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Open research questions in Reproductive Physiology in Livestock

37 unresolved questions extracted from the limitations and future-work sections of 535 Reproductive Physiology in Livestock papers in our library. Each links back to the study that raised it.

What the literature leaves open

  • Future investigations should include larger and more diverse dairy populations, additional longitudinal 3291 doi: 10.14202/vetworld.2026.3282-3294 sampling points, controlled experimental designs, and independent validation cohorts. Multivariable analyses are needed to determine whether Hp independently predicts reproductive outcomes after adjustment for parity, milk yield, body condition score, nutritional status, and environmental variables. Future studies should also establish clinically relevant Hp cutoff values, evaluate rapid field-based Hp assays, and develop predictive models integrating Hp with NEFAs, BHB, PGFM, cortisol, cytokines, oxidative stress markers, and uterine microbiome profiles. Economic analyses are additionally warranted to determine whether Hp-guided reproductive monitoring can improve herd fertility and profitability under commercial dairy production conditions.

    Longitudinal integrated evaluation of metabolic, inflammatory, biochemical, and hormonal biomarkers identifies haptoglobin as a key indicator of reproductive performance in Friesian-Holstein cows with endometritis under tropical Indonesian dairy conditionsia · 2026 · DOI

  • Introduction Heat stress (HS) impairs fertility in dairy cattle, yet the molecular mechanisms underlying uterine dysfunction remain poorly understood.

    Heat stress modulates extracellular vesicles miRNA cargo in bovine uterine fluid and uterine function · 2026 · DOI

  • Given the substantial effects of stress on productivity and animal welfare, there is increasing interest in developing more effective approaches for monitoring, predicting, and mitigating stress responses in cattle. This has highlighted the importance of future research directions and continued technological advancement in this field. Improving current understanding of stress physiology in cattle will likely require integration of emerging technologies, interdisciplinary research approaches, and more standardized evaluation systems. Recent technological developments have made it possible to monitor stress-associated physiological and behavioral changes with greater precision and in real time. Wearable sensors, biometric monitoring devices, and remote sensing systems can continuously measure parameters such as body temperature, heart rate variability, activity patterns, and feeding behavior [60,61]. When combined with advanced analytical approaches, including machine learning algorithms, these data streams may help 13 ACCEPTED 293 294 295 296 297 298 299 300 301 302 303 304 305 306 307 308 309 310 311 312 identify stress triggers, predict stress responses, and support early intervention strategies within precision livestock farming systems. Advances in molecular biology and omics technologies are also contributing to improved understanding of stress resilience at the genetic and cellular levels. Genomic and transcriptomic analyses have facilitated identification of genetic markers associated with thermotolerance and adaptation to stress, supporting breeding strategies aimed at improving resilience in cattle populations [42,62]. In particular, studies involving heat shock proteins and genes related to HPA axis regulation have provided valuable insight into the biological mechanisms underlying variation in stress responsiveness among individuals. Despite these advances, important knowledge gaps remain in cattle stress research. The long-term consequences of chronic stress on animal health, productivity, and welfare are still not fully understood, especially under varying environmental and management conditions. In addition, differences in methodology and the lack of standardized biomarkers limit comparisons among studies and reduce the practical application of current findings. Although biomarkers such as cortisol concentrations in hair, blood, or saliva and heat shock protein expression are widely used, additional validation is needed to improve their reliability and sensitivity under different production conditions [6,39,40]. Another important area for future research involves integrating environmental, physiological, and behavioral information into comprehensive models of stress responses. Multidisciplinary approaches that combine animal physiology, immunology, genetics, and data science may help clarify the complex regulatory interactions underlying stress adaptation. Such approaches could also contribute to the development of predictive models capable of supporting management decisions and improving animal 313 welfare outcomes. 314 315 316 317 318 319 From a practical standpoint, translating scientific findings into effective management strategies and policy frameworks remains challenging. Greater collaboration among researchers, veterinarians, industry stakeholders, and policymakers will be necessary to improve communication between research and field application. Development of evidence-based guidelines tailored to specific production systems and environmental conditions may further improve implementation of stress mitigation practices in cattle farming. In addition, investment in infrastructure, training of livestock handlers, and adoption of low- 14 ACCEPTED 320 stress management practices are likely to play important roles in improving both welfare and production 321 efficiency. 322 323 324 325 326 327 328 329 330 331 332 333 334 335 336 337 338 339 340 341 342 343 344 345 Overall, future cattle stress research should continue integrating technological innovation with biological understanding while addressing existing knowledge gaps and improving practical application of research findings. Such efforts will be important for supporting sustainable livestock production systems that prioritize both animal welfare and long-term productivity under increasingly challenging environmental conditions.

