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Metabolic consequences of an energy and protein rich diet can compromise metabolic health of cattle by promoting a pro-inflammatory phenotype. Laminitis is a common clinical sign, but affected metabolic pathways, underlying pathophysiology and causative relationships of a systemic pro-inflammatory phenotype are unclear. Therefore, the aim of this study was to elucidate changes in metabolome profiles of 20 months old Holstein bulls fed a high energy and protein diet and to identify novel metabolites and affected pathways, associated with diet-related laminitis. In a randomized controlled feeding trial using bulls fed a high energy and protein diet (HEP; metabolizable energy [ME] intake 169.0 ± 1.4 MJ/day; crude protein [CP] intake 2.3 ± 0.02 kg/day; calculated means ± SEM; n = 15) versus a low energy and protein diet (LEP; ME intake 92.9 ± 1.3 MJ/day; CP intake 1.0 ± 0.01 kg/day; n = 15), wide ranging effects of HEP diet on metabolism were demonstrated with a targeted metabolomics approach using the AbsoluteIDQ p180 kit (Biocrates Life Sciences). Multivariate statistics revealed that lower concentrations of phosphatidylcholines and sphingomyelins and higher concentrations of lyso-phosphatidylcholines, branched chain amino acids and aromatic amino acids were associated with an inflammatory state of diet-related laminitis in Holstein bulls fed a HEP diet. The latter two metabolites share similarities with changes in metabolism of obese humans, indicating a conserved pathophysiological role. The observed alterations in the metabolome provide further explanation on the underlying metabolic consequences of excessive dietary nutrient intake.

Calf diarrhea is associated with enteric infections, and also provokes the overuse of antibiotics. Therefore, proper treatment of diarrhea represents a therapeutic challenge in livestock production and public health concerns. Here, we describe the ability of a fecal microbiota transplantation (FMT), to ameliorate diarrhea and restore gut microbial composition in 57 growing calves. We conduct multi-omics analysis of 450 longitudinally collected fecal samples and find that FMT-induced alterations in the gut microbiota (an increase in the family Porphyromonadaceae ) and metabolomic profile (a reduction in fecal amino acid concentration) strongly correlate with the remission of diarrhea. During the continuous follow-up study over 24 months, we find that FMT improves the growth performance of the cattle. This first FMT trial in ruminants suggest that FMT is capable of ameliorating diarrhea in pre-weaning calves with alterations in their gut microbiota, and that FMT may have a potential role in the improvement of growth performance. Here, the authors report the results of a longitudinal multi-omics trial of the use of fecal microbiota transplantation (FMT) to ameliorate diarrhea and restore gut microbial composition in 57 growing calves, and find that oral FMT induces alterations in the gut microbiota correlate with the remission of diarrhea and improves the growth performance of the cattle.

Gastrointestinal parasites (GIP) and mineral deficiencies are significant factors affecting health and productivity of free-ranging cattle. Adequate mineral intake, particularly from natural mineral water sources (hora), is vital for immune function, gastrointestinal health, and nutrient absorption. This study aimed to explore the association between GIP burden and serum mineral concentrations in zebu cattle (Bos indicus) routinely accessing hora mineral water in southwestern Ethiopia. A total of 180 fecal samples were collected from cattle across four districts (Bedele, Dabo, Gechi, and Borecha) and analyzed qualitatively and quantitatively to determine parasite presence and fecal egg count. Concurrently, blood samples were collected to evaluate serum mineral concentrations. The overall GIP prevalence was 55.6%, with Strongyle-type nematodes being the dominant GIP group. Gechi district showed the highest prevalence (64.4%) and mean egg per gram (EPG) of 212.8 (p < 0.05). Although, serum mineral concentrations were generally adequate, significant variations were observed across districts. Strong negative correlations (p < 0.05) were observed between EPG and serum concentrations of zinc (Zn), manganese (Mn), iron (Fe), and copper (Cu), indicating that adequate intake of these minerals, potentially sourced from the hora mineral water, may be associated with improved resistance to parasitic infections. These negative correlations were supported by negative binomial regression analysis which identified Zn as the strongest predictor of EPG. Overall, the findings highlight the importance of hora as a natural mineral supplement in its association with lower GIP burden in free-ranging zebu cattle. While this study indicates a correlation between serum mineral concentrations and GIP burden in grazing cattle, controlled experiments are essential to determine the specific effects of individual minerals found in hora on parasite resistance and establish causality.

