent to which the metabolic profile influences LF secretion in milk during the postpartum period remains unclear. The objective of this study was to assess the associations between metabolic status and milk LF contents in Holstein cows (n = 122) in the first twenty days of lactation. Based on the milk LF contents, the cows were categorized into two groups: LF-LOW (≤123 mg/L; n = 81) and LF-HIGH (>123 mg/L; n = 41). Serum indicators of energy and nitrogen metabolism, hepatic function, and selected macro-/microelements were measured; urine electrolytes and net acid–base excretion (U-ABB) were assessed; and milk composition, including somatic cell count (SCC), was determined. LF-HIGH cows showed higher SCC (p = 0.0516) and serum glucose (p < 0.001), together with lower serum triglycerides (p = 0.0101) versus LF-LOW cows. Milk beta-hydroxybutyric acid (BHB) content was lower in the LF-HIGH group (trend, p ≈ 0.062). LF-HIGH also exhibited significantly greater natriuresis (p = 0.0078) and a more negative U-ABB (p < 0.001), indicating higher acid–base load. In conclusion, elevated LF contents during the postpartum period were associated with the activation of local mammary gland immune defence and concurrent compensatory metabolic processes related to NEB, rather than with pronounced alterations in basic milk composition. Milk LF content may therefore be considered as a specific indicator of immunometabolic compensation during the early postpartum period, rather than as a general marker of overall cow health.

Dairy doi: 10.3390/dairy7010008

Authors: Çağım Akbulut Çakır

Kargı Tulum cheese differs from other Tulum cheeses with its unique production and ripening method. No systematic study has yet explored the change in the volatile compounds and fatty acids during the ripening process of Kargı Tulum cheese. The objective of this study was to monitor the change in the fatty acids and volatile compounds of Kargı Tulum cheese at different time points during the production and ripening stages. Fatty acid profile, lipid quality parameters and volatile compound profiles were determined. A principal component analysis (PCA) was performed to determine how the volatile profiles differed across production and ripening stages. During the ripening, short- and medium-chain fatty acids (FAs) increased with notably high levels of butyric acid. Lipid quality parameters, including total saturated FAs (SFAs), atherogenicity index (AI), and thrombogenicity index (TI), remained unchanged throughout ripening. A total of 62 volatile compounds (VOC) were detected. Esters and ketones were the most abundant groups in fresh curds, while carboxylic acids became the dominant group by the end of the ripening process. The total concentration of most VOC increased over time. Butyric acid, hexanoic acid, ethyl hexanoate and acetic acid were the dominant compounds contributing the flavor of the Kargı Tulum cheese. This study presents data on what flavor compounds form and how they change during ripening and can be useful for comparative purposes in future studies on ripened raw milk cheeses.

Dairy doi: 10.3390/dairy7010007

Authors: Renata Alves das Chagas Tatiane Fernandes Ariadne Patrícia Leonardo Agda Costa Valério Núbia Michelle Vieira da Silva Cláudia Andrea Lima Cardoso Rui José Branquinho de Bessa Fernando Miranda de Vargas Junior

This study aimed to evaluate the effect of the supplementation with rumen-protected fat from soybean or palm and thermally processed soybean on the feed intake, digestibility of nutrients, milk production, and milk content of ewes. Twenty-five Pantaneiras ewes, 3–6 years old, 39.8 ± 3.51 kg body weight, and 65 ± 4 days in milk, were distributed into five treatments (5 ewes in each) in a completely randomized design continuous trial, over 56 days. The treatments consisted of daily supplementation with soybean-based rumen-protected fat (SPF; 30 g/d), palm-based rumen-protected fat (PPF; 30 g/d), a blend of soybean and palm rumen-protected fats (Blend; 30 g/d), thermally processed soybean (TPS; 124 g/d), and a control without supplementation. We performed a daily evaluation of feed intake and milk production, and every 14 days, we evaluated the nutrient digestibility, milk composition, and fatty acid profile. The protein and casein content were lower in the SPF treatment. Supplementation with PPF resulted in a higher saturated fatty acid content, while supplementation with TPS resulted in higher monounsaturated and polyunsaturated fatty acid contents. The supplementation with SPF resulted in higher milk fatty acid functionality. Feeding ewes SPF or TPS enhanced nutrient intake and digestibility, leading to increased milk production and an improved milk fatty acid profile. In contrast, supplementation with PPF resulted in a less favorable fatty acid composition.

Dairy doi: 10.3390/dairy7010006

Authors: Giovanni Niero Marta Pozza Massimo De Marchi

Iodine deficiency is the leading preventable cause of neurological damage worldwide. Dairy foods represent an important dietary iodine source. This study aimed to assess iodine concentration in milk, ricotta cheese, and yogurt, and to evaluate their contribution toward the recommended daily iodine intake. Whole pasteurized milk (WM; n = 12), partially skimmed pasteurized milk (PM; n = 21), skimmed pasteurized milk (SM; n = 7), ricotta cheese (RC; n = 26), whole yogurt (WY; n = 13), and low-fat yogurt (LY; n = 15) were purchased in local stores. Samples were analyzed through inductively coupled plasma mass spectrometry for iodine quantification. After removing outliers, the final dataset comprised 11 WM, 19 PM, 7 SM, 26 RC, 13 WY and 15 LY samples. Data were investigated through a mixed model with iodine concentration as the dependent variable, product type as fixed effect, and brand as random effect. Low-fat yogurt exhibited the greatest estimated iodine concentration (293.76 µg/kg), while SM and WM exhibited the lowest (211.92 and 197.63 µg/kg, respectively). Based on these results, a serving of milk (250 g) would provide 31.82–39.08% of the average daily iodine requirement, a serving of ricotta (125 g) 21.66%, and a yogurt jar (125 g) 21.54–24.11%. These findings confirm the nutritional relevance of dairy products as primary iodine sources.

Dairy doi: 10.3390/dairy7010005

Authors: Liga Marcinkoniene Inga Ciprovica

An elevated somatic cell count (SCC) affects the physicochemical characteristics of milk, altering its renneting ability and ultimately impacting the yield and quality of the cheese produced. This study aims to analyse the variations in the composition of Latvian goat milk and its technological properties in relation to SCC. Individual goat milk samples (n = 240) were collected from one of the largest goat farms in Latvia during 2019 and 2020. Latvian, Saanen, and cross-breed goat milk samples from different lactations were tested for their chemical composition (fat, protein, lactose, dry matter, and SCC), fat-to-protein ratio, freezing point, and curd firmness. Samples were collected during different lactation periods in order to analyse the seasonal effect on milk quality. The results demonstrated that milk samples from goats with lower SCCs (Group I) exhibited the highest fat (3.34%), lactose (4.56%), and dry matter (11.28%) concentrations and fat-to-protein ratios (1.02). Curd firmness decreased progressively from Group I to Group IV, fluctuating between Latvian Native (1.96–1.47 N), Saanen (1.91–1.59 N), and cross-breed (1.66–1.58 N) goat milk samples. A significantly lower (p = 0.030) curd firmness (1.56 N) was determined in the Group IV goat milk samples. Seasonal fluctuations in milk composition were observed in relation to curd firmness, which peaked in late lactation milk (3.85 N), although minor fluctuations were observed in protein concentrations (3.23% to 3.30%) across the sampling periods (2019 and 2020). These findings highlight the necessity of SCC monitoring in Latvian goat milk to ensure appropriate quality for milk processing.

Dairy doi: 10.3390/dairy7010004

Authors: Gamze Güneş Çağım Akbulut Çakır

Using raw milk in cheesemaking poses several risks and often requires higher salt levels. Gel brine is a promising brining method to reduce salt and to prevent excessive softening, yet it was not employed to raw milk cheese before. In this study, the impact of ripening in gel brine—prepared by adding selected thickeners (gelatin and carrageenan) to a 12% salt brine—on the composition, proteolysis, texture, and microbiological properties of raw milk cheese was examined over 120 days. The aim was to assess the potential of gel brine to shorten the ripening time of raw milk cheese at a relatively low salt concentration while maintaining acceptable quality parameters. Response surface methodology was used to determine the optimum ripening time and thickener concentrations required to achieve target microbial counts, proteolysis, and moisture levels. The addition of stabilizers did not significantly influence the overall composition of the cheese, except for salt in dry matter. Stabilizers also limited the increase in trichloroacetic acid-soluble nitrogen (TCA-SN) during storage and led to a marked reduction in Escherichia coli counts. Texture profile analysis results were significantly affected (p < 0.05). The optimum conditions were estimated as 0.9% carrageenan, 0.8% gelatin, and 35 days of ripening.

