AAVS_14_5_1025-1043

Introduction

Mastitis remains one of the most prevalent and economically detrimental diseases in the dairy industry, as it directly reduces milk yield, increases somatic cell count (SCC), and compromises milk quality and economic value (Salvador et al., 2025). Beyond financial losses, mastitis poses significant challenges to animal welfare, dairy sustainability, and food safety (Mylostyvyi and Izhboldina, 2025; Verdon and Beggs, 2024). Despite advances in herd management, mastitis control continues to rely predominantly on antibiotic therapy, raising long-term concerns regarding antimicrobial resistance and the presence of antibiotic residues in milk products (Abdelfattah et al., 2025; Nguyen et al., 2023). These limitations highlight the urgent need for preventive strategies grounded in physiological and metabolic resilience rather than reactive treatment.

Over the past decades, the conceptual understanding of mastitis has evolved from a predominantly bacterial infection paradigm toward a multifactorial disorder influenced by host physiology, metabolic status, and environmental stressors (Liu et al., 2025; Saco and Bassols, 2023). Among the emerging mechanisms, oxidative stress has gained increasing recognition as a key component in mastitis pathophysiology. Oxidative stress arises from an imbalance between reactive oxygen species (ROS) production and antioxidant defense capacity. High milk production, mammary inflammatory responses, and metabolic pressure during lactation collectively contribute to increased ROS generation, which can damage mammary epithelial cells, disrupt cellular integrity, and impair immune competence.

A growing body of experimental and observational studies has reported associations between elevated oxidative stress biomarkers and increased SCC, as well as a higher incidence of subclinical mastitis in high-producing dairy cattle (Agustinho et al., 2021; Cohen-Zinder et al., 2025; Gheller et al., 2023; Khan et al., 2024; Liu et al., 2025; Mazur et al., 2019; Mendoza-Martínez et al., 2024; Okkema and Grandin, 2021; Shen et al., 2025; Wang et al., 2024; Zeweld and Tarekegn, 2025). These studies collectively suggest that oxidative stress may not only occur as a consequence of inflammatory processes but may also contribute to the amplification of mammary tissue dysfunction. However, it is important to note that only a subset of the included studies directly quantified oxidative stress biomarkers, indicating that the proposed mechanistic relationships should be interpreted with caution.

Oxidative stress is closely interconnected with the nutritional and metabolic status of dairy cattle. Deficiencies in energy, minerals, and micronutrients have been associated with reduced activity of key antioxidant enzymes, including superoxide dismutase, glutathione peroxidase, and catalase. Such impairments can weaken redox homeostasis and compromise immune responsiveness, thereby increasing susceptibility to both clinical and subclinical mastitis. Accordingly, nutritional management has emerged as a strategic preventive approach aimed at maintaining antioxidant capacity and supporting udder health (Barbato et al., 2019; Capper and Cady, 2020; Chase and Fortina, 2023; Mazur et al., 2019).

Minerals and trace elements such as zinc, selenium, copper, manganese, and chromium function as essential cofactors of antioxidant enzymes and modulators of immune signaling pathways (Ianni et al., 2019a, b). Zinc plays a critical role in maintaining epithelial barrier integrity and immune cell function, whereas selenium contributes to antioxidant defense through its role in glutathione peroxidase activity. Inadequate supply of these trace elements, particularly under conditions of high metabolic demand, may lead to impaired immune function, elevated SCC, and increased susceptibility to mastitis. These interactions highlight the interconnected roles of metabolic status, oxidative balance, and immune function in shaping mastitis risk.

In addition to mineral supplementation, herbal-based feed additives and natural bioactive compounds have attracted increasing attention as non-antibiotic alternatives for mastitis control (Cohen-Zinder et al., 2025; Shen et al., 2025; Song et al., 2023; Zeweld and Tarekegn, 2025). Phytogenic feed additives contain bioactive compounds with antioxidant, anti-inflammatory, and immunomodulatory properties that may help mitigate oxidative stress and support metabolic stability (Mendoza-Martínez et al., 2024). The combined use of trace minerals and phytogenic compounds has been suggested to exert complementary effects by enhancing antioxidant capacity, improving nutrient utilization efficiency, and supporting mammary tissue resilience against infectious challenges (Agustinho et al., 2021; Khan et al., 2024). Nevertheless, the magnitude and consistency of these responses remain variable across studies.

