Changes in Parameters Registered by Innovative Technologies in Cows with Subclinical Acidosis.

The hypothesis of this study was that there were changes in biomarkers registered by innovative technologies in cows with subclinical acidosis. The aim of this study was to identify changes in the in-line milk fat-to-protein ratio and cow feeding behaviors such as reticulorumen pH, reticulorumen temperature, cow activity, and water intake with subclinical acidosis. From a total of 98 cows, 59 cows were selected to meet the following criteria (2 or more lactations, with 31 days in milk (DIM)). The selected animals were separated into two groups based on general clinical examination and reticulorumen pH: the subclinical acidosis group (SCA, n = 23) and the healthy group (HC, n = 36). During the diagnosis of subclinical acidosis and following the clinical examination of the healthy group using the BROLIS HerdLine system, the daily averages of milk yield (kg/day), milk fat (%), milk protein (%), and the milk fat-to-protein ratio were recorded. Simultaneously, by using Smaxtec technology, reticulorumen parameters and cow activity, including pH, temperature (°C), rumination time (minutes/day), and water intake (hours/day), were registered. Changes in parameters measured using innovative technologies were able to identify cows with subclinical acidosis. Cows with subclinical acidosis had a lower reticulorumen pH by 18.8% (p < 0.0001), a decreased milk yield by 10.49% (p < 0.001), a lower milk fat-to-protein ratio by 11.88% (p < 0.01), and a decreased rumination time by 6.59% (p < 0.01). However, the activity of these cows was higher by 57.19% (p < 0.001) compared to healthy cows. From a practical point of view, we suggest that veterinarians and farmers track parameters such as reticulorumen pH, milk yield, milk fat-to-protein ratio, rumination time, and activity for the identification of subclinical acidosis.

Zoonotic linkage and environmental contamination of Methicillin-resistant Staphylococcus aureus (MRSA) in dairy farms: A one health perspective.

Methicillin-Resistant Staphylococcus aureus (MRSA) is a ubiquitous public health challenge, with its prevalence in human, animal, and environmental interfaces posing significant concerns. This study aimed to characterize and detect the zoonotic linkages of MRSA within the cow-environment-human interfaces in dairy farms to address the One Health perspective. A comprehensive investigation, involving 636 samples (an equal number of raw milk and cow nasal swab samples, along with varying numbers of human nasal swab and environmental samples), revealed an overall MRSA prevalence of 13.4% (n = 271/636). Notably, environmental samples exhibited the highest prevalence (19.3%), emphasizing the potential role of farm surroundings in MRSA transmission, while the lowest prevalence was found in raw milk at 11.8% (n = 31/263). The prevalence in cow nasal swabs and human nasal swabs was 13.3% (n = 35/263) and 15.1% (n = 8/53), respectively. Multiplex PCR analysis revealed the presence of different Staphylococcal enterotoxin (SEa, SEb, SEc, and SEd), and exfoliative toxin-producing genes (Eta, Etb) within the MRSA isolates underlining their potential to induce public health threats. All MRSA isolates exhibited complete resistance to Oxacillin (100%) and Amoxicillin (100%), while the highest sensitivity was observed for Vancomycin (85.8%). Furthermore, these MRSA strains demonstrated varying degrees of resistance to other commonly used antimicrobial drugs, including Cefoxitin (75.3%), Ceftarolin (71.2%), Sulfamethoxazole-Trimethoprim (63.5%), Ciprofloxacin (60%), and Gentamicin (49.5%). Detection of MRSA in cow, human, and environmental samples within the same farm vicinity highlights the risk of zoonotic transmission of MRSA from cows to humans through environmental interfaces. Phylogenetic analysis of the mecA gene in MRSA isolates from all sources within the same farm revealed a high similarity index (>84%) among them suggesting a shared evolutionary origin. Moreover, the MRSA isolates from milk samples showed a close evolutionary relationship with isolates from Kenya and Brazil, while the isolates from humans and the environment displayed noticeable resemblance to isolates from several Asian countries. The findings emphasize the importance of collaborative efforts under the One Health framework to address this multifaceted issue and ensure the safety of our food supply and public health.

Exploring positive welfare measures: preliminary findings from a prototype protocol in loose housing dairy cattle farms.