Dairy doi: 10.3390/dairy7010003

Authors: Konstantinos V. Arsenopoulos Athanasios I. Gelasakis Elias Papadopoulos

Although best known as a major parasite of sheep and goats, the blood-feeding abomasal nematode Haemonchus contortus can also infect cattle and buffaloes under the mixed-grazing Mediterranean conditions prevalent in Greece. The objectives of this study were as follows: (i) to determine the prevalence of H. contortus infections in dairy and beef cattle and buffaloes in Greece through an abattoir survey, (ii) to evaluate potential host- and farm-related risk factors including age, sex, management system, cattle productive orientation, and the co-existence of cattle and buffaloes on the occurrence of haemonchosis, and (iii) to assess the likelihood of detecting homozygous benzimidazole (BZ)-resistant H. contortus in large ruminant populations in relation to these determinants. A total of 213 abomasa (115, 55, and 43 from dairy, beef cattle, and buffaloes, respectively) were examined. A structured questionnaire provided additional animal- and farm-level information. Haemonchus-like helminths were collected and molecularly identified at the species level by amplifying a 321 bp fragment of the internal transcribed spacer 2 region of nuclear DNA. An allele-specific multiplex PCR, targeting codon 200 of the β-tubulin gene, was applied to detect BZ-resistant alleles. The prevalence of H. contortus infection was 21.2% in cattle and 69.8% in buffaloes. In cattle, multivariable analysis revealed that mixed-species farming (i.e., farms where cattle were the primary species and buffaloes were kept in smaller numbers), productive orientation, and slaughter age were significant predictors of increased H. contortus infection. Controversially, none of these factors were significantly associated with infection in buffaloes. Finally, multivariable modelling suggested that resistance patterns varied by host species, being more prevalent in intensively managed, older cattle, yet less common among older buffaloes and in herds where both species coexisted.

Dairy doi: 10.3390/dairy7010002

Authors: Harald Pothmann Michael Mitterer Florian Flicker Maryam Sahebi Vitezslav Havlicek Urban Besenfelder Alexander Tichy Marc Drillich

Hyperketonemia (HYK), defined by blood beta-hydroxybutyrate (BHB) ≥ 1.2 mmol/L, is described as a significant risk factor for cows developing postpartum (pp) diseases and impaired reproductive performance. The goal of the present study was to observe metabolic challenges in transition cows and to identify systemic markers reflecting HYK associated with lessened reproductivity. Fifty-four Simmental cows were monitored, revealing approximately 30% prevalence of HYK at the early pp period on 7, 14, or 28 days in milk (DIM). We assessed the dry matter intake, rumination time (RT), serum liver activity index, non-esterified fatty acids (NEFAs), acute phase proteins, and uterine and oviductal health. Elevated NEFA and reduced RT 14 days antepartum were a good predictor for HYK at 7 DIM. Hyperketonemia at 14 DIM resulted in higher milk yield compared with controls. We could neither detect differences in uterine health nor in reproductive key performance parameters between hyperketonemic and control cows, whereby the proportion of polymorphonuclear neutrophils in oviductal epithelia was significantly lower in hyperketonemic cows 14 DIM. We conclude that elevated concentrations of BHB in HYK 7, 14, or 28 DIM indicated energy supply to support physiological metabolic adaptations and lactation and that, in the absence of excessive inflammation during the transition period, HYK was not a risk factor for impaired fertility.

Dairy doi: 10.3390/dairy7010001

Authors: André Neves Moritz Pfeiffer Rajan Dhakal Svenja Woudstra Volker Krömker Hanne Hansen Ricardo Vieira

We evaluated the effects of rapeseed oil (RSO, rich in oleic, linoleic, and linolenic acids) and a commercial fat source (BOVI-LM, rich in palmitic and stearic acids) on the growth performance and blood biochemical parameters of dairy calves during the pre- and post-weaning phases. Eighteen crossbred bull calves were randomly allocated in a randomized complete block design to three dietary treatments (Control, RSO, and BOVI-LM) during the milk-replacer phase (Phase 1, days 0–28). Due to technical issues, BOVI-LM supplementation was discontinued in Phase 2 (starter-feed phase, days 35–77), leaving only Control (n = 12) and RSO (n = 6). Energy supply was calculated to support target average daily gains of 0.6 (Phase 1) and 1.0 kg (Phase 2). Body weight, clinical health, and blood samples for biochemical analysis were monitored throughout the trial. No treatment effect on body weight or blood biochemical parameters was detected (main effect of treatment: p ≥ 0.18 for all analytes). In contrast, phase and time effects were significant for most biochemical parameters (p < 0.05), reflecting the metabolic transition from a milk-based to a solid-feed diet. Aspartate aminotransferase activity increased over time (p < 0.001), whereas glucose concentrations decreased post-weaning (time × phase: p = 0.020). The treatment × phase interaction was non-significant for all variables (p ≥ 0.13), supporting the absence of detectable carry-over effects and justifying the pooling of former BOVI-LM calves into the Control group in Phase 2.

Dairy doi: 10.3390/dairy6060072

Authors: Mohammad Mortazavi Cristiane Tomaluski Elizangela Marino Julia Feliciano Jeniffer Alvarado-Castro Ingred Rocha de Oliveira Nathalia Carvalho Carla Bittar

This study examines the critical role of whole milk or milk replacer as a liquid diet (LD) with 15% solids in combination with different physical forms of starter as a solid diet (SD), on performance, health, and behavior of pre-weaned calves. Sixty male Holstein calves were used in a 2 × 2 factorial design, and randomly distributed into the following treatments: Whole milk powder diluted to 12.5% of solids and enriched with 25 g/L of milk replacer to achieve 15% solids, associated with either micropelleted stater (WM+micro) or texturized stater (WM+text); milk replacer diluted to 15% solids associated with either micropelleted stater (MRmicro) or texturized stater (MRtext). Starter intake and, consequently, total DMI were higher in the MRtext treatment compared to WM+micro. Calves fed texturized starter showed higher DMI, starter intake time, and rumination time. Calves in the WM+Text group showed greater ADG compared with MR treatments, regardless of starter type. Calves fed WM+ presented a lower number of days with fecal score ≥2, and the first day of diarrhea occurred at older ages. Calves fed MR showed more health challenges but similar feed efficiency with WM+, while texturized starter increased intake, eating duration, and rumination compared with micropelleted starter.

Dairy doi: 10.3390/dairy6060071

Authors: Renáta Szabó Erika Veres Csilla Albert Éva Laslo László Gyenge Rozália Veronika Salamon

Cheese consumption is steadily increasing worldwide, with a growing interest in cheese enriched with bioactive substances, including antioxidants. This study investigated the impact of adding blackcurrant wine to the curd (IC), enriching the curd with blackcurrant wine by soaking and ripening in salted blackcurrant wine (IOC), and cheese soaked and ripened in blackcurrant wine with 5% (w/w) NaCl (OC). The curd and added wine weight ratio (1.5:1, 3:1) effects were also studied. Physicochemical (dry matter, polyphenol content, antioxidant activity, radical-scavenging activity, anthocyanin content like delphinidin-3-rutoside and cyanidin-3-rutoside, ethanol content), microbiological, and sensory properties of the cheeses were evaluated. The results indicated that a week of soaking is sufficient to achieve the maximum antioxidant capacity and polyphenol content of the cheese. From a technological and sensory point of view, a 1.5:1 ratio of blackcurrant wine to curd was better. The maximum transfer rate of delphinidin-3-rutoside from wine was the most pronounced in IOC samples (20.44%). Blackcurrant wine inhibited the growth of lactic acid bacteria, and a longer soaking time can hinder the ripening process of cheese. During tasting, among the treated cheese, IC samples received the highest average acceptance scores for appearance, texture, creaminess, flavor, saltiness, bitterness, freshness and overall impressions.

