Ramanathan Kasimanickam, Priunka Bhowmik, John Kastelic, Joao Ferreira, Vanmathy Kasimanickam
Read at the Multidisciplinary Digital Publishing Institute DOI: 10.3390/ani15192841
Abstract
Postpartum uterine diseases such as metritis and endometritis impair reproductive performance and cause substantial economic losses in dairy cows worldwide. The multifactorial etiology, involving polymicrobial infections and complex host immune responses, poses diagnostic and therapeutic challenges. Traditional treatments rely on antibiotics, e.g., cephalosporins like ceftiofur and cephapirin, with broad-spectrum efficacy. However, emerging antimicrobial resistance, biofilm formation by pathogens such as Trueperella pyogenes, Fusobacterium necrophorum, and Escherichia coli, and bacterial virulence factors have reduced effectiveness of conventional therapies. Advances in systems biology, particularly proteomics, metabolomics, and microRNA (miRNA) profiling, have provided unprecedented insights into the molecular mechanisms underpinning uterine disease pathophysiology. Proteomic analyses reveal dynamic changes in inflammatory proteins and immune pathways, whereas metabolomics highlight shifts in energy metabolism and bacterial-host interactions. Furthermore, miRNAs have critical roles in post-transcriptional gene regulation affecting immune modulation, inflammation, and tissue repair, and also in modulating neutrophil function and inflammatory signaling. Uterine inflammation not only disrupts local tissue homeostasis but also compromises early embryo development by altering endometrial receptivity, cytokine milieu, and oocyte quality. Integration of multi-omics approaches, combined with improved diagnostics and adjunct therapies-including micronutrient supplementation and immunomodulators-offers promising avenues for enhancing disease management and fertility in dairy herds. This review synthesizes current knowledge on proteomics, metabolomics, and miRNAs in postpartum uterine diseases and highlights future directions for research and clinical applications.
Ramanathan Kasimanickam, Joao Ferreira, Vanmathy Kasimanickam
Read at the Multidisciplinary Digital Publishing Institute DOI: 10.3390/ani15192841
Abstract
Postpartum metritis in dairy cows compromises reproductive performance and leads to substantial economic losses. This study investigated the molecular mechanisms underlying metritis by integrating high-throughput circulating microRNA (miRNA) profiling with systems-level bioinformatics. Previously, 30 differentially expressed miRNAs, 16 upregulated and 14 downregulated, were identified in metritis-affected cows compared to healthy controls. Building on these findings, this study predicted miRNA target genes and constructed regulatory networks involving miRNAs, mRNAs, circRNAs, lncRNAs, and snRNAs, alongside protein-protein interaction networks. Functional annotation and KEGG pathway analysis revealed that upregulated miRNAs influenced genes involved in immune activation, apoptosis, and metabolism, while downregulated miRNAs were associated with angiogenesis, immune suppression, and tissue repair. Hub genes such as AKT3, VEGFA, and HIF1A were central to immune and angiogenic signaling, whereas UBE3A and ZEB1 were linked to immune inhibition. Interferon-stimulated genes (e.g., ISG15, RSAD2, CXCL chemokines) were shown to regulate solute carriers, contributing to immune dysregulation. Key pathways included PI3K-Akt, NF-κB, JAK-STAT, insulin resistance, and T cell receptor signaling. Noncoding RNAs such as NEAT1, KCNQ1OT1, and XIST, along with miRNAs like bta-miR-15b and bta-miR-148a, emerged as pro-inflammatory regulators, while bta-miR-199a-3p appeared to exert immunosuppressive effects. These findings offer new insights into the complex regulatory networks driving metritis and suggest potential targets for improving fertility in dairy cows.
Olivia Truitt, Emma Thrower, Amber Itle, Tammy Edmonds, Rachel Claus-Walker, Erick Vergara Barrios, Kaitlin Mirkin, Treat Hurst, Minden Buswell, Caio Figueiredo, Craig McConnel
Abstract
Secure Milk Supply (SMS) plans for enhanced biosecurity provide a business continuity strategy for dairy producers in the event of a foot-and-mouth disease outbreak. During the summers of 2022, 2023, and 2024, Washington State University veterinary and undergraduate students created SMS plans for 101 premises representing 70 dairies within Washington. This project’s success was dependent upon the collaborative efforts of Northwest Dairy Association field managers who proved to be the bridge between the students and producers. Producers reflected that they appreciated having students as an intermediary between them and state officials, preferring to support education rather than regulation.
