Pharmacokinetics of amikacin after intravenous, intra-articular, and combined intravenous and intra-articular administration in healthy neonatal foals.

BACKGROUND: Pharmacokinetics of amikacin administered IV to neonatal foals are described, but little data are available regarding the plasma concentrations contributed by concurrent intra-articular (IA) administration. HYPOTHESIS/OBJECTIVES: Compare the pharmacokinetics of amikacin when the total dose is administered IV compared to being divided between IV and IA routes of administration in neonatal foals and predict the plasma concentrations from various combined IV and IA dosing regimens. ANIMALS: Eight healthy neonatal foals. METHODS: Foals received 3 amikacin treatment protocols: (1) IV-only (25 mg/kg q24h IV), (2) concurrent IV and IA (16.7 mg/kg q24h IV and 8.3 mg/kg q24h into 1 tarsocrural joint), and (3) IA-only (8.3 mg/kg q24h into 1 tarsocrural joint). Protocols were administered for 3 days beginning at 7, 14, and 21 days of age. Plasma concentrations ≥53 µg/mL at 30 minutes were considered therapeutic for isolates with intermediate susceptibility. RESULTS: Foal age was a significant variable. The IV-only protocol met or exceeded the 30-minute plasma concentrations considered therapeutic (mean µg/mL [95% confidence interval, CI]) in 7- to 9-day-old (54.0 [52.2-56.9]), 14- to 16-day-old (58.1 [55.2-61.0]), and 21- to 23-day-old (66.6 [63.7-69.6]) foals. Concurrent IV and IA protocol did not reach the 30-minute concentration considered therapeutic in 7- to 9-day-old foals (46.5 [43.6-49.4]) but did in 14- to 16-day-old (62.9 [60.0-65.8]) and 21-to 23-day-old (62.6 [59.7-65.6]) foals. CONCLUSIONS AND CLINICAL IMPORTANCE: Concurrent IV and IA administration of amikacin produces 30-minute plasma concentrations considered therapeutic in foals 14 to 23 days old, but concentrations observed in younger foals might be below those considered therapeutic for isolates with intermediate susceptibility to amikacin.

Nebulized glycosylated caffeic acid phenylether ester attenuation of environmental particulate-induced airway inflammation in horses.

The objective of this study was to determine the extent that nebulized glycosylated caffeic acid phenylether ester-4-O-alpha-D-glucopyranoside (G-CAPE) attenuates particulate-induced airway inflammation in healthy horses. Our hypothesis was that nebulization with G-CAPE would result in improved respiratory scores, higher arterial oxygen partial pressure, and less inflammatory airway infiltrates in horses with induced airway inflammation, compared with untreated controls. Five healthy adult horses were housed inside a climate controlled, closed barn on straw bedding and fed ad lib moldy grass hay for 16 days to induce airway inflammation. An experimental crossover study was performed in which animals were treated with 200 mg G-CAPE dissolved in 45 mL of 10% triethanolamine (G-CAPE group) or 45 mL of 10% triethanolamine (CONTROL group), and clinical respiratory scoring, arterial blood gases, and bronchoalveolar lavages (BALs) were collected at predetermined time points up to 24 h post nebulization. While the mean neutrophil percentage decreased in treated horses compared to controls (9.3 ± 2.0 and 16.9 ± 2.4, respectively) at 6 hours post treatment (t = 6 h), the difference did not achieve statistical significance (p = 0.1154). Blood gas analysis did not differ significantly between groups. There was a significant difference in the mean respiratory scores of G-CAPE-treated horses between baseline and at 1-h post treatment (from 3.2 ± 0.7 to 1.6 ± 0.7, p = 0.0013). This study demonstrates that a single nebulized dose of G-CAPE decreased clinical respiratory scores 1 h post administration and decreased BAL percentage of neutrophils 6 h post administration in horses with particulate induced airway inflammation. This compound shows promise as an anti-inflammatory and warrants further investigation.

Pharmacokinetic Evaluation of a Cannabidiol Supplement in Horses.