Introduction: Following the increasing interest about the development of indicators of positive welfare and affective state in farm animals, the aim of this research is to present some preliminary results on the application of a prototype protocol based exclusively on positive welfare measures and to suggest potential benefits that can promote positive welfare. Methods: The protocol was applied in 20 loose housing dairy cattle farms (6 on deep litter with straw, 14 in cubicles) and included only indicators of positive welfare and emotional states: feeding and resting synchronization, rumination during resting, comfortable lying postures, no visible eye white, relaxed ear postures, percentage of cow contacts with humans in the Avoidance Distance test. Potential benefits in terms of housing, feeding and management were then related to these variables (Mann-Whitney U test). Qualitative Behavior Assessment (QBA) was also carried out and analyzed by Principal Component Analysis to explore the effect of factors that were not evenly distributed in our sample (number of feed distributions, access to pasture, presence of paddock or environmental enrichments, automatic milking systems). Results: When hay was included in the diet, higher feeding synchronization (93.7 ± 1.6 vs. 52.2 ± 4.7%; p < 0.01), percentage of cows with relaxed ear postures (35.8 ± 5.4 vs. 15.5 ± 2.1%; p < 0.01) and percentage of cows with no visible eye white (55.9 ± 17.0 vs. 36.6 ± 4.1%; n.s.) were recorded. A higher level of feeding synchronization was observed also when the feeding places/cow ratio was > 1 (72.1 ± 9.9 vs. 53.8 ± 5.8%), although differences were not significant (p = 0.14). Deep litter had a more positive effect than cubicles on comfort at resting, with a significantly higher percentage of ruminating cows (65.8 ± 10.2 vs. 34.2 ± 3.7%; p < 0.01), a higher percentage of cows with no visible eye white (55.6 ± 9.9 vs. 33.1 ± 3.7%; p < 0.05) and a higher percentage of cows in a more comfortable posture, with stretched legs (14.3 ± 5.1 vs. 5.6 ± 1.6%; p = 0.09). QBA highlighted the most positive emotional state in the only farm that allowed access to pasture. Conclusions: This study represents a first attempt to apply a protocol for on-farm welfare evaluation based exclusively on the use of positive welfare indicators and provides suggestions on possible benefits (e.g., deep litter, feeding places/cow ratio > 1, hay in the diet and access to pasture) to enhance dairy cattle welfare.

Grape seed oil in the diet of primiparous Jersey cows before and after parturition: effects on performance, health, rumen environment, and milk quality.

The objective of the study was to determine whether adding grape seed oil (GSO) to the diet of primiparous Jersey breeds during the transition period would improve animal health by measuring effects on the rumen environment, serum biochemistry, oxidative response, and the composition and quality of milk. We used 14 Jersey heifers, weighing an average of 430 kg and 240 days of gestation. The animals were divided into two groups and offered a basal diet, including GSO in the concentrate for the GSO group (dose of 25 mL per animal day) and the same dose of soybean oil (SO) for the control group. The animals were allocated and maintained in a compost barn system, receiving an anionic diet (pre-partum) and a diet for postpartum lactating animals. Dry matter intake (DMI), milk production, serum biochemistry, serum and milk oxidative stability, ruminal fluid and milk fatty acid profile, milk qualitative aspects, and ruminal parameters such as pH, bacterial activity, and protozoan count were evaluated. The addition of GSO had a positive effect on the health of the cows, especially on the oxidative stability of the cows, by increasing total thiols (P = 0.03), higher plasma ferric reducing capacity (FRAP) (P = 0.01), and total antioxidant capacity (TAC) (P = 0.01). In the oxidative stability of the milk produced by the treated animals, there was also an increase in TAC (P = 0.05) and FRAP (P = 0.03). Discreet changes were observed in the ruminal environment with a decreasing trend in pH (P = 0.04) but an increase in bacterial activity (P = 0.05) and protozoa counts (P = 0.07) in cows that consumed the additive. GSO consumption affected the fatty acid profile in milk, increasing saturated fatty acids (SFA) (P = 0.05) and reducing unsaturated fatty acids (UFA) (P = 0.03). The oil did not affect milk production or efficiency in the postpartum period. Based on this information, it is concluded that the addition of GSO positively affects the cow's antioxidant system.

Digital and Precision Technologies in Dairy Cattle Farming: A Bibliometric Analysis.

The advancement of technology has significantly transformed the livestock landscape, particularly in the management of dairy cattle, through the incorporation of digital and precision approaches. This study presents a bibliometric analysis focused on these technologies involving dairy farming to explore and map the extent of research in the scientific literature. Through this review, it was possible to investigate academic production related to digital and precision livestock farming and identify emerging patterns, main research themes, and author collaborations. To carry out this investigation in the literature, the entire timeline was considered, finding works from 2008 to November 2023 in the scientific databases Scopus and Web of Science. Next, the Bibliometrix (version 4.1.3) package in R (version 4.3.1) and its Biblioshiny software extension (version 4.1.3) were used as a graphical interface, in addition to the VOSviewer (version 1.6.19) software, focusing on filtering and creating graphs and thematic maps to analyze the temporal evolution of 198 works identified and classified for this research. The results indicate that the main journals of interest for publications with identified affiliations are "Computers and Electronics in Agriculture" and "Journal of Dairy Science". It has been observed that the authors focus on emerging technologies such as machine learning, deep learning, and computer vision for behavioral monitoring, dairy cattle identification, and management of thermal stress in these animals. These technologies are crucial for making decisions that enhance health and efficiency in milk production, contributing to more sustainable practices. This work highlights the evolution of precision livestock farming and introduces the concept of digital livestock farming, demonstrating how the adoption of advanced digital tools can transform dairy herd management. Digital livestock farming not only boosts productivity but also redefines cattle management through technological innovations, emphasizing the significant impact of these trends on the sustainability and efficiency of dairy production.

