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ABSTRACT: The objectives of this study were to determine if weaning would induce behavioral and physiological indicators of a negative affective state, and if supplementation of inactivated Lactobacillus helveticus (ILH) to dairy calves would reduce those indicators of negative affect during weaning. Male Holstein calves (n = 23) were enrolled in the study on d 1 of life. The calves were housed in individual pens in 1 of 4 rooms for the 42-d study. Calves began a step-down weaning from 9 L/d of milk replacer (MR), at 150 g of MR powder/L, on d 35 and received 6 L/d on d 35 to 36, 3 L/d on d 37 to 38, and 0.4 L/d on d 39 to 42. The MR was divided between 3 meals/d until the last 0.4 L/d phase, which was divided between 2 meals/d. Calves had ad libitum water access throughout the study and calf starter from d 28 onward. Within room, calves were assigned to 1 of 2 treatments: (1) control (CON; n = 11) and (2) 5 g of ILH/d split over and mixed into the 0800 h and 2000 h milk feedings from d 3 to 42 (ILH; n = 12). Lying behavior was recorded using HOBO data loggers from d 21 to 41. On d 33, 37, and 41, infrared eye images were taken to determine maximum eye temperature (MET), saliva samples were collected to determine cortisol concentration, and play assessments were conducted to quantify play behavior. On d 34, 38, and 42, blood samples were collected to determine blood serotonin concentration, whereas on d 38 and 39, calves were tested with a cognitive task. A subset of calves (n = 5/treatment) were euthanized to collect gut and brain tissue samples for serotonin concentration on d 43. Weaning resulted in fewer (d 37–41; tendency: d 36), but longer (d 38–41; tendency: d 37), lying bouts and reduced play (d 41), although no changes in lying time, MET, saliva cortisol, nor blood serotonin were detected with initiation of weaning. Supplementation of ILH was associated with less lying time throughout the study, and reduced play duration and higher salivary cortisol during weaning. No differences in lying bouts, play count, blood and tissue (colon, ileum, prefrontal cortex and brain stem) serotonin concentration, and time to complete the cognitive task were detected between the treatments. Overall, weaning induced behavioral changes indicative of negative affective state, and some behavioral differences were observed with ILH supplementation both before and during weaning, with some physiological changes observed during weaning.

ABSTRACT: The gastrointestinal microbial consortium in dairy cattle is critical to determining the energetic status of the dairy cow from birth through her final lactation. The ruminant's microbial community can degrade a wide variety of feedstuffs, which can affect growth, as well as production rate and efficiency on the farm, but can also affect food safety, animal health, and environmental impacts of dairy production. Gut microbial diversity and density are powerful tools that can be harnessed to benefit both producers and consumers. The incentives in the United States to develop Alternatives to Antibiotics for use in food-animal production have been largely driven by the Veterinary Feed Directive and have led to an increased use of probiotic approaches to alter the gastrointestinal microbial community composition, resulting in improved heifer growth, milk production and efficiency, and animal health. However, the efficacy of direct-fed microbials or probiotics in dairy cattle has been highly variable due to specific microbial ecological factors within the host gut and its native microflora. Interactions (both synergistic and antagonistic) between the microbial ecosystem and the host animal physiology (including epithelial cells, immune system, hormones, enzyme activities, and epigenetics) are critical to understanding why some probiotics work but others do not. Increasing availability of next-generation sequencing approaches provides novel insights into how probiotic approaches change the microbial community composition in the gut that can potentially affect animal health (e.g., diarrhea or scours, gut integrity, foodborne pathogens), as well as animal performance (e.g., growth, reproduction, productivity) and fermentation parameters (e.g., pH, short-chain fatty acids, methane production, and microbial profiles) of cattle. However, it remains clear that all direct-fed microbials are not created equal and their efficacy remains highly variable and dependent on stage of production and farm environment. Collectively, data have demonstrated that probiotic effects are not limited to the simple mechanisms that have been traditionally hypothesized, but instead are part of a complex cascade of microbial ecological and host animal physiological effects that ultimately impact dairy production and profitability.

ABSTRACT: This graduate student literature review provides an examination of the ontological adaptations of the calf's immune system and how the intestinal microbiota influences calf immune function in health and disease. Within dairy rearing systems, various nutritional and management factors have emerged as critical determinants of development influencing multiple physiological axes in the calf. Furthermore, we discuss how multiple pre- and postnatal maternal factors influence the trajectory of immune development in favor of establishing regulatory networks to successfully cope with the new environment, while providing early immune protection via immune passive transfer from colostrum. Additionally, our review provides insights into the current understanding of how the intestinal microbiota contributes to the development of the intestinal and systemic immune system in calves. Lastly, we address potential concerns related to the use of prophylactic antimicrobials and waste milk, specifically examining their adverse effects on intestinal health and metabolic function. By examining these factors, we aim to better understand the intricate relationship between current management practices and their long-term effect on animal health.