Cannabidiol (CBD) products have gained popularity among horse owners despite limited evidence regarding pharmacokinetics. The purpose of this study was to describe the pharmacokinetic profile of multiple doses of an orally administered cannabidiol product formulated specifically for horses. A randomized 2-way crossover design was used. Seven horses received 0.35 or 2.0 mg/kg CBD per os every 24 hours for 7 total doses, separated by a 2-week washout. Plasma CBD and delta-9-tetrahydrocannabinol (THC) were quantified using liquid chromatography-tandem mass spectrometry (LC-MS/MS) daily through day 10, then on day 14 after beginning CBD administration. On the final day of CBD administration, plasma CBD and THC were quantified at multiple times. After administration of 0.35 mg/kg of CBD, the Cmax of CBD was 6.6 ± 2.1 ng/mL while Tmax was 1.8 ± 1.2 hour, whereas the Cmax for THC was 0.7 ± 0.6 ng/mL with a Tmax of 2.5 ± 1 hour. After administration of 2.0 mg/kg of CBD, the Cmax of CBD was 51 ± 14 ng/mL with a mean Tmax of 2.4 ± 1.1 hour and terminal phase half-life of 10.4 ± 6 hour, whereas the Cmax of THC was 7.5 ± 2.2 ng/mL with a Tmax of 2.9 ± 1.1 hour. Oral administration of a cannabidiol product at 0.35 mg/kg or 2.0 mg/kg once daily for 7 days was well-tolerated. Based on plasma CBD levels obtained, dose escalation trials in the horse evaluating clinical efficacy at higher mg/kg dose rates are indicated.

Safety and efficacy of cold atmospheric plasma for the sterilization of a Pasteurella multocida-contaminated subcutaneously implanted foreign body in rabbits.

OBJECTIVE: To determine whether a stainless steel implant sterilized with a novel cold atmospheric plasma sterilization (CAPS) device adversely affects local tissues in rabbits and whether CAPS was as effective as steam sterilization with an autoclave to inactivate Pasteurella multocida. ANIMALS: 31 healthy New Zealand White rabbits. PROCEDURES: Steam-autoclaved stainless steel implants inoculated with P multocida underwent a second steam autoclave sterilization (AIA) or CAPS (AICAPS). One AIA implant and 3 AICAPS implants were randomly placed subcutaneously at 4 sites in 21 rabbits (84 implants). These rabbits were monitored daily for 5 days for evidence of systemic illness and local tissue reactions at the implantation sites and then euthanized. Samples were taken from each implant site for bacterial culture and histologic examination. RESULTS: Cultures of samples obtained from all sites were negative for bacterial growth. No significant difference was observed in mean skin thickness or erythema between AIA and AICAPS implant sites on any observed day. Also, individual histologic grades for the epidermis, dermis, subcutis, and muscle and total histologic grade were not significantly different between AIA and AICAPS implant sites. CONCLUSIONS AND CLINICAL RELEVANCE: Cold atmospheric plasma sterilization was noninferior to steam sterilization of P multocida-contaminated stainless steel implants in the rabbits in the present study. However, studies of the efficacy of CAPS for inactivation of other important bacteria are needed.

Subtourniquet pressures generated by application of wide-rubber tourniquets in standing, sedated horses.

OBJECTIVE: To determine the influence of location and movement on subtourniquet pressure (STP) generated by application of a wide-rubber tourniquet (WRT) on equine limbs. STUDY DESIGN: Randomized experimental cross-over design. ANIMALS: Six standing, sedated horses. METHODS: Horses were sedated with detomidine hydrochloride (0.2 µg/kg IV), and 4 investigators applied WRTs to the antebrachium (AB), gaskin (GK), and midmetacarpus (MC) of each horse in a predetermined, randomized order. Subtourniquet pressure was consequently measured at 10-minute intervals (T0, T10, T20, T30) for 30 minutes. Indirect systolic blood pressure (SBP) was measured presedation, postsedation, and throughout the tourniquet application period. Target STP was established as SBP + 100 mm Hg. Limb movements at each location were classified as none, low, moderate, or high, on the basis of counts and magnitude. RESULTS: Mean STP did not change with time (P = .93) and exceeded SBP by 163 mm Hg (95% CI 122-205), 185 mm Hg (95% CI 156-214), and 402 mm Hg (95% CI 351-454) at the AB, GK, and MC, respectively. Mean STP at each location exceeded the target STP in 59 of 70 (81%) of the trials. Limb movements affected STP generated by tourniquets at the AB (P = .04) and MC (P < .0001) but not at the GK (P = .67). CONCLUSION: Wide-rubber tourniquets applied at the AB, GK, and MC generated STP >100 mm Hg above SBP for 30 minutes in standing, sedated horses. CLINICAL SIGNIFICANCE: Wide-rubber tourniquets as applied in this study can achieve and maintain the current recommended STP (SBP + 100 mm Hg) for equine IV regional limb perfusion. Number and magnitude of limb movement can decrease STP over time, potentially reducing the efficacy of a WRT.

