Effects of dietary camelina, flaxseed, and canola oil supplementation on transepidermal water loss, skin and coat health parameters, and plasma prostaglandin E2, glycosaminoglycan, and nitric oxide concentrations in healthy adult horses.

Camelina oil is derived from a low-input, high-yield crop and, in comparison to many other dietary fat sources currently used in equine diets, provides a greater amount of α-linolenic acid [ALA; (n-3)], than linoleic acid [LA; (n-6)]. However, no research exists assessing the effects of feeding camelina oil to horses in contrast to other commonly used oils. The objective of this study was to compare the effect of supplementing camelina oil to that of flaxseed and canola oil supplementation, on outcomes related to skin and coat health in horses. Thirty adult horses [23 mares, 7 geldings; 14.9 years ±â€…5.3 years; 544 ±â€…66 kg body weight (BW) (mean ±â€…SD)] underwent a 4-week wash-in period consuming hay and sunflower oil. Following the wash-in period, horses were blocked by location, age, and BW, and assigned to one of three treatment oils for 16 weeks (370 mg oil/kg BW): camelina (CAM), canola (OLA), or flaxseed (FLX) oil. Blood samples were collected and plasma prostaglandin E2 (PGE2; ELISA), nitric oxide (NO; Griess Reaction), and glycosaminoglycan (GAG; DMMB) concentrations were measured on weeks 0 (n = 30), 14 (n = 24), and 16 (n = 30). On weeks 0, 2, 4, 8, and 16, transepidermal water loss (TEWL) was measured pre- and post-acetone application using a VapoMeter (n = 26), and a 5-point-Likert scale was used to assess skin and coat characteristics on the side and rump of the horses (n = 30). All data were analyzed with repeated measures ANOVA using PROC GLIMMIX in SAS. Independent of treatment, coat color, and quality increased from baseline. There were no differences in the outcomes assessed between the horses supplemented camelina oil and those supplemented canola or flaxseed oil. These results suggest that independent of treatment, all oil supplements improved coat color and quality in horses. This provides indication that camelina oil is comparable to existing plant-based oil supplements in supporting skin and coat health and inflammation in horses.

Horses cannot produce omega-3 α-linolenic acid or omega-6 linoleic acid in the body, and as a result, these fatty acids are required in the diet. Camelina oil contains a lower ratio of omega-6 to omega-3 fatty acids (1:1.8) in comparison to alternative fat ingredients commonly included in many horse diets, such as soybean oil (1:0.12). Omega-3 fatty acids from flaxseed oil or marine-based oils can support skin and coat health and lower inflammation in horses; however, there is a lack of research investigating camelina oil supplementation and its benefits in horses. Thus, the objective of this study was to determine the effects of camelina oil on skin and coat health in horses. Horses were supplemented with sunflower oil for 4 weeks before being assigned to one of three treatment oils (camelina, canola, or flaxseed) for 16 weeks. Skin barrier function was assessed by measuring the transepidermal water loss of the chest, inner elbow, withers, and rump. Blood markers, including prostaglandin E2, nitric oxide, and glycosaminoglycan, were measured. Skin and coat parameters, including shine, softness, hair quality, color intensity, and moisture, were assessed using a 5-point scale on the rump and side of the horses. No differences in transepidermal water loss, blood markers, or skin and coat parameters were observed among treatments. Our results suggest that camelina oil is comparable to existing oil supplements in supporting skin and coat health and inflammation in horses.

A proof of principle study investigating the effects of supplemental concentrated brewer's yeast on markers of gut permeability, inflammation, and fecal metabolites in healthy non-challenged adult sled dogs.