Dairy doi: 10.3390/dairy6060070

Authors: Renata Alves Chagas Tatiane Fernandes Cristiane Rebouças Barbosa Jessica de Carvalho Pantoja Samuel Rodrigues Navarro Marcus Vinicius Morais de Oliveira Cláudia Andréa Lima Cardoso Fernando Miranda de Vargas

This study aimed to evaluate the supplementation of aqueous extract of Moringa oleifera (AEMO) as a natural ruminal modulator to improve the lactation performance of ewes. The AEMO was prepared by chopping Moringa oleifera leaves and diluting them in distilled water (163.3 g DM/L). Twelve ewes were used in a replicated 3 × 3 Latin square, with periods of 14 days (assessments on the last five days of each period). Treatments were as follows: 20 mL of water as Control, 20 mL of AEMO (20-AEMO), and 40 mL of AEMO (40-AEMO). Ewes were milked twice a day (7:30 a.m. and 2:30 p.m.). Diet corresponds to grain mix (at 3% of BW) and hay ad libitum. We determined the intake, digestibility, fermentative measurements, metabolic measurements, and milk production and composition. Intake and digestibility were not affected by AEMO. Milk yield and the concentrations of fat, protein, and lactose were numerically lower in ewes supplemented with 20-AEMO. A linear decrease in milk protein yield was observed when the highest extract level (40-AEMO) was used. Ruminal pH did not differ among treatments; however, there was a tendency for reduced acetate and increased propionate concentrations, which corresponded with a non-significant numerical decrease in methane estimates in 40-AEMO group. Blood and urinary parameters were not affected by AEMO supplementation. Inclusion of Moringa extracts as an additive in lactating ewes diet does not affect intake and nutrient digestibility, but tends to affect ruminal fermentation and microbial synthesis, with possible changes in methane emission estimation, and impair milk protein production. Therefore, we recommend studies with different extract concentrations to investigate possible effects on rumen fermentation and the synthesis of milk compounds.

Dairy doi: 10.3390/dairy6060069

Authors: Gabriel Vasilakis Antonios Georgoulakis Eleni Dalaka Georgios Bekiaris Ilias Diamantis Dimitris Karayannis Maria-Eleftheria Zografaki Panagiota Diamantopoulou Emmanouil Flemetakis Georgios Theodorou Ioannis Politis Seraphim Papanikolaou

Microbial bioconversion of agro-industrial by-products into high-value-added metabolites such as polysaccharides or lipids serves a dual purpose: mitigating environmental pollution through waste reduction and supporting the development of novel bioproducts. In this study, a non-conventional, poorly studied Cryptococcus albidus strain was initially assessed for its ability to grow on semi-defined media containing lactose, glycerol, or glucose under three distinct nitrogen availability conditions at C/N equal to 20, 80, and 160 mol/mol in shake flask cultures. The goal was to evaluate biomass production and synthesis of valuable metabolites under these conditions. C. albidus demonstrated robust growth on all commercial carbon sources, particularly under nitrogen-rich conditions, producing more than 25.0 g/L of microbial biomass with a high intracellular polysaccharide content (>45%, w/w). Additionally, mannitol production was detected in cultures with glycerol and glucose (9.1 and 13.1 g/L, respectively), especially after nitrogen depletion. Subsequently, C. albidus and a Cutaneotrichosporon curvatus strain were batch-cultivated using pretreated secondary cheese whey (SCW) as a carbon-rich waste substrate. When cultivated on SCW, both yeast strains partially metabolized lactose and produced polysaccharide-rich biomass, dominated by β-glucans (>29% of total biomass), compounds known for their functional and bioactive properties. The cellular polysaccharides (cPS extracted from C. albidus exhibited cytotoxic effects against cancer cells, suggesting their potential use as biological response modifiers. In contrast, the cPS from C. curvatus did not affect cell viability, indicating their promise as ingredients for applications in the food, feed, pharmaceutical, or cosmetic sectors.

Dairy doi: 10.3390/dairy6060068

Authors: Giulia Sala Matteo Castelli Chiara Orsetti Arianna Cervelli Giovanni Armenia Francesca Bonelli

Ultrasonography has been proposed as a complementary tool for the evaluation of udder health, yet limited information exists on its application for measuring the glandular cistern area during mastitis. This study aimed to investigate modifications of the glandular cistern area in clinical (CM) and subclinical mastitis (SCM) compared with contralateral healthy quarters, and to assess the reliability of manual and automated ultrasonographic measurements. A longitudinal study was conducted on 42 Italian Holstein cows, comprising 26 SCM and 16 CM quarters, each paired with contralateral healthy controls. Ultrasound examinations were performed at diagnosis (T0), 24 h (T1), and 5 days (T5). Cisternal areas were measured in transversal and longitudinal sections using both manual and ImageJ-guided methods. Intra- and inter-operator reliability was assessed using Intraclass Correlation Coefficients (ICCs). Statistical analyses included two-way mixed ANOVA with post-hoc Bonferroni correction. Mastitic quarters tended to show smaller cisternal areas compared with contralateral healthy quarters, with significant differences observed between contralateral healthy and CM quarters (p < 0.05), but not between contralateral healthy and SCM or between SCM and CM quarters. Temporal trends differed significantly among groups (interaction effect, p < 0.05). Both manual and automated measurements demonstrated excellent intra- and inter-operator reliability, with ICCs consistently > 0.95 across pathology groups and time points. Ultrasonography of the glandular cistern is a feasible, reproducible, and reliable method under field conditions. Clinical mastitis is associated with a measurable reduction in cisternal area, while changes in subclinical mastitis appear less evident. The high reproducibility of measurements, including automated analysis, supports the use of this technique may contribute to improve the use of ultrasound also for the udder heath.

Dairy doi: 10.3390/dairy6060066

Authors: Danilo D. Millen Gercino F. Virgínio Fernanda F. Alves Charles G. Schwab Sergio Calsamiglia

Improving nitrogen efficiency in dairy cattle requires a better understanding of the dietary and ruminal factors that regulate nitrogen partitioning. This meta-analysis evaluated the effects of ruminal pH and dietary characteristics on microbial nitrogen (MN), non-microbial non-ammonia nitrogen (NANMN), and NAN flows to the small intestine in lactating cows. A dataset was assembled from 44 peer-reviewed in vivo studies (163 data points), with dietary intake and ruminal variables standardized across trials. Mixed linear models were developed for each N fraction, and the relative contribution of each predictor to the explained variance was assessed using semipartial coefficients of determination (pR2). Efficiency of microbial protein synthesis (EMPS), rumen undegraded protein intake (RUPI), and organic matter truly digested in the rumen (OMTDR) were the most relevant predictors of NANMN and NAN. Although the ruminal pH itself was not statistically significant in the models, the dietary components that influenced pH, starch concentration, physically effective fiber, and RUP supply were strongly associated with nitrogen flow profiles. Nitrogen utilization was not affected by ruminal pH, but rather by the combination of fermentable substrates and the supply of rumen-degradable and undegraded protein.

Dairy doi: 10.3390/dairy6060067

Authors: Adrián Ponce de León-Door Pedro González-Pérez Guadalupe I. Olivas Francisco Javier Molina-Corral Jesús Cristian Amaro-Hernández David R. Sepulveda

The incorporation of Arthrospira platensis into dairy products offers health benefits but is limited by its undesirable aroma and flavor. This study evaluated three deodorization strategies—adsorption by activated carbon, extraction with ethanol, and fermentation with Saccharomyces cerevisiae—to improve the sensory profile of A. platensis and enhance its acceptability in yogurt. Deodorized powders were characterized for proximal and volatile composition and used to formulate yogurts at five concentrations (0.5–2.5% w/v). Texture, aroma volatile profile, and sensory attributes were assessed after yogurt production, while shelf-life quality attributes were monitored over 29 days of refrigerated storage. Yogurts containing fermented A. platensis showed higher sensory scores (>8.7/10), the greatest purchase intent (>71.4%), and improved texture, remaining acceptable at an addition level of 2.5%. In contrast, yogurts with untreated or carbon-activated treated A. platensis were only acceptable at low addition levels (≤0.5%) due to off-flavors and textural issues. Ethanol effectively reduced aldehydes and ketones (such as Safranal and β-Ionone), while fermentation eliminated pyrazines and generated desirable alcohols and acids (such as 1-Pentanol and Butanoic acid). These findings highlight fermentation as a promising strategy to deodorize A. platensis and improve its integration into dairy matrices, enabling the development of functional yogurts with enhanced sensory quality and nutritionally relevant microalgae content.

