Cardiac vagal tone, plasma cortisol, and dehydroepiandrosterone response to an ACTH challenge in lame and nonlame dairy cows.

We studied the adrenocortical and vagal tone responses to a single ACTH challenge in lame (n = 9) vs nonlame (n = 9) dairy cows. Cows were paired according to parity, days in milk, and milk yield. Plasma cortisol and dehydroepiandrosterone concentrations and cardiac vagal tone response (high-frequency component of heart rate variability) were compared after intravenous ACTH administration. Baseline, minimum or maximum, amplitude of the response and area under the response curve were compared. No difference was detected between groups in the cortisol response. Dehydroepiandrosterone was irresponsive to ACTH treatment, and concentrations did not differ between lame and nonlame cows. Vagal tone decreased in response to the ACTH treatment. High frequency component of heart rate variability was lower in the lame group at all sampling times. Lameness was associated with delayed return to baseline. We concluded that the adrenal response capacity is not influenced by lameness, which supports the concept of lameness being a chronic intermittent rather than a chronically persistent stressor. Dehydroepiandrosterone concentrations were not proven to be useful indicators of hypothalamus-pituitary axis dysfunction in cattle. A decreased vagal contribution to heart rate variability-possibly coupled with increased sympathetic modulation-was observed in lame cows, which suggests that lameness affects the mechanisms underlying the action of ACTH on cardiovascular activity.

Serum metabolic fingerprinting of pre-lameness dairy cows by GC-MS reveals typical profiles that can identify susceptible cows.

The objectives of this study were to identify metabolite fingerprints in the serum related to amino acid (AA), carbohydrate, and lipid metabolism in transition dairy cows at -8 and -4 wks prior to parturition, at +2 wks postpartum during lameness diagnosis as well as at +4 and +8 wks after parturition. All cases of lameness occurred at around +2 wks after parturition. Out of 100 dairy cows included in this nested case-control study only 6 pregnant multiparous (parity: 3.0 ± 0.6, Mean ± SEM) Holstein dairy cows with lameness only and 20 healthy control cows (CON) were selected for serum GC-MS metabolomics analysis. All cows selected were not injured mechanically and had similar parity (3.3 ± 0.6) and body condition score (BCS). A total of 29 metabolites were identified and quantified in the serum. Results showed that 18 and 15 metabolites differentiated pre-lame cows from CON ones at -8 and -4 wks prior to parturition. Ten metabolites were found altered at the week of lameness diagnosis. Of note: pre-lame cows were characterized by greater concentrations of several amino acids including Gly, Leu, Phe, Ser, Val, D-mannose, Myo-inositol, and phosphoric acid (PA) at -8 and -4 wks prior to lameness and at the week of lameness diagnosis. At +4 wks after parturition 11 metabolites were altered in lameness cows, and at +8 wks there were 13 metabolites that differentiated the two groups. The high accuracy of the top 6 metabolites at -8 wks prior to parturition or approximately 9-11 wks before lameness diagnosis (Glu, Orn, Phe, Ser, Val, and PA) and another 5 metabolites at -4 wks before parturition, or approximately 5-7 wks before lameness diagnosis (Leu, Orn, Phe, Ser, and D-mannose) suggest that those metabolites may serve as potential monitoring biomarkers of lameness prior to lameness diagnosis. Data also showed multiple alterations during the week of lameness as well as at +4 and +8 wks postpartum suggesting lame cows are not metabolically healthy several weeks after the incidence of lameness. SIGNIFICANCE: Lameness is one of the top three health issues of dairy cows in Canada that influences early culling of dairy cows. Despite a few efforts, there is scarcity of data regarding metabolic alterations that precede, associate, and follow lameness. We investigated whether alterations in the metabolite signatures prior, during, and after development of lameness can be used to screen cows for susceptibility to lameness, characterize lameness from the metabolic prospective, and predict the outcome of this economically important health issue of dairy cows. The results demonstrate typical metabotypes as shown by increased serum concentrations of Val, Gly, Ser, Leu, Phe, D-mannose, myo-inositol, and phosphoric acid at -8 and -4 wks prior to parturition (or -6 to -10 wks prior to occurrence of lameness) and at the week of lameness diagnosis.