W.M. Coelho Jr., H.F. Monteiro, C.C. Figueiredo, B. Mion, J.E.P. Santos, R.S. Bisinotto, F. Peñagaricano, P. Vahmani, E.S. Ribeiro, and F.S. Lima
Read at ScienceDirect.com DOI: 10.3168/jds.2024-25454
Abstract
Feed efficiency is critical in dairy farming, affecting production costs and environmental sustainability. The development of the trait residual feed intake (RFI) has provided an opportunity to select dairy cows that are more efficient in converting nutrients into milk. Note that RFI requires individual daily intake records, which are typically collected on a limited number of research farms. In this context, the identification of biomarkers that can be used to identify and select more feed-efficient cows is of great interest. As such, this study aimed to identify ruminal and serum biomarkers associated with RFI in mid-lactation Holstein cows. A selected subset of 24 out of 454 Holstein cows was used in this study. This subset was strategically selected to represent extremes of least feed-efficient (LFE; n = 12, RFI = 2.44) and most feed-efficient (MFE; n = 12, RFI = −2.69) cows with no difference in the 3 energy sinks, namely BW change, metabolic BW, and energy secreted in milk. Rumen fluid and serum samples were collected between 60 and 90 DIM. Rumen fluid samples were collected using an oro-esophageal tubing procedure. Serum samples were used to measure fatty acids using a 2-step assay. The fatty acid methyl ester was assessed using solid-phase extraction and quantified using the chromatographic peak area and internal standard-based calculations. Ruminal ammonia nitrogen was measured using a phenol-hypochlorite assay, and serum urea was measured using a commercial ELISA kit. Cows in the MFE group had higher ruminal ammonia nitrogen concentrations than cows in the LFE group. There were no differences in serum urea concentration between MFE and LFE cows. Serum fatty acid concentrations differed between groups, with myristic acid (C14:0), palmitic acid (C16:0), cis-heptadecenoic acid (cis-9–17:1), stearic acid (C18:0), and total SFA having greater concentrations in the MFE group than in the LFE group. The total PUFA concentration was lower in the MFE group than in the LFE group. A model incorporating C14:0, C16:0, palmitoleic acid (trans-9-C16:1), anteiso-heptadecanoic acid plus palmitoleic acid (C17:0+trans-13-C16:1), oleic acid (cis-9-C18:1), cis-vaccenic acid (cis-11-C18:1), petroselinic acid (cis-12-C18:1), C18:0, linoleic acid (C18:2n-6), dihomo-γ-linolenic acid (C20:3n-6), cis-MUFA, n-6 PUFA, total PUFA, total SFA, and other or unknown fatty acids was used to assess goodness-of-fit for RFI and showed an adjusted R2 of 0.74. When ruminal ammonia nitrogen was added to the previous model, the adjusted R2 improved to 0.84. Our findings provide evidence that ruminal ammonia nitrogen and serum fatty acids are associated with RFI, thus suggesting that these metabolites might be helpful in identifying more feed-efficient dairy cows.
A. Martelo Pereira, F.N.S. Pereira, P.R. Menta, E.B. de Oliveira, J.G. Prim, F.S. Lima, V.S. Machado, K.N. Galvão, and C.C. Figueiredo
Read at ScienceDirect.com DOI: 10.3168/jds.2025-26288
Abstract
The objective of this study was to evaluate the differences in milk production, reproductive performance, and culling associated with antimicrobial therapy and clinical cure of metritis in dairy cows. In this study, data from 2 randomized controlled trials consisting of 900 Holstein cows from 5 dairy farms in California, Florida, and Texas were used. Cows were examined for metritis (fetid, watery, reddish-brownish vaginal discharge; VD) at 4, 7, and 10 DIM (study d 0). Cows diagnosed with metritis (n = 900) were randomly assigned (regardless of fever) to receive ceftiofur (CEF = 457) or to remain untreated (NT = 443). Clinical cure of metritis was assessed at study d 13 ± 1 based on the visual evaluation of the VD; cows that underwent clinical cure displayed clear, mucopurulent, or purulent discharge (cured = 710), whereas cows with clinical cure failure had persistent fetid, watery, reddish-brownish VD (not cured = 190). The data were analyzed considering antimicrobial treatment, cure status on d 13 ± 1, and their interaction. Risk of pregnancy and culling at 300 DIM were analyzed using logistic regression, time to pregnancy and culling by 300 DIM were analyzed using Cox’s proportional hazard, and milk production within 300 DIM was analyzed using ANOVA with repeated measures. Antimicrobial treatment and cure status were included as fixed effects and farm as a random effect in all models. Both randomized trials that generated data used in this study reported a greater risk of clinical cure associated with CEF treatment. Differences in risk of pregnancy at 300 DIM were associated with cure status (cured = 70.9% vs. not cured = 61.7%), but not with antimicrobial treatment or antimicrobial treatment by cure interaction. The risk of culling at 300 DIM was not associated with cure status, antimicrobial treatment, or antimicrobial treatment by cure interaction. Reduced median time to pregnancy was associated with cure status (cured = 134 vs. not cured = 154 d) but not with antimicrobial treatment or antimicrobial treatment by cure interaction. No interaction of antimicrobial treatment by cure status was observed on milk production; however, interactions between antimicrobial treatment, parity, and month of lactation and cure, parity, and month of lactation were observed. Differences in milk production between multiparous CEF and NT cows were observed in the third month of lactation. Conversely, differences in milk production associated with cure status were observed for primiparous and multiparous cows in the first and second months of lactation, whereas only multiparous cows in the eighth, ninth, and tenth months of lactation presented differences in milk production. Clinical cure of metritis was positively associated with milk production and reproduction, regardless of antimicrobial therapy, warranting further investigation regarding selective therapy of metritis.
