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.

Dairy doi: 10.3390/dairy6040037

Authors: Larissa V. F. Cruz Ruan R. Daros André Ostrensky Cristina S. Sotomaior

To study behavioral and productive factors to detect changes that may indicate and predict clinical mastitis cure, Holstein dairy cows (n = 60), in an automatic milking system (AMS) and equipped with behavioral monitoring collar, were monitored from the diagnosis of clinical mastitis (D0) until clinical cure. The parameters collected through sensors were feeding activity, milk electrical conductivity (EC), milk yield, Mastitis Detection Index (MDi), milk flow, and number of gate passages. Clinical mastitis cases (n = 22) were monitored and divided into cured cases (n = 14) and non-cured cases within 30 days (n = 8), paired with a control case group (n = 28). Cows were assessed three times per week, and cure was determined when both clinical assessment and California Mastitis Test (CMT) results were negative in three consecutive evaluations. Mixed generalized linear regression was used to assess the relationship between parameters and clinical mastitis results. Mixed generalized logistic regression was used to create a predictive model. The average clinical cure time for cows with clinical mastitis was 11 days. Feeding activity, gate passages, milk yield, milk flow, EC, and the MDi were associated with cure. The predictive model based on data from D0 showed an Area Under the Curve of 0.89 (95% CI = 0.75–1). Sensitivity and specificity were 1 (95% CI = 1–1) and 0.63 (95% CI = 0.37–0.91), respectively. The predictive model demonstrated to have good internal sensitivity and specificity, showing promising potential for predicting clinical mastitis cure within 14 days based on data on the day of clinical mastitis diagnosis.

Dairy doi: 10.3390/dairy6040036

Authors: Rita Inácio Ana Gomes Jorge Saraiva

Serra da Estrela cheese (a raw ewe’s milk) ripened for 45 days was treated at 600 MPa for 6 min (P1) and at 450 MPa for 6 (P2) and 9 min (P3) and kept in refrigerated storage for 15 months. Lactobacillus and Lactococcus viable cell numbers were reduced in 3.2–3.6 and 2.7–3.6 log cycle units, respectively. Lower reductions were verified for total aerobic mesophilic and Enterococcus viable cell numbers in cheeses treated at 450 MPa (2.4–2.5 and 1.2 log reductions, respectively). In HPP cheeses, yeasts and moulds were below the enumeration limit up to 6 months of storage, but at 15 months 3.6–4.2 log cfu/g were quantified in all cheeses, while Enterobacteriaceae were inactivated to below the quantification limit. The increment of pressure treatment caused a greater impact on the microbiota’s viability than the increase in time under pressure. During storage, minor total colour differences were determined for HPP P3 cheese surface relative to control cheeses, ChC, at 45 days of ripening. HPP can thus be a good process to apply after cheese manufacture, since it offers a good potential to render raw-milk cheese microbiologically safer, with minimal changes in quality.

Dairy doi: 10.3390/dairy6040035

Authors: Bence Tarr János Tőzsér István Szabó András Revoly

Milk quality has a growing importance for farmers as component-based pricing becomes more widespread. Food quality and precision manufacturing techniques demand consistent milk composition. Udder health, general cow condition, environmental factors, and especially feed composition all influence milk quality. The large volume of routinely collected milk data can be used to build prediction models that estimate valuable constituents from other measured parameters. In this study, casein was chosen as the target variable because of its high economic value. We developed a multiple linear-regression model and a feed-forward neural network model to estimate casein content from twelve commonly recorded milk traits. Evaluated on an independent test set, the regression model achieved R2 = 0.86 and RMSE = 0.018%, with mean bias = +0.003% and slope bias = −0.10, whereas the neural network improved performance to R2 = 0.924 and RMSE = 0.084%. In silico microgreen inclusion from 0% to 100% of dietary dry matter raised the predicted casein concentration from 2.662% to 3.398%, a relative increase of 27.6%. To extend practical applicability, a simulation module was created to explore how microgreen supplementation might modify milk casein levels, enabling virtual testing of dietary strategies before in vivo trials. Together, the predictive models and the microgreen simulation form a cost-effective, non-invasive decision-support tool that can accelerate diet optimization and improve casein management in precision dairy production.

Dairy doi: 10.3390/dairy6040034

Authors: Pablo M. Roskopf Alejandra Cuatrin Matías Stangaferro Gino Storani Emmanuel Angeli Gustavo J. Hein Eloy E. Salado

This study evaluated the effects of supplementing early-lactation Holstein cows with rumen-protected omega-3 fatty acids (calcium salts) on productive and reproductive performance. Thirty-six multiparous cows were randomly assigned to one of two treatments from 21 ± 2 days before calving to 105 ± 5 days in milk (DIM): a Control group (C) or an Omega-3-supplemented group (O-3), receiving a blend of linseed and fish oil (60:40). Both groups were fed isoenergetic diets, with ground corn as the control supplement. Total dry matter and net energy intake did not differ between treatments. A treatment-by-time interaction was observed for milk yield, with cows in the O-3 group producing more milk than controls at specific time points. Additionally, O-3 cows had higher overall protein yield and improved feed efficiency. No differences were found in body weight, BCS, glucose, insulin, IGF-1, or urea concentrations, but a tendency toward higher plasma NEFA and BHBA concentrations and lower energy balance was observed in the O-3 group. Supplementation increased plasma cholesterol and progesterone concentrations and was associated with a higher proportion of cows being pregnant at 130 DIM. These findings suggest that omega-3 supplementation may improve specific aspects of lactational performance and reproductive efficiency without compromising metabolic status.