The transition period represents a critical phase in the dairy production cycle, characterized by profound physiological and metabolic adaptations. During this stage, dairy cattle frequently experience negative energy balance, increased lipid mobilization, and elevated oxidative stress, all of which can impair immune function and increase susceptibility to inflammatory and infectious diseases, including mastitis (Gheller et al., 2023; Singh, 2020; Singh et al., 2023). These dynamics underscore the importance of targeted nutritional strategies during the transition period to maintain metabolic stability and reduce disease risk.

Mastitis development in dairy cattle is increasingly understood as the result of complex interactions among metabolic status, oxidative balance, immune responses, and pathogen pressure. To illustrate these relationships, a conceptual framework is presented in Figure 1. This framework summarizes interactions reported in the literature and serves as a conceptual guide for interpreting the relationships discussed in this review. It is important to emphasize that this framework represents a conceptual synthesis rather than a validated causal model derived directly from the present analysis.

Therefore, this review aims to synthesize current scientific evidence on the role of nutritional strategies in modulating oxidative stress and reducing mastitis risk in dairy cattle. Although numerous studies have explored nutritional approaches to mastitis prevention, the available evidence remains fragmented and methodologically heterogeneous, with substantial variation in study design, biomarkers, and intervention strategies. In contrast to previous reviews, this study provides a systematic synthesis that integrates metabolic status, oxidative balance, and immune responses within a unified conceptual framework, while explicitly accounting for heterogeneity and studies reporting both positive and neutral outcomes. This approach is intended to provide a more balanced and mechanistically informed understanding of how nutritional strategies may influence mastitis susceptibility in dairy cattle.

Materials and Methods

Study design and reporting guidelines

This study was conducted as a Systematic Literature Review (SLR). The review protocol was prospectively registered on the Open Science Framework (OSF) (DOI: 10.17605/OSF.IO/W5RCH) to ensure transparency and methodological rigor. All methodological steps, including the search strategy, eligibility criteria, and data extraction procedures, were predefined prior to study initiation to minimize bias and enhance reproducibility. The review was designed and reported in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) 2020 guidelines.

Research question and scope of the review

The primary research question guiding this review was: “What feeding and nutritional strategies have been reported to reduce mastitis risk or improve udder health in dairy cattle?”

The review focused on nutritional interventions that directly or indirectly influenced mastitis-related outcomes, particularly through mechanisms involving immune modulation, reduction of oxidative stress, decreased somatic cell count (SCC), and improvements in milk quality and composition.

The primary outcomes of interest were mastitis incidence (clinical and subclinical) and SCC. Secondary outcomes included oxidative stress biomarkers, immune response parameters, antioxidant enzyme activity, and indicators of udder health. The nutritional strategies examined included mineral and trace mineral supplementation, antioxidant nutrients, phytogenic or herbal-based feed additives, and nutritional management during lactation and the transition period. This review focuses exclusively on preventive nutritional strategies and does not include studies evaluating nutritional treatments for active clinical mastitis. Studies were included if they reported outcomes related to mastitis risk, subclinical mastitis (e.g., somatic cell count), or udder health indicators.

Literature search strategy

A systematic literature search was conducted in four electronic databases: Scopus, Web of Science, PubMed, and ScienceDirect. Articles published between 2015 and 2025 were considered to capture recent developments in nutritional strategies and emerging insights into oxidative stress and immune modulation in dairy cattle. Only peer-reviewed articles published in English were included.

The search strategy employed combinations of keywords structured using Boolean operators. A representative search string used in Scopus was as follows: (“mastitis” AND (“dairy cattle” OR “dairy cows”)) AND (“nutrition” OR “feeding strategies” OR “mineral supplementation” OR “trace elements” OR “zinc” OR “selenium” OR “antioxidant” OR “oxidative stress” OR “phytogenic feed additives” OR “herbal supplementation”). The search syntax was adapted for each database according to its indexing system, controlled vocabulary (e.g., MeSH terms in PubMed), and search interface. The full search strings for each database are provided in the Supplementary Table 1 to ensure transparency and reproducibility. In addition, backward reference searching was performed by screening the reference lists of eligible articles to identify potentially relevant studies not captured in the initial database search

Inclusion and exclusion criteria

Eligibility criteria were predefined prior to the literature search.

The inclusion criteria were as follows:

Original research articles.
Studies conducted in dairy cattle.
Investigations evaluating feeding or nutritional interventions.
Studies reporting mastitis-related parameters, including clinical or subclinical mastitis incidence, somatic cell count (SCC), udder health indicators, immune responses, or oxidative stress biomarkers.
Availability of full-text articles.

The exclusion criteria included:

Review articles, conference proceedings, editorials, or short communications.
Studies conducted in non-dairy livestock species.
Research unrelated to nutritional interventions.
Articles not reporting outcomes relevant to mastitis or udder health.