ABSTRACT: This study aimed to characterize the development of systemic and colon tissue resident B and γδ T cells in newborn calves from birth until weaning. At birth, calves have limited capacity to initiate immune responses, and the immune system gradually matures over time. Gamma delta (γδ) T cells are an important lymphocyte subset in neonatal calves that confer protection and promote immune tolerance. A total of 36 newborn calves were enrolled in a longitudinal study to characterize how systemic and colon tissue resident B and γδ T cells develop from birth until weaning. Blood and colon biopsy samples were collected on d 2, 28, and 42 to determine the proportions of various B and γδ T cell subsets by flow cytometry. We classified γδ T cells into different functional subsets according to the level of expression intensity of the coreceptors WC1.1 (effector function) and WC1.2 (regulatory function). Furthermore, naive B cells were classified based on the expression IgM receptor, and activation state was determined based on expression of CD21 and CD32, 2 receptors with opposing signals involved in B cell activation in early life. Additional colon biopsy samples were used for 16S sequencing, and microbial diversity data are reported. At birth, γδ T cells were the most abundant lymphocyte population in blood, accounting for 58.5% of the lymphocyte pool, after which the proportions of these cells declined to 38.2% after weaning. The proportion of γδ T cells expressing WC1.1 decreased by 50% from d 2 to d 28, whereas no change was observed in the expression of WC1.2. In the colon, there was a 50% increase of γδ T cells after weaning and the proportion of WC1.2+ γδ T cells doubled from d 28 to 42. The proportion of IgM+ B lymphocytes in blood increased from 23.6% at birth to 30% after weaning, were the proportion of B cells expressing CD21 increased by 25%, while the proportion of B cells expressing CD32 decreased by 30%. While no changes were observed for the overall proportion of IgM+ B lymphocytes in the colon, there was a 6-fold increase in the proportion of CD21+ B cells from pre- (d 28) to postweaning (d 42). Microbial diversity increased from d 2 of life to 28 and declined abruptly after weaning. The reduction in microbial diversity during weaning was negatively correlated with the increase in all γδ T cell subsets and CD21+ B cells. These data suggest that developmental adaptations after birth coordinate expansion of γδ T cells to provide early systemic protection, as well as to steer immune tolerance, while B cells mature over time. Additionally, the increase of colonic γδ T cells on d 42 suggests a protective role of these cells during weaning.

ABSTRACT: This study was designed to develop a protocol for repeated intramuscular indomethacin injections to replicate leaky-gut-like symptoms in male Holstein calves to model and study the detrimental effects of leaky gut on gut tissue function and inflammatory response. A generalized randomized block design was used to evaluate how repeated indomethacin intramuscular injections affected the development of leaky gut in 18 male Holstein calves. Animals were enrolled at 3 ± 1 d of life, and after 21 d of adaptation, they were randomly assigned to 1 of 3 treatments consisting of intramuscular saline or indomethacin injections every 12 h for 48 h: (1) control (CTL), saline injection, (2) low intramuscular indomethacin (INDO-L) dosed at 1.2 mg/kg of body weight (BW), and (3) high intramuscular indomethacin (INDO-H) dosed at 2.4 mg/kg of BW. During the challenge, milk intake, starter intake, fecal scores, and rectal temperature were measured daily, and BW was measured at the beginning and at the end of the challenge. Plasma samples were used to measure the recovery of markers of intestinal permeability before and after the challenge by dosing lactulose, d-mannitol, and chromium-EDTA. In addition, several cytokines were measured in plasma during the challenge. Calves were dissected at the end of the challenge to obtain tissue and digesta samples from the gastrointestinal tract and liver. No treatment differences were observed for starter and milk intakes, fecal scores, BW, and rectal temperature. The difference in marker concentrations between pre and post challenges was higher for INDO calves compared with CTL calves in the case of lactulose and chromium-EDTA. In addition, chemokine ligand 2 and 4 and IL-6 were higher for INDO-H calves compared with CTL. Both doses of indomethacin resulted in reductions in villus length and surface area in the distal jejunum and ileum and reductions in crypt depth and width in the colon. We showed that repeated indomethacin injections over a 48-h period induced leaky-gut-like symptoms in a region-specific manner, affecting mainly the distal section of the intestine. This outcome was characterized by histomorphological changes in the distal jejunum, ileum, and colon and by increased gut permeability. Interestingly, changes in liver morphology and immune function also occurred, possibly due to the increased translocation of foreign antigens breaching the epithelial cell wall. The leaky gut challenge model described here could be used to improve understanding of the pathogenesis of intestinal disorders in cattle and provide a reliable alternative for testing feed additives with intestinal health benefits.