Viral testing of 18 consecutive cases of equine serum hepatitis: A prospective study (2014-2018).

BACKGROUND: Three flaviviruses (equine pegivirus [EPgV]; Theiler's disease-associated virus [TDAV]; non-primate hepacivirus [NPHV]) and equine parvovirus (EqPV-H) are present in equine blood products; the TDAV, NPHV, and EqPV-H have been suggested as potential causes of serum hepatitis. OBJECTIVE: To determine the prevalence of these viruses in horses with equine serum hepatitis. ANIMALS: Eighteen horses diagnosed with serum hepatitis, enrolled from US referral hospitals. METHODS: In the prospective case study, liver, serum, or both samples were tested for EPgV, TDAV, NPHV, and EqPV-H by PCR. RESULTS: Both liver tissue and serum were tested for 6 cases, serum only for 8 cases, and liver only for 4 cases. Twelve horses received tetanus antitoxin (TAT) 4-12.7 weeks (median = 8 weeks), 3 horses received commercial equine plasma 6-8.6 weeks, and 3 horses received allogenic stem cells 6.4-7.6 weeks before the onset of hepatic failure. All samples were TDAV negative. Two of 14 serum samples were NPHV-positive. Six of 14 serum samples were EPgV-positive. All liver samples were NPHV-negative and EPgV-negative. EqPV-H was detected in the serum (N = 8), liver (N = 4), or both samples (N = 6) of all 18 cases. The TAT of the same lot number was available for virologic testing in 10 of 12 TAT-associated cases, and all 10 samples were EqPV-H positive. CONCLUSIONS AND CLINICAL IMPORTANCE: We demonstrated EqPV-H in 18 consecutive cases of serum hepatitis. EPgV, TDAV, and NPHV were not consistently present. This information should encourage blood product manufacturers to test for EqPV-H and eliminate EqPV-H-infected horses from their donor herds.

Normal reference intervals and the effects of sample handling on dynamic viscoelastic coagulometry (Sonoclot) in healthy adult horses.

OBJECTIVE: To determine reference intervals and the effect of sample agitation and rest time on Sonoclot analysis in healthy adult horses. DESIGN: Original prospective study. SETTING: University veterinary medical teaching hospital. ANIMALS: Sixty healthy adult horses. INTERVENTIONS: Blood was collected for assessment of complete blood count, serum biochemical analysis, and Sonoclot analysis. MEASUREMENTS AND MAIN RESULTS: Horses were determined to be healthy based upon physical examination, CBC, and serum biochemistry analysis. Blood was analyzed in a glass bead-containing cuvette using the Sienco Sonoclot analyzer following 2 rest periods (30 mins and 240 min) and with 2 sample handling interventions (agitated and nonagitated), to obtain values for clot rate, time-to-peak, activated clotting time, and platelet function. This study failed to detect a significant difference when a rest time of 30 minutes was compared with 240 minutes, but based on wide limits of agreement the 2 rest times were not considered interchangeable. Agitation at both rest times significantly affected all Sonoclot analyses leading to changes indicative of hypercoagulability. CONCLUSIONS: Sample agitation and rest time should be taken into consideration when developing preanalytical guidelines for Sonoclot analysis in horses. Calculated reference intervals were relatively wide. Further research is needed to evaluate the clinical utility of Sonoclot analysis in horses.

Efficacy of the early administration of valacyclovir hydrochloride for the treatment of neuropathogenic equine herpesvirus type-1 infection in horses.