Dairy doi: 10.3390/dairy6040033

Authors: Anita Fleming Philip Wescombe Pablo Gregorini

The objective of this work was to explore the effect of taxonomically and phytochemically rich swards, as opposed to ‘status quo’ monoculture of ryegrass and white clover swards, on animals and milk, by assessing the metabolomic profile of plant and milk samples. The results of this study suggest that metabolomic profiles and metabolism are altered by dietary diversity and grazing management. Several metabolites associated with enhanced consumer health were elevated in milk from cows that were grazed in functionally diverse swards as opposed to monoculture of ryegrass and white clover swards. The results show the value of metabolomics for the identification of metabolites associated with a ‘pasture-fed diet’, which is considered beneficial to human health compared with grain-fed milk products. Metabolomics also provide evidence that grazing management and the presence of key forages may be more beneficial for market differentiation of milk products that can enhance consumer health than maximisation of sward species diversification.

Dairy doi: 10.3390/dairy6040032

Authors: Miguel Mazorra-Manzano Angelica Martínez-García María Torres-Llanez Juan Ramírez-Suárez Yolanda López-Franco Francisco Brown-Bojórquez José Teutle-Paredes María Lugo-Sánchez

Requesón, a Mexican whey cheese, has a short shelf life due to its high moisture content, near-neutral pH, and the limited preservation infrastructure of the artisanal cheese sector. Therefore, the development of requesón powder provides an innovative pathway to enhance market potential and expand its applications. This study aimed to evaluate the techno-functional properties of requesón powder produced through air-convective drying and to assess the protective effects of two natural gums, mesquite gum and guar gum, at concentrations of 0.25 and 0.5 g/L. Thermal dehydration significantly affected (p < 0.05) water holding capacity, swelling capacity, and hardness of the reconstituted powder. Although gum addition did not significantly enhance water holding capacity, it moderately improved texture and led to notable increases in swelling capacity (21–34%) and emulsifying capacity (11–20%) at high concentrations (p < 0.05). Structural analyses using X-ray diffraction and electron microscopy revealed that thermal dehydration induced protein aggregation and reduced microporosity, impairing rehydration performance compared to requesón powder obtained by lyophilization. These findings suggest that requesón powder production is a promising strategy for valorizing whey and extending the applications of this traditional cheese as a functional food ingredient.

Dairy doi: 10.3390/dairy6040031

Authors: Malaiporn Wongkaew Bow Tinpovong Aekarin Inpramoon Pikulthong Chaimongkol Auengploy Chailangka Sureerat Thomya Nuttinee Salee

Consumer scrutiny of fat content in foods is becoming a notable trend in health concerns. This study aims to develop a novel functional low-fat goat feta cheese by utilizing polydextrose (PDX) and inulin as dietary fiber-based fat replacers to improve its overall characteristics. The physicochemical and textural properties, along with consumer acceptance, of the feta cheese were evaluated across three fat levels (full-fat [FFC], reduced-fat [RFC], low-fat [LFC]) and three fibers: PDX, inulin, and their combination. The intercorrelation of all characteristics was assessed through principal component analysis and Pearson’s correlation. Fat reduction significantly altered the cheese’s visual properties, increasing lightness and the total color difference, which inversely correlated with a* and b* values. Lower-fat cheeses exhibited decreased pH and increased lactic acid, with salinity playing a crucial role in both lactic acid development and texture. Under Scanning Electron Microscopy (SEM), PDX yielded a cheese matrix with a finer pore structure than inulin or the combined fibers. Lower-fat cheeses exhibited greater hardness, with PDX resulting in the highest hardness among the fiber treatments. Crucially, the RFC with PDX was as well-received by consumers as the FFC. These findings not only empower goat farmers and cheese entrepreneurs to increase their product value for niche market but also contribute to sustainability by providing a healthier food option for functional benefits.

Dairy doi: 10.3390/dairy6040030

Authors: Erçin Aydoğan Vasıf Kubilay Ayrancı Pelin Salum Zafer Erbay

In this study, Malatya cheeses were produced using cow’s milk, sheep’s milk, and a cow–sheep milk mixture (1:1), were stored in brine solutions, and samples from both the cheeses and their brines were collected and analyzed at 0, 30, 90, and 180 days of storage to investigate the impact of the milk type and storage time on the cheese characteristics. Cheese made from cow’s milk exhibited a lower fat content (14.5%), whereas cheese made from sheep’s milk had a lower protein content (17.5%). During storage, salt and ash contents increased. Water-soluble nitrogen (WSN) and trichloroacetic acid-soluble nitrogen (TCASN) levels decreased during the first 90 days of storage, followed by a subsequent increase. Cow’s milk cheese demonstrated higher ripening extension index (REI) values, indicating early-stage proteolysis, whereas sheep milk cheese showed higher ripening depth index (RDI) values, reflecting more advanced ripening. The total concentration of volatile compounds in the headspace increased over time, rising from 576.7–1060.2 to 5795.1–7360.1 µg/kg dry matter by day 180 of storage, with acids being the dominant volatile group in both quantity and diversity. Free fatty acids (FFAs) were the predominant volatiles and branched-chain acids and alcohols associated with proteolysis were particularly notable in cow’s milk cheeses. Additionally, the transfer of proteins and volatile compounds into the brine increased throughout the storage period. Overall, storage time significantly influenced the cheese characteristics, while milk type also played a role, albeit to a lesser extent.

