Responses to an intra-articular lipopolysaccharide challenge following dietary supplementation of Saccharomyces cerevisiae fermentation product in young horses

Dietary intervention may be a valuable strategy to optimize the intra-articular environment in young horses to prolong their performance career. To test the hypothesis that dietary supplementation of a Saccharomyces cerevisiae fermentation product would reduce markers of joint inflammation and increase markers of cartilage metabolism following a single inflammatory insult, Quarter Horse yearlings (mean ± SD; 9 ± 1.0 mo) were balanced by age, sex, body weight (BW), and farm of origin and randomly assigned to the following treatment groups: 1.25% BW/d (dry matter basis) custom-formulated concentrate only (CON; n = 9) or concentrate top-dressed with 21 g/d S. cerevisiae fermentation product (SCFP; n = 10) for 98 d. Horses had ad libitum access to Coastal bermudagrass hay. On day 84, one randomly selected radial carpal joint from each horse was injected with 0.5 ng lipopolysaccharide (LPS) solution. The remaining carpal joint was injected with sterile lactated Ringer’s solution as a contralateral control. Synovial fluid obtained before supplementation (day 0) and on day 84 at preinjection hour 0 and 6, 12, 24, 168, and 336 h postinjection was analyzed for prostaglandin E₂ (PGE₂), carboxypropeptide of type II collagen (CPII), and collagenase cleavage neopeptide (C2C) by commercial assays. Rectal temperature, heart rate, respiration rate, carpal surface temperature, and carpal circumference were recorded prior to each sample collection and for 24 h postinjection. Data were analyzed using linear models with repeated measures. From day 0 to 84, synovial C2C declined (P ≤ 0.01) and the CPII:C2C ratio increased (P ≤ 0.01) in all horses with no effect of diet. In response to intra-articular LPS, synovial PGE₂ increased by hour 6 (P ≤ 0.01) and returned to baseline by hour 336; CPII increased by hour 12, remained elevated through hour 168 (P ≤ 0.01), and returned to baseline by hour 336; and C2C increased by hour 6 (P ≤ 0.01) but did not return to baseline through hour 336 (P ≤ 0.01). Post-intra-articular injection, PGE₂ levels were lower in SCFP than CON horses (P = 0.01) regardless of injection type. Synovial CPII and the CPII:C2C ratio demonstrated stability during the LPS challenge in SCFP compared with CON horses (P ≤ 0.01). Clinical parameters were not influenced by diet but increased in response to repeated arthrocentesis (P ≤ 0.01). Dietary SCFP may favorably modulate intra-articular inflammation following an acute stressor and influence cartilage turnover in young horses.     

Metabolic responses to oral tryptophan supplementation before exercise in horses

This study was conducted to evaluate the effects of oral tryptophan (Trp) supplementation on exercise capacity and metabolic responses in horses. Three horses had to perform an exercise test: a 15-min warm-up followed by a 60-min walk (1.7 m/s, W1), a 10-min trot (3.1 m/s, T1), a second 60-min walk (1.7 m/s, W2), a second 10-min trot (3.1 m/s, T2) and a final 30-min walk (1.7 m/s, W3) until the horses were unwilling to continue. The horses exercised on a treadmill at a 6% incline and with a constant draught load of 40 kg (0.44 kN). Two hours before exercise horses were given 50 g Trp (9.8-10.7 g Trp/100 kg BW) by nasogastric tube. A control exercise test was conducted without Trp. During the control test, one horse was able to finish the final 30-min walk (W3), whereas two horses finished W3 after Trp administration. Higher plasma Trp levels after Trp administration did not change significantly during exercise (Trp: start exercise, 524 +/- 41 micromol/l; end exercise 547 +/- 20 micromol/l; control: start exercise, 70 +/- 10 micromol/l; end exercise, 58 +/- 21 micromol/l). After Trp supplementation, blood lactate concentrations were significantly lower after the first and second trotting periods. Free fatty acids in plasma increased during exercise without any treatment-related differences. Although experimental plasma Trp levels were seven times higher than the control levels, Trp supplementation had no effect on exercise performance and metabolic responses to draught load exercise.     