Although full-text availability was defined as an inclusion criterion, 21 potentially eligible studies could not be retrieved despite repeated access attempts and were therefore excluded at the full-text screening stage. This limitation is acknowledged as part of the study selection process.

Study selection process

All retrieved records were imported into reference management software, and duplicate entries were identified and removed prior to screening. After duplicate removal, studies were screened in three sequential stages: title screening, abstract screening, and full-text assessment.

The screening process was conducted independently by two reviewers to minimize selection bias. To ensure consistency, a subset of records was cross-checked by both reviewers at each screening stage, and any discrepancies were discussed and resolved through consensus. This iterative process was applied throughout the screening procedure to maintain alignment in eligibility decisions and to enhance the reliability of study selection.

The study selection process followed PRISMA 2020 guidelines and was documented using a PRISMA flow diagram, illustrating the number of records identified, screened, excluded, and ultimately included in the final synthesis, along with reasons for exclusion at the full-text stage.

Quality assessment and risk of bias

The methodological quality and risk of bias of the included studies were evaluated using the SYRCLE risk-of-bias tool developed for animal intervention studies. This tool assesses potential bias across several domains, including selection bias, performance bias, detection bias, attrition bias, reporting bias, and other potential sources of bias.

Each study was independently assessed by two reviewers, and any discrepancies were resolved through discussion until consensus was achieved. In accordance with SYRCLE guidelines, each domain was judged as “low risk,” “high risk,” or “unclear risk” of bias.

Importantly, studies that did not explicitly report key methodological details such as randomization procedures, allocation concealment, or blinding were not assumed to be at low risk but were instead classified as having an “unclear” or “high” risk of bias, depending on the extent of missing information. This approach was applied to avoid overestimation of study quality.

Overall study quality was then categorized as low, moderate, or high risk of bias based on the combined assessment across domains. Several domains were frequently rated as “unclear” due to insufficient reporting, particularly in relation to randomization and blinding procedures, which are commonly underreported in animal nutrition studies.

The results of the risk-of-bias assessment were considered during data interpretation and are presented in Supplementary Table 2 to ensure transparency and reproducibility.

Data extraction

Data extraction was conducted using a structured data extraction form to ensure consistency and completeness. Extracted information included author names and publication year, country of study, study design, animal characteristics, type of nutritional intervention, dosage and duration of supplementation, evaluated parameters, and primary outcomes related to mastitis prevention and udder health. The extracted information was summarized in a study characteristics table to facilitate comparison across the included studies.

Data synthesis

Due to substantial heterogeneity in study designs, types of nutritional interventions, dosages, duration of supplementation, and outcome measures, a quantitative meta-analysis was not considered appropriate. The included studies varied considerably in terms of experimental conditions, animal characteristics, and reported endpoints, which limited the feasibility of statistical pooling and effect size comparison.

Therefore, the data were synthesized using a structured narrative synthesis approach. Studies were grouped according to the type of nutritional strategy, including mineral and trace mineral supplementation, antioxidant nutrients, phytogenic feed additives, and nutritional management during the transition period.

Within each category, findings were systematically compared based on reported outcomes, including somatic cell count (SCC), mastitis incidence, oxidative stress biomarkers, immune response indicators, and milk production parameters. Both positive and non-significant results were considered to provide a balanced interpretation of the evidence.

The synthesis aimed to identify recurring patterns, potential biological mechanisms, and inconsistencies across studies, while also highlighting gaps in the current literature. Particular attention was given to the relationships among metabolic status, oxidative balance, and immune function in the context of mastitis susceptibility in dairy cattle.

RESULTS

Descriptive summaries of included studies

Study selection based on PRISMA

The study selection process followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) 2020 guidelines, and the complete workflow is presented in the PRISMA flow diagram (Figure 2).

The initial literature search conducted across four electronic databases (Scopus, Web of Science, PubMed, and Science Direct) using predefined keyword combinations identified a total of 811 records. After removing 104 duplicate records, 707 articles remained for title and abstract screening.

During the screening stage, 637 records were excluded because they did not meet the predefined inclusion criteria. The primary reasons for exclusion included lack of relevance to nutritional interventions (n= 318), studies conducted in non-dairy species (n= 152), absence of mastitis-related outcome parameters (n= 109), and non-original study types such as reviews, conference proceedings, or editorials (n= 58).

Following the screening process, 70 reports were sought for retrieval for full-text evaluation. Of these, 21 reports could not be retrieved due to restricted access, unavailable full texts, or incomplete database records, and were therefore excluded prior to eligibility assessment.