Dairy doi: 10.3390/dairy6030029

Authors: Ja’far Al-Khaza’leh Belal S. Obeidat

Utilizing agro-industrial by-products to feed livestock is crucial for environmental protection and, simultaneously, lowering production and feeding costs. In light of these aims, in this study, two trials were conducted to evaluate the impact of jojoba meal (JJM) on Awassi ewes’ milk properties, nutrient intake, digestibility, and the pre-weaning growth of their nursing lambs. In the first trial, 22 Awassi ewes were divided equally between two experimental diets at random (11 ewes per diet): 0% JJM (CON) and 15% JJM (JJM15) of dietary dry matter (DM). In the second trial, 10 ewes were randomly selected (5 ewes/diet) to assess nutritional digestibility and N balance. The results of trial 1 showed that neutral detergent fiber (NDF), acid detergent fiber (ADF), and ether extract (EE) intake values were greater (p ≤ 0.02) in the JJM15 group compared to the CON group. The total gain and average daily gain (ADG) of the lambs in the JJM15 group were significantly higher compared to those of the CON group (p < 0.05). The average milk yield of nursing ewes was similar between the two dietary groups (p ˃ 0.05). The percentage and yield of solids-not-fat (SNF) in the JJM15 group were significantly higher compared to the CON group. The other milk components, including protein, lactose, fat, and total solids yields, were similar between the dietary groups (p ˃ 0.05). The milk production cost was significantly lower for the JJM15 diet than the CON diet (p = 0.004). Triglyceride levels were lower (p = 0.001) in the JJM15 diet group than in the CON diet group. In trial 2, nutrient digestibility and N balance were not affected (p ≥ 0.073) by the consumption of JJM15. In conclusion, JJM at 15% of dietary DM resulted in no negative effects on the health, performance, or milk quality of Awassi ewes. Most significantly, by lowering the cost of production, our results appear to indicate that this dietary supplement improves economic efficiency.

Dairy doi: 10.3390/dairy6030028

Authors: Mahmoud H. Emam Abdelmonem Abdallah Elise Shepley Luciano S. Caixeta

Hyperketonemia (HYK) is a common disorder in high-producing dairy cows, resulting in significant economic losses. Defined by elevated beta-hydroxybutyrate (BHB; ≥1.2 mmol/L) without clinical signs, HYK is often considered a gateway disease, predisposing cows to other metabolic and infectious problems. Our objective was to investigate the association between previous lactation risk factors and both BHB concentration and HYK status during the first week postpartum in the subsequent lactation. A retrospective study was conducted using previously collected blood samples from 2336 Holstein multiparous dairy cows from 7 dairy herds, where BHB concentration was measured during the first week postpartum. Data from the previous lactation were extracted from electronic farm records. Log-transformed BHB concentrations and HYK status were each modeled using separate linear mixed models. Both models included the same set of risk factors—lactation, previous lactation total times bred, dry length period, previous lactation days in milk, previous lactation days open, previous lactation days carried calf, previous lactation peak milk production, previous lactation total milk production, previous lactation total milk fat, and previous lactation total milk protein—to investigate their association with these outcomes. Potential confounding variables were offered to the models, and stepwise backward elimination was used to determine which covariates to retain. Significant associations were detected between BHB concentration and dry period length (DDRY), lactation number (LACT), previous lactation total milk protein (TOTP), and previous lactation days open (PDOPN). Inclusive, significant associations were detected between HYK status and previous lactation total milk production (PTOTM), DDRY, LACT, TOTP, and PDOPN. Our results suggest that a dry period longer than 60 days, days open exceeding 130 days, being in their third or greater lactation, and each additional 1000 kg of milk produced in the previous lactation are associated with an increased risk of having high BHB and HYK in the first week postpartum in the subsequent lactation.

Dairy doi: 10.3390/dairy6030027

Authors: Nora Idalia Diaz-Negrete Gabriela Ramos Clamont Montfort Cristina Cueto-Wong Silvia Gabriela López-Fernández Rene Renato Balandrán-Quintana

This study evaluated two techniques for encapsulating Lactiplantibacillus plantarum using bovine lactoferrin and sodium alginate. The first method involved a layer-by-layer (LbL) coating of lactoferrin and alginate directly onto individual cells, using three layers of these electrolytes. The second method focused on encapsulating the probiotics in calcium alginate miniaturized beads, followed by a lactoferrin coating (AAcL). Encapsulation efficiency was measured at 52.7% for the LbL method and 32.6% for AAcL. Encapsulation was confirmed through zeta potential changes and scanning electron microscopy (SEM) micrographs. After freeze drying, the LbL technique showed a 2.67 log CFU decrease in survival rates, whereas the AAcL method resulted in a 3.77 log CFU decline. Nonencapsulated probiotics experienced a reduction of 5.8 log CFU. In storage at −20 °C, the LbL method led to a 32% decrease in survival after 30 days and 41% after 90 days, while the AAcL method showed a decline of 15% after 30 days and 28% after 90 days. Both techniques preserved 75% of the initial L. plantarum population under simulated gastrointestinal conditions. Overall, these methods effectively protected the probiotic from environmental stress.