Yeast-derived ß-glucans impact immunity, though their effects on gut permeability and inflammation are less understood. Most research has investigated other components of the yeast cell wall, such as the prebiotic mannan- and fructo-oligosaccharides. The objective of this study was to assess the effects of feeding a concentrated yeast product on markers of inflammation (serum amyloid A [SAA] and haptoglobin [Hp]) and oxidative status (malondialdehyde [MDA]), fecal products of fermentation, and gut permeability. Nineteen privately owned domestic Siberian huskies, and one Alaskan husky (9 females: 5 intact, 4 spayed; 11 males: 3 intact, 8 neutered), with an average age of 4.8 ± 2.6 yr and body weight (BW) of 25.6 ± 4.1 kg, were used in this study. Dogs were blocked and randomly allocated to one of two diet groups. Ten dogs received a dry extruded diet. The other 10 received the same diet top dressed with yeast for a daily ß-glucan dose of 7 mg/kg BW for 10 wk. Fecal collection, for evaluation of fecal metabolites, and scoring occurred weekly. Gut permeability was assessed using the chromium-labeled ethylenediamine tetra-acetic acid (Cr-EDTA) and iohexol markers prior to the initiation of dietary treatment and after 10 wk of treatment. Blood samples were collected premarker administration and 0.5, 1, 2, 3, 4, 5, and 6 h postadministration. Fasting concentrations of SAA, Hp, and MDA were measured on weeks -1, 2, 4, and 8. Incremental area under the curve (I-AUC) was calculated for serum iohexol and Cr-EDTA concentrations. All data were analyzed using PROC GLIMMIX of SAS with dog as random effect, and week as fixed effect and repeated measure. Dogs receiving treatment tended to have decreased I-AUC of Iohexol (P = 0.10) and Cr-EDTA (P = 0.06) between baseline and cessation of treatment compared to the change over time in I-AUC for control (Ctl) dogs. Treatment dogs had lower Hp concentrations (P ≤ 0.05) than Ctl. There were no differences between treatments for SAA and MDA concentrations (P > 0.05). Fecal arabinose concentrations were greater in treatment (Trt) dogs (P ≤ 0.05) compared to Ctl, though no other fecal metabolites were affected by treatment. There was no difference in the relative frequency of defecations scored at any fecal score between Trt and Ctl dogs, and mean score did not differ between groups (P > 0.10). These data suggest that concentrated brewer's yeast may have the potential to reduce gut permeability without impacting inflammatory status and markers of health in adult dogs.

This study evaluated the effects of concentrated brewer's yeast on gut health in dogs. Nineteen Siberian Huskies and one Alaskan husky were blocked and randomly allocated to one of two groups. Treatment dogs received a yeast supplement for 10 wk, while control dogs received no supplement. Dogs were administered two markers to assess intestinal permeability prior to start of treatment and following 10 wk of treatment. Blood samples were collected and analyzed for markers of inflammatory status (serum amyloid A [SAA] and Haptoglobin [Hp]) and oxidative status (serum malondialdehyde [MDA]). Fecal samples were collected weekly to assess fecal score as well as fecal metabolite concentrations. Intestinal permeability was reduced in treatment dogs following treatment, and no change was observed in the control group. Treatment dogs had lower Hp concentrations than control (Ctl), but there were no differences between treatments for SAA and MDA. Fecal arabinose concentrations were significantly greater in the treatment group when compared to control. There were no differences in the relative frequency of defecations scored at any fecal score between treatment and Ctl dogs, nor did mean score differ between the groups. This study suggests that concentrated brewer's yeast may reduce gut permeability and inflammation without detrimentally impacting markers of health in adult dogs.

Vascular Ehlers-Danlos Syndrome: Treatment of a Complex Abdominal Wound with Vitamin C and Mesenchymal Stromal Cells.

ABSTRACT: Vascular Ehlers-Danlos syndrome (EDSv) can present with life-threatening surgical complications. The article describes the case of a patient with EDSv who developed total abdominal wound dehiscence and multiple enterocutaneous fistulas. Treatment with IV allogeneic mesenchymal stromal cells (MSCs) and high-dose vitamin C was trialed with success. Near-complete wound healing of the abdominal dehiscence with a 94% reduction in the size of the wound bed occurred. Maturation of the enterocutaneous fistulas also ensued.There is no current consensus on the management of large cutaneous wounds in EDSv. This article discusses the pathophysiology of wound healing with regard to nutrition requirements and growth factors with special reference to collagen deficits in EDSv. A potential therapy with IV vitamin C supplementation and MSCs is proposed following the patient's positive outcome. Medium-dose MSCs and high-dose IV vitamin C may offer significant benefits to complex and problematic wounds.

Effects of a nutraceutical supplement on gastrointestinal health in racing standardbreds.