OBJECTIVE To determine whether prophylactic administration of valacyclovir hydrochloride versus initiation of treatment at the onset of fever would differentially protect horses from viral replication and clinical disease attributable to equine herpesvirus type-1 (EHV-1) infection. ANIMALS 18 aged mares. PROCEDURES Horses were randomly assigned to receive an oral placebo (control), treatment at detection of fever, or prophylactic treatment (initiated 1 day prior to viral challenge) and then inoculated intranasally with a neuropathogenic strain of EHV-1. Placebo or valacyclovir was administered orally for 7 or 14 days after EHV-1 inoculation or detection of fever (3 horses/group). Effects of treatment on viral replication and clinical disease were evaluated. Plasma acyclovir concentrations and viremia were assessed to determine inhibitory concentrations of valacyclovir. RESULTS Valacyclovir administration decreased shedding of virus and viremia, compared with findings for control horses. Rectal temperatures and clinical disease scores in horses that received valacyclovir prophylactically for 2 weeks were lower than those in control horses. The severity of but not the risk for ataxia was decreased by valacyclovir administration. Viremia was decreased when steady-state trough plasma acyclovir concentrations were > 0.8 µg/mL, supporting the time-dependent activity of acyclovir. CONCLUSIONS AND CLINICAL RELEVANCE Valacyclovir treatment significantly decreased viral replication and signs of disease in EHV-1-infected horses; effects were greatest when treatment was initiated before viral inoculation, but treatment was also effective when initiated as late as 2 days after inoculation. During an outbreak of equine herpesvirus myeloencephalopathy, antiviral treatment may be initiated in horses at various stages of infection, including horses that have not yet developed signs of viral disease.

Effects of tourniquet number and exsanguination on amikacin concentrations in the radiocarpal and distal interphalangeal joints after low volume intravenous regional limb perfusion in horses.

OBJECTIVE: To determine the influence of a dual tourniquet technique and limb exsanguination on amikacin concentrations in the synovial fluid of the radiocarpal joint (RCJ) and distal interphalangeal joint (DIPJ) after low volume, cephalic intravenous regional limb perfusion (IVRLP). STUDY DESIGN: Randomized cross-over design. ANIMALS: Six healthy adult horses. METHODS: One gram of amikacin in 6 mL of 0.9% NaCl was infused via cephalic IVRLP in 6 standing, sedated horses using 4 techniques: proximal pneumatic tourniquet (P), proximal pneumatic tourniquet with exsanguination (PE), proximal pneumatic and distal Esmarch tourniquet (PD), and proximal pneumatic with distal Esmarch tourniquet and exsanguination (PDE). Amikacin concentrations were measured in RCJ and DIPJ synovial fluid samples, collected just before perfusion (time 0), and at 15 and 30 minutes (before tourniquet release) after perfusion. RESULTS: Synovial fluid amikacin concentrations achieved in the RCJ were higher with techniques PD and PDE than those achieved with techniques P and PE 15 and 30 minutes after perfusion (P < .0001). Synovial fluid amikacin concentrations in the DIPJ were higher with techniques P and PE than those achieved with techniques PD and PDE at 15 minutes (P = .0002) and were higher than technique PDE at 30 minutes after perfusion (P < .0001). CONCLUSION: Low volume (10 mL) cephalic IVRLP should be combined with the placement of 2 tourniquets (proximal and distal to the carpus) to achieve therapeutic amikacin concentrations in the RCJ. Exsanguination prior to low volume IVRLP does not alter synovial fluid amikacin concentrations.

Effect of Regional Intravenous Limb Perfusate Volume on Synovial Fluid Concentration of Amikacin and Local Venous Blood Pressure in the Horse.

OBJECTIVE: To determine the effect of volume of amikacin perfusate for intravenous regional limb perfusion (IVRLP) via the cephalic vein in standing, sedated horses on (1) amikacin concentrations in the synovial fluid of the radiocarpal joint (RCJ) and distal interphalangeal joint (DIPJ) and, (2) amikacin concentration in the systemic circulation, and (3) regional intravenous pressure. STUDY DESIGN: Randomized cross-over design. ANIMALS: Six adult horses. METHODS: Each horse received IVRLP using 4 perfusate volumes (10, 30, 60 & 120 ml) in random order, after a minimum of 1 week washout. After application of a pneumatic tourniquet, IVRLP with 1 g of amikacin in 0.9% NaCl was performed. Synovial fluid from the RCJ and DIPJ, and systemic and regional venous blood were sampled, and regional blood pressure was measured, immediately before perfusion (time 0), and 15 and 30 minutes after perfusion but before tourniquet release. RESULTS: No difference was observed in the mean amikacin concentration of synovial fluid for the 4 perfusate volumes (P>.09). For all volumes, mean amikacin concentration for DIPJ synovial fluid was higher than for RCJ (P<.0001). The mean amikacin concentration in DIPJ synovial fluid was therapeutic for resistant pathogens using the 10, 60, and 120 mL volumes but the mean amikacin concentration for RCJ synovial fluid was not therapeutic for resistant pathogens with any perfusate volume. All volumes resulted in an immediate increase in mean regional intravascular pressure after perfusion (P<.0001) but was not different across the 4 perfusate volumes. CONCLUSION: Cephalic IVRLP of 1 g of amikacin diluted to a volume of 10-120 mL with 0.9% NaCl will achieve amikacin concentrations therapeutic for resistant pathogens in the synovial fluid from the DIPJ. Concentrations below therapeutic levels for resistant pathogens are reached in the synovial fluid from the RCJ.