Dairy doi: 10.3390/dairy6030026

Authors: Ryotaro Miura Taiki Izumi Yuriko Wada Yujiro Hagita Kosuke Iga Natsumi Sobu Akiya Hirase Osamu Yoneyama Yo-ichi Miyake Tsuyoshi Tajima Tadaharu Ajito Shingo Haneda Motozumi Matsui

The combined effects of human chorionic gonadotropin (hCG) treatment on Day 5 (Day 0 = the day of artificial insemination: AI) and intravaginal progesterone device (IVPD) treatment from Day 5 to 19 on the conception rate and plasma sex steroid hormone were examined in lactating dairy cows. In Experiment 1, cows were divided into a non-treatment group (n = 181), untreated; an IVPD group (n = 74), with an IVPD inserted into the vagina from Day 5 to 19; and an hCG + IVPD group (n = 87), with 3000 IU hCG administered intramuscularly on Day 5 and an IVPD inserted into the vagina from Day 5 to 19. The conception rates of the hCG + IVPD group, which underwent ≤3 AIs (52.8%) and >3 AIs (73.3%), and the IVPD group, which underwent >3 AIs (63.0%), were significantly higher than that of the non-treatment group, which underwent ≤3 AIs (34.2%). In Experiment 2, blood samples were collected from the non-treatment group (n = 6), the IVPD group (n = 6), and the hCG + IVPD group (n = 7) on days 5, 8, 12, and 15 from estrus to measure plasma progesterone (P4) and total estrogen (E) concentrations. The plasma P4 concentration of the hCG + IVPD group tended to be higher than that of the non-treatment group on Day 15; however, plasma E concentrations were not different among groups. These results suggest that the conception rate was improved by hCG + IVPD treatment regardless of AI number because of higher plasma P4 concentrations later in the estrous cycle.

Dairy doi: 10.3390/dairy6030025

Authors: Bruna Samara dos Santos Rekowsky Lorena Brandão Rocha Martinez Fernandez Katherine Gutierrez Alzate Carini Aparecida Lelis Marcelo Resende de Souza Carolina Oliveira de Souza Bruno Dutra da Silva Carlos Adam Conte-Júnior Carla Paulo Vieira José Givanildo da Silva Marion Pereira da Costa

Buffalo milk is a rich source of precursor fatty acids for bioactive compounds and provides an optimal environment for bacterial growth. This study aimed to isolate and select lactic acid bacteria strains with potential to conjugated linoleic acid (CLA) production for technological application in fermented buffalo milk. Fifty-eight strains were isolated from raw milk, kefir, artisanal cheese, kombucha, and jaboticaba juice and tested for CLA biosynthesis. In milk fermentation, selected strains with linoleic acid (LA) conversion rates ranging from 65.66% to 21.86% were L. paraplantarum, L. plantarum, P. pentosaceus, and L. fermentum. The highest viability average values between 11.85 and 11.15 Log CFU/mL were observed after 8 h of fermentation for the L. plantarum, control L. plantarum, and L. fermentum treatments, while it took 10 h of fermentation for L. paraplantarum and P. pentosaceus to reach a stationary phase, with pH stabilizing at 4.60 ± 0.1 after 30 h. Despite L. paraplantarum showing the highest in vitro CLA production (0.99 mg/mL), in buffalo milk, all strains similarly produced c9t11 CLA, with no detectable t11c12 CLA. P. pentosaceus and L. fermentum showed a fatty acid profile with higher PUFA content, especially in CLA and MUFA, related to a lower degree of atherogenicity (IA) and thrombogenicity index (ThI). These findings boost understanding of dairy (raw milk, artisanal cheese, and milk kefir) and non-dairy substrates (kombucha and jaboticaba juice) as reservoirs for functional bacteria and highlight buffalo milk as a matrix for diversification of naturally enriched fermented dairy products.

Dairy doi: 10.3390/dairy6030024

Authors: María Carmen Sánchez Eduardo Soria Arancha Llama-Palacios Franco Almirón Alberto Valdés Alejandro Cifuentes Marta Hernández María José Ciudad Luis Collado

This study explores the microbiota and metabolite profile of raw cow’s milk, analyzing the correlation between them and their potential impact on consumer health, focusing on factors like cow’s feeding regimen (conventional vs. grazing), the herd’s geographical location (coastal vs. inland), and the thermal stability of raw milk samples. To achieve this, raw milk quality was assessed through mesophilic bacterial count and thermal stability using the 70° and 80° ethanol test. Metataxonomic analysis was performed using Illumina 16S rRNA hypervariable region sequencing, and untargeted metabolomics was conducted using UHPLC-Q/TOF MS/MS, followed by multivariate correlation analysis. All samples met quality standards. A total of 1258 prokaryotic ASVs were identified, spanning 48 bacterial phyla and one archaeal phylum, with 909 genera and 349 unidentified ASVs. Additionally, 70 metabolites were identified, including essential amino acids, vitamins, and bioactive compounds with antimicrobial, anti-inflammatory, and prebiotic properties. Significant correlations between bacterial diversity and metabolite profiles were observed. Feeding regimen and geographical location influence microbial composition and metabolite abundance, while thermal stability was linked to specific metabolites like oleamide and pyridoxal. These findings suggest that microbiota-derived metabolites can enhance the nutritional and functional value of milk. Further research is needed to understand how environmental and processing factors influence these dynamics.