Correlation between synovial fluid cytokine concentrations and limb function in normal dogs and in dogs with lameness from spontaneous osteoarthritis.

OBJECTIVE: To determine the relationship between synovial biomarker concentrations and severity of lameness and to assess the ability to differentiate normal from osteoarthritic joints with synovial biomarker concentrations. STUDY DESIGN: Prospective clinical study. SAMPLE POPULATION: Twelve hounds with no evidence of osteoarthritis (OA) and 27 client-owned dogs with unilateral lameness and joint pain in a single joint from naturally occurring OA. METHODS: Enrollment in the OA group required a history of lameness, radiographic evidence of OA on orthogonal joint radiographs, and ≥6% gait asymmetry between contralateral limbs. The concentrations of 14 synovial OA biomarkers in synovial samples obtained after gait analysis were measured with enzyme-linked immunosorbent assays and compared between normal and OA joints. RESULTS: Concentrations of monocyte chemoattractant protein (MCP)-1, substance P, interleukin (IL)-6, IL-8, KC-like, matrix metalloproteinase (MMP)-1, and MMP-3 were greater (P ≤ .05) in OA than in normal joints. The concentrations of bradykinin and tissue inhibitors of metalloproteinase-4 were decreased in OA compared with normal joints. Monocyte chemoattractant protein 1 was identified as the most accurate marker to distinguish OA from normal joints. No correlation was detected between any OA biomarker concentration, individually or in combination, and severity of gait asymmetry at the walk. CONCLUSION: Differences in proinflammatory and anti-inflammatory biomarkers were detected between OA and normal joints, but no relationship was identified between biomarker concentrations and gait asymmetry in dogs with OA. CLINICAL IMPACT: This information will help guide future studies to elucidate how factors such as disease chronicity, severity, and etiology affect these relationships.

Chondronecrosis with osteomyelitis in broilers: Further defining a bacterial challenge model using the wire flooring model.

Lameness in broiler chickens is a significant animal welfare and financial issue. Bacterial chondronecrosis with osteomyelitis (BCO) leading to lameness can be enhanced by rearing young broilers on wire flooring. Using the wire floor system, we identified Staphylococcus agnetis as the predominant isolate in BCO of the proximal tibiae and femora, and blood of lame broilers. Administration of S. agnetis isolates in water can induce lameness. We now report that the wire floor system increases bacterial translocation into the blood stream. We have also determined that approximately 105 CFU/mL is the minimum effective dose in the drinking water and that challenge at 10, 20, or 30 days of age produces similar incidences of lameness. BCO isolates of S. agnetis are much more effective than other Staphylococcus species and can overwhelm the protective effects of some commercial probiotics. Finally, we also demonstrated that the BCO lameness induced by administration of S. agnetis in the drinking water is transmissible to unchallenged broilers in the same pen.

An investigation into blood microbiota and its potential association with Bacterial Chondronecrosis with Osteomyelitis (BCO) in Broilers.

Bacterial chondronecrosis with osteomyelitis (BCO) is a common cause of lameness in commercial broiler chickens worldwide. BCO represents substantial production loss and welfare issues of chickens. The bacterial species or communities underlying BCO pathogenesis still remain to be fully characterized. To gain insights on blood microbiota in broilers and its potential association with BCO, blood samples collected from healthy (n = 240) and lame (n = 12) chickens were analyzed by deep sequencing of 16S RNA genes. The chicken blood microbiota were dominated by Proteobacteria (60.58% ± 0.65) followed by Bactroidetes (13.99% ± 0.29), Firmicutes (11.45% ± 0.51), Actinobacteria (10.21% ± 0.37) and Cyanobacteria (1.96% ± 0.21) that constituted 98.18% (± 0.22) of the whole phyla. The bacterial communities consist of 30-40 OTUs in the blood of broiler chickens, regardless of ages and other environmental or host conditions, and the blood microbiomes of BCO chickens were largely distinct from those of healthy chickens. In addition, Linear discriminant analysis (LDA) effect size (LEfSe) method revealed that Staphylococcus, Granulicatella, and Microbacterium were significantly enriched in BCO chickens as compared to healthy chickens. The results from this study have significant implications in understanding blood microbiota present in broiler chickens and its potential role in BCO pathogenesis.