Dairy doi: 10.3390/dairy6060065

Authors: Ermioni Meleti Michalis Koureas Athanasios Manouras Persephoni Giannouli Eleni Malissiova

Bioactive peptides (BAPs) from dairy products have garnered increasing attention as natural agents with health-promoting properties, including antihypertensive, antioxidant, antimicrobial, immunomodulatory, opioid, and antidiabetic activities. This systematic review synthesizes research published between 2014 and 2024, retrieved from Scopus and PubMed, and selected according to PRISMA guidelines. A total of 192 studies met the inclusion criteria, collectively reporting over 3200 distinct peptides, with antihypertensive sequences, predominantly angiotensin-converting enzyme (ACE) inhibitors, constituting the largest category (n = 1237). β-casein was the principal precursor across bioactivities, followed by αs1-casein, β-lactoglobulin, and α-lactalbumin. Peptides were primarily produced via enzymatic hydrolysis, microbial fermentation, and gastrointestinal digestion, with peptide profiles influenced by the type of milk, microbial strains, and processing conditions. While cow’s milk remained the dominant source, investigations into goat, sheep, camel, buffalo, and donkey milk revealed species-specific biopeptides. Recent advances in proteomics have enhanced peptide identification and bioactivity prediction, enabling the discovery of novel sequences. These findings underscore the significant potential of dairy-derived BAPs as functional food components and nutraceutical ingredients, while highlighting the need for further in vivo validation, bioavailability studies, and broader exploration of underrepresented milk sources.

Dairy doi: 10.3390/dairy6060064

Authors: Eron Bajrami Gaiping Wen Sarah M. Grundmann Robert Ringseis Denise K. Gessner Klaus Eder

Previous studies have demonstrated that high-yielding dairy cows experience endoplasmic reticulum (ER) stress in the liver during early lactation. To date, most insights into the role of ER stress in metabolism and disease pathophysiology have been derived from rodent and human models. In dairy cattle, however, the specific impact of ER stress on metabolic pathways and its contribution to disease development remain insufficiently characterized. The objective of this study was therefore to investigate the molecular effects of ER stress using a bovine liver cell model (BFH12 cells). ER stress was induced by incubation with Tunicamycin (TM) and Thapsigargin (TG). Molecular responses to ER stress were assessed via a whole-genome array analysis and PCR targeting genes involved in selected metabolic pathways. Incubation with both ER stress inducers resulted in a marked upregulation of genes associated with the unfolded protein response (UPR) within a 4 to 24-h time frame, indicative of the production of robust ER stress in these cells. Unexpectedly, treatment with TM led to a downregulation of numerous genes involved in lipid biosynthesis, including those related to lipogenesis and cholesterol synthesis. Furthermore, incubation with TM and TG induced upregulation of genes involved in fatty acid oxidation and was accompanied by a reduction in intracellular triglyceride concentrations. Genes associated with inflammatory responses were upregulated by both TM and TG, whereas genes encoding antioxidant enzymes were downregulated. Genes involved in ketogenesis did not exhibit a consistent pattern of regulation. Overall, several effects of ER stress previously described in rodent models could not be replicated in this bovine liver cell system. Extrapolating these findings to dairy cows suggests that while ER stress may contribute to hepatic inflammation, it is unlikely to play a significant role in the development of hepatic lipidosis or ketosis.

Dairy doi: 10.3390/dairy6060063

Authors: João M. Santos João M. Garcia João Dias João C. Martins Nuno Alvarenga Elsa M. Gonçalves Daniela Freitas Karina Silvério Jaime Fernandes Sandra Gomes Manuela Lageiro Miguel Potes José Jasnau Caeiro

Improving the energy efficiency of traditional production methods while preserving their cultural and economic value is a challenge aligned with the Sustainable Development Goals of the 2030 agenda. Refrigeration during cheese maturation is particularly energy-intensive, contributing significantly to greenhouse gas emissions and operating costs. An approach to make traditional cheese production more sustainable, through the development of a prototype ripening chamber with a natural refrigerant-based refrigeration system powered by renewable energy was studied. A dedicated system based on an Internet of Things architecture was developed using low-cost sensors, microcontroller units, and single-board computers to enable real-time measurement and monitoring of environmental variables and energy consumption throughout the ripening process. A comparative analysis was conducted using ewe’s milk cheese, produced and ripened with Protected Designation of Origin conditions, in both the prototype and the conventional chambers over four weeks, quantifying energy consumption and evaluating product quality. Results demonstrate the technical feasibility of energy efficient and sustainable refrigeration systems, as well as the possibility of retrofitting installed cheese ripening chambers with affordable IoT monitoring systems, while maintaining traditional cheese quality standards.

Dairy doi: 10.3390/dairy6060062

Authors: Simona Rinaldi Michela Contò Carlo Boselli Giuliano Palocci Angelo Citro Sebastiana Failla

Sialic acids (SIAs) are bioactive compounds present in glycoproteins and glycolipids, playing key roles in human physiology. N-acetylneuraminic acid (Neu5Ac), the only sialic acid synthesized by humans, contributes to neurodevelopment and immune function, whereas N-glycolylneuraminic acid (Neu5Gc), acquired from animal-derived foods, is potentially harmful, showing pro-inflammatory properties. Accordingly, a high Neu5Ac/Neu5Gc ratio represents an important nutritional indicator. This study quantified the two main forms of SIA in five commercial cheese types (mozzarella, stracchino, caciotta, robiola, ricotta) produced from cow (C), buffalo (B), and goat (G) milk, as well as in laboratory cheesemaking from 1000 g of milk from each species. Neu5Gc concentrations were significantly higher in G cheeses (p < 0.001), while Neu5Ac was more abundant in C cheeses (p < 0.05). Consequently, the Neu5Ac/Neu5Gc ratio was markedly higher in C and B cheeses (24.11 and 21.57, respectively) than in G (1.29), reflecting species-specific glycosylation patterns. Among cheese types, ricotta produced from whey exhibited the highest SIA concentrations among all species, followed by robiola, mozzarella and stracchino. In contrast, caciotta showed the lowest SIA levels. The laboratory cheesemaking trial confirmed that most SIAs remained bound to whey proteins. These findings highlighted the combined effect of milk species and cheesemaking on sialylation, with potential nutritional implications.

Dairy doi: 10.3390/dairy6050061

Authors: Cristina Manis Mattia Casula Margherita Chessa Nicoletta P. Mangia Pierluigi Caboni

Fermented dairy products such as yogurt, kefir, and traditional cheeses are increasingly consumed worldwide for their nutritional and probiotic properties. Lipidomic profiling provides valuable insights into microbial-driven biochemical changes during fermentation. In this study, we performed a comprehensive untargeted lipidomic analysis of sheep milk and Gioddu, a traditional Sardinian fermented dairy product. Using UHPLC-QTOF-MS platform, we observed that fermentation significantly reshaped the lipidome. Gioddu samples showed higher levels of phosphatidylethanolamines (PE) and lysophosphatidylethanolamines (LPE), together with a pronounced reduction in sphingolipids (glucosylceramides, ceramides, sphingomyelins) and glycerophospholipids (phosphatidylinositols, phosphatidylserines, phosphatidylcholines) compared to sheep milk. These findings align with known enzymatic activities of lactic acid bacteria (LAB), including phospholipases A1 and A2, phosphatidylinositol-specific phospholipase C (PI-PLC), and sphingomyelinase. Fermentation also affected triglycerides, with reduced levels of FA 18:1-containing species, suggesting the selective lipolysis of monounsaturated fatty acids by microbial lipases. Complementary metabolomic profiling revealed reduced levels of simple sugars such as galactose and inositol in Gioddu samples, consistent with their use as primary carbon sources during early fermentation. Conversely, a marked accumulation of carboxylic acids (succinic, malic, hydroxyisovaleric, hydroxyglutaric, glyceric) was revealed, reflecting enhanced microbial fermentative activity. Increased levels of amino acids, including alanine, serine, proline, and ethanolamine, further highlighted active proteolysis and membrane remodeling driven by LAB metabolism. These findings show that LAB enzymes play a key role in modifying the lipidome of fermented dairy products, highlighting their metabolic flexibility and potential impact on nutritional and health properties. This integrated approach sheds new light on the metabolic plasticity of fermentative processes and underscores the value of omics-based tools in understanding traditional food systems.