Dairy doi: 10.3390/dairy6030023

Authors: Agni M. N. Corrêa Camila S. da Silva Marco A. S. Gama Luciana F. P. Soares Andreia F. de Souza Michelle C. B. Siqueira Elizabeth Q. L. de Vasconcelos Victor J. L. Galeano Robert E. Mora-Luna Thayane V. M. Santos Adriano H. N. Rangel Marcelo de A. Ferreira

The present study explored the combined effects of CC (Opuntia stricta [Haw.] Haw.) and full-fat corn germ (FFCG) as a source of supplementary PUFA on milk fatty acid (FA) composition of dairy goats fed elephant grass (EG). Twelve Saanen goats were used in a replicated 4 × 4 Latin square design in a 2 × 2 factorial arrangement of treatments (GC or FFCG as energy sources, and the inclusion or not of CC in the diet as a partial substitute for EG). The proportions of various milk FAs were influenced by CC, FFCG, or both. Significant interactions between CC and FFCG were noted for most trans-C18:1 and CLA isomers. Specifically, including CC in the FFCG-supplemented diet increased the levels of trans-11 C18:1 and cis-9, trans-11 CLA in milk fat, whereas these isomers were unchanged or slightly reduced with CC in the GC diet. Similar patterns were observed for C18:2 n-6, while C16:0 increased with CC in the GC diet. Ratios of trans-C18:1/C18:0 and trans-11 C18:1/C18:0 were notably higher when CC was included in the FFCG-supplemented diet. These findings suggest that CC inhibits the last step of rumen biohydrogenation in dairy goats, enriching milk with trans-11 C18:1 and cis-9, trans-11 CLA when supplementary PUFA is provided in the diet.

Dairy doi: 10.3390/dairy6030022

Authors: Burim Ametaj

This review highlights a paradigm shift in our understanding of hypocalcemia during milk fever by introducing the concept of the Calci-Inflammatory Network. Traditionally viewed as a pathological deficiency necessitating rapid correction (e.g., through calcium borogluconate infusions or dietary adjustments like dietary cation-anion difference), periparturient hypocalcemia is reinterpreted here as an adaptive, protective response. Within this new framework, reduced circulating calcium levels may help temper systemic inflammation by limiting lipopolysaccharide (LPS) aggregation and curbing excessive macrophage activation. The review discusses how calcium signaling, the calcium-sensing receptor (CaSR), and immune cell functions adapt under hypocalcemic conditions to modulate inflammatory processes. This integrated perspective not only redefines the role of hypocalcemia but also proposes the Calci-Inflammatory Network as a novel concept through which we can understand how changes in calcium homeostasis mitigate inflammatory cascades—potentially lowering the incidence of periparturient diseases and enhance overall cow health and farm productivity. Future research should investigate the long-term effects of hypocalcemia, the environmental influences on this Calci-Inflammatory Network, and their collective impact on disease susceptibility and inflammation.

Dairy doi: 10.3390/dairy6030021

Authors: Marianna Karela Lambros Sakkas Evangelia Zoidou Golfo Moatsou Konstantina Milosi Ekaterini Moschopoulou

The acid whey derived during the production of Greek yoghurt, i.e., yoghurt acid whey (YAW), is considered as environmental pollutant due to its low pH and high lactose content. YAW may be obtained by centrifugation or ultrafiltration or filtration through cloth bags (traditional method) of the initial yoghurt, methods that could affect its composition. In the last years, efforts have been made to improve its composition using membrane technology. In the present work, the composition of 35 different YAW samples from Greek Yoghurt dairies was studied. The YAW samples were grouped according to the way of production and the kind of milk in the case of the traditional method. The results showed that both the kind of milk and the way of yoghurt staining affected its composition. Ovine YAW derived from traditional straining was richer in lactic acid and calcium than bovine YAW. Moreover, the composition of bovine YAW varied among the different ways of straining, and this affected its behaviour during membrane processing. Nanofiltration of three representative bovine YAW samples and one ovine YAW sample at their natural pH, i.e., pH 4.5, and at 25 °C removed the lactic acid at a range from 40 to 55%, and the monovalent cations > 60% and retained lactose and galactose at percentages > 95% and 80% respectively.

Dairy doi: 10.3390/dairy6030020

Authors: Michail Sabino Moroz Camila Cecilia Martin Ruan Rolnei Daros

There are few studies on how dairy × beef crossbred calves perform during the pre-weaning phase compared to dairy calves. This observational study evaluated birth weight, average daily gain (ADG), and disease occurrence in Angus × Holstein (Ang × Hol) crossbred and Holstein calves reared under the same conditions on a commercial dairy farm. Retrospective data from 379 calves (290 Holstein females; 89 Ang × Hol crossbreds: 46 males, 43 females) born between January 2022 and August 2023 were analyzed. Variables included dam parity, calving type, birth weight, colostrum Brix levels, serum total protein (STP), mortality, disease occurrence, ADG, and weaning weight. Statistical analysis used linear and logistic regression models. Ang × Hol male calves had higher odds of assisted calving. Male and female Ang × Hol calves had greater birth weights than Holstein calves, with males being the heaviest. No differences in STP were observed. Ang × Hol calves (both sexes) showed higher ADG than Holsteins but did not differ from each other. Holstein calves had higher odds of diarrhea (OR: 2.95, 95% CI: 1.63–5.35), while Bovine Respiratory Disease (BRD) incidence was similar across groups. Overall, Ang × Hol crossbred calves demonstrated superior growth and lower diarrhea risk under the same management conditions.