M. Nehme Marinho, M.C. Perdomo, B. Souza Simões, A. Husnain, U. Arshad, C.C. Figueiredo, P.M. Peixoto, H.F. Monteiro, B. Anderson, and J.E.P. Santos.
Read at ScienceDirect.com DOI: 10.3168/jds.2025-26900
Abstract
Dietary microbial additives often benefit production performance in dairy cows. The objectives were to determine the effects of microbial additives on rumen microbial diversity, total-tract digestibility, incorporation of dietary N into milk N, and serum metabolites. One-hundred and 17 cows at 61 d postpartum (31–87 d) were blocked by parity group and pretreatment ECM yield. Within block, cows were randomly assigned to receive 100 g of corn meal containing no microbial additive (CON; n = 40), 100 g of corn meal containing 5 g of a mixture of Clostridium beijerinckii and Pichia kudriavzevii (G1; 4 × 107 cfu of C. beijerinckii and 1 × 109 cfu of P. kudriavzevii; n = 38), or 100 g of corn meal containing 5 g of a mixture of C. beijerinckii, P. kudriavzevii, Butyrivibrio fibrisolvens, and Ruminococcus bovis (G2; 4 × 107 cfu of C. beijerinckii, 1 × 109 cfu of P. kudriavzevii, 1 × 108 cfu of B. fibrisolvens, and 1 × 108 cfu of R. bovis; n = 39). Treatments were top-dressed for 140 d. Total-tract digestibility and rumen microbial composition were evaluated on d 61 and 124 of the experiment. Orthogonal contrasts compared the effect of microbial additive (MA) and the type of microbial additive (TMA). On experimental d 61, supplementing MA did not influence rumen microbial diversity, although cows fed G2 had a less diverse microbiome than those fed G1. On d 124, both MA and TMA reduced evenness, which resulted in lesser microbial α-diversity in cows supplemented with MA than those in CON. Minor differences were observed in rumen microbial β-diversity, except for multiparous cows on d 124 in which treatment tended to affect diversity because 16 amplicon sequence variant (ASV) groups representing phylogenetically related ASV differed among treatments. Two bacteria supplemented in G2, also native to the rumen, were identified, B. fibrisolvens and R. bovis, but treatment did not affect their abundance. Cows fed MA had increased total short-chain fatty acid concentration and partition of dietary N into milk N on d 61, but not on d 124. On d 61, feeding MA increased milk N secretion by 10 g/d (187 g/d vs. 193 g/d vs. 201 g/d), and incorporation of dietary N into milk N by 1.7 percentage units (CON = 30.8% vs. G1 = 31.2% vs. G2 = 33.7%). An interaction between TMA and parity affected the estimated microbial N production on d 124 because of the differences between G1 and G2 in primiparous cows (G1 = 302 g/d vs. G2 = 264 g/d). Type of MA affected digestibility on d 124, but the effect differed with parity. Feeding G1 increased digestibility of DM, OM, and CP in primiparous cows by 2.7, 2.8, and 4.7 percentage units, but not in multiparous cows. Collectively, supplementing diets of dairy cows with MA resulted in minor effects on the composition of the ruminal microbiota, total-tract digestibility, and use of dietary N. Present findings might not fully explain differences in production performance previously observed with the same MA added to the diet of cows.
F.N.S. Pereira, A. Martelo Pereira, K.N. Galvão, R.S. Bisinotto, and C.C. Figueiredo
Read on Oxford Academic, Biology of Reproduction, Volume 112, Issue 5, DOI: 10.1093/biolre.ioaf038
Abstract
This study investigated differences in uterine and serum metabolome associated with clinical cure failure of metritis in dairy cows. Metritis was diagnosed in lactating Holstein cows from two Florida dairies and defined by the presence of fetid, watery, reddish-brown vaginal discharge from 4 to 12 days postpartum (dpp). Cows with metritis (n = 24) were paired with cows without metritis of similar parity and dpp (n = 24). On the day of metritis diagnosis (day 0), all cows with metritis received antimicrobial therapy. The continued presence of the fetid, watery, reddish-brown discharge on day 5 (n = 16) was defined as clinical cure failure, whereas clinical cure was defined by its absence (n = 8). Metabolome analyses of uterine lavage (days 0 and 5) and serum samples (day 0) were conducted using untargeted gas chromatography time-of-flight mass spectrometry. Normalized data were analyzed using partial least squares–discriminant analysis and ANOVA, adjusting P-values for multiple comparisons. Differences in the uterine metabolome on day 0 associated with clinical cure failure were linked to carbohydrate, amino acid, and lipid metabolism. Greater concentrations of arachidonic acid, ribose, and glutaric acid were associated with clinical cure failure, suggesting a greater degree of tissue lesion and inflammation. No differences in the serum metabolome were associated with cure failure. No differences in uterine metabolome were associated with clinical cure failure on day 5. The findings suggest that clinical cure failure is associated with a greater uterine inflammatory process that did not persist until cure assessment day.