Lameness in Cattle-Etiopathogenesis, Prevention and Treatment.

The aim of this review was to analyse the health problem of lameness in dairy cows by assessing the health and economic losses. This review also presents in detail the etiopathogenesis of lameness in dairy cattle and examples of its treatment and prevention. This work is based on a review of available publications. In selecting articles for the manuscript, the authors focused on issues observed in cattle herds during their clinical work. Lameness in dairy cattle is a serious health and economic problem around the world. Production losses result from reduced milk yield, reduced feed intake, reproductive disorders, treatment costs, and costs associated with early culling. A significant difficulty in the control and treatment of lameness is the multifactorial nature of the disease; causes may be individual or species-specific and may be associated with the environment, nutrition, or the presence of concomitant diseases. An important role is ascribed to infectious agents of both systemic and local infections, which can cause problems with movement in animals. It is also worth noting the long treatment process, which can last up to several months, thus significantly affecting yield and production. Given the high economic losses resulting from lameness in dairy cows, reaching even >40% (depending on the scale of production), there seems to be a need to implement extensive preventive measures to reduce the occurrence of limb infections in animals. The most important effective preventive measures to reduce the occurrence of limb diseases with symptoms of lameness are periodic hoof examinations and correction, nutritional control, and bathing with disinfectants. A clean and dry environment for cows should also be a priority.

Unraveling metabolic stress response in dairy cows: genetic control of plasma biomarkers throughout lactation and the transition period.

Breeding animals able to effectively respond to stress could be a long-term, sustainable, and affordable strategy to improve resilience and welfare in livestock systems. In the present study, the concentrations of 29 plasma biomarkers were used as candidate endophenotypes for metabolic stress response in single-SNP, gene- and haplotype-based GWAS using 739 healthy lactating Italian Holstein cows and 88,271 variants. Significant genetic associations were found in all the 3 GWAS approaches for plasma γ-glutamyl transferase concentration on BTA17, for paraoxonase on BTA4, and for alkaline phosphatase and zinc on BTA2. On these chromosomes, single-SNP and gene-based chromosome-wide association studies were performed, confirming GWAS findings. The signals identified for paraoxonase, γ-glutamyl transferase, and alkaline phosphatase were in proximity of the genes coding for them. The heritability of these 4 biomarkers ranged from moderate to high (from 0.39 to 0.54). Plasma biomarkers are known to undergo large changes in concentration during metabolic stress in the transition period, with an inter-individual variability in the rate of change and recovery time. Genetics may account in part for these differences. To assess this, we studied a subset of 139 periparturient cows homozygous at 3 SNPs known to be respectively associated with concentration of plasma ceruloplasmin, paraoxonase and γ-glutamyl transferase. We compared the immune-metabolic profile measured in plasma at -7, +5 and +30 d relative to calving between groups of opposite homozygotes. A significant effect of the genotype was found on paraoxonase and γ-glutamyl transferase plasma concentration at all the 3 time points. No evidence for genotype effect was detected for ceruloplasmin. Understanding the genetic control underlying metabolic stress response may suggest new approaches to foster resilience in dairy cows.

Diacylglycerol O-acyltransferase (DGAT) isoforms play a role in peridroplet mitochondrial fatty acid metabolism in bovine liver.

Hepatocellular lipid accumulation characterizes fatty liver in dairy cows. Lipid droplets (LD), specialized organelles that store lipids and maintain cellular lipid homeostasis, are responsible for the ectopic storage of lipids associated with several metabolic disorders. In recent years, non-ruminant studies have reported that LD-mitochondria interactions play an important role in lipid metabolism. Due to the role of diacylglycerol acyltransferase isoforms (DGAT1 and DGAT2) in LD synthesis, we explored mechanisms of mitochondrial fatty acid transport in ketotic cows using liver biopsies and isolated primary hepatocytes. Compared with healthy cows, cows with fatty liver had massive accumulation of LD and high protein expression of the triglyceride (TAG) synthesis-related enzymes DGAT1 and DGAT2, LD synthesis-related proteins perilipin 2 (PLIN2) and perilipin 5 (PLIN5), and the mitochondrial fragmentation-related proteins dynamin-related protein 1 (DRP1) and fission 1 (FIS1). In contrast, factors associated with fatty acid oxidation, mitochondrial fusion and mitochondrial electron transport chain complex were lower compared with those in the healthy cows. In addition, transmission electron microscopy revealed significant contacts between LD-mitochondria in liver tissue from cows with fatty liver. Compared with isolated cytoplasmic mitochondria, expression of carnitine palmitoyl transferase 1A (CPT1A) and DRP1 was lower, but mitofusin 2 (MFN2) and mitochondrial electron transport chain complex was greater in isolated peridroplet mitochondria from hepatic tissue of cows with fatty liver. In vitro data indicated that exogenous free fatty acids (FFA) induced hepatocyte LD synthesis and mitochondrial dynamics consistent with in vivo results. Furthermore, DGAT2 inhibitor treatment attenuated the FFA-induced upregulation of PLIN2 and PLIN5 and rescued the impairment of mitochondrial dynamics. Inhibition of DGAT2 also restored mitochondrial membrane potential and reduced hepatocyte reactive oxygen species production. The present in vivo and in vitro results indicated there are functional differences among different types of mitochondria in the liver tissue of dairy cows with ketosis. Activity of DGAT2 may play a key role in maintaining liver mitochondrial function and lipid homeostasis in dairy cows during the transition period.