Dairy doi: 10.3390/dairy6020019

Authors: Barjam Hasanllari Benjamin P. Willing Liang Li Xian Luo Burim N. Ametaj

The dairy industry relies on the health and well-being of dairy cows for the optimal production of milk and dairy products. Mastitis, a prevalent and economically burdensome disease characterized by udder inflammation, poses significant challenges to dairy farmers worldwide. In this study, we employed peptidomics to explore the peptide profiles of dry secretions collected from dairy cows at specific intervals during the dry-off period. We hypothesized that alterations in peptide composition during the dry period may influence pathogen proliferation and immune cell functioning, thereby impacting mastitis susceptibility. Our objectives were to investigate the following: (i) differences in peptide composition and alterations between healthy cows and those with subclinical mastitis, potentially serving as biomarkers for early mastitis detection and offering insights into udder bioprocesses; (ii) variations in peptide profiles between the early (day 2) and later (day 21) stages of the dry-off period during both health conditions. Dry secretions were collected from 16 udder quarters of 8 cows at two defined time points—Day 2 (D2) and Day 21 (D21)—during the dry period. Our results revealed distinct peptide patterns between healthy and subclinical mastitis cows, as well as temporal variations in peptide profiles throughout the dry-off period. A total of 1235 peptides, originating from 59 distinct proteins (primarily β-casein), were detected across the four groups: subclinical mastitis day 21 (SCM-D21), subclinical mastitis day 2 (SCM-D2), healthy day 21 (H-D21), and healthy day 2 (H-D2). Furthermore, 56 out of the 1235 peptides identified in total matched known functional peptides, with a total of 17 different functions including antimicrobial, antioxidant, and immunomodulatory, suggesting their potential roles in mastitis pathogenesis and mammary gland physiology. Comparative analyses revealed changes in the levels of these functional peptides across the four different groups, suggesting their potential roles in regulating immune responses, oxidative stress, inflammatory processes, and other biological activities during subclinical mastitis and the dry-off period. These findings provide valuable insights into mastitis detection, management strategies, and dairy cow health monitoring, offering promising avenues for enhancing milk quality and dairy industry sustainability.

Dairy doi: 10.3390/dairy6020018

Authors: Mark Slattery Mary Garvey

The prevention of human infectious diseases associated with waterborne pathogens is reliant on the effective disinfection of water supplies by drinking water treatment plants and adequately maintained distribution networks. For decades, the chlorination of water has safeguarded public health, where chlorine is broadly applied in both water disinfection and food production facilities, including the dairy industry, from farm to fork. The identification of chlorine disinfection byproducts in water supplies and dairy food produce is of great concern, however, due to their cytotoxic, genotoxic, mutagenic, teratogenic, and potential endocrine-disrupting activity. The association between the trihalomethanes (THMs) and haloacetic acids (HAAs) and tumour formation is documented and has led to the implementation of maximum contaminant levels enforced by the European Union. Furthermore, chlorine resistance in bacterial species is associated with multidrug resistance in clinically relevant pathogens, where antibiotic- and biocidal-resistant genes are also environmental pollutants. Increasing the concentration of chlorine to surmount this resistance will ultimately lead to increasing concentrations of byproducts in both water and food products, exceeding the EU requirements. This article provides insight into chlorine DBPs as a toxicological public health risk and the relationship between chlorine resistance and antibiotic resistance in microbes relevant to dairy food production.

Dairy doi: 10.3390/dairy6020017

Authors: Mirka Thorsteinsson Samantha Joan Noel Peter Lund Martin Riis Weisbjerg Anne Louise Frydendahl Hellwing Mette Olaf Nielsen

This pilot study investigated whether reductions in enteric CH4 emissions could be obtained without affecting dry matter intake (DMI) when iodoform was mixed into total mixed rations (TMRs). The experiment consisted of four periods of 14 d with four rumen-cannulated Holstein dairy cows. In the pre-period, no iodoform was added to TMR, while either 8, 16, or 20 mg iodoform/kg DM was added to TMR in the remaining periods in a change-over design. However, the experiment was not balanced across treatments and periods due to unexpected animal responses in the second period. Dry matter intake and gas exchange were measured the last 3 d in each period using respiration chambers. Rumen grab samples were collected for microbial analyses on d 14. Dry matter intake was unaffected by the addition of iodoform to TMR at or below 20 mg/kg DMI. Methane and H2 yields (g/kg DMI) quadratically decreased (up to 46%) and increased (up to 1127%), respectively, with the increasing dose. This pilot study indicated that CH4 reductions can be obtained with an addition of up to 20 mg iodoform/kg DM in the diets of dairy cows without affecting DMI. However, high iodine concentration in iodoform limits its use in commercial herds within the EU.