Dairy doi: 10.3390/dairy6050060

Authors: Renata López Valentina Villarroel Víctor Neira Carolina Aguayo Karina Saadi Katherinne Orozco Pedro Abalos Patricio Retamal

Bovine tuberculosis (bTB) is mainly caused by Mycobacterium bovis, which affects cattle, leading to significant economic losses. In Chile, the vaccination with the M. bovis Bacillus Calmette-Guérin (BCG) strain has been implemented in dairy herds with high prevalence of bTB. This study evaluated non-specific protection associated with BCG on the detection of pathogen-associated genes (nsp5, stx1, stx2, invA, IS1081) and mortality related to diarrhea and pneumonia in calves. A total of 186 calves from a commercial dairy farm were enrolled and grouped as vaccinated (n = 96) and non-vaccinated (n = 90). The BCG Russia strain (2–5 × 105 UFC) was inoculated subcutaneously within the first 30 days after birth. Animals were monitored through fecal sampling at 3 and 6 months of age for molecular detection of gene sequences. A logistic regression analysis showed differences in detection rates of the stx1 sequence at 3 months, with a higher risk for the non-vaccinated individuals (OR 2.91, CI 1.42–5.94, p = 0.03) and for those born in the cold season (OR 9.55, CI 2.02–45.11, p = 0.004). A Kaplan–Meier survival analysis showed a significant difference in deaths in vaccinated calves compared with non-vaccinated animals (p = 0.018), suggesting that BCG confers non-specific protection during the first 3 months after birth, in field conditions.

Dairy doi: 10.3390/dairy6050059

Authors: Jessica Vidlund Agricola Odoi Russell Zaretzki Chika C. Okafor

The dairy industry faces challenges from mastitis, which affects milk quality. Somatic Cell Counts (SCCs) are key indicators of udder health, subclinical mastitis presence, and legal thresholds. However, limited research has explored geographic disparities and temporal patterns in SCCs across the USA, despite their critical role in informing targeted herd management strategies, optimizing policy interventions, and ensuring consistent milk quality standards. This study aimed to examine temporal trends and geographic disparities in median weighted SCCs (mwSCCs) across USA states. This study analyzes SCC data using records from the Dairy Herd Improvement Association across 42–45 states between 2011 and 2023. State-level differences in mwSCCs were examined, with temporal changes assessed using percent differences between 2011 and 2023. Moran’s I and Local Indicators of Spatial Association (LISA) were used to identify spatial clusters of states with high and low mwSCCs. The mwSCCs decreased by 24.8%, from 234,000 cells/mL in 2011 to 176,000 cells/mL in 2023, with significant reductions in Virginia and Georgia, while Tennessee and South Carolina had minimal declines. However, Texas, California, and Colorado saw increases, with Colorado rising by 147.9%. Spatial clustering revealed high mwSCCs in the southeast and low levels in the northeast, highlighting the need for region-specific strategies.

Dairy doi: 10.3390/dairy6050058

Authors: Sergiu Pădureţ

Milk products are a diverse group of foods and important sources of essential nutrients, including high-quality proteins, fatty acids, vitamins, and minerals. Among their key quality attributes, texture is particularly critical, as it strongly influences consumer perception and overall product quality. Numerous devices and techniques have been developed to evaluate the texture of milk products, most of which rely on mechanical tests such as puncture, compression, shearing, creep, and relaxation. Instrumental evaluations are essential for correlating physical measurements with sensory perceptions, yet several challenges limit their reliability. Inconsistencies in testing protocols—such as reporting force versus penetration depth versus force versus time; variations in testing temperature, sample shape and dimensions; probe geometry; compression depth; and container size for semisolid samples contribute to discrepancies across studies. Additionally, many studies omit these critical methodological details, reducing reproducibility and comparability. This review systematically examines the current methods used to assess dairy product texture, identifies gaps and challenges in standardization, and provides guidance to support future research aimed at obtaining accurate, reproducible, and meaningful texture measurements.

Dairy doi: 10.3390/dairy6050057

Authors: Austen Ashfield Michael Wallace Claire Jack

Profitable dairy farming requires continuous appraisal and adaptation of production systems in response to changing market and agricultural policy conditions. Geopolitical and climate events have exemplified the exposure of farm incomes to the increased volatility associated with often-global market factors. In this context, bio-economic models can be a useful tool for researchers seeking to understand the financial resilience of different production systems to these changing circumstances. The AFBI Dairy Systems Model is presented and used to simulate the impacts of alternative price scenarios for Northern Ireland-based dairy systems. The whole farm model consists of four interdependent components, comprising farm system, animal nutrition, feed supply and financial sub models. The model is used to evaluate how fluctuations in milk, concentrate, fertiliser, contractor, and electricity prices, as well as interest rate changes, affect three distinct production systems. The financial performance of all systems was sensitive to variations in milk and concentrate prices but relatively insensitive to variations in fertiliser, contractor, and electricity prices and interest rate changes. The profitability of a low-output system was less exposed to variations in prices. In contrast, a high-output system was more exposed to price variations. However, a medium-input system was the most profitable across the majority of price scenarios investigated.

Dairy doi: 10.3390/dairy6050056

Authors: Carlos Dias Pereira Klaudia Hodowaniec Karolina Kucz Katarzyna Szkolnicka David Gomes Arona Pires

This research work evaluated the application of whey-based edible coatings to cheeses. Coatings were prepared with a bioprotective culture (BC) containing Lacticaseibacillus paracasei and Lacticaseibacillus rhamnosus alone, or in conjunction with thyme essential oil (TEO). The samples containing the BC or the BC plus TEO were compared with cheeses without coating, with cheeses with whey-based coatings without BC or TEO, and with cheeses treated with natamycin. The cheeses were evaluated regarding their physicochemical, microbiological, and sensory properties. All cheeses produced were classified as full-fat (≥45–60% fat in dry matter—FDM) and semihard (>54–<63% moisture in the defatted cheese—MDC), with an exception made for the control cheese, which presented lower levels of MDC, graded as hard (>49–<56% MDC). Most of the parameters evaluated presented significant differences between samples and as a result of ripening time. Regarding color parameters, it was observed that, after ripening, the external color of the samples with the whey coating presented higher lightness values (L*), higher a* values, and lower b* values. These differences clearly resulted from the white color imparted by the coating. Significant differences were also observed with respect to the texture parameters of the cheeses. The samples containing the BC or the BC plus TEO presented higher values for hardness and chewiness. In what concerns the microbiological evaluation, in all cases, lactic acid bacteria counts increased from log 7.5–8 CFU/g on the first day to ca. log 10 CFU/g at the end of the ripening period. Yeast and mold counts were significantly lower in samples containing the BC or the BC plus TEO, with values of ca. log 3 CFU/g and log 2.5 CFU/g, respectively. These values are like those obtained in samples with natamycin, with 1–2 log cycles below those of cheeses without treatment. However, the use of BC and BC plus TEO had a negative impact on the sensory properties of cheeses. Future work should evaluate the synergistic effect of different BCs and EOs.

Dairy doi: 10.3390/dairy6050055

Authors: Julia Maria Pires de Matos Vitral Igor Lima de Paula Paola Fazollo Lazzarini Thomas Henle Uwe Schwarzenbolz Alan Frederick Wolfschoon-Pombo Ítalo Tuler Perrone Luiz Fernando Cappa de Oliveira Rodrigo Stephani

Doce de leite (DL) is a product originating in South America, particularly in countries such as Brazil, Argentina, and Uruguay. The product is widely used for direct consumption and industrial applications and can be added with thickening agents to modify its rheological properties. This study aimed to produce DL with the addition of three different components (skimmed milk powder—SMP, whey protein concentrate—WPC, and microparticulated whey protein—MWP) in two different concentrations (2.1 and 4.2%). The composition of the products aligned with Mercosur regulations, except for the fat content, where the result was slightly lower. The HMF (5-hydroxymethylfurfural) concentration in the samples was also determined, showing no statistically significant differences between products. The products with SMP showed a higher browning index and color saturation compared to WPC and MWP formulations. The addition of MWP increased the hardness (from 10.2 to 467.8 g), gumminess (from 8.2 to 362.1 g), and viscosity (from 6.7 to 18.3 Pa.s) of the DL compared to the control product (SMP). These findings suggest that adding MWP can modulate the rheological and textural characteristics of such products without significantly altering their proximate composition, offering an alternative to using thickeners in this type of product.