Changes on the rumen microbial community composition in dairy cows subjected to an acidogenic diet.

In modern breeding systems, cows are subjected to many stress factors. Animals fed with a high-grain diet may have a decreased rumen pH, which would lead to subacute ruminal acidosis syndrome. The aim of this study was to investigate the evolution of microbial community composition in cows undergoing a dietary stress challenge. Twelve cows were subjected to a challenge period consisted in a rapid change of ration, from a normal (45.4:54.6 forage: concentrate) to a high-grain content diet (24.8:75.2 forage: concentrate) to induce sub-acute ruminal acidosis. Individual rumen fluid content samples were collected before (T0), and during the challenge (T3, T14, T28). DNA from rumen contents was extracted, purified, and sequenced to evaluate Bacterial populations and sequencing was performed on Illumina MiSeq. The effect of animal conditions on rumen microbial community was quantified through a linear mixed model. The acidogenic diet created 2 main clusters: ruminal hypomotility (RH) and milk fat depression (MFD). The microbial composition did not differ in T0 between the 2 groups, while during the challenge Ruminococcus spp., Treponema spp., Methanobrevibacter spp., and Methanosphaera spp. concentrations increased in RH cows; Succinivibrio spp. and Butyrivibrio spp. concentrations increased in MFD cows. Prevotella spp. and Ruminococcus spp., were negatively correlated, while Christenellaceae family were positively correlated with both Methanobrevibacter spp. and Methanosphaera spp. Moreover, the same diet affected differently cows' microbiota composition, underlying the impact of the host effect. Other studies are necessary to deepen the relationship between microbiota composition and host.

Udder health outcomes in Irish herds participating in cellcheck dry cow consults.

The CellCheck Dry Cow Consult (DCC) was developed by the CellCheck Technical Working Group to enable farmers to engage with their nominated vet to develop farm-specific selective dry cow therapy (SDCT) plans, where appropriate. This study evaluated the effect of the DCC on farmer decision-making around dry cow therapy, and the udder health impact of implementing SDCT, in study herds over the 2019 and 2020 dry periods. The DCC was a 3-h consult, delivered and funded as part of the Targeted Advisory Service on Animal Health (TASAH). Herds that completed a DCC were invited to register for a Dry Cow Review the following year. The combined data set for analysis across both years comprised of 439 herds and 25,357 cows. Available herd size ranged from 25 to 800. The median SCC of cows dried off with teat sealant only was 47,000 cells/ml before drying off in 2019 and 48,000 cells/ml at first milk recording in 2020, and 43,000 cells/ml before drying off in 2020 and 39,000 cells/ml at first milking recording in 2021. Following both the 2019 and 2020 dry periods, cows tended to converge toward a similar median SCC early in the following lactation, irrespective of prior dry cow treatment strategy. The uptake of SDCT was greater in Review herds, with 21% of cows receiving teat sealant only in 2020, compared with 16.3% of cows in herds participating in a Consult for the first time in 2020. At an individual cow level, in both years dry period new infection rate (NIR) was approximately 2.7% higher for cows treated with teat sealant only, than for those treated with both dry cow antibiotic tubes and teat sealant, and 1.2% higher than cows treated with antibiotic only. Regardless of treatment, there was a significant association between increasing parity and the risk of a dry period new infection. Increasing herd size had a statistically significant effect on the risk of dry period new infection rates. At a herd level, there was no statistically significant increase in NIR when SDCT was used compared with herds where blanket dry cow therapy was used. While not without risk, SDCT can be successfully implemented in Irish herds; however, constant attention to hygiene and management is essential. Though there are challenges to face, facilitating continued farmer education and engagement with professional guidance will be important.

Pregnancy risk in beef and dairy cows after supplementing semen with transforming growth factor beta-1 at the time of artificial insemination.