Dairy doi: 10.3390/dairy6020016

Authors: Sophie A. Mahendran Richard E. Booth D. Claire Wathes Nicola Blackie

Use of pair housing for preweaning calves has shown positive benefits in terms of health and production, with it also being thought to offer social support in times of stress such as when calves are disbudded. Calf disbudding through thermal cautery is a common procedure, with recognized physiological and behavioral implications. This study’s objective was to utilize continuous monitoring accelerometers to compare the impact of disbudding on the behavior of pair and individually housed calves. Data from an all-year-round dairy herd in the southwest of England were collected via an accelerometer (ABS breeder tag, Genus, Cheshire, UK) fitted to a forelimb of a convenience subset of calves (n = 265) within two weeks of age. Data were collected between March and December 2020 and covered the period 7 days prior to disbudding until 30 days post disbudding. Calves underwent thermal cautery disbudding by farm staff using local anesthesia and a systemic nonsteroidal anti-inflammatory drug. Data from 205 calves (125 pair-housed and 80 individually housed) were included in analysis through generalized estimating equations models. The calf activity (p = 0.98), number of lying bouts (p = 0.40), and lying time per day (p = 0.75) were not associated with the housing treatment of the calves. Calf activity was associated with the day compared to disbudding (p < 0.001), with both housing treatments having decreased activity in the days following disbudding, taking approximately 14 days to return to pretreatment levels.

Dairy doi: 10.3390/dairy6020015

Authors: Raja Buatig Miriam Clegg Nicholas Michael Maria-Jose Oruna-Concha

Cow’s milk is an important part of the human diet, primarily due to its high nutritional content, particularly proteins and fats. The processing of milk enhances its safety while modifying its composition and structure, influencing bioactive peptide release, which impacts protein quality, digestion, and allergenicity. Recently, filtered milk, which undergoes pasteurisation and microfiltration, has become available in UK supermarkets, offering a longer shelf life than pasteurised milk. This study aimed to evaluate the effects of microfiltration on the protein structure of cow’s milk, compared with pasteurisation, including analysis of the peptide profile released after in vitro gastrointestinal digestion of commercially available semi-skimmed filtered and pasteurised cow’s milk. Dynamic light scattering analysis revealed that the Z-average particle size of filtered milk was significantly (p < 0.05) larger than that of pasteurised milk across all brands. Additionally, filtered milk exhibited a significantly (p < 0.05) lower free thiol concentration compared with pasteurised milk, indicating structural modifications in proteins. Confocal laser scanning microscopy (CLSM) further highlighted heterogeneities in the distribution of fat and protein, with filtered milk samples showing increased interaction between fat globules and proteins. After gastrointestinal digestion, the numbers of peptides released from filtered milk exceeded those of peptides released from pasteurised milk by an average of 5%. These results provide new insights into the potential impact of microfiltration on the microstructure of milk. Further investigations using advanced analytical techniques are needed to assess the implications of these changes on protein bioavailability and human health.

Dairy doi: 10.3390/dairy6020014

Authors: Ruohan Li Yuzhuo Wang Changhui Li Jiaxiang Huang Qingkun Zeng Ling Li Pan Yang Pengjie Wang Min Chu Jie Luo Fazheng Ren Hao Zhang

Colostrum is a nutrient-dense food rich in proteins, immune modulators, and growth factors essential for neonatal development. Its lipids serve as a key energy source and facilitate cellular functions. While yak colostrum (YC) and buffalo colostrum (BC) exhibit high nutritional value, their lipid compositions remain unclear. This study employed lipidomics to analyze and compare the lipid profiles of YC and BC with cow colostrum (CC), identifying 872 lipids across 33 subclasses. Differential analysis revealed 137, 100, and 131 lipids, with significant expression differences between YC vs. CC, BC vs. CC, and YC vs. BC, respectively. YC exhibited higher levels of α-linolenic acid and docosahexaenoic acid, suggesting a potential role in enhancing neurodevelopment and cognitive function. In contrast, the upregulation of specific lipid components in BC, such as phosphatidylethanolamine (PE) and phosphatidylinositol (PI), coupled with the downregulation of specific lysophosphatidylcholine (LPC) and lysophosphatidylethanolamine (LPE), indicated a potential benefit for lipid metabolism and inflammatory regulation. These findings suggest that YC may be particularly suitable for neonates requiring enhanced energy support, while BC may offer advantages in lipid metabolism modulation. The study provides critical insights into the distinct lipid compositions of YC and BC, laying a scientific foundation for the development of tailored nutritional supplements. These results also hold significant implications for the dairy industry, driving innovation and optimization of colostrum-based products to meet diverse nutritional demands.

Dairy doi: 10.3390/dairy6020013

Authors: Yining Meng Wen Zhu Shitong Han Hui Jiang Jie Chen Zhou Zhou Xiaoli Hao Tianle Xu Aijian Qin Zhangping Yang Shaobin Shang Yi Yang

Pseudomonas aeruginosa is a ubiquitous environmental bacterium that causes a variety of infections in humans and animals. Although antibiotic resistance in livestock has been extensively documented, continuous surveillance remains crucial for tracking emerging resistance trends and assessing control measures. During 2017 and 2018, 234 strains of P. aeruginosa were identified from 1063 strains of pathogenic and nonpathogenic bacteria isolated from raw milk of healthy and mastitis cows. In this study, 132 convenience P. aeruginosa isolates were recovered and tested for antimicrobial susceptibility and the presence of antimicrobial resistance genes and virulence factors. Antimicrobial susceptibility testing revealed that these P. aeruginosa isolates were resistant to three (gentamicin, tobramycin, and ceftazidime) out of eight antibiotics. Real-time PCR targeting 21 antibiotic resistance genes indicated that aminoglycoside modifying enzyme (AME) gene ant(3″)-I was most frequently identified in both antimicrobial-resistant and -susceptible P. aeruginosa isolates, followed by aac(6′)-II and aac(6′)-Ib. The β-lactamase encoding gene, blaPDC, was mainly identified in susceptible P. aeruginosa isolates. Virulence factors screening revealed the presence of exoS, exoT, exoU, pyo, aprA, toxA, plcH, algD, lasB, lasI, lasR, rh1L, and rh1R in resistant isolates, with the detection rates ranging from 16.7% to 88.9%. Additionally, next-generation sequencing was conducted on three resistant isolates to validate these findings. This study showed the antibiotic resistance of P. aeruginosa in raw milk samples from large-scale dairy farms in Jiangsu and Shandong provinces, China.