ABSTRACT: The objectives of this study were to examine the effects of pelleted starter diets differing in starch and neutral detergent fiber (NDF) content when fed differing levels of milk replacer (MR) on nutrient digestibility, whole gastrointestinal tract fermentation, pH, and inflammatory markers in dairy calves around weaning. Calves were randomly assigned to 1 of 4 dietary treatments (n = 12 per treatment) in a 2 × 2 factorial design based on daily MR allowance and amount of starch in pelleted starter (SPS): 0.691 kg of MR per day [dry matter (DM) basis] with starter containing low or high starch (12.0% and 35.6% starch on DM basis, respectively), and 1.382 kg of MR per day (DM) with starter containing low or high starch. All calves were housed in individual pens with straw bedding until wk 5 when bedding was covered. Calves were fed MR twice daily (0700 and 1700 h) containing 24.5% crude protein (DM) and 19.8% fat (DM), and had access to pelleted starter (increased by 50 g/d if there were no refusals before weaning and then 200 g/d during and after weaning) and water starting on d 1. Calves arrived between 1 and 3 d of age and were enrolled into an 8-wk study, with calves undergoing step-down weaning during wk 7. Starting on d 35, an indwelling pH logger was inserted orally to monitor rumen pH until calves were dissected at the end of the study in wk 8. Higher SPS calves showed an increase in rumen pH magnitude (1.46 ± 0.07) compared with low SPS calves (1.16 ± 0.07), a decrease in rumen pH in wk 8 (high SPS: 5.37 ± 0.12; low SPS: 5.57 ± 0.12), and a decrease in haptoglobin in wk 8 (high SPS: 0.24 ± 0.06 g/L; low SPS: 0.49 ± 0.06 g/L). The majority of differences came from increased starter intake in general, which suggests that with completely pelleted starters the differences in starch and NDF do not elicit drastic changes in fermentation, subsequent end products, and any resulting inflammation in calves around weaning.

This research aimed: (i) to evaluate on-farm (FARM data) multiparametric models developed under controlled experiment (INRAE data) and based on non-invasive indicators to detect subacute ruminal acidosis (SARA) in dairy cows. We also aimed to recover high discrimination capacity, if needed, by (ii) building new models with combined INRAE and FARM data; and (iii) enriching the models increasing from 2 to 5 indicators per model. For model enrichment, we focused on indicators determinable on-farm by quick and inexpensive routine analysis. Fifteen commercial dairy farms were selected to cover a wide range of SARA risk. In each farm, four Holstein early-lactating healthy primiparous cows were selected based on their last on-farm recording of milk yield and somatic cell count analysis. Cows were equipped with a reticulo-rumen pH sensor. The pH kinetics were analysed over a subsequent 7-day period. Relative pH indicators were used to classify cows with or without SARA. Milk, blood, faeces, and urine were collected for analysis of the indicators included in the models developed by Villot et al. (2020) on INRAE data that were externally evaluated using FARM data. Then, new models based on the same indicators were developed combining INRAE and FARM data to test whether a possible loss in performance was due to a limited validity domain of model by Villot et al (2020). Finally, the models developed combining INRAE and FARM data were adapted to the on-farm application and enriched by increasing indicators from 2 to 5 per model using linear discriminant analysis and leave-one-out cross-validation. The sensitivities (true-positive rate) in external evaluation on FARM data were substantially lower than those from cross-validation by Villot et al. (2020) (range: 0.1–0.75 vs 0.79–0.96, respectively), and the specificities (true-negative rate) showed a larger range with lower minimum values (range: 0.18–1.0 vs 0.62–0.97, respectively). The sensitivities of new models developed combining INRAE and FARM data ranged from 0.63 to 0.77. Models involving blood cholesterol, β-hydroxybutyrate, haptoglobin, milk and blood urea, and models involving milk fat/protein ratio, dietary starch proportion, and milk fatty acids had the highest performances, whereas models including sieved faecal residues and urine pH had the lowest. Enriching models to three indicators per model improved sensitivity and specificity, but the inclusion of more indicators was less or not effective. Larger field trials are required to validate our results and to increase variability and validity domain of models.