Relationship of skeletal muscle inflammation with obesity and obesity-associated hyperinsulinemia in horses.

Local (skeletal muscle and adipose) and systemic inflammation are implicated in the development of obesity-associated insulin resistance in humans. In horses, obesity is neither strongly nor consistently associated with systemic inflammation. The role of skeletal muscle inflammation in the development of insulin dysregulation (insulin resistance or hyperinsulinemia) remains to be determined. We hypothesized that skeletal muscle inflammation is related to obesity-associated hyperinsulinemia in horses. Thirty-five light-breed horses with body condition scores (BCSs) of 3/9 to 9/9 were studied, including 7 obese, normoinsulinemic (BCS ≥ 7, resting serum insulin < 30 µIU/mL) and 6 obese, hyperinsulinemic (resting serum insulin ≥ 30 µIU/mL) horses. Inflammatory biomarkers were evaluated in skeletal muscle biopsies and plasma. Relationships between markers of inflammation and BCS were evaluated. To assess the role of inflammation in obesity-associated hyperinsulinemia, markers of inflammation were compared among lean or ideal, normoinsulinemic (L-NI); obese, normoinsulinemic (O-NI); and obese, hyperinsulinemic (O-HI) horses. Skeletal muscle and plasma tumor necrosis factor alpha (TNFα) concentrations were negatively correlated with BCS. When comparing inflammatory markers among groups, skeletal muscle TNFα was lower in the O-HI group than in the O-NI or L-NI groups. In horses, neither skeletal muscle nor systemic inflammation appears to be positively related to obesity or obesity-associated hyperinsulinemia.

L'inflammation locale (muscle squelettique et tissu adipeux) et systémique sont impliquées dans le développement de la résistance à l'insuline associée à l'obésité chez l'humain. Chez les chevaux, l'obésité n'est pas fortement ou de manière constante associée avec l'inflammation systémique. Le rôle de l'inflammation des muscles squelettiques dans le développement de la dérégulation de l'insuline (résistance à l'insuline ou hyper-insulinémie) reste à être déterminé. Nous avons émis l'hypothèse que chez les chevaux l'inflammation des muscles squelettiques est reliée à l'hyper-insulinémie associée à l'obésité. Trente-cinq chevaux de race légère avec des pointages de condition corporelle (PCCs) variant de 3/9 à 9/9 ont été étudiés, incluant sept chevaux obèses, normo-insulinémique (PCC ≥ 7, insuline sérique au repos < 30 µUI/mL) et six chevaux obèses, hyper-insulinémique (insuline sérique au repos ≥ 30 µUI/mL). Les biomarqueurs de l'inflammation ont été évalués dans des biopsies de muscles squelettiques et le plasma. Les relations entre les marqueurs de l'inflammation et le PCC ont été évaluées. Pour apprécier le rôle de l'inflammation dans l'hyper-insulinémie associée à l'obésité, les marqueurs de l'inflammation ont été comparés parmi les chevaux élancés ou idéal, normo-insulinémique (L-NI); les chevaux obèses, normo-insulinémique (O-NI); et les chevaux obèses, hyperinsulinémique (O-HI). Les concentrations du facteur nécrosant des tumeurs alpha (TNFα) étaient corrélées négativement avec le PCC. Lors de la comparaison des marqueurs de l'inflammation entre les groupes, la concentration de TNFα dans les muscles squelettiques était plus basse dans le groupe O-HI que dans les groupes O-NI ou L-NI. Chez les chevaux, ni l'inflammation systémique ou celle des muscles squelettiques ne semblent reliées positivement à l'obésité ou à l'hyper-insulinémie associée à l'obésité.(Traduit par Docteur Serge Messier).