The remaining 49 reports were successfully retrieved, assessed for eligibility, and included in the final qualitative synthesis.

Overall, the included studies represent approximately 6.04% of the 811 records initially identified. The complete study selection process, including identification, screening, eligibility, and inclusion stages, is presented in Figure 2.

Although full-text availability was defined as an inclusion criterion, 21 potentially eligible studies could not be retrieved despite repeated access attempts. This limitation is acknowledged and may have influenced the completeness of the evidence synthesis and the overall interpretation of findings.

The characteristics of the included studies are summarized in Supplementary Table 1, which provides detailed information regarding study design, animal species, nutritional interventions, and evaluated outcomes.

The methodological quality and risk of bias of the included studies were evaluated using the SYRCLE risk-of-bias tool for animal intervention studies. The detailed results of the risk-of-bias assessment are presented in Supplementary Table 2. Overall, studies were classified as having low to moderate risk of bias, with several domains frequently rated as “unclear” due to limited reporting of methodological details such as randomization and blinding procedures.

Mineral and trace elements in mastitis prevention

The synthesized evidence indicates that mineral and trace element supplementation represents one of the most frequently investigated nutritional strategies associated with udder health and mastitis-related indicators in dairy cattle. Across the included studies, the majority reported that improved mineral status was associated with reductions in somatic cell count (SCC), enhanced metabolic stability, and improved immune-related parameters (De et al., 2023; Ianni et al., 2019b; Rodrigues et al., 2024). However, considering the heterogeneity in study design, supplementation protocols, and baseline nutritional conditions, the strength and consistency of these associations should be interpreted cautiously.

Among the trace elements evaluated, selenium has received the most consistent attention due to its central role in antioxidant defense through glutathione peroxidase (GSH-Px) activity. Most studies reported associations between improved selenium status and reductions in SCC or inflammatory indicators (Ianni et al., 2019b; Wang et al., 2021). Reductions in SCC reported across studies ranged from approximately 10% to over 60%, depending on supplementation dose, duration, and baseline mineral status. However, several studies found no statistically significant effects, highlighting variability in outcomes.

The variability in selenium responses suggests that supplementation effects are highly context-dependent. Studies reporting limited or no improvement were often associated with adequate baseline mineral status or shorter experimental durations, whereas stronger responses were observed under conditions of deficiency or increased metabolic stress (Ianni et al., 2019b; Wang et al., 2021). In addition, some studies reported negative correlations between serum selenium concentrations and SCC, together with increased glutathione peroxidase activity and reduced inflammatory mediators such as interleukin-6 following supplementation (Ding et al., 2025; Wang et al., 2021). However, only a limited number of studies directly assessed oxidative processes at the mammary tissue level, and therefore these mechanistic pathways should be interpreted as biologically plausible rather than definitive causal relationships.

The high prevalence of selenium deficiency reported in both cows and calves, together with low selenium concentrations in forage in certain production systems, suggests that mineral imbalances may be influenced by agroecological conditions (Andělová et al., 2024; Stybayev et al., 2025). Under such conditions, selenium supplementation has been associated not only with improvements in udder health indicators but also with broader reproductive and metabolic outcomes, including improved natality rates in some herds (Lembovičiūtė et al., 2025). Nevertheless, responses remain dependent on baseline nutritional status and management conditions (Żarczyńska et al., 2017).

In addition to selenium, other trace minerals such as zinc and copper, together with antioxidant vitamins such as vitamin E, have also been investigated for their roles in maintaining mammary epithelial integrity and supporting immune function (Agustinho et al., 2021; Ianni et al., 2019a; Kegley et al., 2016; Khan et al., 2024). Zinc contributes to epithelial barrier function and leukocyte activity, whereas copper participates in enzymatic antioxidant systems. Several studies reported reductions in SCC following combined mineral and vitamin supplementation, although responses were not uniform across studies (Germon and Martins, 2023; Goyal et al., 2024; Nudda et al., 2023). indicating potential interactions between mineral balance, diet composition, and physiological stage.

The association between negative energy balance (NEB), metabolic stress, and oxidative stress further supports the interpretation of subclinical mastitis as a systemic metabolic–immunological disorder rather than a purely localized infection (Agustinho et al., 2021; Khan et al., 2024). High-concentrate diets during early lactation have been reported to increase metabolic load and oxidative stress, potentially influencing immune- and antioxidant-related gene expression (Ayemele et al., 2021). These metabolic conditions may increase susceptibility to inflammatory responses or pathogen colonization, suggesting that nutritional strategies targeting metabolic stability and antioxidant balance may indirectly support udder health.