    Unraveling the complexity of stress responses in cattle with a focus on immune function · 2026 · DOI

  • Progesterone (P4) exerts important vascular and immunomodulatory effects that influence platelet (PLT) activation and serotonin (5-HT) handling across mammalian species; nevertheless, its role in modulating PLT physiology during diestrus in mares remains poorly defined.

    Progesterone-Dependent Changes in Platelet Activation Without Morphological Variation in Diestrus Mares · 2026 · DOI

  • The referenced work by Tsuchiya et al. (reference 19) demonstrates that liver transcriptome response to periparturient changes depends on postpartum subacute ruminal acidosis occurrence, but the current study does not explicitly examine whether dietary interventions (rumen-protected nutrients as mentioned in references 16-18) or specific nutritional states modulate the identified transcriptional signatures during the periparturient transition.

    Liver transcriptome dynamics in Holstein cows during the periparturient transition · 2026 · DOI

  • While enriched GO biological process, molecular function, and cellular component terms are listed in Tables S4-S6 for up-regulated and down-regulated genes plus the red module, the paper does not specify integration of these transcriptomic findings with proteomics, metabolomics, or hepatic enzyme activity measurements to establish mechanistic links between transcriptional changes and actual metabolic phenotypes during the periparturient transition.

    Liver transcriptome dynamics in Holstein cows during the periparturient transition · 2026 · DOI

  • The study used only 11 samples (as shown in Fig. S1 PCA plot) for downstream transcriptomic analysis of periparturient liver dynamics; validation of the identified differentially expressed genes and enriched GO terms across independent cohorts with larger sample sizes and diverse Holstein populations at different metabolic stress levels remains unaddressed.

    Liver transcriptome dynamics in Holstein cows during the periparturient transition · 2026 · DOI

  • Table S7 indicates comparison of major periparturient liver transcriptome RNA-seq studies exists, but the excerpt does not specify which methodological differences (sampling timepoints, cow breed diversity, lactation number, dietary treatments) across these studies need harmonization or systematic evaluation to improve reproducibility of liver transcriptome dynamics findings in Holstein periparturient transitions.

    Liver transcriptome dynamics in Holstein cows during the periparturient transition · 2026 · DOI

  • Hesperidin metabolites were detected in adipose tissue but not in liver, kidney, or mammary tissue; the tissue-specific distribution, accumulation kinetics, and local bioactivity of hesperetin and its conjugates across metabolically active organs during the periparturient period require investigation.

    Hesperidin alleviates systemic inflammation and oxidative stress by remodeling adipose tissue lipid metabolism in periparturient dairy cows · 2026 · DOI

  • The enrichment of glycerophospholipid and sphingolipid metabolism pathways in adipose tissue and serum lipid profiles was identified through pathway analysis, but the specific roles of individual phosphatidylcholine (PC) and phosphatidylethanolamine (PE) species in mediating hesperidin's anti-inflammatory effects were not functionally interrogated.

    Hesperidin alleviates systemic inflammation and oxidative stress by remodeling adipose tissue lipid metabolism in periparturient dairy cows · 2026 · DOI

  • While the study detected correlations between differential ceramide species (Cer(d18:1/24:0), Cer(d18:1/16:0), Cer(d18:1/18:0)) in adipose tissue and serum with antioxidant and inflammatory indicators, the specific enzymes regulating ceramide synthesis and degradation (e.g., serine palmitoyltransferase, ceramidase) were not measured, leaving the regulatory mechanisms of sphingolipid remodeling unclear.

    Hesperidin alleviates systemic inflammation and oxidative stress by remodeling adipose tissue lipid metabolism in periparturient dairy cows · 2026 · DOI

  • The mechanistic pathway by which hesperidin suppresses NLRP3 inflammasome activation (IL-1β, IL-18, caspase-1, ASC downregulation) in adipose tissue was inferred but not directly validated through inflammasome reconstitution assays or NLRP3 pathway component knockdown studies in dairy cow adipocytes.