Quantitative assessment of two methods of tiludronate administration for the treatment of lameness caused by navicular syndrome in horses.

OBJECTIVE: To determine effects of 2 tiludronate administration protocols on measures of lameness in horses with navicular syndrome (NS). ANIMALS: 12 horses with bilateral forelimb NS. PROCEDURES: Horses were randomly assigned to receive tiludronate (1 mg/kg), diluted in 5 L of isotonic electrolyte solution and delivered through a jugular vein catheter (systemic treatment group; n = 6), or tiludronate (0.1 mg/kg), diluted with saline (0.9% NaCl) solution to a total volume of 35 mL and delivered into the lateral digital vein of each forelimb with an IV regional limb perfusion (IVRLP) technique (IVRLP group; 6). Mean peak vertical ground reaction force (pVGRF) measured with a stationary force plate and subjective lameness scores (SLSs) were recorded before (day -1) and at predetermined time points after tiludronate administration on day 0. Mean pVGRFs (standardized as percentage body weight of force) and mean SLSs for the most lame forelimb and for both forelimbs of horses in each group were compared with day -1 values to determine treatment effect. RESULTS: Mean pVGRF for both forelimbs and for the most lame forelimbs of systemically treated horses were significantly increased on days 120 and 200, compared with day -1 results. No significant difference in mean pVGRF was observed for IVRLP-treated horses. The SLSs were not improved at any time point following systemic treatment and were improved only on day 120 following IVRLP. CONCLUSIONS AND CLINICAL RELEVANCE: Tiludronate (1 mg/kg, IV) as a single systemic treatment appeared to be beneficial for horses with NS, but no horses were judged as sound during the study period. Additional research on IVRLP with tiludronate is needed before this method can be recommended.

Comparison of echocardiographic measurements in elite and nonelite Arabian endurance horses.

OBJECTIVE: To determine whether echocardiographic variables differed between successful (elite) and less successful (nonelite) Arabian endurance horses. ANIMALS: 34 Arabian horses that competed in endurance racing. PROCEDURES: Horses were assigned to either an elite or nonelite group on the basis of results of a previous competition, and a standardized echocardiographic examination was performed on each horse within 1 to 4 weeks after that competition. Multivariable logistic regression with backward stepwise elimination was used to create a prediction model for the determination of horse status (elite or nonelite) as a function of the measured echocardiographic variables. RESULTS: The elite and nonelite groups consisted of 23 and 11 horses, respectively. One horse in the nonelite group had a frequent ventricular dysrhythmia that could have negatively affected its performance and rider's safety, whereas none of the horses in the elite group had remarkable cardiac abnormalities. The left ventricular internal diameter during systole and diastole and left ventricular mass and stroke volume were significantly greater for horses in the elite group, compared with those for horses in the nonelite group. The final logistic regression model correctly predicted the horse status for all of the horses in the elite group and 8 of 11 horses in the nonelite group. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that heart size was significantly associated with performance for Arabian endurance horses in a manner similar to findings for Thoroughbred and Standardbred racehorses in active competition.

Antibodies reactive to Ehrlichia spp. are common in Oklahoma horses.

Abstract Tick infestations and infection with tick-borne agents are commonly recognized in horses in North America, but equine infection with true Ehrlichia spp. has not been described. To determine the degree to which horses in the south-central United States are naturally exposed to and infected with tick-borne disease agents, serum samples were collected at random (n=240) or from horses with active tick infestations (n=73) and tested by immunofluorescence antibody assay (IFA) and/or enzyme-linked immunosorbent assay (ELISA) for evidence of antibodies reactive to Ehrlichia spp., Anaplasma spp., and Borrelia burgdorferi. Positive samples were further evaluated by species-specific serology for antibodies reactive to E. canis and E. chaffeensis, and whole blood samples were tested by PCR for evidence of infection with E. canis, E. chaffeensis, E. ewingii, and an E. ruminantium-like organism referred to as the Panola Mountain Ehrlichia. Antibodies reactive to Ehrlichia spp. were identified in 8.75% (21/240) of the randomly acquired samples and 24.7% (18/73) of the serum samples from tick-infested horses, but species-specific ELISA and PCR failed to confirm exposure to or infection with any known Ehrlichia spp. Antibodies to Anaplasma spp. (5/313; 1.6%) and B. burgdorferi (3/313; 1.0%) were uncommon. These data suggest that horses in the south-central United States are likely exposed to a novel Ehrlichia sp. Further research is needed to identify the etiologic agent responsible for the serologic activity seen and to determine the clinical significance, if any, of this finding.