Rift Valley fever (RVF) is a zoonotic disease caused by Rift Valley fever virus (RVFV) found in Africa and the Middle East. Outbreaks can cause extensive morbidity and mortality in humans and livestock. Following the diagnosis of two acute human RVF cases in Kabale district, Uganda, we conducted a serosurvey to estimate RVFV seroprevalence in humans and livestock and to identify associated risk factors. Humans and animals at abattoirs and villages in Kabale district were sampled. Persons were interviewed about RVFV exposure risk factors. Human blood was tested for anti-RVFV IgM and IgG, and animal blood for anti-RVFV IgG. 655 human and 1051 animal blood samples were collected. Anti-RVFV IgG was detected in 78 (12%) human samples; 3 human samples (0.5%) had detectable IgM only, and 7 (1%) had both IgM and IgG. Of the 10 IgM-positive persons, 2 samples were positive for RVFV by PCR, confirming recent infection. Odds of RVFV seropositivity were greater in participants who were butchers (odds ratio [OR] 5.1; 95% confidence interval [95% CI]: 1.7-15.1) and those who reported handling raw meat (OR 3.4; 95% CI 1.2-9.8). No persons under age 20 were RVFV seropositive. The overall animal seropositivity was 13%, with 27% of cattle, 7% of goats, and 4% of sheep seropositive. In a multivariate logistic regression, cattle species (OR 9.1; 95% CI 4.1-20.5), adult age (OR 3.0; 95% CI 1.6-5.6), and female sex (OR 2.1; 95%CI 1.0-4.3) were significantly associated with animal seropositivity. Individual human seropositivity was significantly associated with animal seropositivity by subcounty after adjusting for sex, age, and occupation (p < 0.05). Although no RVF cases had been detected in Uganda from 1968 to March 2016, our study suggests that RVFV has been circulating undetected in both humans and animals living in and around Kabale district. RVFV seropositivity in humans was associated with occupation, suggesting that the primary mode of RVFV transmission to humans in Kabale district could be through contact with animal blood or body fluids.

Background Tick infestation is one of the main challenges in tropical beef cattle production, leading to significant economic losses. Knowledge of the molecular factors underlying natural tick resistance in cattle contributes to genetic selection through the identification of biomarkers that can be used to accurately identify animals resistant to ticks. Although several genes associated with resistance to ticks have been identified, the molecular mechanisms underlying tick resistance are yet to be elucidated. This study investigated the biological processes, pathways, and key proteins involved in the resistance to the tick Rhipicephalus (Boophilus) microplus in a tropically adapted beef cattle breed. Tick resistance was evaluated in 162 Caracu cows. Blood samples were collected from a subset of 16 extreme animals, including eight with a high tick load (SUS) and eight with a low tick load (RES), for proteomic analysis by LC-MS/MS. Results A total of 172 and 34 proteins were exclusively identified in plasma samples from the SUS and RES groups, respectively. In addition, 14,034 proteins were detected in the blood plasma of both groups, of which 51 and 101 proteins were significantly increased in plasma samples of the SUS and RES groups, respectively. Among the top 20 proteins with the highest absolute log-fold change values, those encoded by the RNASE1 , TNS2 , NOXO1 , ZNRF3 , APOA4 , KMT2B , RPS6KA5 , PON1 , C4BPA , SETD2 , HP , TMEM63A , MAST2 , and SETD1B genes were highlighted based on their functions that may contribute to a response to tick infestation. Functional enrichment analysis revealed 36 biological processes, 35 molecular functions, and 16 pathways to be significant ( P  < 0.05), highlighting those related to hemostasis, vesicular transport, cell proliferation and migration, calcium, actin, lipids, scavenger receptors, hydrogen peroxide, tyrosine, and insulin-like growth factor, which may contribute to tick resistance. In addition, PPI network analysis revealed several proteins involved in complement and coagulation systems, hematopoiesis, and immune response as important nodes, based on their centrality and edges. Conclusions The identification of differentially abundant proteins between RES and SUS animals, as well as their relationships and roles in key biological processes and molecular pathways detected, contribute to improving our understanding of the mechanisms underlying tick resistance in naturally adapted cattle breeds. Furthermore, the differentially abundant proteins detected in this study are potential biomarkers for the response to R. microplus infestation.