Our objective was to determine if the addition of a concentrated human recombinant transforming growth factor beta-1 (TGF) to bovine semen at the time of AI would result in increased risk of pregnancy in beef and dairy cows. Suckled beef cows (n = 1,132) in 11 herds across 2 states and lactating dairy cows (n = 2,208) in one organic-certified herd were enrolled. Beef cows received fixed-time AI (FTAI) following a 7 d CO-Synch + controlled internal drug release estrous synchronization protocol. Dairy cows were inseminated following observation of natural estrus expression. Cows received either no treatment as a control (CON) or 10 ng of TGF in 10 µL added through the cut-end of a thawed straw of semen immediately prior to AI. At the time of FTAI of beef cows, the mean ±â€…SD age was 5.0 ±â€…2.4 yr, BCS was 5.3 ±â€…0.7, and days postpartum was 78.2 ±â€…15.5 d. The overall pregnancy risk (PR) in beef cows was 55.2% to AI and 90.5% season-long. PR in beef cows was not affected (P = 0.27) by the addition of TGF (53.1% vs. 58.1%). Furthermore, there was no difference (P = 0.88) for season-long PR in beef cows that received TGF (91.2% vs. 91.5%). At the time of insemination of dairy cows, the mean ±â€…SD lactation was 3.0 ±â€…1.3 lactations, BCS was 2.9 ±â€…0.3, days in milk was 115.6 ±â€…56.6 d, and cows had received 2.4 ±â€…1.5 inseminations/cow. The overall pregnancy risk to AI in dairy cows was 23.1%. PR to AI for dairy cows was not affected (P = 0.32) by addition of TGF (22.0% vs. 23.8%). In conclusion, PR to AI was not affected by addition of TGF to thawed semen immediately prior to AI in beef or dairy cows.

Seminal plasma is the fluid portion of the ejaculate that is routinely removed or significantly diluted when preparing semen for artificial insemination. Seminal plasma has been shown to elicit changes to the tissues of the uterus at the time of insemination that improves pregnancy outcomes in rodents and swine. Here, we supplemented the molecule of seminal plasma, transforming growth factor beta-1, to semen at the time of artificial insemination in an attempt to improve pregnancy rates in beef and dairy cattle. In total, 3,340 cows were inseminated; half received no treatment, and the other half received a supplementation of transforming growth factor beta-1. We found that supplementing transforming growth factor beta-1 did not improve the pregnancy rate in beef or dairy cattle. We conclude that the pregnancy rate was not affected by the supplementation of transforming growth factor beta-1 to semen at the time of insemination. Future studies should consider the effects of transforming growth factor beta-1 on other pregnancy outcomes, such as calving rate, birth weight, and postnatal growth.

Insights into Effects of Combined Capric and Lauric Acid on Rumen Bacterial Composition.

This study used next-generation sequencing to assess the impact of combined capric acid (C10) and lauric acid (C12) on the ruminal bacterial composition. Eight Holstein cows were randomly assigned to two groups using a cross-over design. The cows were fed two silage-based diets with the addition of either 100 g of stearic acid per cow per day (control), or 50 g of capric acid and 50 g of lauric acid per cow per day (C10 + C12). On day 18, 250 mL of rumen fluid was collected from each cow, and DNA was isolated, amplified, and sequenced. Treatment did not alter bacterial diversity indices, the relative abundance of archaea, nor the fiber-degrading microorganisms, except for a decrease in Fibrobacter (from 2.9% to 0.7%; p = 0.04). The relative abundance of Prevotellaceae decreased (from 39.9% to 29.6%; p = 0.009), which is notable because some members help to efficiently utilize ammonia by releasing it slowly into the rumen. Furthermore, the relative abundance of Clostridia increased (from 28.4% to 41.5%; p = 0.008), which may have aided the increased ammonia-nitrogen levels in the rumen, as this class contains hyperammonia-producing members. Our study reveals alterations in bacterial abundances with implications for rumen ammonia levels, offering insights into potential strategies for modulating rumen fermentation processes and methane production in ruminant livestock.

Pregnancy Rates of Holstein Friesian Cows with Cavitary or Compact Corpus Luteum.

Cavitary corpora lutea are commonly observed during the estrous cycle in bovines. Since the quality of the corpus luteum (CL) is extremely important during embryo transfer when embryos are implanted into the recipient, the ultrasonographic examination of the CL is becoming more and more important in terms of the outcome of the procedure. In the present study, a total of 2477 ultrasonographic transrectal diagnoses were performed, and data were collected between the years of 2018 and 2020 in a large-scale Holstein Friesian dairy farm in Hungary. In 91.1% (n = 2257) and in 8.9% (n = 220) of the cases, compact CLs and cavitary CLs, respectively, were diagnosed at pregnancy diagnosis. The presence of a cavitary CL on the ovary at pregnancy diagnosis increased the odds of remaining open after pregnancy by 21 times compared to the presence of a compact CL (OR = 21.0, p < 0.001) in the cows. The presence of cavitary CL was not influenced either by month or season. Ovarian cysts were detected in 196 cases (8.0%) in the examined animals. The presence of a cavitary CL decreased by 9 times when an ovarian cyst was also diagnosed (OR = 9.0, 1.6% vs. 9.5%, p < 0.001). The presence of an ovarian cyst decreased the odds of established pregnancy by 81 times (OR = 81.1, p < 0.001). Based on our results, the presence of a cavitary CL between days 31 and 42 after artificial insemination is associated with a smaller chance of conception in Holstein Friesian cows. The presence of an ovarian cyst decreases the occurrence of cavitary CL and the chance of conception.

Microbiome and metabolome analyses of milk and feces from dairy cows with healthy, subclinical, and clinical mastitis.