Amino acid supplementation improves muscle mass in aged and young horses

The objective of the study was to evaluate the effect of supplementary AA on the ability to support muscle mass in aging horses. Sixteen horses of light horse type were used in a 2 x 2 factorial arrangement of treatments with two age groups [< or = 10 yr (average = 9.1 +/- 0.29 yr) and > or = 20 yr (average = 22.4 +/- 0.87 yr)] and two diet groups [no supplementation (N) or supplementary lysine and threonine (S; 20.0 and 15 g/d, respectively)]. Horses were fed the diets for 14 wk and received regular light exercise throughout the study. Body weight, BCS, and venous blood samples were taken every 2 wk. Plasma was analyzed for total protein, albumin, creatinine, urea N (PUN), and an AA profile, including 3-methyl histidine (3MH) and sulfur AA. Photographs of the horses taken at the start and at the end of the experiment were used to assign a subjective muscle mass score from 1 to 5 (1 = lowest to 5 = highest). There was no difference in BW caused by diet; however, the S-group horses tended (P = 0.064) to gain more weight (6.91 +/- 2.3 kg), and in fact, the N-group horses lost weight (- 11.76 +/- 5.2 kg) during the experiment. Repeated measures analysis revealed that BCS was lower for the aged vs. the young horses (P = 0.001) as well as for the S- vs. the N-group horses (P = 0.026). Subjective muscle mass scores were not different at the start of the experiment but were greater (P = 0.047) for the S-group horses (3.77 +/- 0.13) at the end of the experiment compared with the N-group horses (3.28 +/- 0.14). Plasma creatinine was greater (P = 0.032), and PUN was lower (P = 0.027), for S-group horses compared with N-group horses. Initial 3MH concentrations were not different; however, at the end of the experiment, 3MH was lower for the S-group horses (P = 0.016) compared with the N-group horses. Plasma lysine and threonine concentrations were greater for S-group horses at the end of the experiment than for N-group horses (P = 0.023 and 0.009, respectively). Both 3MH and PUN concentrations were negatively correlated to lysine (R2 = 0.57 and 0.65, respectively) and threonine intake (R2 = 0.56 and 0.60, respectively) at the end of the study. These data suggest that horses receiving supplementary AA were able to maintain muscle mass better than those without supplementation, regardless of age, as evidenced by the improvement in muscle mass scores, lower BCS with no difference in BW, greater creatinine, and lower 3MH and PUN concentrations in the S-group horses.     

Intra-articular injections for joint disease in horses.

Lameness remains an important source of reduced performance in many types of horses. Pain referable to joints is among the most common causes, and intra-articular injections remain a common and important means of addressing lameness referable to joints in horses.     

Effect of longeing and glucosamine supplementation on serum markers of bone and joint metabolism in yearling quarter horses.

The effect of longeing and glucosamine supplementation on known biological markers of joint disease was studied in yearling quarter horses. Twenty-one yearling quarter horses were randomly assigned to one of 4 treatments: 1) longeing (longeing 20 min daily) supplement control (LN); 2) longeing/glucosamine (LG); 3) walking (mechanical walker for 120 min daily (WN)); and 4) walking/glucosamine (WG). Oral glucosamine was administered at 5.5 g b.i.d. weeks 1-4, 3.5 g b.i.d. during weeks 5-6, and 2.0 g b.i.d. during weeks 7-8. Serum was obtained weekly for 8 wk and analyzed for keratan sulfate and osteocalcin concentrations. Walked horses receiving glucosamine showed slight elevation in serum keratan sulfate compared to controls (P = 0.04). Glucosamine or longeing exercise had no significant effect (6 > or = 0.08) on serum osteocalcin concentrations. Under these conditions, longeing and/or glucosamine supplementation did not significantly alter serum concentrations of keratan sulfate or osteocalcin.     