Crimean-Congo Hemorrhagic Fever (CCHF) is a tick-borne zoonosis of major public health concern, not only because of its potential for severe outcomes in humans, but also due to its endemic presence in many regions and its expanding geographic distribution. We report on the first serological survey conducted in mainland France to detect antibodies against the Crimean-Congo Hemorrhagic Fever Virus (CCHFV) in domestic and wild fauna, and provides critical insights into the virus’s circulation. We analyzed 8,609 cattle sera and 2,182 wildlife sera collected across the French Mediterranean region from 2008 to 2022, using enzyme-linked immunosorbent assays (ELISA) and pseudo-plaque reduction neutralization tests (PPRNT) for antibody detection and confirmation. Seropositivity was detected in both cattle (2.04%) and wildlife (2.25%), with higher rates observed in specific regions including the Pyrénées-Orientales and Hautes-Pyrénées. These findings reveal spatial clusters of CCHFV circulation and suggest the existence of enzootic transmission cycles involving local tick vectors and animal hosts. Our multivariate analysis identified key factors that influence seropositivity, including animal age, habitat characteristics, and potential wildlife interactions. The presence of natural open habitats and coniferous forests was significantly associated with higher seropositivity in cattle, while sex and geographical variability played a role in wildlife seroprevalence. These findings highlight the importance of environmental and anthropogenic factors in shaping the dynamics of CCHFV transmission. This work demonstrates that CCHFV is actively circulating in parts of mainland France, emphasizing the need for enhanced surveillance and integrated approaches to monitor zoonotic pathogens. It also raises questions about the role of additional tick vectors, such as Hyalomma lusitanicum , in the transmission cycle. These results advance our understanding of CCHF epidemiology and offer valuable guidance for public health strategies to mitigate the risks associated with this emerging disease.

Background/Aims: Dairy cows with ketosis are characterized by oxidative stress and hepatic damage. The aim of this study was to investigate hepatic oxidative stress and the apoptotic status of ketotic cows, as well as the underlying apoptosis pathway. Methods: The blood aspartate aminotransferase (AST), alanine aminotransferase (ALT), glutamate dehydrogenase (GLDH) and gamma-glutamyl transferase (GGT) activities and the haptoglobin (HP), serum amyloid A (SAA) and serum apoptotic cytokeratin 18 neo-epitope M30 (CK18 M30) concentrations were determined by commercially available kits and ELISA kits, respectively. Liver histology, TUNEL and Oil red O staining were performed in liver tissue samples. TG contents were measured using an enzymatic kit; Caspase 3 assays were carried out using the Caspase 3 activity assay kit; oxidation and antioxidant markers were measured using biochemical kits; apoptosis pathway were determined by qRT-PCR and western blot. Results: Ketotic cows displayed hepatic fat accumulation. The hepatic malondialdehyde (MDA) content was significantly increased, but the activities of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were markedly decreased in ketotic cows compared with control cows, indicating that ketotic cows displayed severe oxidative stress. Significantly higher serum levels of the hepatic damage markers AST, ALT, GGT and GLDH were observed in ketotic cows than in control cows. The blood concentration of the apoptotic marker CK18 M30 and the number of TUNEL-positive cells in the liver of ketotic cows were 1.19- and 2.61-fold, respectively, higher than the values observed in control cows. Besides, Caspase 3 activity was significantly increased in the liver of ketosis cows. Importantly, the levels of phosphorylated c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (p38MAPK) were significantly increased but the level of phosphorylated extracellular signal-regulated kinase1/2 (ERK1/2) was markedly decreased, which further promoted tumor protein 53 (p53) expression and inhibited nuclear factor E2-related factor 2 (Nrf2) expression. The apoptosis-related molecules p21, MDM2, Caspase 3, Caspase 9 and Bax were expressed at significantly higher levels in ketotic cows than in healthy cows, whereas the anti-apoptosis molecule Bcl-2 was expressed at significantly lower levels. Conclusions: Based on these results, ketotic cows display severe hepatic oxidative stress. The hepatic MAPK-p53-Nrf2 apoptotic pathway is over induced and partially mediated apoptotic damage in the liver.