Mastitis is commonly recognized as a localized inflammatory udder disease induced by the infiltration of exogenous pathogens. In the present study, our objective was to discern fecal and milk variations in both microbiota composition and metabolite profiles among three distinct groups of cows: healthy cows, cows with subclinical mastitis and cows with clinical mastitis. The fecal microbial community of cows with clinical mastitis was significantly less rich and diverse than the one harbored by healthy cows. In parallel, mastitis caused a strong disturbance in milk microbiota. Metabolomic profiles showed that eleven and twenty-eight molecules exhibited significant differences among the three groups in feces and milk, respectively. Similarly, to microbiota profile, milk metabolome was affected by mastitis more extensively than fecal metabolome, with particular reference to amino acids and sugars. Pathway analysis revealed that amino acids metabolism and energy metabolism could be considered as the main pathways altered by mastitis. These findings underscore the notable distinctions of fecal and milk samples among groups, from microbiome and metabolomic points of view. This observation stands to enhance our comprehension of mastitis in dairy cows.

Comparative analysis of computer vision algorithms for the real-time detection of digital dermatitis in dairy cows.

Digital dermatitis (DD) is a bovine claw disease responsible for ulcerative lesions on the planar aspect of the hoof. DD is associated with massive herd outbreaks of lameness and influences cattle welfare and production. Early detection of DD can lead to prompt treatment and decrease lameness. Computer vision (CV) provides a unique opportunity to improve early detection. The study aims to train and compare applications for the real-time detection of DD in dairy cows. Eight CV models were trained for detection and scoring, compared using performance metrics and inference time, and the best model was automated for real-time detection using images and video. Images were collected from commercial dairy farms while facing the interdigital space on the plantar surface of the foot. Images were scored for M-stages of DD by a trained investigator using the M-stage DD classification system with distinct labels for hyperkeratosis (H) and proliferations (P). Two sets of images were compiled: the first dataset (Dataset 1) containing 1,177 M0/M4H and 1,050 M2/M2P images and the second dataset (Dataset 2) containing 240 M0, 17 M2, 51 M2P, 114 M4H, and 108 M4P images. Models were trained to detect and score DD lesions and compared for precision, recall, and mean average precision (mAP) in addition to inference time in frame per second (FPS). Seven of the nine CV models performed well compared to the ground truth of labeled images using Dataset 1. The six models, Faster R-CNN, Cascade R-CNN, YOLOv3, Tiny YOLOv3, YOLOv4, Tiny YOLOv4, and YOLOv5s achieved an mAP between 0.964 and 0.998, whereas the other two models, SSD and SSD Lite, yielded an mAP of 0.371 and 0.387 respectively. Overall, YOLOv4, Tiny YOLOv4, and YOLOv5s outperformed all other models with almost perfect precision, perfect recall, and a higher mAP. Tiny YOLOv4 outperformed all other models with respect to inference time at 333 FPS, followed by YOLOv5s at 133 FPS and YOLOv4 at 65 FPS. YOLOv4 and Tiny YOLOv4 performed better than YOLOv5s compared to the ground truth using Dataset 2. YOLOv4 and Tiny YOLOv4 yielded a similar mAP of 0.896 and 0.895, respectively. However, Tiny YOLOv4 achieved both higher precision and recall compared to YOLOv4. Finally, Tiny YOLOv4 was able to detect DD lesions on a commercial dairy farm with high performance and speed. The proposed CV tool can be used for early detection and prompt treatment of DD in dairy cows. This result is a step towards applying CV algorithms to veterinary medicine and implementing real-time DD detection on dairy farms.

Analysis of differences in the rumen microbiome and metabolic function in prepartum dairy cows with different body condition scores.

BACKGROUND: The rumen is a crucial digestive organ for dairy cows. The rumen microbiota assists in the digestion of plant feed through microbe-mediated fermentation, during which the plant feed is transformed into nutrients for the cow's use. Variations in the composition and function of the rumen microbiome affect the energy utilization efficiency of dairy cows, which is one of the reasons for the varying body condition scores (BCSs). This study focused on prepartum Holstein dairy cows to analyze differences in rumen microbiota and metabolites among cows with different BCSs. Twelve prepartum dairy cows were divided into two groups, low BCS (LBCS, BCS = 2.75, n = 6) and high BCS (HBCS, BCS = 3.5, n = 6), to explore differences in microbial composition and metabolites. RESULTS: In the HBCS group, the genera within the phylum Firmicutes exhibited stronger correlations and greater abundances. Phyla such as Firmicutes, Patescibacteria, Acidobacteriota, Euryarchaeota, and Desulfobacterota, in addition to most of their constituent microbial groups, were significantly more abundant in the HBCS group than in the LBCS group. At the genus level, the abundances of Anaerovibrio, Veillonellaceae_UCG_001, Ruminococcus_gauvreauii_group, Blautia, Eubacterium, Prevotellaceae_YAB2003_group, Schwartzia, and Halomonas significantly increased in the HBCS group. The citrate cycle, involved in carbohydrate metabolism, exhibited a significant enrichment trend, with a notable increase in the abundance of its key substrate, citrate, in the HBCS group. This increase was significantly positively correlated with the differential bacterial genera. CONCLUSION: In this study, prepartum dairy cows with higher BCS exhibited greater abundance of Firmicutes. This study provides theoretical support for microbiological research on dairy cows with different BCSs and suggests that regulating the rumen microbiome could help maintain prepartum dairy cows within an optimal BCS range.