Analysis of Mineral Elements, Metabolism, and Inflammation Indexes in the Plasma of Dairy Cows Suffering from Different Degrees of Lameness.

In the plasma of dairy cows with 1-5 points of lameness, the mineral elements [calcium (Ca), iron (Fe), copper (Cu), zinc (Zn), iodine (I), selenium (Se), molybdenum (Mo), and chromium (Cr)], the energy metabolic indicators [triglyceride (TG), glucose (Glu), total cholesterol (CHO), nonesterified fatty acids (NEFA), ß-hydroxybutyrate (BHBA), lactate (LA), and blood urea nitrogen (BUN)], and inflammatory indexes [bovine haptoglobin (BoHp), histamine (HIS), and immunoglobulin G (IgG)] were measured, respectively. Furthermore, the correlations of the measured indicators with the degrees of lameness were analyzed. The results showed that in the plasma of dairy cows with 2/3-5 points of lameness, for the mineral elements' levels, Ca, Cu, I, Se, and Fe significantly decreased, Cr significantly increased, and Mo showed a decreasing trend; for levels of the energy metabolism indicators, NEFA and BHBA significantly decreased, BUN and LA significantly increased, and Glu, CHO, and TG showed an increasing trend; for inflammation indexes, the concentrations of HIS, BoHp, and IgG all significantly increased; and further analysis indicated that the Mo, Fe, NEFA, BUN, BHBA, IgG, Ca, and Se had a significant correlation with the degrees of lameness. Resulting data revealed the changes of mineral elements, metabolism, and inflammation indexes in the plasma of dairy cows suffering from different degrees of lameness, which will provided basic knowledge for in-depth understanding of lameness in dairy cows.

Oxidative stress and imbalance of mineral metabolism contribute to lameness in dairy cows.

The objective of this study was to investigate correlations between oxidative stress, metabolism of mineral elements, and lameness in dairy cows. Forty multiparous Chinese Holstein dairy cows were selected and divided into two groups (healthy vs lame, n = 20) by gait score. The experiment lasted for 60 days and samples of hair, blood, and hoof were collected at days 0, 30, and 60 of experiment period, individually. Compared with healthy cows, elevation of MDA, CTX-II, COMP levels, and GSSG/GSH ratio together with depletion of SOD and MT levels in the serum were revealed in lame cows. Simultaneously, significant decreased contents of Zn, Cu, and Mn in the serum, hair, and hoof samples were shown in lame cows, but there was no obvious difference in contents of P, Mg, and Ca (except hoof Ca) in the serum, hair, and hoof between healthy and lame cows. In addition, histological examination and the hardness test demonstrated a poor hoof quality in lame cows. In summary, oxidative stress is implicated in the pathogenesis of lameness caused by imbalance of nutrients (especially selective minerals promoting healthy hoof growth) in dairy cows.

Gene biomarkers in peripheral white blood cells of horses with experimentally induced osteoarthritis.

OBJECTIVE: To use microarray analysis to identify genes that are differentially expressed in horses with experimentally induced osteoarthritis. ANIMALS: 24 horses. PROCEDURES: During arthroscopic surgery, a fragment was created in the distal aspect of the radiocarpal bone in 1 forelimb of each horse to induce osteoarthritis. At day 14 after osteoarthritis induction, horses began exercise on a treadmill. Blood and synovial fluid samples were collected before and after surgery. At day 70, horses were euthanized and tissues were harvested for RNA analysis. An equine-specific microarray was used to measure RNA expression in peripheral WBCs. These data were compared with mRNA expression (determined via PCR assay) in WBCs, cartilage, and synovium as well as 2 protein biomarkers of cartilage matrix turnover in serum and synovial fluid. RESULTS: A metalloproteinase domain-like protein decysin-1 (ADAMDEC1), glucose-regulated protein (GRP) 94, hematopoietic cell signal transducer (HCST), Unc-93 homolog A (hUNC-93A), and ribonucleotide reductase M2 polypeptide (RRM2) were significantly differentially regulated in WBCs of horses with osteoarthritis, compared with values prior to induction of osteoarthritis. There was correlation between the gene expression profile in WBCs, cartilage, and synovium and the cartilage turnover proteins. Gene expression of ADAMDEC1, hUNC-93A, and RRM2 in WBCs were correlated when measured via microarray analysis and PCR assay. CONCLUSIONS AND CLINICAL RELEVANCE: Expression of ADAMDEC1, GRP94, HCST, hUNC-93A, and RRM2 was differentially regulated in peripheral WBCs obtained from horses with experimentally induced osteoarthritis. Gene expression of ADAMDEC1, hUNC-93A, and RRM2 in peripheral WBCs has the potential for use as a diagnostic aid for osteoarthritis in horses.