Recent studies also indicate that nutritional interventions may influence mammary health at the molecular level. For example, inulin supplementation has been reported to modify the profile of milk small extracellular vesicle-associated microRNAs (sEV-miRNAs) involved in inflammatory signaling pathways and milk synthesis regulation (Yu et al., 2023). Additionally, microbial-based feed additives have been associated with improvements in feed intake and metabolic balance, which may indirectly contribute to reductions in SCC (Kalińska et al., 2023; Prapaiwong et al., 2023; Rodrigues et al., 2024). However, most molecular evidence originates from controlled experimental conditions rather than large-scale field studies, and therefore requires further validation.

From an immunological perspective, mammary defense mechanisms involve complex interactions between innate and adaptive immune responses. Early responses to infection include cytokine signaling, bioactive lipid mediators, and complement system activation (Kegley et al., 2016). Within this context, essential trace minerals may play dual roles by supporting host immune function while also serving as resources for certain bacterial pathogens, including Staphylococcus aureus, which possesses specialized metal acquisition systems (Carlson et al., 2020; Magro et al., 2023). The dynamics of nutritional immunity in ruminants remain incompletely understood, and extrapolation from non-ruminant models should be approached cautiously.

Overall, the available evidence suggests that optimization of mineral balance, energy status, and antioxidant supply may contribute to improved udder health and reduced mastitis risk in dairy cattle. However, the magnitude and consistency of these effects remain context-dependent and influenced by factors such as baseline nutritional status, lactation stage, dietary formulation, and herd management conditions. Therefore, mineral-based nutritional interventions should be considered as components of integrated mastitis prevention strategies rather than as standalone solutions.

Herbal-based feed additives and natural antioxidants

Herbal and natural antioxidant-based feed additives have received increasing attention as potential non-antibiotic alternatives for mastitis control in dairy production systems. The reviewed studies generally report associations between phytogenic feed additives and improvements in udder health parameters, particularly reductions in somatic cell count (SCC) and improvements in antioxidant status. However, the magnitude and consistency of these responses vary across studies due to differences in additive composition, dosage, experimental design, and physiological status of the animals.

An integrated conceptual model illustrating potential pathways through which herbal and natural antioxidant feed additives may influence oxidative balance, immune responses, and mammary gland health is presented in Figure 3. This model synthesizes relationships reported in the literature and should be interpreted as a conceptual framework rather than an experimentally validated causal model.

As illustrated in Figure 3, the proposed relationships highlight recurring patterns identified across studies and provide a structured interpretation of how phytogenic interventions may influence mastitis-related parameters.

The use of phytogenic additives has been explored as a preventive nutritional strategy, particularly in response to concerns regarding antimicrobial resistance, production sustainability, and milk functional quality (Barcelo-Villalobos et al., 2025; Cunha et al., 2023; Fantini et al., 2024; Kholif et al., 2021; Yu et al., 2023). Across studies, these additives were more consistently associated with improvements in udder health indicators and antioxidant status than with increases in milk yield (Fraga et al., 2024; Prapaiwong et al., 2023; Prodanović et al., 2025). Suggesting a primary role in supporting physiological stability rather than directly enhancing production performance.

From a biological perspective, modulation of redox homeostasis is frequently proposed as a central mechanism underlying these effects (Mazur et al., 2019). Several studies reported reductions in oxidative stress markers, including reactive oxygen species (ROS), lipid peroxidation products, and antioxidant indices, following supplementation (Ayemele et al., 2021; Mazur et al., 2019). These responses were often accompanied by reductions in SCC; however, the magnitude of these effects ranged from modest to moderate and varied substantially across studies.

Importantly, only a limited number of studies directly assessed oxidative processes within mammary tissue. Therefore, the proposed mechanistic links between antioxidant supplementation and mastitis-related outcomes should be interpreted as biologically plausible rather than definitive.

Responses to phytogenic supplementation were not uniform. Several studies reported limited or non-significant changes in oxidative biomarkers or SCC, particularly under conditions of low baseline oxidative stress or modest supplementation levels (Cunha et al., 2023; Kotsampasi et al., 2018; Nudda et al., 2023). This variability indicates that the effectiveness of phytogenic additives is highly context-dependent and influenced by baseline physiological status, additive composition, and management conditions.

Furthermore, SCC alone may not fully reflect mastitis status, as it can be influenced by non-infectious factors such as metabolic stress, stage of lactation, and physiological adaptation. Therefore, interpretation of SCC responses should be considered alongside other indicators of udder health and immune function.