Dairy doi: 10.3390/dairy6050054

Authors: Marinela Enculescu Ioana Nicolae Dinu Gavojdian

This pilot study evaluated the effects of cannabidiol (CBD) oil supplementation on growth performance, stress biomarkers, and haematological profiles in dairy calves undergoing the weaning transition. Nineteen Holstein calves were divided into two paternal-sibling groups: a CBD-supplemented experimental group (n = 10) and a CON-control group (n = 9). The CBD group received 5 mL/head/day of CBD oil for the first two days (pre-weaning), followed by 10 mL/head/day for three consecutive days post-weaning. Body weight increased significantly over time in both groups (p = 0.000); nevertheless, no significant differences were observed between groups (p = 0.173) or for the group × time interaction (p = 0.929), indicating that CBD did not affect overall growth trajectory. However, a significant group × time interaction (p = 0.006) for average daily gains in the CBD group was observed. Serum cortisol concentrations were significantly lower in CBD-supplemented calves at Day 0 and +2 days, compared to the CON group, indicating a transient anti-stress effect (p = 0.043 for group effect). At +5 days, cortisol levels in the CBD group increased, surpassing control values, though this difference was not significant. A trend-level group × time interaction (p = 0.067) suggested a distinct temporal cortisol response in CBD-treated calves. Immune cell counts (LYM, MON, NEU) showed no significant differences, though monocyte levels trended lower in CBD calves at early time points. Platelet indices revealed a significant reduction in mean platelet volume (p = 0.047) and stable PDWc and plateletcrit values in the CBD group, suggesting modulation of inflammatory status. Alanine aminotransferase levels increased over time with a significant group effect (p = 0.014), indicating a mild hepatic response, while glucose and alkaline phosphatase remained within physiological ranges. These findings suggest that short-term CBD supplementation may transiently modulate stress and inflammatory responses during weaning, with potential benefits for physiological resilience. However, rebound endocrine effects and hepatic sensitivity highlight the need for further research to refine dosing strategies and assess long-term safety in dairy production systems.

Dairy doi: 10.3390/dairy6050053

Authors: Chenyang Li Xiaoyang Chen Tingting Fang Jie Gao Guangyong Zhao Xianhong Gu

Stereotypic behaviors are common in farm animals and often signal poor welfare. Tongue-rolling is the most prevalent stereotypic behavior in cows. In this study, we compared the plasma and milk composition of 16 high-frequency tongue-rolling cows (HTR group) and 16 non-stereotypic cows (CON group). All cows were primiparous cows. Biochemical tests, plasma metabolomics, and milk proteomics revealed higher plasma triiodothyronine levels in HTR cows, and lower levels of αs1-casein, β-casein, κ-casein, and lactoferrin in their milk. Multi-omics analyses identified 103 differential metabolites and 73 differential proteins, including various GTP-binding proteins, with the Ras signaling pathway being significantly upregulated in the HTR cows. GO enrichment analysis highlighted significant changes in molecular function, particularly related to GTP/GDP-binding proteins. Additionally, HTR cows exhibited elevated cellular metabolic activity. These findings suggest that high-frequency tongue-rolling is associated with altered endocrine and metabolic profiles, disrupted milk protein synthesis, and impaired immune function potential. The reduction in key milk proteins and lactoferrin may negatively impact milk quality and immune defense. Further research is needed to clarify the causal relationship between these physiological changes and tongue-rolling, providing insights into the underlying mechanisms of stereotypic behaviors in dairy cows and their implications for animal welfare and milk production.

Dairy doi: 10.3390/dairy6050052

Authors: Andrés Grajales-Zuleta Sandra Estrada Andrea Hermosa Isidra Recio Beatriz Miralles Mar Villamiel

Cheese products worldwide have gained protected designation of origin status in many instances, yet this food group also has the highest reported fraud rates. Quesillo Caquetá is the first Colombian cheese to acquire a protected designation of origin, but still there is a lack of information regarding its composition. In this study, a compositional analysis was performed to establish a set of characteristic parameters to aid the identification of the authenticity of Quesillo Caquetá. Physicochemical analysis, mineral composition determination, carbohydrate, fatty acid, and peptide profiles were conducted on 29 samples of Quesillo Caquetá made with milk from the northern, southern, and central regions of the province of Caquetá. The results revealed 7 minerals, 3 carbohydrates, 19 fatty acids, and 45 peptides (21 peptides from bovine αs1-casein and 24 peptides from bovine β-casein). This suggests that Quesillo Caquetá is a significant source of sodium, calcium, phosphorus, and monounsaturated fatty acids such as oleic acid, omega-3, and omega-6, as well as some peptides that match sequences with antihypertensive, immunomodulatory, antioxidant, and antimicrobial activity reported in the literature. The specificity of the fatty acid and peptide profiles can become a valuable tool for identifying the authenticity of Quesillo Caquetá against possible imitations in the market.

Dairy doi: 10.3390/dairy6050051

Authors: Ondrej Hanušovský Milan Šimko Michal Rolinec Branislav Gálik Mária Kapusniaková Stanislava Drotárová Matúš Džima Luboš Zábranský Miroslav Juráček

Total Mixed Ration (TMR) particle size significantly impacts dairy cow health and productivity. This study investigated the effects of TMR particle size tertiles on rumen pH, dry matter intake (DMI), and milk characteristics in Simmental cows by continuous pH monitoring (Moonsyst Ltd., Kilkenny, Republic of Ireland) and particle separation by 19, 8, 4 mm sieves and pad using the Wasserbauer particle separator, along with regular milk and DMI measurements. Data were analyzed by IBM SPSS 26.0 with ANOVA, Pearson correlations and statistically significant differences between tertiles by post hoc Tukey HSD test were performed (p < 0.05). Tertiles by frequency analysis were used to categorize particle size proportions into three groups, each containing an equal number of observations. Principal component analysis (PCA) and heatmaps by SRplot were generated. Moderate particle size distributions (second tertiles of 19 mm, 8 mm, 4 mm sieves, and pad as the fraction of TMR particles that pass through the all sieves and are collected in the bottom pan) optimized rumen pH stability, reducing time below 6.2 (SARA risk) or above 6.8, and correlated with milk β-hydroxybutyrate (BHB), oleic acid, and acetone levels. Moreover, milk production was maximized with a combination of coarser (19 mm and 8 mm, third tertiles) and finer (4 mm, first tertile) particles, milk fat peaked in both the finest pad fraction (third tertile) and coarsest larger sieves (first tertiles), and milk protein in the first tertiles of 19 mm and 8 mm sieves. Similarly, DMI positively correlated with coarser particles, but sometimes negatively with milk quality. In addition, PCA showed fine particle groups clustering with higher milk fat-to-protein ratios, somatic cell counts, and urea. In conclusion, mid-range TMR particle sizes (second tertiles) consistently provided the most benefits across ruminal, metabolic, and production parameters, underscoring TMR structure as a crucial precision feeding tool.

Dairy doi: 10.3390/dairy6050050

Authors: Karina Džermeikaitė Monika Šidlauskaitė Ramūnas Antanaitis Lina Anskienė

The convergence of genomic selection and artificial intelligence (AI) is redefining precision breeding in dairy cattle, enabling earlier, more accurate, and multi-trait selection for health, fertility, climate resilience, and economic efficiency. This review critically examines how advanced genomic tools—such as genome-wide association studies (GWAS), genomic breeding values (GEBVs), machine learning (ML), and deep learning (DL) models to accelerate genetic gain for complex, low heritability traits. Key applications include improved resistance to mastitis and metabolic diseases, enhanced thermotolerance, reduced enteric methane emissions, and increased milk yield. We discuss emerging computational frameworks that combine sensor-derived phenotypes, omics datasets, and environmental data to support data-driven selection decisions. Furthermore, we address implementation challenges related to data integration, model interpretability, ethical considerations, and access in low-resource settings. By synthesizing interdisciplinary advances, this review provides a roadmap for developing AI-augmented genomic selection pipelines that support sustainable, climate-smart, and economically viable dairy systems.

Dairy doi: 10.3390/dairy6050049

Authors: Melanie Schären-Bannert Benno Waurich Fanny Rachidi Adriana Wöckel Wolf Wippermann Julia Wittich Guntram Hermenau Erik Bannert Peter Hufe Detlef May Sven Dänicke Hermann Swalve Alexander Starke

Managing transition cows and preventing diseases related to this period is challenging due to the latter’s multifactorial nature. The aim of this applied observational case study is to illustrate and discuss the different aspects involved and provide an approach to analysis and the resulting management solutions using a real-life case within a 500-cow herd. The initial assessment, involving the collection of data on the level of production, animal health and behaviour, and metabolic indicators, as well as management and housing key indicators, revealed key risk factors, including overcrowding, suboptimal feeding strategies, inadequate water supply, and insufficient disease monitoring. These factors contributed to increased cases of metabolic disorders such as hypocalcemia (annual incidence 7.8%), excessive lipomobilisation, and displaced abomasum (annual incidence 5.2%). A holistic approach combining feeding adjustments, disease monitoring, facility improvements, and long-term management strategies was implemented to address these challenges. Short-term interventions, such as optimizing the dietary cation–anion balance and enhancing disease detection protocols, led to noticeable improvements. However, structural constraints and external factors, such as extreme weather conditions (heat stress) and economic limitations, created significant hurdles in achieving immediate and sustained success. The farm ultimately opted for infrastructural improvements, including a new transition cow facility, to provide a long-term solution to these recurring issues. This case highlights the complexity of transition cow management, demonstrating that long-term success depends on continuous monitoring, interdisciplinary collaboration, and adaptability in response to evolving challenges in dairy production.