Studies on "stiffness of extremities disease" in the yak (Bos mutus).

Wild yak (Bos mutus) are affected by a disorder known colloquially as "stiffness of extremities disease," characterized by emaciation, lameness, stiffness in the gait, enlargement of the costochondral junctions, and abnormal curvature in the long bones. Results from preliminary epidemiologic and clinical observations suggested that this was a local, nutritional and metabolic disease associated with some mineral deficiency. Our objective was to determine the possible relationship between this disease and phosphorus (P) deficiency. We found that P concentrations in forage samples from affected areas were significantly lower than were those from unaffected areas, and the mean calcium:P ratio in the affected forage was 14:1. Phosphorus concentrations of blood, bone, teeth, and hair from affected yak were also significantly lower than were those from reference yak. Serum P levels of affected animals were much lower than were those of reference yak, whereas serum alkaline phosphatase levels were significantly higher than were those from reference yak. The P deficiency disease could be cured with supplement of disodium hydrogen phosphate (Na(2)HPO(4)). We conclude that the disease is mainly caused by P deficiency in forage.

The relationship between serum biotin and oxidant/antioxidant activities in bovine lameness.

Serum biotin concentrations, erythrocyte superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), reduced glutathione (GSH) and plasma thiobarbituric acid reactive substances (TBARS) were measured in 36 dairy cows, 18 of them were healthy and served as control. In the 18 cows with lameness problems, there were 5 cows with interdigital necrobacillosis, 5 cows with subsolar abscessation, 2 cows with solar ulcers, 2 cows with white line disease, 2 cows with chronic laminitis and 2 cows with septic arthritis. The degree of lameness was estimated to be slight in 3 cows, moderate in 11 cows and severe in 4 cows. Plasma fibrinogen levels and TBARS concentrations were increased significantly (P≤0.05) in lame cows compared to control group. The antioxidant enzymes GSH-Px, and CAT concentrations were increased significantly (P≤0.05) in lame cows. The level of reduced glutathione and the activity of SOD were significantly decreased in affected cows compared to healthy ones. Serum biotin levels in healthy cows ranged from 2.25 to 3.5ng/ml while in lame cows, biotin levels ranged from 1.17 to 2.3ng/ml. Biotin levels correlated positively with blood GSH (r=0.870, P≤0.05), (r=0.735, P≤0.05) and with GSH-Px (r=0.539, P≤0.05), (r=0.637, P≤0.05) and with SOD (r=0.637, P≤0.05), (r=0.449, P≤0.05) and with catalase (r=0.533, P≤0.05), (r=0.585, P≤0.05) in both healthy and lameness affected subjects, respectively.

Effects of extracorporeal shock wave therapy and polysulfated glycosaminoglycan treatment on subchondral bone, serum biomarkers, and synovial fluid biomarkers in horses with induced osteoarthritis.