Plant-derived bioactive compounds, including polyphenols, flavonoids, saponins, and terpenoids, have been associated with immunomodulatory effects (Nudda et al., 2023; Stobiecka et al., 2023). Some studies reported reductions in circulating oxidative markers and changes in immune-related parameters, such as neutrophil counts and globulin concentrations (Cunha et al., 2023). However, these effects were not consistently observed across all studies and appear to depend on the specific phytochemical profile of the additives.

Interactions between phytogenic additives and rumen metabolism may also contribute to observed responses. Improvements in nutrient digestibility and volatile fatty acid (VFA) production have been reported in some studies (Prapaiwong et al., 2023). Suggesting modulation of rumen microbial activity. These changes may indirectly influence metabolic stability and mammary gland resilience.

Alterations in milk fatty acid profiles, including increased proportions of polyunsaturated fatty acids (PUFA), omega-3, and monounsaturated fatty acids (MUFA), have also been reported following phytogenic supplementation (Dokou et al., 2023; Renna et al., 2020). However, some studies identified trade-offs between milk composition and production performance, indicating dose-dependent and diet-dependent responses (Niu et al., 2023).

Emerging evidence suggests that nutritional interventions may influence mammary physiology through molecular regulatory mechanisms, including extracellular vesicles and microRNAs. For example, modifications in milk small extracellular vesicle-derived microRNAs (sEV-miRNAs) have been reported following dietary supplementation (Yu et al., 2023). However, these findings are based on a limited number of experimental studies and require further validation under field conditions.

Within the context of sustainable dairy production, phytogenic additives have been proposed as complementary strategies to reduce reliance on antibiotics. Some studies also reported improvements in nitrogen utilization efficiency and reductions in milk urea concentrations (Prodanović et al., 2025), which may contribute to environmental sustainability (Han et al., 2024). Nevertheless, the overall strength of evidence remains constrained by heterogeneous study designs and limited long-term validation.

Overall, the available evidence suggests that herbal and natural antioxidant feed additives may contribute to improved udder health through multiple interacting pathways involving oxidative balance, immune modulation, and metabolic regulation. However, given the variability across studies, these additives should be considered as complementary components within integrated herd management strategies rather than standalone interventions.

Nutritional management factors affecting mastitis

The included studies indicate that nutritional management is closely associated with mastitis-related outcomes through its influence on metabolic status, immune function, and oxidative balance in dairy cattle (Haq et al., 2024; Kelly et al., 2024; Rial et al., 2023). However, the magnitude and consistency of these effects varied across studies depending on dietary composition, physiological stage, and herd management conditions.

The summary visual matrix (Figure 4) was constructed based on qualitative extraction and synthesis of reported outcomes from the included studies. Each study was evaluated for its reported effects on metabolic balance, oxidative status, and immune responses, and the matrix summarizes recurring patterns observed across studies. The figure does not represent a quantitative scoring system but rather a structured synthesis of evidence derived from the systematic review process.

As illustrated in Figure 4, the matrix provides a simplified overview of complex interactions across studies, facilitating comparison of different nutritional strategies within a unified framework. Energy balance during early lactation was frequently identified as an important factor associated with mastitis susceptibility, although responses varied depending on management and physiological conditions.

Energy balance during early lactation was consistently identified as a key factor influencing mastitis susceptibility. Several studies reported that negative energy balance (NEB) was associated with impaired immune function and increased susceptibility to inflammatory conditions (Cheng et al., 2022; Goyal et al., 2024). Nutritional strategies aimed at improving energy intake, including appropriate concentrate supplementation and the use of buffering agents, were associated with improvements in rumen fermentation, increased volatile fatty acid (VFA) production, and enhanced nutrient digestibility, which were in some cases accompanied by reductions in somatic cell count (SCC).

However, responses to high-concentrate diets were not uniform across studies. While some studies reported improved production performance and reduced SCC, others indicated increased metabolic stress and oxidative challenges, particularly in animals with higher physiological demands. This variability suggests that dietary strategies must be adapted to animal-specific and production-specific conditions.

Feed additives have also been evaluated as part of nutritional management strategies. Several studies reported associations between phytogenic or microbial-based additives and improvements in rumen fermentation, nutrient utilization, and SCC (Prapaiwong et al., 2023; Rodrigues et al., 2024). For example, supplementation with citrus by-product hydrolysates was associated with improved digestibility and ruminal fermentation parameters, accompanied by reductions in SCC (Prapaiwong et al., 2023). These findings suggest that nutritional interventions may influence udder health indirectly through metabolic pathways rather than direct antimicrobial effects.