Dairy doi: 10.3390/dairy6020012

Authors: Mete Deniz Mutlu Hilal Kanmaz Büşra Kaya Ali Adnan Hayaloğlu

In this study, the total phenolic and anthocyanin contents, antioxidant activity, color, pH, serum separation, water holding capacity (WHC), rheology, texture and viscosity of cranberry (Cornus mas L.)-enriched yogurt were determined. The addition of cranberries (5–15%) to yogurt resulted in a proportional increase in antioxidant activity, total anthocyanin and phenolic contents. In yogurt samples to which cranberries were added, the WHC increased, while the serum separation values decreased. Due to the red color of the cranberry fruits, the L* (lightness) and b* (yellowness) values decreased, and the a* (redness) values increased (p < 0.05). The sensory evaluation showed that the 10% (w/w) cranberry-added yogurt had the highest general acceptability score when compared to the other samples. Also, it was found that the addition of 10% (w/w) cranberries to the yogurt samples contributed positively to the physicochemical (textural properties, rheological behavior, color and serum separation) and biochemical (antioxidant activity, phenolics and anthocyanins) properties of the samples. The addition of cranberries to yogurt influenced the growth of microbial populations. The number of starter bacteria (counts for Lactobacillus delbrueckii subsp. bulgaricus) in the yogurt samples with cranberries was slightly lower than in the control sample; but was at an acceptable level. E. coli and coliform bacteria were not detected in either the control yogurt sample or the samples with added cranberries. In conclusion, the addition of 10% (w/w) cranberries to yogurt can be recommended in order to achieve acceptable physical and sensory properties as well as the enrichment of yogurt with nutritional and functional aspects.

Dairy doi: 10.3390/dairy6020011

Authors: Emília Maria França Lima Maria Eduarda Marques Soutelino Adriana Cristina de Oliveira Silva Uelinton Manoel Pinto Svetoslav Dimitrov Todorov Ramon da Silva Rocha

This study aims to clarify the use of Limosilactobaillus reuteri (Lmb. reuteri) in dairy products, emphasizing its main characteristics and limitations through a comprehensive literature review. Lmb. reuteri, previously classified as Lactobacillus reuteri, is a lactic acid bacterium (LAB) generally present in the gastrointestinal tracts of humans and other animals, such as sheep, chickens, and rodents. Lmb. reuteri was reclassified as part of the genus Limosilactobacillus in April 2020, reflecting advancements in biomolecular research that identified distinct metabolic and biochemical characteristics among strains. This species is an important producer of reuterin, an antimicrobial compound facilitated through glycerol fermentation via specific enzymatic pathways. In addition, selected strains of Lmb. reuteri can be considered probiotic bacteria with numerous health benefits and that lead to well-being improvements. It is consistently related to improvements in gut health, immune function enhancement, and cholesterol reduction. Furthermore, its application in dairy products has gained prominence and is increasingly reported in the literature due to its technological and sensory benefits. Despite the challenges of its incorporation into the dairy matrix, largely due to the need to supplement these products, it has already demonstrated significant effects on several dairy products’ technological, sensory, and quality characteristics. Future research should address challenges like strain-specific efficacy and regulatory hurdles for the application of Lmb. reuteri in foods.

Dairy doi: 10.3390/dairy6020010

Authors: Caroline Barroso dos Anjos Pinto Uwe Schwarzenbolz Thomas Henle Alan Frederick Wolfschoon-Pombo Ítalo Tuler Perrone Rodrigo Stephani

Doce de leite is a caramel-like confection, mainly produced in several Latin American countries, with increasing popularity worldwide. This overview outlines nine distinct industrial technologies for the production of doce de leite: (1) total batch manufacturing process; (2) batch manufacturing system with fractionated mix addition; (3) manufacturing with pre-concentration in a vacuum evaporator and finishing in an open pan; (4) manufacturing with pre-concentration in a vacuum evaporator, finishing in an open pan, and lactose micro-crystallization; (5) continuous flow manufacturing with total concentration in a vacuum evaporator and a viscosity control holding tank (hot well); (6) manufacturing with total concentration in a vacuum evaporator and sterilization in an autoclave system; (7) manufacturing with sucrose pre-caramelization and a total batch system; (8) manufacturing in colloidal mill without an evaporation process; and (9) manufacturing based of doce de leite bars with a sucrose crystallization stage. We conducted a literature review to gather data on the discussed processes and their principal characteristics, which may be pertinent to doce de leite manufacturers. The choice of a specific process will depend on the desired doce de leite characteristics, the type of doce de leite to be produced, and the manufacturing company’s requirements. When properly integrated, these technologies contribute to efficient and profitable production, yielding high-quality products with appropriate chemical, microbiological, and sensory characteristics at an industrial scale.