OBJECTIVE: To evaluate effects of extracorporeal shock wave therapy (ESWT) and polysulfated glycosaminoglycan treatment (PSGAGT) on subchondral bone (SCB), serum biomarkers, and synovial fluid biomarkers in horses with induced osteoarthritis. ANIMALS: 24 healthy 2- to 3-year-old horses. PROCEDURES: An osteochondral fragment was created on the distal aspect of the radial carpal bone in 1 middle carpal joint of each horse. Horses were randomly allocated to receive local application of ESWT (days 14 and 28; n = 8), PSGAGT (IM, q 4 d for 28 days; 8), or a sham ESWT probe (placebo; days 14 and 28; 8). Serum biomarkers were measured every 7 days, and synovial fluid biomarkers were measured every 14 days. Bone density was measured by use of computed tomography on days 0 and 70, and microdamage and bone formation variables were compared among groups at the end of the study (day 70). RESULTS: There was no significant effect of ESWT or PSGAGT on any bone variable. Serum osteocalcin concentration was significantly greater in horses that received ESWT, compared with placebo-treated horses, and serum concentration of the C-terminal telopeptide of type I collagen was significantly higher in horses that received ESWT, compared with placebo- and PSGAG-treated horses. Concentrations of the synovial fluid epitope CS846 were significantly higher in joints with osteoarthritis treated with ESWT CONCLUSIONS AND CLINICAL RELEVANCE: Treatment of osteoarthritis with ESWT had no effect on SCB but did induce increases in serum biomarkers indicative of bone remodeling. Treatment of osteoarthritis with PSGAG had no effect on SCB or biomarkers.

Influence of lameness on follicular growth, ovulation, reproductive hormone concentrations and estrus behavior in dairy cows.

The objective of this study was to examine the effect of a chronic stressor, lameness, on reproductive parameters. Seventy cows 30-80 days post-partum were scored for lameness and follicular phases synchronized with GnRH followed seven days later by prostaglandin (PG). Fifteen Lame animals did not respond to GnRH ovarian stimulation. Milk progesterone for 5 days prior to PG was lower in the remaining Lame cows than Healthy herdmates. Fewer Lame cows ovulated (26/37 versus 17/18; P = 0.04) and the interval from PG to ovulation was shorter in Lame cows. In Subset 1 (20 animals), the LH pulse frequency was similar in ovulating animals (Lame and Healthy) but lower in Lame non-ovulators. An LH surge always preceded ovulation but lameness did not affect the interval from PG to LH surge onset or LH surge concentrations. Before the LH surge, estradiol was lower in non-ovulating cows compared to those that ovulated and estradiol concentrations were positively correlated with LH pulse frequency. In Subset 2 (45 cows), Lame ovulating cows had a less intense estrus than Healthy cows, although Lame cows began estrus and stood-to-be-mounted earlier than Healthy cows. In conclusion, we have identified several parameters to explain poor fertility in some chronically stressed animals. From 30 to 80 days post-partum, there was a graded effect that ranged from 29% Lame cows with absence of ovarian activity, whereas another 21% Lame cows failed to express estrus or ovulate a low estrogenic follicle; in 50% cows, many reproductive parameters were unaffected by lameness.

Development of a lameness model in sheep for assessing efficacy of analgesics.

OBJECTIVE: To develop a lameness model to assess the efficacy of analgesics for alleviating pain, swelling and systemic signs of inflammation in sheep. PROCEDURES: The response to subcutaneous injection of 0.1 or 0.2 mL turpentine in a forelimb pastern (n = 4 ewes per dose) was examined at 0, 3, 6, 24, 48 and 72 h. In a second experiment, responses were measured at 0, 2, 4, 6, 8, 10, 12 and 24 h in ewes receiving 0.1 mL turpentine ± meloxicam 1 mg/kg IV at 0 h (n = 6 per group). Responses measured included forceplate pressure, skin temperature, limb circumference, nociception, leucocyte count, neutrophil : lymphocyte ratio, haptoglobin and daily feed intake. RESULTS: Turpentine injection caused a decrease in weight borne on the treated limb, increased skin temperature, increased sensitivity at the injection site and leucocytosis by 2 h and increased limb circumference by 4 h. Weight borne and sensitivity of the injected limb returned to control levels after around 24 h, whereas tissue swelling, elevated skin temperature and elevated haptoglobin levels persisted for at least 72 h. Treatment with meloxicam improved weight borne by and tolerance to pressure exerted on the turpentine-injected limb. CONCLUSIONS: The local and systemic signs of inflammation and pain, temporary reduction in function of the affected limb and partial amelioration of some of these changes by the dose of meloxicam used here suggest that injection of turpentine in the lower forelimb provides a suitable model for examining the efficacy of analgesics for alleviation of pain and inflammation in sheep.