BACKGROUND: Feed additives that accelerate gastrointestinal transit time may help support normal gastrointestinal function in horses at risk for impaction colic. Previous research has demonstrated significant stimulatory effect of a hemp-based nutraceutical product (Gs Formula; GF) on contractility of gastric smooth muscle and gastrin production in vitro. OBJECTIVES: To quantify effects of GF on indicators of GIT transit time and tight junction proteins. STUDY DESIGN: Randomized placebo-controlled cross-over study. METHODS: Eight Standardbreds were administered 200 plastic beads by nasogastric tube before (baseline; BL) and after receiving a diet containing GF (CON: 0 g/day, LO: 160 g/day or HI: 480 g/day) for 28 days. Total manure collection occurred every 2 hours for 72 hours after bead administration. Outcome measures included GIT transit time, faecal dry matter (DM), water intake, and complete biochemistry and haematology screens. RESULTS: There was no effect of GF on GIT transit time. Faecal output was significantly lower in LO and HI horses than CON horses after 28 days on the supplement. HI horses have significantly lower rouleaux formation and lower faecal DM on Day 28 compared with BL. GF also produced changes in electrolytes associated with pH balance, which may indicate a role for GF as an alkalinizing compound in exercising horses. Clinical pathology results support the safety of GF up to 480 g/day for 28 days with no adverse effects being observed in haematology or biochemistry results. MAIN LIMITATIONS: Future studies on GF should focus on evaluating effect of GF on gastrointestinal transit in horses with naturally or experimentally delayed gastrointestinal motility, and its effect on exercise performance and onset of fatigue. CONCLUSIONS: GF may help support normal gastrointestinal function in horses at risk for impaction colic by reducing faecal DM and rouleaux formation.

A Botanical-Based Equine Nutraceutical Reduces Gastric Smooth Muscle Contractile Force In Vitro.

The objective of this study was to evaluate the effect of a botanical-based equine nutraceutical on contractility of gastric smooth muscle in vitro. Gastric ulcers are prevalent in performance horses and negatively impact horse welfare. Gastric hypermotility has been positively associated with the development of gastric ulceration in nonequine species, and reduction of hypermotility may be protective against their development. Stomachs from 12 pigs processed for food at a provincially inspected abattoir were collected within 1 hour of slaughter. Explants of nonglandular gastric tissue were prepared and suspended in a tissue bath, attached to a force transducer, in the presence or absence of a simulated digest extract of the nutraceutical. Tissue was stimulated to contract using increasing doses of acetylcholine. Peak and mean contractile force over 1 and 2 minutes after exposure to acetylcholine were measured. Exposure of gastric smooth muscle to the nutraceutical significantly reduced contractility of the tissue. These data provide support for the use of this nutraceutical to reduce contractility of nonglandular gastric smooth muscle and may indicate a protective effect of this nutraceutical in horses with mechanically induced gastric ulcers. Future studies are needed to clarify the role of gastric hypermotility on development of equine gastric ulcers and to determine the effect of this nutraceutical on equine gastric contractility and ulcerogenesis in vivo.

A biological extract of turmeric (Curcuma longa) modulates response of cartilage explants to lipopolysaccharide.

BACKGROUND: Turmeric is commonly used as a dietary treatment for inflammation, but few studies have evaluated the direct effect of turmeric on cartilage. The purpose of this study was to characterize cartilage explants' inflammatory responses to lipopolysaccharide in the presence of a simulated biological extract of turmeric. METHODS: Turmeric was incubated in simulated gastric and intestinal fluid, followed by inclusion of liver microsomes and NADPH. The resulting extract (TURsim) was used to condition cartilage explants in the presence or absence of lipopolysaccharide. Explants were cultured for 96 h (h); the first 24 h in basal tissue culture media and the remaining 72 h in basal tissue culture media containing TURsim (0, 3, 9 or 15 µg/mL). Lipopolysaccharide (0 or 5 µg/mL) was added for the final 48 H. media samples were collected immediately prior to lipopolysaccharide exposure (0 h) and then at 24 and 48 h after, and analyzed for prostaglandin E2 (PGE2), glycosaminoglycan (GAG), and nitric oxide (NO). Explants were stained with calcein-AM for an estimate of live cells. Data were analyzed using a 2-way repeated measures (GAG, PGE2, NO) or 1-way ANOVA without repeated measures (viability). Significance accepted at p < 0.05. RESULTS: TURsim significantly reduced PGE2, NO and GAG, and calcein fluorescence was reduced. CONCLUSIONS: These data contribute to the growing body of evidence for the utility of turmeric as an intervention for cartilage inflammation.