Dairy doi: 10.3390/dairy6050048

Authors: Yu Wang Kuan Chen Shulin Yuan Jianying Liu Jianchao Guo Yongqing Guo

Methane (CH4) is the second largest greenhouse gas (GHG) after carbon dioxide (CO2), and ruminant production is an important source of CH4 emissions. Among the six types of livestock animal species that produce GHGs, cattle (including beef cattle and dairy cows) are responsible for 62% of livestock-produced GHGs. Compared to beef cattle, continuous lactation in dairy cows requires sustained energy intake to drive rumen fermentation and CH4 production, making it a key mitigation target for balancing dairy production and environmental sustainability. Determining how to safely and efficiently reduce CH4 emissions from dairy cows is essential to promote the sustainable development of animal husbandry and environmental friendliness and plays an important role in improving feed conversion, reducing environmental pollution, and improving the performance of dairy cows. Combined with the factors influencing CH4 emissions from dairy cows and previous research reports, this paper reviews the research progress on reducing the enteric CH4 emissions (EMEs) of dairy cows from the perspectives of the CH4 generation mechanism and emission reduction strategies, and it summarizes various measures for CH4 emission reduction in dairy cows, mainly including accelerating genetic breeding, improving diet composition, optimizing feeding management, and improving fecal treatment. Future research should focus on optimizing the combination of strategies, explore more innovative methods, reduce EME without affecting the growth performance of dairy cows and milk safety, and scientifically and effectively promote the sustainable development of animal husbandry.

Dairy doi: 10.3390/dairy6040047

Authors: Tom R. Angel Ben Barber Rachel Hayton Sophie A. Mahendran

Use of sedation for disbudding is common practice in a number European countries, with United Kingdom (UK) practices adopting its use. This study assessed the effects of disbudding with and without xylazine sedation on growth rates and calf health on a UK calf rearing unit. Data was collected from 485 dairy crossed with beef breed calves between April and August 2024 from a single calf rearing unit in England. Calves were purchased from multiple farms across the UK and arrived on site at approximately 21 days of age. Calves were disbudded—and, in the case of male calves, surgically castrated—at approximately three weeks after arrival on farm. Pens of calves were assigned to undergo disbudding with (SED, n = 238) or without (CTL = 234) xylazine sedation at a dose of 0.2 mg/kg administered intramuscularly. Calves from both groups were provided with local anaesthetic (procaine hydrochloride) as a cornual nerve block and a non-steroidal anti-inflammatory drug (meloxicam). While other studies have demonstrated some behavioural and physiological indicators of pain to be reduced with sedation, this study found that calves in the SED group had a reduced daily liveweight gain (DLWG) of 0.14 kg/day in the short term (mean 20 days) following disbudding (p < 0.001), but there was no difference in growth rates in the medium-term (mean 43 days) post-disbudding (p = 0.30). Some of this difference could be explained by the slightly higher DLWG pre-disbudding in the CTR group, and it is likely that the physiological impacts of sedation accounted for the rest of this difference. This initial reduction in DLWG following disbudding with sedation should be considered by vets, especially on farms where growth rates may already be compromised. In the sedated calves, 19.3% exhibited either some movement or entry into sternal recumbency. Specifically, a light plane of sedation with calves entering sternal recumbency was associated with a reduction in DLWG of 0.89 kg/day compared to 0.98 kg/day for those that remained in lateral recumbency throughout (p = 0.008). The light plane of sedation may have created additional calf stress, impacted feeding behaviours, and impinged welfare, with further work needed to establish the reasons for insufficient sedation. There was no difference in the number of post-disbudding treatment outcomes between calves disbudded with and without sedation (p = 0.97).

Dairy doi: 10.3390/dairy6040046

Authors: Beatriz Danieli Ana Luiza Bachmann Schogor Jardel Zucchi André Thaler Neto

This study aimed to examine the relationship between a high somatic cell count (SCC) in cows and milk quality during the hot season in different breeds. Milk samples from 500 cows in the hot season and 431 in the cold season of 2022 were collected across 39 farms in Santa Catarina, Brazil. The samples were analyzed for SCC, milk composition, and physical attributes. Cows were also evaluated for udder depth, udder clearance, teat-end condition, and leg and udder cleanliness. Based on the SCC levels, cows were categorized as low (≤200,000 cells/mL), medium (>200,000 and ≤615,000), or high (>615,000). Data were analyzed by ANOVA with a statistical model that included the effects of the SCC class, season, days in milk, parity, genetic group, and the interaction of the SCC level and season. The results showed that cows with a high SCC produced less milk with lower component levels but higher chloride content. Milk from the hot season had lower acidity and reduced component levels. The impact of SCC on the physical traits of milk did not vary with season. Furthermore, cows with deeper udders and lower udder clearance were more likely to have a high SCC, regardless of genetics. Both a high SCC and hot temperatures independently compromised milk yield and quality, thereby increasing the risk of culling. Therefore, improving udder conformation and avoiding cows with deep udders may help to reduce SCC levels.

Dairy doi: 10.3390/dairy6040045

Authors: Leonardo Leite Cardozo Deise Aline Knob Pauline Thais dos Santos Angela Pelizza Ana Paula Mori Mauricio Camera Sandra Maria Ferraz Marcella Zampoli de Assis André Thaler Neto

Maintaining teat-end integrity in dairy cows is essential to preventing intramammary infections (IMIs) in dairy cows, yet the relationship between hyperkeratosis, teat conformation, and mammary health remais underexplored. This study evaluated the relationship between teat-end hyperkeratosis, teat conformation traits, microbial colonization, and somatic cell count (SCC) in milk from 170 cows on ten commercial dairy farms in Santa Catarina, Brazil. During two farm visits, milk and teat-end swab samples from paired teats (one with hyperkeratosis, one without) were analyzed for microbial growth and SCC. SCC data were transformed into somatic cell scores (SCS). Results showed no significant association between hyperkeratosis and mastitis microorganisms, although environmental microorganisms tended to be more frequent in hyperkeratotic teats (p = 0.0778). Major microorganisms in milk were significantly associated with higher SCC (p = 0.0132). No relationship was observed between teat conformation traits and hyperkeratosis. These findings suggest that hyperkeratosis may subtly influence the teat canal to environmental bacterial colonization, underscoring the need for improved milking management practices to minimize hyperkeratosis and associated mastitis risks.

Dairy doi: 10.3390/dairy6040044

Authors: Rogério D’Antonio Pires Aline Moreira Borowsky Tobias Alves e Silva Giovanna Canela Ruiz Castro Evangelista Carla Maris Machado Bittar Carlos Humberto Corassin

The diet of dairy calves can be contaminated with mycotoxins, posing a potential risk to animal health. This case study report aimed to make the first assessment of the presence of multiple mycotoxins in concentrates fed to dairy calves in Brazil. A total of 19 concentrate samples intended for dairy calves were analyzed using liquid chromatography coupled with mass spectrometry. Aflatoxins, deoxynivalenol, and T-2 toxin were not detected in any samples, whereas fumonisins B1 (FB1) and B2 (FB2) were present in 100% of the samples, with mean concentrations of 2750.1 μg/kg and 834.9 μg/kg, respectively. Zearalenone (ZEN) was detected in 36.8% of samples, with a mean concentration of 929.9 μg/kg. Significant correlations were observed between FB1 and FB2 (ρ = 0.978; p < 0.001) and between FB2 and ZEN (ρ = 0.735; p = 0.05). While the physical form of the concentrate did not influence (p > 0.05) mycotoxin concentrations, a trend was observed for FB1 (ρ = −0.417; p = 0.07) and FB2 (ρ = −0.395; p = 0.09). These findings highlight the frequent occurrence of Fusarium mycotoxins, likely due to pre-harvest contamination, emphasizing the potential risk of additive or synergistic effects in dairy calves.