Association between body condition profiles, milk production and reproduction performance in Holstein and Normande cows.

Body-condition dynamics are known to affect the different steps of reproduction in cattle (cyclicity, estrus expression, fertilization, embryo development). This has led to a widespread idea that there is an ideal-target optimal body condition, but no clear profile has yet been identified. Here we investigated the relationships between body condition score (BCS) profiles and reproductive performance in dairy cows. Data were from Holstein or Normande herds in 6 French experimental farms. In the Holstein breed, we discriminated 4 BCS profiles based on combining BCS at calving (Low: around 2.6 points, or High: around 3.3 points) with BCS loss after calving (Moderate (M): ≤ 1.0 points, or Severe (S): > 1.0 points). The Low-M profile mostly included multiparous cows with higher milk yield and lower reproductive performance than cows in the 3 other profiles. Low-M cows that experienced abnormal ovarian activity had lower reproductive performance than their profile-mates. Moreover, 67% of Low-M cows kept the same profile at the following lactation. The High-S profile mostly included primiparous cows with lower milk yield and higher reproductive performance than cows in other profiles. In High-S cows, higher milk yields correlated to higher risk of failure to calf on first insemination. Moreover, 38% of High-S cows kept the same profile at the following lactation, and none changed to Low-M. The other 2 BCS profiles (Low-S and High-M) were intermediate in terms of milk yield and reproductive performance. In Normande, we discriminated 3 BCS profiles based on combining BCS at calving (Low: around 2.6 points, or High: around 3.5 points) with BCS loss after calving (Flat (F): flat with no loss, Moderate (M): around 0.5 points, or Severe (S): around 1.0 point). The Low-M and High-S profiles included cows with similar performance, even though High-S-profile cows showed better but not significantly different milk yield and reproduction performance. The High-F profile included cows that were more likely to experience abnormal ovarian activity and fail at first insemination than cows in other profiles. More than 50% of Normande cows with 2 successive lactations kept in the same BCS profile at the next lactation. Even though a low BCS at calving combined with severe BCS loss (more than 1 point) after calving was found to increase reproductive failure, there was no evidence of an optimal BCS profile for reproduction in dairy cows, and reproductive success or failure is multifactorial.

Altered bile acid and correlations with gut microbiome in transition dairy cows with different glucose and lipid metabolism status.

The transition from pregnancy to lactation is critical in dairy cows. Among others, dairy cows experience a metabolic stress due to a large change in glucose and lipid metabolism. Recent studies revealed that bile acids (BA), besides being involved in both the emulsification and solubilization of fats during intestinal absorption, can also affect the metabolism of glucose and lipids, both directly or indirectly by affecting the gut microbiota. Thus, we used untargeted and targeted metabolomics and 16S rRNA sequencing approaches to investigate the concentration of plasma metabolites and BA, the composition of the rectum microbial community, and assess their interaction in transition dairy cows. In Experiment 1, we investigated BA and other blood parameters and gut microbiota in dairy cows without clinical diseases during the transition period, which can be seen as well adapted to the challenge of changed glucose and lipid metabolism. As expected, we detected an increased plasma concentration of ß-hydroxybutyrate (BHBA) and nonesterified fatty acids (NEFA) but decreased concentration of glucose, cholesterol, and triglycerides (TG). Untargeted metabolomic analysis of the plasma revealed primary BA biosynthesis was one of the affected pathways, and was consistent with the increased concentration of BA in the plasma. A correlation approach revealed a complex association between BA and microbiota with the host plasma concentration of glucose and lipid metabolites. Among BA, chenodeoxycholic acid derivates such as glycolithocholic acid, taurolithocholic acid, lithocholic acid, taurochenodeoxycholic acid, and taurodeoxycholic acid were the main hub nodes connecting microbe and blood metabolites (such as glucose, TG, and NEFA). In Experiment 2, we investigated early postpartum dairy cows with or without hyperketonemia (HPK). As expected, HPK cows had increased concentration of NEFA and decreased concentrations of glucose and triglycerides. The untargeted metabolomic analysis of the plasma revealed that primary BA biosynthesis was also one of the affected pathways. Even though the BA concentration was similar among the 2 groups, the profiles of taurine conjugated BA changed significantly. A correlation analysis also revealed an association between BA and microbiota with the concentration in plasma of glucose and lipid metabolites (such as BHBA). Among BA, cholic acid and its derivates such as taurocholic acid, tauro α-muricholic acid, and taurodeoxycholic acid were the main hub nodes connecting microbe and blood metabolites. Our results indicated an association between BA, intestinal microbe, and glucose and lipid metabolism in transition dairy cows. These findings provide new insight into the adaptation mechanisms of dairy cows during the transition period.