Effect of oral antioxidant supplementation on blood antioxidant status in trained thoroughbred horses

The oxidant/antioxidant equilibrium of trained thoroughbred horses (n = 40) was assessed on three occasions during a period of three months under field conditions by blood antioxidant markers analysis, i.e. plasma ascorbic acid (AA), plasma antioxidant capacity of water-soluble components (ACW), whole blood (GSH) and oxidised (GSSG) glutathione, plasma alpha-tocopherol and beta-carotene, plasma antioxidant capacity of lipid-soluble components (ACL), red blood cell superoxide dismutase (SOD) and glutathione-peroxidase activity (GPx) and plasma trace-elements, i.e. selenium (Se), copper (Cu), zinc (Zn). A control group of ten horses receiving a placebo and an antioxidant group of 30 horses orally supplemented with an antioxidant mixture were randomly formed. An antioxidant imbalance was observed after three months in the control group, reflected by a significant decrease in GSH, SOD, GPx, Se (P < 0.05) and a significant increase in GSSG (P < 0.05). The antioxidant supplement prevented GPx and Se decrease and significantly increased ACW, alpha-tocopherol, beta-carotene and ACL (P < 0.05). Significant sex- or age-related differences were found for AA, ACW, alpha-tocopherol, SOD, GPx and Se, and there were significant correlations between ACW-AA, ACL-alpha-tocopherol, GPx-Se, CPK-Se, CPK-alpha-tocopherol and CPK-Cu. This field study has shown that trained thoroughbred horses undergo significant changes of several blood antioxidant markers and that oral antioxidant supplementation might partially counterbalance these changes by improving the hydrophilic, lipophilic and enzymatic antioxidant blood capacity.     

Effects of dietary antioxidant supplementation before and after oral acetaminophen challenge in cats

OBJECTIVE: To determine effects of lipoic acid, vitamin E, and cysteine before and after oxidant challenge in cats. ANIMALS: 24 sexually intact adult cats. PROCEDURE: Cats were allocated into 4 equal groups. For 25 weeks, group A was fed a control dry diet and groups B, C, and D received this diet supplemented with vitamin E (2200 U/kg [dry matter basis {DMB}]) plus cysteine (9.5 g/kg [DMB]), lipoate (150 mg/kg [DMB]), or all 3 antioxidants together, respectively. Weights were measured every 3 days and venous blood obtained every 5 weeks for CBC; serum biochemical analyses; lymphocyte blastogenesis; thiobarbituric acid reactive substances concentration; and concentrations of plasma protein carbonyl, 8-OH d-guanosine, blood glutathione, plasma amino acid, lipoate, and dihydrolipoate. At 15 weeks, all cats received acetaminophen (9 mg/kg, PO, once), clinical effects were observed, and methemoglobin concentrations were measured. RESULTS: Lymphocyte blastogenesis increased transiently in group C and D cats. After acetaminophen administration, all groups had transient increases in methemoglobin within 4 hours and mild, brief facial edema; group C had decreased glutathione concentration and increased 8-OH d-guanosine concentration versus controls; and protein carbonyl concentration increased least for group B. Plasma lipoate and dihydrolipoate concentrations peaked by week 10 for groups C and D. Conclusions and Clinical Relevance-Lipoate, vitamin E, and cysteine did not have synergistic effects. Lipoate supplementation (150 mg/kg [DMB]) did not act as an antioxidant but appeared to enhance oxidant effects of acetaminophen. Vitamin E plus cysteine had protective effects.     

Dietary supplementation of a Saccharomyces cerevisiae fermentation product attenuates exercise-induced stress markers in young horses