High-starch diets (HSDs) fed to high-producing ruminants are often responsible for rumen dysfunction and could impair animal health and production. Feeding HSDs are often characterized by transient rumen pH depression, accurate monitoring of which requires costly or invasive methods. Numerous clinical signs can be followed to monitor such diet changes but no specific indicator is able to make a statement at animal level on-farm. The aim of this pilot study was to assess a combination of non-invasive indicators in dairy cows able to monitor a HSD in experimental conditions. A longitudinal study was conducted in 11 primiparous dairy cows fed with two different diets during three successive periods: a 4-week control period (P1) with a low-starch diet (LSD; 13% starch), a 4-week period with an HSD (P2, 35% starch) and a 3-week recovery period (P3) again with the LSD. Animal behaviour was monitored throughout the experiment, and faeces, urine, saliva, milk and blood were sampled simultaneously in each animal at least once a week for analysis. A total of 136 variables were screened by successive statistical approaches including: partial least squares-discriminant analysis, multivariate analysis and mixed-effect models. Finally, 16 indicators were selected as the most representative of a HSD challenge. A generalized linear mixed model analysis was applied to highlight parsimonious combinations of indicators able to identify animals under our experimental conditions. Eighteen models were established and the combination of milk urea nitrogen, blood bicarbonate and feed intake was the best to detect the different periods of the challenge with both 100% of specificity and sensitivity. Other indicators such as the number of drinking acts, fat:protein ratio in milk, urine, and faecal pH, were the most frequently used in the proposed models. Finally, the established models highlight the necessity for animals to have more than 1 week of recovery diet to return to their initial control state after a HSD challenge. This pilot study demonstrates the interest of using combinations of non-invasive indicators to monitor feed changes from a LSD to a HSD to dairy cows in order to improve prevention of rumen dysfunction on-farm. However, the adjustment and robustness of the proposed combinations of indicators need to be challenged using a greater number of animals as well as different acidogenic conditions before being applied on-farm.

Although hay is the foundation of most equine diets, horse owners rarely ask for biochemical analysis and the routine practice is to choose hay based on its ‘perceived‘ nutritional value. The present study aimed at exploring the relationship between sensory properties as perceived by sight, touch and smell, and the nutritional value of hay measured by biochemical analysis using a ‘free sorting task’ method. Fifty-four non-expert participants were asked individually to: (1) observe 21 hays samples, (2) group together hays that they perceived as similar for each of the three modalities (hay appearance, odour or texture) and (3) characterize each formed group with a maximum of five descriptive terms. For each modality, results were recorded in a contingency matrix (hays × terms) where only terms cited at the minimum five times for at least one sample, were kept for data analysis. A correspondence analysis (CA) was performed on the contingency matrix to plot both samples and descriptive terms on a χ2 metric map. Then, a Hierarchical Ascending Classification (HAC) was performed on the coordinates of samples in the CA space. Clusters were identified by truncating the HAC tree-diagrams. The attributes that defined the best resulting clusters were identified by computing their probability of characterizing a cluster. Correlations were computed between each biochemical parameter on one hand, and the first two dimensions of the CA map on the other. Finally, correlations between the values of each hay on the first dimension of the three CA maps (appearance, odour and texture) were computed. Hedonic descriptive terms were primarily used for describing odour and texture modalities. For describing hay appearance, participants spontaneously used visual cues referring to colour or aspect. Based on the tree-diagrams resulting from the HAC, 3, 5 and 2 groups were clustered, respectively for appearance, odour and texture description. Digestible energy was correlated to the first dimension on the three CA maps, whereas CP was correlated to the first dimension of the CA appearance map only. While NDF value was correlated to the first and second dimensions on the CA odour map only, ADF content was correlated to the first dimension on the three CA maps. Non-fibre carbohydrates were correlated to the first dimension of the CA appearance map only. The similarity-based approach which is part of the standard toolbox of food sensory evaluation by untrained consumers was well adapted to animal feeds evaluation by non-experts.