Protection against LPS-induced cartilage inflammation and degradation provided by a biological extract of Mentha spicata.

BACKGROUND: A variety of mint [Mentha spicata] has been bred which over-expresses Rosmarinic acid (RA) by approximately 20-fold. RA has demonstrated significant anti-inflammatory activity in vitro and in small rodents; thus it was hypothesized that this plant would demonstrate significant anti-inflammatory activity in vitro. The objectives of this study were: a) to develop an in vitro extraction procedure which mimics digestion and hepatic metabolism, b) to compare anti-inflammatory properties of High-Rosmarinic-Acid Mentha spicata (HRAM) with wild-type control M. spicata (CM), and c) to quantify the relative contributions of RA and three of its hepatic metabolites [ferulic acid (FA), caffeic acid (CA), coumaric acid (CO)] to anti-inflammatory activity of HRAM. METHODS: HRAM and CM were incubated in simulated gastric and intestinal fluid, liver microsomes (from male rat) and NADPH. Concentrations of RA, CA, CO, and FA in simulated digest of HRAM (HRAMsim) and CM (CMsim) were determined (HPLC) and compared with concentrations in aqueous extracts of HRAM and CM. Cartilage explants (porcine) were cultured with LPS (0 or 3 microg/mL) and test article [HRAMsim (0, 8, 40, 80, 240, or 400 microg/mL), or CMsim (0, 1, 5 or 10 mg/mL), or RA (0.640 microg/mL), or CA (0.384 microg/mL), or CO (0.057 microg/mL) or FA (0.038 microg/mL)] for 96 h. Media samples were analyzed for prostaglandin E2 (PGE2), interleukin 1beta (IL-1), glycosaminoglycan (GAG), nitric oxide (NO) and cell viability (differential live-dead cell staining). RESULTS: RA concentration of HRAMsim and CMsim was 49.3 and 0.4 microg/mL, respectively. CA, FA and CO were identified in HRAMsim but not in aqueous extract of HRAM. HRAMsim (> or = 8 microg/mL) inhibited LPS-induced PGE2 and NO; HRAMsim (> or = 80 microg/mL) inhibited LPS-induced GAG release. RA inhibited LPS-induced GAG release. No anti-inflammatory or chondroprotective effects of RA metabolites on cartilage explants were identified. CONCLUSIONS: Our biological extraction procedure produces a substance which is similar in composition to post-hepatic products. HRAMsim is an effective inhibitor of LPS-induced inflammation in cartilage explants, and effects are primarily independent of RA. Further research is needed to identify bioactive phytochemical(s) in HRAMsim.

Evaluation of inflammatory responses induced via intra-articular injection of interleukin-1 in horses receiving a dietary nutraceutical and assessment of the clinical effects of long-term nutraceutical administration.

OBJECTIVE: To evaluate inflammatory responses induced via intra-articular recombinant human interleukin (IL)-1beta treatment in horses receiving a dietary nutraceutical (DN; composed of mussel, shark cartilage, abalone, and Biota orientalis lipid extract) and assess the clinical effects of long-term DN administration. ANIMALS: 22 healthy horses. PROCEDURES: 12 horses were fed 0, 15, 45, or 75 mg of DN (3 horses/treatment) daily for 84 days. General health and clinicopathologic variables were monitored at intervals. Ten other horses received 0 or 15 g of DN/d (5 horses/treatment) for 29 days (beginning day -14). One intercarpal joint in each horse was injected twice with IL-1beta (10 and 100 ng on days 0 and 1, respectively), and the contralateral joint was similarly injected with saline (0.9% NaCl) solution. Synovial fluid prostaglandin E(2) (PGE(2)), sulfated glycosaminoglycan (GAG), nitric oxide (NO), and protein concentrations and leukocyte counts were analyzed before and at intervals after injections. RESULTS: Administration of the DN (up to 75 g/d) to horses for 84 days did not induce any adverse effects. In the other experiment, synovial fluid PGE(2), GAG, and protein concentrations and leukocyte count increased after intra-articular injections of IL-1beta (compared with effects of saline solution injections) in horses that received no DN; NO concentration was not affected. In horses that were fed the DN, intra-articular IL-1beta injections did not induce significant increases in synovial fluid PGE(2) and GAG concentrations. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that administration of the DN may be useful in preventing inflammation associated with arthritis and degenerative joint disease in horses.