Unveiling the Regulatory Role of SIRT1 in Oxidative Stress Response of bovine mammary cells.

High-yield dairy cows typically undergo intense cellular metabolism, leading to oxidative stress in their mammary tissues. Our study found that these high-yield cows had significantly elevated levels of hydrogen peroxide (H2O2), lipoperoxidase, and total antioxidant capacity in their blood, compared with ordinary cows. This increased oxidative stress is associated with heightened expression of genes such as GCLC, GCLM and SIRT1 and proteins such as SIRT1 in the mammary tissue of high-yield cows. MAC-T cells were stimulated with H2O2 at a concentration equal to the average H2O2 level in the serum of ethically high-yielding cows, as detected by an assay kit. Our observations revealed that short-term exposure (12 h) to H2O2 upregulated the expression of SIRT1 gene and protein. It also increased gene expression for SOD2, CAT, GCLC, GCLM, PGC-1α, and NQO1, elevated the phosphorylation of AMPK, and enhanced protein expression of PGC-1α, NQO1, Nrf2, and HO-1, while reducing the phosphorylation of NF-κB. Additionally, short-term H2O2 stimulation resulted in increased total antioxidant capacity, SOD, GSH, and CAT levels in the mammary epithelial cells of dairy cows. In contrast, prolonged exposure to H2O2 (24 h) yielded opposite results, indicating reduced antioxidant capacity. Further investigation showed that SIRT1 inhibitor (EX 527) could reverse the enhanced cellular antioxidant capacity triggered by short-term oxidative stress. However, it is crucial to note that while 12 h H2O2 stimulation improved antioxidant capacity, reactive oxygen species (ROS) and malondialdehyde (MDA) levels inside the cell gradually increased over time, suggesting greater damage under long-term stimulation. Conversely, the SIRT1 activator (SRT 2104) could reverse the reduced cellular antioxidant capacity caused by long-term oxidative stress and significantly inhibit the accumulation of ROS and MDA. Notably, SRT 2104 demonstrated similar effects in MAC-T cells during lactation. In summary, SIRT1 plays a crucial role in regulating the antioxidant capacity of mammary epithelial cells in dairy cows. This discovery provides valuable insights into the antioxidant mechanisms of mammary cells, which can serve as a theoretical foundation for future mammary health strategies.

Multiomics analysis revealed that the metabolite profile of raw milk is associated with lactation stage of dairy cows and could be affected by variations in the ruminal microbiota.

The nutritional components and quality of milk are influenced by the rumen microbiota and its metabolites at different lactation stages. Hence, rumen fluid and milk samples from 6 dairy cows fed the same diet were collected during peak, early mid- and later mid-lactation. Untargeted metabolomics and 16S rRNA sequencing were applied for analyzing milk and rumen metabolites, as well as rumen microbial composition, respectively. The levels of lipid-related metabolites, L-glutamate, glucose-1-phosphate and acetylphosphate in milk exhibited lactation-dependent attenuation. Maltol, N-acetyl-D-glucosamine, and choline, which are associated with milk flavor or coagulation properties, as well as L-valine, lansioside-A, clitocine and ginsenoside-La increased significantly in early mid- and later mid-lactation, especially in later mid-lactation. The obvious increase in rumen microbial diversities (Ace and Shannon indices) were observed in early mid-lactation compared with peak lactation. Twenty-one differential bacterial genera of the rumen were identified, with Succinivibrionaceae_UCG-001, Candidatus Saccharimonas, Fibrobacter, and SP3-e08 being significantly enriched in peak lactation. Rikenellaceae_RC9_gut_group, Eubacterium_ruminantium_group, Lachnospira, Butyrivibrio, Eubacterium_hallii_group, and Schwartzia were most significantly enriched in early mid-lactation. In comparison, only 2 bacteria (unclassified_f__Prevotellaceae and Prevotellaceae_UCG-001) were enriched in later mid-lactation. For rumen metabolites, LPE(16:0), L-glutamate and L-tyrosine had higher levels in peak lactation, whereas PE(17:0/0:0), PE(16:0/0:0), PS(18:1(9Z)/0:0), L-phenylalanine, dulcitol, 2-(methoxymethyl)furan and 3-phenylpropyl acetate showed higher levels in early mid- and later mid-lactation. Multiomics integrated analysis revealed that a greater abundance of Fibrobacter contributed to phospholipid content in milk by increasing ruminal acetate, L-glutamate and LysoPE(16:0). Prevotellaceae_UCG-001 and unclassified_f_Prevotellaceae provide substrates for milk metabolites of the same category by increasing ruminal L-phenylalanine and dulcitol contents. These results demonstrated that milk metabolomic fingerprints and critical functional metabolites during lactation, and the key bacteria in rumen related to them. These findings provide new insights into the development of functional dairy products.