Subacute ruminal acidosis (SARA) is usually characterized by abnormal and intermittent drops in rumen pH. Nevertheless, high individual animal variability in rumen pH and the difference in measurement methods for pH data acquisition decrease the sensitivity and accuracy of pH indicators for detecting SARA in ruminants. The aim of this study was to refine rumen pH indicators in long-term SARA based on individual dairy cow reticulo-rumen pH kinetics. Animal performances and rumen parameters were studied weekly in order to validate SARA syndrome and rumen pH was continuously measured using reticulo-rumen sensors. In total, 11 primiparous dairy cows were consecutively fed two different diets for 12 successive weeks: a control diet as low-starch diet (LSD; 13% starch for 4 weeks in period 1), an acidotic diet as high-starch diet (HSD; 32% starch for 4 weeks in period 2), and again the LSD diet (3 weeks in period 3). There was a 1-week dietary transition between LSD and HSD. Commonly used absolute SARA pH indicators such as daily average, area under the curve (AUC) and time spent below pH

This research aimed: (i) to evaluate on-farm (FARM data) multiparametric models developed under controlled experiment (INRAE data) and based on non-invasive indicators to detect subacute ruminal acidosis (SARA) in dairy cows. We also aimed to recover high discrimination capacity, if needed, by (ii) building new models with combined INRAE and FARM data; and (iii) enriching the models increasing from 2 to 5 indicators per model. For model enrichment, we focused on indicators determinable on-farm by quick and inexpensive routine analysis. Fifteen commercial dairy farms were selected to cover a wide range of SARA risk. In each farm, four Holstein early-lactating healthy primiparous cows were selected based on their last on-farm recording of milk yield and somatic cell count analysis. Cows were equipped with a reticulo-rumen pH sensor. The pH kinetics were analysed over a subsequent 7-day period. Relative pH indicators were used to classify cows with or without SARA. Milk, blood, faeces, and urine were collected for analysis of the indicators included in the models developed by Villot et al. (2020) on INRAE data that were externally evaluated using FARM data. Then, new models based on the same indicators were developed combining INRAE and FARM data to test whether a possible loss in performance was due to a limited validity domain of model by Villot et al (2020). Finally, the models developed combining INRAE and FARM data were adapted to the on-farm application and enriched by increasing indicators from 2 to 5 per model using linear discriminant analysis and leave-one-out cross-validation. The sensitivities (true-positive rate) in external evaluation on FARM data were substantially lower than those from cross-validation by Villot et al. (2020) (range: 0.1-0.75 vs 0.79-0.96, respectively), and the specificities (true-negative rate) showed a larger range with lower minimum values (range: 0.18-1.0 vs 0.62-0.97, respectively). The sensitivities of new models developed combining INRAE and FARM data ranged from 0.63 to 0.77. Models involving blood cholesterol, β-hydroxybutyrate, haptoglobin, milk and blood urea, and models involving milk fat/protein ratio, dietary starch proportion, and milk fatty acids had the highest performances, whereas models including sieved faecal residues and urine pH had the lowest. Enriching models to three indicators per model improved sensitivity and specificity, but the inclusion of more indicators was less or not effective. Larger field trials are required to validate our results and to increase variability and validity domain of models., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

High-starch diets (HSDs) fed to high-producing ruminants are often responsible for rumen dysfunction and could impair animal health and production. Feeding HSDs are often characterized by transient rumen pH depression, accurate monitoring of which requires costly or invasive methods. Numerous clinical signs can be followed to monitor such diet changes but no specific indicator is able to make a statement at animal level on-farm. The aim of this pilot study was to assess a combination of non-invasive indicators in dairy cows able to monitor a HSD in experimental conditions. A longitudinal study was conducted in 11 primiparous dairy cows fed with two different diets during three successive periods: a 4-week control period (P1) with a low-starch diet (LSD; 13% starch), a 4-week period with an HSD (P2, 35% starch) and a 3-week recovery period (P3) again with the LSD. Animal behaviour was monitored throughout the experiment, and faeces, urine, saliva, milk and blood were sampled simultaneously in each animal at least once a week for analysis. A total of 136 variables were screened by successive statistical approaches including: partial least squares-discriminant analysis, multivariate analysis and mixed-effect models. Finally, 16 indicators were selected as the most representative of a HSD challenge. A generalized linear mixed model analysis was applied to highlight parsimonious combinations of indicators able to identify animals under our experimental conditions. Eighteen models were established and the combination of milk urea nitrogen, blood bicarbonate and feed intake was the best to detect the different periods of the challenge with both 100% of specificity and sensitivity. Other indicators such as the number of drinking acts, fat:protein ratio in milk, urine, and faecal pH, were the most frequently used in the proposed models. Finally, the established models highlight the necessity for animals to have more than 1 week of recovery diet to return to their initial control state after a HSD challenge. This pilot study demonstrates the interest of using combinations of non-invasive indicators to monitor feed changes from a LSD to a HSD to dairy cows in order to improve prevention of rumen dysfunction on-farm. However, the adjustment and robustness of the proposed combinations of indicators need to be challenged using a greater number of animals as well as different acidogenic conditions before being applied on-farm.