Concurrent use of veterinary drugs and herbal medicines in racing standardbreds.

Standardbred trainers from 1 racetrack and 7 off-track training facilities were surveyed to determine the most common drugs, and prevalence of concurrent herb administration. Furosemide (on-track) and anti-inflammatory drugs (off-track) were the most common drugs administered. Among horses on-track, 9.8% received herbs compared with 13.8% off-track horses; 67% and 58% of these horses, respectively, received concurrent drugs.

Effects of simulated digests of Biota orientalis and a dietary nutraceutical on interleukin-1- induced inflammatory responses in cartilage explants.

OBJECTIVE: To test the hypothesis that simulated digests of Biota orientalis (BO) and a dietary nutraceutical (DN; composed of mussel, shark cartilage, abalone, and BO seed lipid extract) inhibit prostaglandin E2 (PGE2), nitric oxide (NO), and glycosaminoglycan (GAG) production in interleukin (IL)-1-stimulated cartilage explants. SAMPLE POPULATION: Cartilage tissue from 12 pigs. PROCEDURES: Articular cartilage explants were conditioned with a simulated digest of BO (BOsim) or DN (DNsim) at concentrations of 0, 0.06, or 0.18 mg/mL or indomethacin (INDOsim; 0 or 0.02 mg/mL) for 72 hours. Control explants received digest vehicle only. Explants were or were not stimulated with recombinant human-IL-1beta (10 or 0 ng/mL) during the final 48 hours of culture. Concentrations of PGE2, GAG, and NO in media samples (mPGE2,mGAG, and mNO concentrations, respectively) were analyzed, and explant tissue was stained fluorochromatically to determine chondrocyte viability. Treatment effects during the final 48-hour culture period were analyzed. RESULTS: IL-1 increased mPGE2, mGAG, and mNO concentrations in control explants without adversely affecting cell viability. Treatment with INDOsim blocked PGE2 production and increased mNO concentration in IL-1-stimulated and unstimulated explants and increased mGAG concentration in unstimulated explants. Treatment with DNsim (0.06 and 0.18 mg/mL) reduced mPGE2 concentration in IL-1-stimulated and unstimulated explants, reduced mNO concentration in IL-1-stimulated explants, and increased mNO concentration in unstimulated explants. Treatment with 0.18 mg of DNsim/mL increased cell viability in the presence of IL-1. In IL-1-stimulated explants, BOsim (0.06 and 0.18 mg/mL) reduced mPGE2 concentration, but 0.18 mg of BOsim/mL increased cell viability. CONCLUSIONS AND CLINICAL RELEVANCE: Effects of IL-1 on cartilage explants in vitro were modulated by DNsim and BOsim.

Low-dose ginseng (Panax quinquefolium) modulates the course and magnitude of the antibody response to vaccination against equid herpesvirus I in horses.

The purpose of this study was to determine if ginseng fed at low levels enhances a horse's antibody response to vaccination against Equid herpesvirus 1 (EHV-1). For 28 d, 5 horses received ground, powdered ginseng (35 mg/kg body weight, 1.7 mg/kg total ginsenosides) in molasses as a carrier, and 5 received molasses only. On day 14, each horse was vaccinated against EHV-1. The time course of the antibody response to vaccination was significantly altered in the horses receiving ginseng, a clinically relevant increase in antibody titer being observed by postvaccination day 2 compared with day 6 in the control horses. The horses receiving ginseng also had a significant decrease in serum levels of sodium and a significant increase in serum levels of potassium. No adverse effects of ginseng treatment were identified by hematologic and blood biochemistry profiles. Thus, low-dose dietary supplementation with ginseng in healthy horses may be a useful adjunct to vaccination.

Anti-inflammatory and chondroprotective effects of nutraceuticals from Sasha's Blend in a cartilage explant model of inflammation.