Mitigation of exercise-induced stress is of key interest in determining ways to optimize performance horse health. To test the hypothesis that dietary supplementation of a Saccharomyces cerevisiae fermentation product would decrease markers of exercise-induced stress and inflammation in young horses, Quarter Horse yearlings (mean ± SD; 9 ± 1 mo) were randomly assigned to receive either no supplementation (CON; n = 8) or 21 g/d S. cerevisiae fermentation product (10.5 g/feeding twice daily; SCFP; n = 10) top-dressed on a basal diet of custom-formulated grain as well as ad libitum Coastal bermudagrass hay. After 8 wk of dietary treatments, horses underwent a 2-h submaximal exercise test (SET) on a free-stall mechanical exerciser. Serum was collected before dietary treatment supplementation (week 0), at week 8 pre-SET, and 0, 1, and 6 h post-SET and analyzed for concentrations of cortisol and serum amyloid A (SAA) by commercial enzyme-linked immunosorbent assay (ELISA) and for cytokine concentrations by commercial bead-based ELISA. Data were analyzed using linear models with repeated measures in SAS v9.4. From week 0 to 8 (pre-SET), serum cortisol decreased (P = 0.01) and SAA did not change, but neither were affected by diet. Serum concentrations of all cytokines decreased from week 0 to 8 (P ≤ 0.008), but granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor, and interleukin-8 (IL-8) decreased to a greater extent in CON than in SCFP horses (P ≤0.003). In response to the week 8 SET, serum cortisol increased in all horses (P < 0.0001) but returned to pre-SET levels by 1 h post-SET in horses receiving SCFP. At 6 h post-SET, cortisol concentrations in CON horses returned to pre-SET concentrations, whereas cortisol declined further in SCFP horses to below pre-SET levels (P = 0.0002) and lower than CON (P = 0.003) at that time point. SAA increased at 6 h post-SET in CON (P < 0.0001) but was unchanged through 6 h in SCFP horses. All cytokines except G-CSF increased in response to the SET (P < 0.0001) but showed differing response patterns. Concentrations of IL-1β, IL-6, and tumor necrosis factor-alpha were lesser (P ≤ 0.05), and concentrations of G-CSF and IL-18 tended to be lesser (P ≤ 0.09) in SCFP compared with CON horses throughout recovery from the SET. In summary, 8 wk of dietary supplementation with 21 g/d of SCFP may mitigate cellular stress following a single, prolonged submaximal exercise bout in young horses.     

Influence of procyanidin supplementation on the immune responses of broilers challenged with lipopolysaccharide

In the present study, the effect of dietary procyanidin (PCA, from pine needles) supplementation on the innate immunity of broilers were investigated. The experiment was designed as a 2 × 4 factorial arrangement (eight cages / treatment; six birds (one‐day‐old) / cage) with dietary PCA concentrations (0, 0.05, 0.075 and 0.1%) and two immune treatments (injection of lipopolysaccharide (LPS) (0.5 mg/kg body weight) or saline). LPS was dissolved in sterile 9 g/L (w/v) NaCl solution at 16, 18, 20 days of age to mimic immune stress. The remaining birds were injected with saline as a placebo. The results indicated that, prior to LPS challenge, the PCA diet had no significant effect on bird growth performance. The injection of LPS was also not associated with any significant changes in poultry performance. LPS injection increased the activity of nitrogen oxides (NOx) and the concentrations of inflammatory cytokines (interferon‐γ (IFN‐γ), interleukin‐1β (IL‐1β), IL‐2, IL‐4, IL‐6 and IL‐10) in serum; dietary PCA decreased these concentrations (P < 0.05) in the PCA 0.1% group, further illustrating the immune effect of PCA. In conclusion, PCA supplementation has a beneficial effect on LPS challenge, which may be associated with the inhibition of the secretion of cytokines and decrease in the proinflammatory marker NOx.     

Endotoxemia in horses: protection provided by antiserum to core lipopolysaccharide

An equine antiserum to core lipopolysaccharide was produced by inoculation of 6 horses with a boiled cell bacterin made from the J-5 mutant of Escherichia coli O111:B4. The antiserum immunoglobulin G titer to J-5 mutant E coli, as determined by enzyme-linked immunosorbent assay, was 1:15,006. Pooled serum prepared before inoculation (preimmune serum) had a J-5 immunoglobulin G titer of 1:350. The J-5 antiserum was tested for its protective efficacy in sublethal endotoxemia in 14 horses. Four horses served as nontreated controls and were given nothing before endotoxin challenge exposure (10 micrograms/kg of body weight, IV). Pooled preimmune serum (3 ml/kg, IV) was administered to 5 horses and J-5 antiserum (3 ml/kg, IV) was administered to 5 other horses 2 to 15 hours before endotoxin challenge exposure. During the 24 hours postendotoxin challenge exposure, endotoxemia was accompanied by significant (P less than 0.05) time-related changes in temperature, heart rate, pulse character, respiratory rate and character, capillary refill time, mucous membrane color, fecal composition, attitude, PCV, total plasma protein, WBC count, platelet count, plasma fibrinogen, prothrombin time, activated partial thromboplastin time, fibrinolytic degradation products, plasma glucose, and plasma lactate in all horses. There were no apparent treatment vs time interactions (P greater than 0.05). Two horses (1 control and 1 given J-5 antiserum) died suddenly from unknown causes immediately after endotoxin challenge exposure. Seemingly, equine antiserum to core lipopolysaccharide did not provide protection from the adverse effects of experimental endotoxemia produced by bolus IV infusion of 10 micrograms of endotoxin/kg.     