Dairy doi: 10.3390/dairy6040043

Authors: Giovanna Lomolino Stefania Zannoni Mara Vegro Alberto De Iseppi

Consumer interest in vegetarian, ethical, and clean-label foods is reviving the use of plant-derived milk coagulants. Cardosins from Cynara cardunculus (“thistle”) are aspartic proteases with strong clotting activity, yet their technological impact in cheese remains under-explored. This study compared a commercial thistle extract (PC) with traditional bovine rennet rich in chymosin (AC) during manufacture and 60-day ripening of Caciotta cheese. Classical compositional assays (ripening index, texture profile, color, solubility) were integrated with scanning electron microscopy, three-dimensional surface reconstruction, and descriptive sensory analysis. AC cheeses displayed slower but sustained proteolysis, yielding a higher and more linear ripening index, softer body, greater solubility, and brighter, more yellow appearance. Imaging revealed a continuous protein matrix with uniformly distributed, larger pores, consistent with a dairy-like sensory profile dominated by milky and umami notes. Conversely, PC cheeses underwent rapid early proteolysis that plateaued, producing firmer, chewier curds with lower solubility and darker color. Micrographs showed a fragmented matrix with smaller, heterogeneous pores; sensory evaluation highlighted vegetal, bitter, and astringent attributes. The data demonstrate that thistle coagulant can successfully replace animal rennet but generates cheeses with distinct structural and sensory fingerprints. The optimization of process parameters is therefore required when targeting specific product styles.

Dairy doi: 10.3390/dairy6040042

Authors: Fabiana Ribeiro Caldara Jéssica Lucilene Cantarini Buchini Rodrigo Garófallo Garcia

Background: Environmental enrichment is a promising strategy to improve the welfare of dairy goats and sheep. However, studies in this field remain scattered, and its effects on productivity are unclear. Objectives: To evaluate the effects of environmental enrichment on behavioral, physiological, and productive parameters in dairy goats and sheep. Data sources: Scopus and Web of Science were searched for studies published from 2010 to 2025. Study eligibility criteria: Experimental or observational peer-reviewed studies comparing enriched vs. non-enriched housing in dairy goats or sheep, reporting on welfare or productivity outcomes. Methods: This review followed PRISMA 2020 guidelines and the PICO framework. Two independent reviewers screened and extracted data. Risk of bias was assessed with the SYRCLE tool. Results: Thirteen studies were included, mostly with goats. Physical, sensory, and social enrichments showed benefits for behavior (e.g., activity, fewer stereotypies) and stress physiology. However, results varied by social rank, enrichment type, and physiological stage. Only three studies assessed productive parameters (weight gain in kids/lambs); none evaluated milk yield or quality. Limitations: Most studies had small samples and short durations. No meta-analysis was conducted due to heterogeneity. Conclusions: Environmental enrichment can benefit the welfare of dairy goats and sheep. However, evidence on productivity is scarce. Long-term studies are needed to evaluate its cost-effectiveness and potential impacts on milk yield and reproductive performance.

Dairy doi: 10.3390/dairy6040041

Authors: Ulises Alvarado Jhon Tacuri Alejandro Coloma Edgar Gallegos Rojas Herbert Callo Cristina Valencia-Sullca Nancy Curasi Rafael Manuel Castillo

Monitoring pH and acidity during yoghurt fermentation is essential for product quality and process efficiency. Conventional measurement methods, however, are invasive and labour-intensive. This study developed artificial neural network (ANN) models to predict pH and titratable acidity during yoghurt fermentation using CIELAB colour parameters (L, a*, b*). Reconstituted milk powder with 12% total solids was prepared with varying protein levels (4.2–4.8%), inoculum concentrations (1–3%), and fermentation temperatures (36–44 °C). Data were collected every 10 min until pH 4.6 was reached. Forty models were trained for each output variable, using 90% of the data for training and 10% for validation. The first two phases of the fermentation process were clearly distinguishable, lasting between 4.5 and 7 h and exceeding 0.6% lactic acid in all treatments evaluated. The best pH model used two hidden layers with 28 neurons (R2 = 0.969; RMSE = 0.007), while the optimal acidity model had four hidden layers with 32 neurons (R2 = 0.868; RMSE = 0.002). The strong correlation between colour and physicochemical changes confirms the feasibility of this non-destructive approach. Integrating ANN models and colourimetry offers a practical solution for real-time monitoring, helping improve process control in industrial yoghurt production.

Dairy doi: 10.3390/dairy6040040

Authors: Hunter R. Ford Joseph Klopfenstein Serkan Ates Sebastiano Busato Erminio Trevisi Massimo Bionaz

The objective of this study was to evaluate the effects of feeding a combination of chicory–plantain silage and supplementing Se yeast on the response of early-lactating ewes to induce subclinical mastitis. Polypay ewes (n = 32) were fed either chicory–plantain silage or grass silage and supplemented with 3.6 mg Se yeast/ewe/day for approximately 2 months prior to the infusion of S. uberis into both mammary glands (i.e., intramammary infection or IMI). The ewes had a typical subclinical mastitis response with an 8-fold increase in milk somatic cell count within 24 h post-IMI, a decrease in milk yield, and changes in all milk components measured. The ewes experienced a mild systemic inflammation post-IMI as determined by an increase in rectal temperature and decrease in feed and water intake and, in blood, by an increase in the concentration of ceruloplasmin, haptoglobin, and myeloperoxidase and a decrease in paraoxonase, Zn, advanced oxidation protein products, and hematocrit with no effect on pro-inflammatory cytokines. No effect of silage type, likely due to a low concentration of secondary compounds, or Se supplementation was detected in response to IMI. In summary, the subclinical mastitis model used was effective in mounting an inflammatory response, although this was mild; however, feeding chicory–plantain silage with a low concentration of secondary compounds and supplementing Se yeast had no significant effect on the response of ewes to mammary infection.

Dairy doi: 10.3390/dairy6040039

Authors: Fabiellen Pereira Sagara Kumara Muhammad Ahsin Lamis Ali Ying Xi Stephan van Vliet Simon Kelly Anita Fleming Pablo Gregorini

This on-farm study evaluated the effects of a regenerative (plant polyculture) as compared to conventional (monoculture) pasture-based New Zealand dairy production system on milk and yoghurt nutraceutical properties and environmental impact. Milk and yoghurt produced by two adjacent regenerative and conventional farms were sampled throughout the year and analyzed for chemical composition, metabolomics, and microbiome. Milk samples were also collected over four consecutive days (one day after herbage sampling) on four occasions throughout lactation: early lactation (October), peak lactation (December/January), mid-lactation (March), and late lactation (May). Overall, the regenerative system had a lower environmental impact while maintaining a similar yield and the same milk composition compared to conventional systems. Furthermore, milk and yoghurt from the regenerative system had a more favourable profile of phytochemical antioxidants with potential positive benefits to human health (anti-inflammatory and antioxidant).

Dairy doi: 10.3390/dairy6040038

Authors: Rafael Vieira de Sousa Jéssica Caetano Dias Campos Gabriel Pagin Danilo Florentino Pereira Aline Rabello Conceição Rubens André Tabile Luciane Silva Martello

Infrared thermography (IRT) is explored as a non-invasive method for indirectly measuring parameters related to animal performance and welfare. This study investigates a feature extraction method termed the “thermal signature” (TS), a descriptor vector derived from the temperature matrix of an animal’s body surface, representing the percentage distribution of temperatures within predefined ranges. The TS, combined with environmental data, serves as a predictor attribute for machine learning-based classifier models to assess heat stress levels. The methodology was applied to a dataset collected from two groups of five dairy calves housed in a climate-controlled chamber and exposed to two artificial heat waves over 13 days. Data, including IRT measurements, respiratory rate (RR), rectal temperature (RT), and environmental variables, were collected five times daily (from 6 a.m. to 10 p.m., every four hours). Classifier models were developed using random forest (RF), support vector machine (SVM), artificial neural network (ANN), and K-nearest neighbor (KNN) algorithms. The RF models based on RR achieved the highest accuracies, 94.1% for two heat stress levels and 80.3% for three heat stress levels, using TS configurations with six temperature ranges. The integration of TS with machine learning-based models demonstrates promising results for developing or enhancing classifiers of heat stress levels in dairy calves.