Oxidative stress was frequently reported as a contributing factor linking metabolic imbalance to mastitis susceptibility (Ayemele et al., 2021; Cohen-Zinder et al., 2025). Nutritional strategies aimed at improving antioxidant status, including supplementation with vitamins and trace minerals, were associated with improvements in immune-related parameters and reductions in oxidative stress indicators in several studies. However, the extent of these effects varied across studies, and direct measurements of oxidative processes at the mammary level were limited.

Mineral balance was also identified as an important component of nutritional management. Some studies reported associations between altered mineral status and mastitis-related outcomes, including changes in calcium and phosphorus levels, which may influence physiological functions such as teat sphincter integrity and immune responsiveness (Okkema and Grandin, 2021). However, the evidence remains variable and dependent on baseline nutritional conditions.

Feed contamination with mycotoxins represents an additional nutritional factor that may influence mastitis risk. Studies have reported associations between mycotoxin exposure and impaired immune function, altered nitrogen metabolism, and increased susceptibility to inflammatory responses (Mukhamadieva et al., 2023). These findings suggest that feed quality management is an important component of mastitis prevention strategies.

Some studies also highlighted the role of integrated herd management approaches, including technological interventions such as Automatic Milking Systems (AMS), which have been associated with improvements in udder health indicators and reductions in SCC under certain conditions (Lage et al., 2024). However, these effects are influenced by management practices and monitoring strategies.

Overall, the evidence suggests that nutritional management influences mastitis risk through multiple interacting pathways involving metabolic balance, oxidative status, and immune function. However, the variability across studies indicates that these effects are context-dependent and should be interpreted within the framework of integrated herd management systems rather than as isolated nutritional interventions.

Discussion

The findings of this systematic review indicate that nutritional strategies are associated with mastitis-related outcomes in dairy cattle, although the magnitude and consistency of these effects vary across studies. Rather than acting through isolated mechanisms, the evidence supports the interpretation that nutritional interventions influence mastitis susceptibility through interconnected pathways involving metabolic status, oxidative balance, and immune function (Ayemele et al., 2021; Mazur et al., 2019).

Across the 49 included studies, a substantial proportion reported associations between nutritional interventions particularly trace mineral supplementation, antioxidant nutrients, phytogenic additives, and transition-period management and improvements in somatic cell count (SCC), oxidative status, and immune-related parameters. Reported reductions in SCC ranged from modest (approximately 5–15%) to more substantial (>50%) in certain controlled studies (Ianni et al., 2019a; Wang et al., 2021), while other studies reported limited or non-significant effects, highlighting the context-dependent nature of these interventions.

While previous reviews have discussed the roles of nutrition and oxidative stress in mastitis, the present study contributes by integrating recent evidence (2015–2025) into a unified framework linking metabolic status, oxidative balance, and immune function. This integrative perspective helps explain inconsistencies across studies and emphasizes that nutritional interventions should be interpreted as modulatory factors within a complex physiological system rather than direct causal solutions for mastitis prevention.

Rather than relying on single-study findings, the present synthesis indicates that trace mineral supplementation particularly selenium has been associated across multiple studies with improvements in antioxidant enzyme activity and reductions in SCC, although these responses vary depending on baseline mineral status and supplementation protocols. Similarly, phytogenic feed additives were associated in several studies with improvements in oxidative and immune-related parameters; however, the magnitude of these effects was inconsistent and influenced by additive composition, dosage, and management conditions (Cunha et al., 2023; Prapaiwong et al., 2023).

Some of the biological mechanisms proposed in the literature suggest that these responses may be mediated through improved rumen fermentation, enhanced nutrient utilization, and modulation of systemic oxidative balance. However, it is important to distinguish that these mechanisms are inferred from experimental and physiological studies and were not consistently measured directly across all included studies. Therefore, these interpretations should be considered as biologically plausible explanations rather than definitive causal pathways.

Substantial heterogeneity was observed among the included studies in terms of experimental design, animal characteristics, supplementation strategies, dosage levels, and outcome measurements. A formal subgroup analysis or meta-analysis was not feasible due to this heterogeneity and the lack of standardized reporting across studies. Key sources of variability included differences in baseline nutritional status, lactation stage, environmental conditions, and types of biomarkers used to assess mastitis and oxidative stress.