Effect of heparin administration on urine protein excretion during the developmental stage of experimentally induced laminitis in horses.

OBJECTIVE: To investigate the effects of heparin administration on urine protein excretion during the developmental stages of experimentally induced laminitis in horses. ANIMALS: 13 horses. Procedures-Horses received unfractionated heparin (80 U/kg, SC, q 8 h; n=7) or no treatment (control group; 6) beginning 3 days prior to induction of laminitis. All horses were given 3 oligofructose loading doses (1 g/kg each) at 24-hour intervals and a laminitis induction dose (10 g of oligofructose/kg) 24 hours following the final loading dose (designated as 0 hours) via nasogastric tube. Serum glucose and insulin concentrations were measured before administration of the first loading dose (baseline) and at 0 and 24 hours; urine protein-to-creatinine (UP:C) ratio was determined at 0 hours and every 4 hours thereafter. Lameness was evaluated every 6 hours, and horses were euthanized when Obel grade 2 lameness was observed. RESULTS: Mean±SD time until euthanasia did not differ significantly between the heparin-treated (28.9±6.5 hours) and control (29.0±6.9 hours) horses. The UP:C ratio was significantly increased from baseline at 20 to 28 hours after induction of laminitis (ie, 4±4 hours before lameness was evident) in control horses but did not change significantly from baseline in heparin-treated horses. Serum glucose or insulin concentration did not change significantly from baseline in either group. CONCLUSIONS AND CLINICAL RELEVANCE: Urine protein excretion increased during the developmental stages of carbohydrate-induced laminitis in horses; administration of heparin prevented that increase, but did not delay onset or decrease severity of lameness.

Serum haptoglobin concentrations in dairy cattle with lameness due to claw disorders.

In cattle, elevated blood serum concentrations of haptoglobin, an acute phase protein, have been demonstrated in association with several diseases, but not with lameness. Serum haptoglobin was measured in 60 Holstein dairy cattle diagnosed with lameness due to four claw disorders, pododermatitis septica (PS; n=41), pododermatitis circumscripta (PC; n=8), interdigital necrobacillosis (IN; n=7), papillomatous digital dermatitis (PDD; n=4). Haptoglobin was measured on day 1 (0-3 days after lameness was observed but before treatment) and on days 3 and 5. A total of 10 healthy cows served as controls (haptoglobin values <1.0 mg/dL). Each of the claw disorders was associated with elevated haptoglobin on day 1 (PS, PC, IN and PDD: 65.9%, 37.5%, 71.4% and 25.0%, respectively). Trimming and antibiotic treatment led to a reduction in the number of PS and IN cows with increased haptoglobin concentrations, respectively (P<0.05), but trimming did not lead to any reduction in cows with PC. The study showed that lameness due to claw disorders can be associated with a systemic acute phase response and elevated serum haptoglobin in dairy cattle. Based on the course of haptoglobin, treatments seemed effective for all claw disorders except for PC.

The effect of metformin on measurements of insulin sensitivity and beta cell response in 18 horses and ponies with insulin resistance.

REASONS FOR PERFORMING STUDY: Laminitis in equids is a very common debilitating disease, and insulin resistance (IR) and hyperinsulinaemia are increasingly recognised as important predisposing factors. Pharmacological modification of IR and hyperinsulinaemia might reduce the risk of laminitis. HYPOTHESIS: Metformin, a drug commonly prescribed for treatment of human IR, may also decrease IR in equids. METHODS: Eighteen horses and ponies with IR and recurrent laminitis were treated with 15 mg/kg bwt metformin per os q. 12 h. Each animal served as its own control by comparing pre- and post treatment proxies for IR, insulin sensitivity (IS) and pancreatic beta cell function while controlling for possible dietary and managemental influences on IR. RESULTS: Evidence of significantly improved IS and decreased pancreatic beta cell secretion was found following metformin treatment. The magnitude of effect was greater at earlier resampling (6-14 days) than at later times (23-220 days). Apparent subjective clinical benefits were good but less favourable than effects on IR. CONCLUSIONS: Metformin is safe and appears to increase IS in equids. POTENTIAL RELEVANCE: Metformin may be indicated as a treatment for IR in equids. Further studies are required to define appropriate selection of subjects warranting therapy, dosing schedule and pharmacokinetics.