    Hesperidin alleviates systemic inflammation and oxidative stress by remodeling adipose tissue lipid metabolism in periparturient dairy cows · 2026 · DOI

  • Hesperidin bioavailability and metabolite accumulation were only measured at day 10 postpartum; the temporal dynamics of hesperetin, hesperetin-7-O-glucoside, and hesperetin-7-O-sulfate accumulation across the entire periparturient period (from late gestation through peak lactation) remain uncharacterized.

    Hesperidin alleviates systemic inflammation and oxidative stress by remodeling adipose tissue lipid metabolism in periparturient dairy cows · 2026 · DOI

  • Many methods have been used to increase the pregnancy rate for repeat breeder cows, such as intrauterine infusion of antibacterial agents or antibiotics, hormonal treatments for oestrus synchronisation and induction of ovulation, and progesterone supplementation or induction of accessory corpus luteum; however, the results were inconsistent between studies.

    Embryo transfer as an option to improve fertility in repeat breeder dairy cows · 2021 · DOI

  • Artificial insemination (AI) in goat is currently limited by the poor fertility obtained following the cervical insemination with frozen-thawed semen.

    Cervical Insemination with Frozen Thawed Semen in Goats at Different Breeding Age · 2019 · DOI

  • Mast cell (MC)-released mediators participate in uterine inflammation and immune response, but their role in postpartum metritis in cows has not been reported.

    Role of mast cells in cow metritis · 2016 · DOI

  • In conclusion, further investigation is required to determine the correlation of the FSHR and LHR genes with early puberty for the improvement of sexual precocity, and it is considered that the STAT5A gene may be used to improve the milk yield and milk yield trails of local cattle breeds, including those indigenous to Turkey.

    Investigation of STAT5A, FSHR, and LHR gene polymorphisms in Turkish indigenous cattle breeds (East Anatolian Red, South Anatolian Red, Turkish Grey, Anatolian Black, and Zavot) · 2015 · DOI

  • Although all of the students in the higher categories successfully localized the cervix, uterus, and ovaries, they had difficulties in interpreting these structures, suggesting that palpation of 200 cows is insufficient to reach a consistent level of expertise.

    Teaching Transrectal Palpation of the Internal Genital Organs in Cattle · 2009 · DOI

  • The MilkBovExp chip representing 90 genes encoding transcription factors expressed in the bovine mammary gland and/or involved in mammary gland signaling pathways was designed for further investigation of impact of gene expression and/or its encoded products on milk traits performance.

    Towards an integrated approach to study SNPs and expression of candidate genes associated with milk protein biosynthesis · 2008 · DOI

  • However, most previous studies have focused on maternal physiology or early cleavage- stage embryos, whereas the potential influence of vitamin D on embryo quality and survival after vitrification has not been investigated.

    Improved cryotolerance of bovine blastocysts by 1α,25-dihydroxyvitamin D<sub>3</sub> supplementation during in vitro culture and post-thaw recovery · 2026 · DOI

  • Furthermore, an independent validation cohort was not included, and uterine microbiome profiling, oxidative stress markers, cytokines, microRNA expression, and adipokine measurements were not investigated.

    Longitudinal integrated evaluation of metabolic, inflammatory, biochemical, and hormonal biomarkers identifies haptoglobin as a key indicator of reproductive performance in Friesian-Holstein cows with endometritis under tropical Indonesian dairy conditionsia · 2026 · DOI

  • Further investigation into the origin and functional roles of pregnanolone are needed to elucidate its contribution to pregnancy and reproductive success in roe deer.

    Pregnanolone appearance coincides with the period of implantation following embryonic diapause in the roe deer · 2026 · DOI

  • These data establish a safety profile in healthy cows and support further investigation of this product in studies focusing on efficacy and microbiome modulation.

    Evaluation of the local tolerance and systemic safety of a novel intravaginal probiotic product in cows · 2026 · DOI

  • The accuracy of the Taidoc BHBA portable analyser has not yet been tested and validated.

    Validation of A Portable Beta-Hydroxybutyric Acid Analyser for The Diagnosis of Subclinical Ketosis in Dairy Cows · 2023 · DOI

  • In addition, insufficient knowledge about the patterns of change in the reactivity of the nervous sexual centers and the corresponding reactions of the body to the administration of hormones reduces their effectiveness, which limits the widespread use.

    Induction and synchronization of oestrus in sheep and goats · 2021 · DOI

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37 open questions have been extracted from the limitations and future-work passages of 535 Reproductive Physiology in Livestock papers in our library. Each one below links back to the study that raised it, so you can read the original claim in context.

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