New Zealand green lipped mussel (NZGLM), abalone (AB), and shark cartilage (SC) are extensively used for treatment of and/or as preventatives for arthritis, despite a relative paucity of scientific evidence for efficacy. This research integrated a simulated digestion protocol with ultrafiltration and cartilage explants to generate new information on the anti-inflammatory and chondroprotective properties of NZGLM, SC, and AB. Each nutraceutical was artificially digested using simulated gastric and intestinal fluids, and the crude digest was ultrafiltered (50 kDa). Each filtrate was applied individually to cartilage explants before the explants were stimulated with IL-1 to induce an acute inflammatory response. Media were collected daily for 48 h and analyzed for prostaglandin E(2) (PGE(2)), glycosaminoglycan (GAG), and nitric oxide (NO), and cartilage tissue was differentially stained to determine the relative proportion of live and dead cells. SC and NZGLM significantly inhibited IL-1-induced PGE(2) synthesis and IL-1-induced GAG release, and AB was an effective inhibitor of IL-1-induced NO production. The three test nutraceuticals affect at least three major pathways involved in the catabolic cycle of arthritis and may prove important treatments and/or preventatives for the pain and degradation associated with this condition. The methodology and results describe a useful model for evaluating dietary nutraceuticals in vitro.

Pilot study investigating the ability of an herbal composite to alleviate clinical signs of respiratory dysfunction in horses with recurrent airway obstruction.

Recurrent airway obstruction (RAO), known previously as chronic obstructive pulmonary disease (COPD), is a debilitating respiratory condition that significantly contributes to lost training days and illness in racehorses. Herbs are becoming increasingly popular for the prophylaxis or treatment of the clinical signs of RAO despite a paucity of research on efficacy and safety. We evaluated the ability of an herbal composite containing garlic, white horehound, boneset, aniseed, fennel, licorice, thyme, and hyssop to reduce the clinical signs of RAO, hypothesizing that the product would safely reduce signs and would improve the inflammatory cell profile within the lungs. The composite was fed to 6 horses with symptomatic RAO for 21 d in a crossover manner. Ventigraphs were used to record respiratory rate and intrapleural pressure; the proportion of inflammatory cells in fluid aspirated from the trachea was determined. Blood biochemical and hematologic screening was conducted to identify possible adverse effects. Treatment with the composite did not result in statistically significant changes in any of the parameters evaluated. A trend to a decrease in respiratory rate (P = 0.1) and an increase in the proportion of macrophages (P = 0.1) was observed in the horses receiving the herbal composite compared with placebo. These data indicate a potential for the herbal composite to safely reduce the elevated respiratory rate in horses with RAO. Future research with a greater number of horses is warranted to further characterize the effect of this product on horses with RAO.

Association of maximum voluntary dietary intake of freeze-dried garlic with Heinz body anemia in horses.

OBJECTIVE: To characterize hematologic and clinical consequences of chronic dietary consumption of freeze-dried garlic at maximum voluntary intake in horses. ANIMALS: 4 healthy sex- and age-matched horses. PROCEDURE: An initial garlic dose (0.05 g/kg, twice daily) was fed to 2 horses in a molasses carrier as part of their normal ration and was gradually increased to maximum voluntary intake (0.25 g/kg, twice daily) over 41 days. Dietary supplementation then continued for a total of 71 days. Two control horses were fed molasses with no garlic with their ration. Blood samples were collected weekly and analyzed for hematologic and biochemical changes, including the presence of Heinz bodies. Recovery of affected blood values was followed for 5 weeks after termination of dietary supplementation with garlic. RESULTS: At a daily dose of > 0.2 g/kg, horses fed garlic developed hematologic and biochemical indications of Heinz body anemia, as characterized by increases in Heinz body score (HBS), mean corpuscular volume (MCV), mean corpuscular hemoglobin, platelet count, and serum unconjugated and total bilirubin concentrations and decreases in RBC count, blood hemoglobin concentration, mean corpuscular hemoglobin concentration, and serum haptoglobin concentration. Recovery from anemia was largely complete within 5 weeks after termination of dietary supplementation with garlic. Heinz body score and MCV remained high at the end of the 5-week recovery period. CONCLUSIONS AND CLINICAL RELEVANCE: Horses will voluntarily consume sufficient quantities of garlic to cause Heinz body anemia. The potential for garlic toxicosis exists when horses are chronically fed garlic. Further study is required to determine the safe dietary dose of garlic in horses.