Bovine respiratory disease (BRD) is a multifactorial disease complex and the leading infectious disease in post-weaned beef cattle. Clinical manifestations of BRD are recognized in beef calves within a high-risk setting, commonly associated with weaning, shipping, and novel feeding and housing environments. However, the understanding of complex host immune interactions and genomic mechanisms involved in BRD susceptibility remain elusive. Utilizing high-throughput RNA-sequencing, we contrasted the at-arrival blood transcriptomes of 6 beef cattle that ultimately developed BRD against 5 beef cattle that remained healthy within the same herd, differentiating BRD diagnosis from production metadata and treatment records. We identified 135 differentially expressed genes (DEGs) using the differential gene expression tools edgeR and DESeq2. Thirty-six of the DEGs shared between these two analysis platforms were prioritized for investigation of their relevance to infectious disease resistance using WebGestalt, STRING, and Reactome. Biological processes related to inflammatory response, immunological defense, lipoxin metabolism, and macrophage function were identified. Production of specialized pro-resolvin mediators (SPMs) and endogenous metabolism of angiotensinogen were increased in animals that resisted BRD. Protein-protein interaction modeling of gene products with significantly higher expression in cattle that naturally acquire BRD identified molecular processes involving microbial killing. Accordingly, identification of DEGs in whole blood at arrival revealed a clear distinction between calves that went on to develop BRD and those that resisted BRD. These results provide novel insight into host immune factors that are present at the time of arrival that confer protection from BRD.

Ketosis often occurs during the postpartum transition period in dairy cows, leading to economic and welfare problems. Previously, ketosis was reported to be associated with hematological and serum biochemical parameters. However, the association between the parameters on the calving date and ketosis during the postpartum transition period remains unclear. This study aimed to investigate this association. Blood samples were collected from the jugular vein of Holstein cows on the calving date and β-hydroxybutyrate was tested once every 3 days (8 times in 21 days). The cows were divided into three groups: non-ketosis, subclinical ketosis, and clinical ketosis. The clinical ketosis group significantly had the highest values of mean corpuscular volume, mean corpuscular hemoglobin, β-hydroxybutyrate, non-esterified fatty acids, and total bilirubin, but the lowest values of red cell distribution width, the counts of white blood cell, monocyte, and eosinophil, albumin, alanine transaminase, lactate dehydrogenase, and amylase. In contrast, the non-ketosis group showed the opposite results ( p  < 0.05). In conclusion, these parameters are associated with the development and severity of ketosis. Our findings suggest that these parameters on the calving date may be useful indicators to identify dairy Holstein cow susceptible to ketosis during the transition period.

The objectives of this study were to 1) establish cow-level critical thresholds for serum concentrations of nonesterified fatty acids (NEFA) and β-hydroxybutyrate (BHBA) to predict periparturient diseases [displaced abomasa (DA), clinical ketosis (CK), metritis and retained placenta, or any of these three], and 2) investigate the magnitude of the metabolites’ association with these diseases within 30 d in milk. In a prospective cohort study of 100 freestall, total mixed ration-fed herds in the northeastern United States, blood samples were collected from approximately 15 prepartum and 15 different postpartum transition animals in each herd, for a total of 2,758 samples. Serum NEFA concentrations were measured in the prepartum group, and both NEFA and BHBA were measured in the postpartum group. The critical thresholds for NEFA or BHBA were evaluated with receiver operator characteristic analysis for all diseases in both cohorts. The risk ratios (RR) of a disease outcome given NEFA or BHBA concentrations and other covariates were modeled with multivariable regression techniques, accounting for clustering of cows within herds. The NEFA critical threshold that predicted any of the 3 diseases in the prepartum cohort was 0.29mEq/L and in the postpartum cohort was 0.57mEq/L. The critical threshold for serum BHBA in the postpartum cohort was 10mg/dL, which predicted any of the 3 diseases. All RR with NEFA as a predictor of disease were >1.8; however, RR were greatest in animals sampled postpartum (e.g., RR for DA=9.7; 95% CI=4.2 to 22.4. All RR with BHBA as the predictor of disease were >2.3 (e.g., RR for DA=6.9; 95% CI=3.7 to 12.9). Although prepartum NEFA and postpartum BHBA were both significantly associated with development of clinical disease, postpartum serum NEFA concentration was most associated with the risk of developing DA, CK, metritis, or retained placenta during the first 30 d in milk.