Changes in synovial fluid and serum biomarkers with exercise and early osteoarthritis in horses.

OBJECTIVE: To discriminate between changes in biomarkers with exercise compared to changes in biomarkers with osteoarthritis (OA) in exercising horses. METHOD: Sixteen, 2-year-old horses were randomly assigned either to an exercise-alone (n=8) or OA-affected (also exercised) (n=8) group. All horses had both mid-carpal joints arthroscoped and OA induced in one mid-carpal joint in the OA-affected joints of OA-affected horses. Two weeks after surgery all horses commenced a strenuous exercise program on a high-speed treadmill. Clinical outcomes and synovial fluid and serum biomarkers, were evaluated weekly. Synovial and serum biomarkers evaluated were epitope CS846 (CS846), epitope CPII (CPII), glycosaminoglycans (GAGs), epitope Col CEQ (Col CEQ) (a marker of type II collagen degradation), type I and II collagen degradation fragments (C1,2C), osteocalcin, C-terminal of bone type I collagen (CTX1), type I collagen (Col I) and (synovial fluid only of cartilage) prostaglandin E2 (PGE2) levels. Horses were euthanized at day 91 and their joints assessed grossly, histopathologically, and histochemically. RESULTS: Exercise induced a significant increase in synovial fluid CS846, CPII, GAG, Col CEQ, C1,2C, osteocalcin and Col I concentrations. There was a significant increase in synovial fluid CS846, CPII, Col CEQ, C1,2C, osteocalcin, Col I and PGE2 concentrations in OA-affected joints compared to exercise-alone joints. The concentration of serum CS846, CPII, GAG, osteocalcin, C1,2C and Col I increased with exercise. For each of these biomarkers there was also a statistically significant increase in serum biomarker levels in OA-affected horses compared to exercise-alone horses. CONCLUSIONS: Six synovial fluid and serum biomarkers were useful in separating early experimental OA from exercise alone but synovial fluid CTX1 and serum Col CEQ and CTX1 were not.

Depressed DHEA and increased sickness response behaviors in lame dairy cows with inflammatory foot lesions.

Lameness is a multifactorial condition influenced by the environment, genetics, management and nutrition. Detection of lameness is subjective and currently limited to visual locomotion observations which lack reliability and sensitivity. The objective of this study was to search for potential biomarkers of inflammatory foot lesions that underlie most cases of lameness in dairy cows, with a focus on the sickness response and relevant endocrine, immune and behavioral changes. Serum and peripheral blood mononuclear cells (PBMC) were collected from eight sound and eight lame high-producing Holstein cows. Immune cell activation was investigated in PBMCs using a candidate gene approach in which the expression of pro-opiomelanocortin, interleukin-1beta, l-selectin, matrix metalloproteinase-9 and glucocorticoid receptor-alpha was measured via quantitative real time-RT-PCR. Endocrine changes were investigated by monitoring serum concentrations of cortisol and dehydroepiandrosterone (DHEA). Additionally, systematic behavioral observations were carried out to characterize a behavioral profile associated with a sickness response typical of this condition. Lame cows showed significantly lower eating (P=0.01) and ruminating (P=0.01) behaviors and higher incidence of self-grooming (P=0.04) compared to sound cows. Lame cows also showed a 23% decrease in serum DHEA (P=0.01) and 65% higher cortisol:DHEA ratio (P=0.06) compared to sound cows. However, no significant differences were found in candidate gene expression between lame and sound cows. In association with sickness behaviors, serum DHEA concentration and cortisol:DHEA ratio are promising objective indicators of inflammatory foot lesions in dairy cattle and may be useful as diagnostic targets for animals in need of treatment.