Background Sphingolipids(SL)are key regulators of inflammatory processes,yet their roles in dairy cows remain poorly understood.This study investigated the effects of inflammation(plasma haptoglobin concentration),ket...Background Sphingolipids(SL)are key regulators of inflammatory processes,yet their roles in dairy cows remain poorly understood.This study investigated the effects of inflammation(plasma haptoglobin concentration),ketosis,and mastitis on plasma SL profiles in Holstein cows sampled seven days postpartum.From a cohort of 427 cows across 25 farms,80 animals were classified into four groups:inflammation(n=20),ketosis(n=19),mastitis(n=21),and healthy controls(n=20).Plasma SL were quantified by targeted HPLC-MS/MS,while cytokines were quantified with a 15-plex bead-based assay.Both univariate and multivariate analyses were applied to assess pathological effects,along with SL ratios and correlations between SL and cytokines.Results Systemic inflammation detected through the haptoglobin measure induced the most pronounced alterations in SL metabolism,characterized by elevated dihydrosphingomyelins(DHSM)and lactosylceramides(Lac Cer),higher C22-24:C16 ratios,and lower unsaturated:saturated ratios in ceramides(Cer)and sphingomyelins(SM).Although total Cer,SM,and the Cer:SM ratio remained unchanged,specific reductions were observed in both Cer and SM in C14,Cer C18:1,SM C16:1,and SM C23:1,whereas SM C25:0 and C26:0 increased.Sphingosine-1-phosphate(So1P)was positively correlated with IL-10 as well as IL-1α and TNFα,while C18-20 Cer correlated positively with multiple pro-inflammatory cytokines and chemokines such as CXCL8 and CCL2.Ketosis induced subtler changes,primarily an increase in plasma DHSM and DHSM:SM ratio(driven by C16:0),an increase in C22-24:C16 DHCer ratio,and a decrease in both Lac So:Lac Cer and unsaturated:saturated ratios in C23-SM.In this group,So1P correlated positively with CXCL8 and CCL2.Moreover C18-20 Cer and DHCer were positively associated with CXCL8,CCL2,CCL3,and CCL4,which also showed correlations with most Lac Cer species.Analysis of chronic mastitis cases yielded a clear separation from controls in multivariate analysis but only minimal changes in SL concentrations and ratios,maybe due to the localized nature of the inflammatory response.Conclusions In summary,heightened inflammatory response in early post-partum is associated with the strongest systemic effects on SL metabolism,followed by ketosis,while mastitis induced only modest alterations.These findings highlight condition-specific patterns of SL regulation postpartum and suggest potential immunometabolic biomarkers of disease.展开更多
Background Subclinical intramammary infection(IMI)represents a significant problem in maintaining dairy cows’health.Disease severity and extent depend on the interaction between the causative agent,environment,and ho...Background Subclinical intramammary infection(IMI)represents a significant problem in maintaining dairy cows’health.Disease severity and extent depend on the interaction between the causative agent,environment,and host.To investigate the molecular mechanisms behind the host immune response,we used RNA-Seq for the milk somatic cells(SC)transcriptome profiling in healthy cows(n=9),and cows naturally affected by subclinical IMI from Proto-theca spp.(n=11)and Streptococcus agalactiae(S.agalactiae;n=11).Data Integration Analysis for Biomarker discov-ery using Latent Components(DIABLO)was used to integrate transcriptomic data and host phenotypic traits related to milk composition,SC composition,and udder health to identify hub variables for subclinical IMI detection.Results A total of 1,682 and 2,427 differentially expressed genes(DEGs)were identified when comparing Prototheca spp.and S.agalactiae to healthy animals,respectively.Pathogen-specific pathway analyses evidenced that Proto-theca’s infection upregulated antigen processing and lymphocyte proliferation pathways while S.agalactiae induced a reduction of energy-related pathways like the tricarboxylic acid cycle,and carbohydrate and lipid metabolism.The integrative analysis of commonly shared DEGs between the two pathogens(n=681)referred to the core-mastitis response genes,and phenotypic data evidenced a strong covariation between those genes and the flow cytometry immune cells(r2=0.72),followed by the udder health(r2=0.64)and milk quality parameters(r2=0.64).Variables with r≥0.90 were used to build a network in which the top 20 hub variables were identified with the Cytoscape cyto-hubba plug-in.The genes in common between DIABLO and cytohubba(n=10)were submitted to a ROC analysis which showed they had excellent predictive performances in terms of discriminating healthy and mastitis-affected animals(sensitivity>0.89,specificity>0.81,accuracy>0.87,and precision>0.69).Among these genes,CIITA could play a key role in regulating the animals’response to subclinical IMI.Conclusions Despite some differences in the enriched pathways,the two mastitis-causing pathogens seemed to induce a shared host immune-transcriptomic response.The hub variables identified with the integrative approach might be included in screening and diagnostic tools for subclinical IMI detection.展开更多