Although hay is the foundation of most equine diets, horse owners rarely ask for biochemical analysis and the routine practice is to choose hay based on its 'perceived' nutritional value. The present study aimed at exploring the relationship between sensory properties as perceived by sight, touch and smell, and the nutritional value of hay measured by biochemical analysis using a 'free sorting task' method. Fifty-four non-expert participants were asked individually to: (1) observe 21 hays samples, (2) group together hays that they perceived as similar for each of the three modalities (hay appearance, odour or texture) and (3) characterize each formed group with a maximum of five descriptive terms. For each modality, results were recorded in a contingency matrix (hays × terms) where only terms cited at the minimum five times for at least one sample, were kept for data analysis. A correspondence analysis (CA) was performed on the contingency matrix to plot both samples and descriptive terms on a χ2 metric map. Then, a Hierarchical Ascending Classification (HAC) was performed on the coordinates of samples in the CA space. Clusters were identified by truncating the HAC tree-diagrams. The attributes that defined the best resulting clusters were identified by computing their probability of characterizing a cluster. Correlations were computed between each biochemical parameter on one hand, and the first two dimensions of the CA map on the other. Finally, correlations between the values of each hay on the first dimension of the three CA maps (appearance, odour and texture) were computed. Hedonic descriptive terms were primarily used for describing odour and texture modalities. For describing hay appearance, participants spontaneously used visual cues referring to colour or aspect. Based on the tree-diagrams resulting from the HAC, 3, 5 and 2 groups were clustered, respectively for appearance, odour and texture description. Digestible energy was correlated to the first dimension on the three CA maps, whereas CP was correlated to the first dimension of the CA appearance map only. While NDF value was correlated to the first and second dimensions on the CA odour map only, ADF content was correlated to the first dimension on the three CA maps. Non-fibre carbohydrates were correlated to the first dimension of the CA appearance map only. The similarity-based approach which is part of the standard toolbox of food sensory evaluation by untrained consumers was well adapted to animal feeds evaluation by non-experts.

Subacute ruminal acidosis (SARA) is usually characterized by abnormal and intermittent drops in rumen pH. Nevertheless, high individual animal variability in rumen pH and the difference in measurement methods for pH data acquisition decrease the sensitivity and accuracy of pH indicators for detecting SARA in ruminants. The aim of this study was to refine rumen pH indicators in long-term SARA based on individual dairy cow reticulo-rumen pH kinetics. Animal performances and rumen parameters were studied weekly in order to validate SARA syndrome and rumen pH was continuously measured using reticulo-rumen sensors. In total, 11 primiparous dairy cows were consecutively fed two different diets for 12 successive weeks: a control diet as low-starch diet (LSD; 13% starch for 4 weeks in period 1), an acidotic diet as high-starch diet (HSD; 32% starch for 4 weeks in period 2), and again the LSD diet (3 weeks in period 3). There was a 1-week dietary transition between LSD and HSD. Commonly used absolute SARA pH indicators such as daily average, area under the curve (AUC) and time spent below pH<5.8 and pH<6 were processed from absolute (raw) daily kinetics. Then signal processing was applied to raw pH values in order to calculate relative pH indicators by filtering and normalizing data to remove inter-individual variability, sensor drift and sensor noise. Normalized AUC, times spent below NpH<-0.3 and NpH<-0.5, NpH range and NpH standard deviation were calculated. Those relative pH indicators were compared with commonly used pH indicators to assess their ability to detect SARA. This syndrome induced by HSD was confirmed by consistent expected changes in milk quality, dry matter intake and acetate : propionate ratio in the rumen, whereas the ruminal concentration of lipopolysaccharide was increased. Commonly used pH SARA indicators were not able to discriminate SARA syndrome due to high animal variability and sensor drift and noise, whereas relative pH indicators developed in this study appeared more relevant for SARA detection as assessed by receiver operating characteristic tests. This work shows that absolute pH kinetics should be corrected for drift, noise and animal variability to produce relative pH indicators that are more robust for SARA detection. These relative pH indicators could be more relevant for identifying affected animals in a herd and also for comparing SARA risk among studies.