Publication bias cannot be entirely excluded, as studies reporting positive outcomes may be more likely to be published. No formal assessment of publication bias (e.g., funnel plot analysis) was conducted, as the heterogeneity of study designs and outcomes precluded quantitative synthesis. This limitation may result in an overestimation of the beneficial effects of nutritional interventions.

From a practical perspective, the economic implications of nutritional interventions remain insufficiently explored. Although several strategies have been associated with improvements in SCC and udder health indicators, limited data are available regarding cost-effectiveness under commercial conditions. As a result, while certain interventions such as chelated mineral supplementation or phytogenic additives appear promising, their economic viability under field conditions remains uncertain.

In addition, although improved nutritional management may contribute to enhanced animal health and resilience, direct evidence linking these strategies to reduced antibiotic usage is limited. Most studies evaluated surrogate indicators such as SCC rather than treatment frequency or incidence of clinical mastitis. Therefore, conclusions regarding antimicrobial reduction should be interpreted cautiously.

Future research should prioritize standardized experimental designs and reporting frameworks. Key biomarkers that should be consistently measured include somatic cell count (SCC), oxidative stress indicators such as malondialdehyde (MDA) and glutathione peroxidase (GSH-Px), and immune parameters including cytokines such as interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α). In addition, studies should incorporate both biological and economic outcome measures to improve the applicability of findings for commercial dairy systems.

While emerging approaches such as metabolomics and transcriptomics may provide deeper mechanistic insights, their application in commercial dairy systems remains limited. Therefore, future research should balance mechanistic exploration with practical feasibility, ensuring that proposed strategies are both scientifically robust and economically viable.

Overall, the findings of this review support the role of nutritional management as a complementary component of mastitis prevention strategies. Rather than functioning as standalone interventions, nutritional strategies should be integrated with broader herd management practices, including hygiene, genetics, and environmental control. Within this integrated framework, optimizing metabolic balance, antioxidant capacity, and immune competence may contribute to improved mammary gland resilience and more sustainable dairy production systems.

Conclusion

Mastitis in dairy cattle is a multifactorial disorder associated with interactions among metabolic imbalance, oxidative stress, and immune dysregulation, particularly during the periparturient transition period. The evidence synthesized in this review supports an integrated interpretation of mastitis involving interactions within the metabolic–oxidative–immune axis; however, these relationships should be considered as associations supported by current evidence rather than fully validated causal mechanisms.

Nutritional strategies, including optimization of energy balance, trace mineral and vitamin supplementation, phytogenic antioxidants, and microbial modulators, have been associated across multiple studies with improvements in metabolic stability, redox balance, and immune-related parameters, as well as reductions in somatic cell count (SCC) under specific conditions. Nevertheless, the magnitude and consistency of these effects varied considerably depending on physiological stage, baseline nutritional status, and management conditions, indicating that responses to nutritional interventions are context-dependent.

Overall, the findings indicate that nutrition represents an important complementary component of mastitis risk management rather than a standalone solution. Effective mastitis control requires integration of nutritional strategies with broader herd health practices, including hygiene, genetics, and environmental management.

Based on the gaps identified in this review, future research should prioritize: (i) longitudinal studies evaluating both subclinical and clinical mastitis outcomes; (ii) standardized measurement of key biomarkers, including SCC, oxidative stress indicators (e.g., malondialdehyde and glutathione peroxidase activity), and immune parameters (e.g., cytokine profiles); and (iii) evaluation of economic feasibility and cost-effectiveness of nutritional interventions under commercial production conditions.

In addition, although nutritional strategies may support improved animal health and resilience, direct evidence linking these approaches to reductions in antibiotic usage remains limited. Therefore, conclusions regarding antimicrobial reduction and sustainability should be interpreted cautiously and require further validation.

Overall, this review highlights the importance of adopting an integrated and context-specific approach to nutritional management, with the aim of improving mammary gland health and supporting more sustainable dairy production systems.

Acknowledgments

The authors thank Politeknik Negeri Tanah Laut (Politala) for supporting the coordination and facilitation of this research.

Novelty Statement

This review synthesizes current evidence on multifunctional nutritional interventions for mastitis prevention by focusing on the interaction between metabolic disorders, oxidative stress, and immune regulation to improve udder health and reduce mastitis risk in dairy cattle.

Author’s Contribution

I.P. conceptualized the review, conducted the litera-ture search, synthesized the data, and prepared the original draft of the manuscript. All authors reviewed, revised, and approved the final version of the manuscript.

Generative AI and AI assisted technology statement

The authors declare that no generative AI and AI assisted technology was used in the creation of this manuscript.

Conflict of interest

The authors have declared no conflict of interest.

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