基金partly funded by APIS-GENEsupported by a grant Bonus QualitéRecherche(BQR)s from ENVT。
文摘Background Sphingolipids(SL)are key regulators of inflammatory processes,yet their roles in dairy cows remain poorly understood.This study investigated the effects of inflammation(plasma haptoglobin concentration),ketosis,and mastitis on plasma SL profiles in Holstein cows sampled seven days postpartum.From a cohort of 427 cows across 25 farms,80 animals were classified into four groups:inflammation(n=20),ketosis(n=19),mastitis(n=21),and healthy controls(n=20).Plasma SL were quantified by targeted HPLC-MS/MS,while cytokines were quantified with a 15-plex bead-based assay.Both univariate and multivariate analyses were applied to assess pathological effects,along with SL ratios and correlations between SL and cytokines.Results Systemic inflammation detected through the haptoglobin measure induced the most pronounced alterations in SL metabolism,characterized by elevated dihydrosphingomyelins(DHSM)and lactosylceramides(Lac Cer),higher C22-24:C16 ratios,and lower unsaturated:saturated ratios in ceramides(Cer)and sphingomyelins(SM).Although total Cer,SM,and the Cer:SM ratio remained unchanged,specific reductions were observed in both Cer and SM in C14,Cer C18:1,SM C16:1,and SM C23:1,whereas SM C25:0 and C26:0 increased.Sphingosine-1-phosphate(So1P)was positively correlated with IL-10 as well as IL-1α and TNFα,while C18-20 Cer correlated positively with multiple pro-inflammatory cytokines and chemokines such as CXCL8 and CCL2.Ketosis induced subtler changes,primarily an increase in plasma DHSM and DHSM:SM ratio(driven by C16:0),an increase in C22-24:C16 DHCer ratio,and a decrease in both Lac So:Lac Cer and unsaturated:saturated ratios in C23-SM.In this group,So1P correlated positively with CXCL8 and CCL2.Moreover C18-20 Cer and DHCer were positively associated with CXCL8,CCL2,CCL3,and CCL4,which also showed correlations with most Lac Cer species.Analysis of chronic mastitis cases yielded a clear separation from controls in multivariate analysis but only minimal changes in SL concentrations and ratios,maybe due to the localized nature of the inflammatory response.Conclusions In summary,heightened inflammatory response in early post-partum is associated with the strongest systemic effects on SL metabolism,followed by ketosis,while mastitis induced only modest alterations.These findings highlight condition-specific patterns of SL regulation postpartum and suggest potential immunometabolic biomarkers of disease.
基金the Ministero delle politiche agricole alimentari,forestali e del turismo(MIPAAF),Rome,Italy.Moreover,the study was conducted within the Agritech National Research Center and received funding from the European Union Next-GenerationEU(PIANO NAZIONALE DI RIPRESA E RESILIENZA(PNRR)-MISSIONE 4 COMPONENTE 2,INVESTIMENTO 1.4-D.D.103217/06/2022,CN00000022).
文摘Background Subclinical intramammary infection(IMI)represents a significant problem in maintaining dairy cows’health.Disease severity and extent depend on the interaction between the causative agent,environment,and host.To investigate the molecular mechanisms behind the host immune response,we used RNA-Seq for the milk somatic cells(SC)transcriptome profiling in healthy cows(n=9),and cows naturally affected by subclinical IMI from Proto-theca spp.(n=11)and Streptococcus agalactiae(S.agalactiae;n=11).Data Integration Analysis for Biomarker discov-ery using Latent Components(DIABLO)was used to integrate transcriptomic data and host phenotypic traits related to milk composition,SC composition,and udder health to identify hub variables for subclinical IMI detection.Results A total of 1,682 and 2,427 differentially expressed genes(DEGs)were identified when comparing Prototheca spp.and S.agalactiae to healthy animals,respectively.Pathogen-specific pathway analyses evidenced that Proto-theca’s infection upregulated antigen processing and lymphocyte proliferation pathways while S.agalactiae induced a reduction of energy-related pathways like the tricarboxylic acid cycle,and carbohydrate and lipid metabolism.The integrative analysis of commonly shared DEGs between the two pathogens(n=681)referred to the core-mastitis response genes,and phenotypic data evidenced a strong covariation between those genes and the flow cytometry immune cells(r2=0.72),followed by the udder health(r2=0.64)and milk quality parameters(r2=0.64).Variables with r≥0.90 were used to build a network in which the top 20 hub variables were identified with the Cytoscape cyto-hubba plug-in.The genes in common between DIABLO and cytohubba(n=10)were submitted to a ROC analysis which showed they had excellent predictive performances in terms of discriminating healthy and mastitis-affected animals(sensitivity>0.89,specificity>0.81,accuracy>0.87,and precision>0.69).Among these genes,CIITA could play a key role in regulating the animals’response to subclinical IMI.Conclusions Despite some differences in the enriched pathways,the two mastitis-causing pathogens seemed to induce a shared host immune-transcriptomic response.The hub variables identified with the integrative approach might be included in screening and diagnostic tools for subclinical IMI detection.