Hemangiosarcoma in 11 young horses

Hemangiosarcoma is a rare neoplasm of horses and hemangiosarcoma in young horses might behave differently than in mature horses. The purpose of this study was to identify the characteristics of hemangiosarcoma occurring in horses < or = 3 years of age. Medical records from 1982 to 2004 were searched for horses < or = 3 years of age with a histopathologic diagnosis of hemangiosarcoma. Eleven records were identified. Thoroughbred and Thoroughbred crosses predominated. Age ranged from 9 days to 3 years. All horses presented with cutaneous or leg swellings or joint effusion. Physical examination findings included tachycardia, fever, and depression. Laboratory abnormalities included anemia (5/11), hyperfibrinogenemia (4/11), hypofibrinogenemia (3/11), thrombocytopenia (2/11), and neutrophilic leukocytosis (1/11). Ultrasonographic and radiographic evaluation was not diagnostic in any case. Antemortem histopathologic diagnosis was obtained in 10 cases. Six of 11 horses were euthanized. Surgical resection was performed in 5 horses, 2 of which were later euthanized. Diagnosis was confirmed histologically at postmortem examination in all euthanized horses. Two cases resolved spontaneously. Early histopathologic diagnosis may allow cure if the mass is localized and amenable to surgical resection. In cases where the horse is medically stable, and masses are not interfering with quality of life, a period of observation may be warranted.     

Intra-articular administration of antibiotics in horses: Justifications,risks, reconsideration of use and outcomes

Antibiotics have been injected intra-articularly by equine veterinarians for decades, either prophylactically when other drugs are administered for osteoarthritis or therapeutically to treat septic arthritis. This route of administration has also more recently gained attention in human orthopaedic clinical practice, particularly as an alternative to systemic antibiotic administration to treat infections following prosthetic arthroplasty. While the rationale for injecting antibiotics intra-articularly has been largely focused on achieving high local drug concentrations, there has been relatively little focus on pharmacokinetic parameters of antibiotics administered by this route, or on the potential for local toxicity. The increasing incidence of antibiotic resistance in veterinary and human medicine prompts reconsideration of off-label antibiotic usage and evaluation of evidence-based dosing strategies. The purpose of this review was to summarise the current literature describing intra-articular antibiotic usage, including specific studies where pharmacokinetics, potential safety and toxicity have been evaluated. This review will advance practitioners’ understanding of the use of intra-articularly administered antibiotics, including the overall pros and cons of the approach.     

Histamine inhalation challenge in normal horses and in horses with small airway disease

A histamine inhalation challenge (HIC) procedure was developed to assess hyperreactive states in horses. Following clinical evaluation, percutaneous lung biopsies were performed on nine light breed mares aged 6 to 15 years. Five horses, with normal small airways, were classified as group A and four subjects with small airway disease (SAD) lesions formed group B. Pulmonary mechanics parameters were monitored following an aerosol of 0.9% saline and every 5 min for up to 30 min after HIC with 0.5% w/v of histamine diphosphate, administered through a face mask for 2.5 min. Tidal volume (VT) and airflow (V) values were obtained with a pneumotachograph. Transpulmonary pressure (delta Ppl) was measured by the esophageal balloon catheter method. Dynamic compliance (Cdyn), total pulmonary resistance (RL), end expiratory work of breathing (EEW) and respiratory rate (f) were calculated by a pulmonary mechanics computer. Group A horses had increases in RL, and decreases in Cdyn whereas horses in group B were hyperreactive and showed greater changes in EEW, Cdyn, and delta Ppl but with a relatively lower variation of RL. One horse in clinical remission from SAD, but with a high biopsy score (group B), and one clinically normal horse belonging to group A showed marked hyperreactivity as shown by increases in EEW, maximum change in delta Ppl and RL and decreases in Cdyn. These results suggest that the HIC described can be used as a method to investigate airway hyperreactivity and SAD in horses.