Development of a double sandwich fluorescent ELISA to detect rattlesnake venom in biological samples from horses with a clinical diagnosis of rattlesnake bite.

Rattlesnake bites in horses are not uncommon and the clinical outcomes are widely variable. Treatment of horses with anti-venom is often cost prohibitive and could have negative consequences; therefore, the development of a quantitative test to determine if anti-venom therapy is indicated would be valuable. The objective of this study was to develop an ELISA to detect rattlesnake venom in biological samples from clinically bitten horses. Nineteen horses were enrolled in the study. Urine was available from 19 horses and bite site samples were available from 9 horses. A double sandwich fluorescent ELISA was developed and venom was detected in 5 of 9 bite site samples and 12 of 19 urine samples. In order to determine if this assay is useful as a guide for treatment, a correlation between venom concentration and clinical outcome needs to be established. For this, first peak venom concentration needs to be determined. More frequent, consistent sample collection will be required to define a venom elimination pattern in horses and determine the ideal sample collection time to best estimate the maximum venom dose. This report describes development of an assay with the ability to detect rattlesnake venom in the urine and at the bite site of horses with a clinical diagnosis of rattlesnake bite.

Antibody responses to natural rattlesnake envenomation and a rattlesnake toxoid vaccine in horses.

Antivenom antibody titers following administration of rattlesnake venom for antivenom production in horses are well documented; however, antivenom antibody titers following natural rattlesnake envenomation in horses are not. Antibody titers produced in response to the commercially available rattlesnake venom vaccine are also not published. Our study objectives were to measure antivenom antibody titers in rattlesnake-bitten horses and compare them to titers in horses vaccinated with the rattlesnake venom vaccine. Additionally, titers were compared in pregnant versus nonpregnant horses to assess the affect of pregnancy on vaccine response and were measured pre- and postsuckle in foals of vaccinated mares to detect passive transfer of vaccine immunoglobulins. Blood samples were collected from 16 rattlesnake-bitten horses. Thirty-six horses (11 pregnant mares, 12 nonpregnant mares, 13 geldings) were vaccinated using a Crotalus atrox venom toxoid vaccine. Blood was collected before administering each vaccination and 30 days following the third vaccination. Blood was collected from foals of vaccinated mares pre- and postsuckle. All serum was assayed for anti-Crotalus atrox venom antibodies using an enzyme-linked immunosorbent assay (ELISA). Rattlesnake-bitten horses had higher (P = 0.001) titers than vaccinated horses. There was no significant difference between titers in vaccinated pregnant versus nonpregnant horses. One mare had a positive titer at foaling, and the foals had positive postsuckle titers. Antivenom antibody titer development was variable following natural envenomation and vaccination, and vaccine-induced titers were lower than natural envenomation titers. Further studies are required to determine if natural or vaccine antivenom antibody titers reduce the effects of envenomation.

Prevalence and species of ticks on horses in central Oklahoma.

Ticks are common on horses, but there is a dearth of contemporary data on infestation prevalence, predominant species, and tick-borne disease agents important in this host. To determine the species of ticks most common on horses and the prevalence of equine exposure to and infection with tick-borne disease agents, ticks and blood samples were collected from 73 horses during May, June, and July of 2010. Adult ticks were identified to species, and antibodies to Ehrlichia spp., Anaplasma spp., and Borrelia burgdorferi were identified using indirect fluorescence antibody assay, a commercial point-of-care enzyme-linked immunosorbent assay, or both. In total, 1,721 ticks were recovered at the majority (85%) of equid examinations. Amblyomma americanum (L.) was the most common tick collected (1,598 out of 1,721; 92.9%) followed by Dermacentor variabilis (Say, 1821) (85 out of 1,721; 4.9%) and Amblyomma maculatum Koch, 1844 (36 out of 1,721; 2.1%); single specimens of Ixodes scapularis Say, 1821 and Dermacentor albipictus (Packard, 1869) were also identified. Antibodies reactive to Ehrlichia spp. were found in 18 out of 73 (24.7%) of horses tested, and were more commonly identified in horses with moderate or high tick infestations than those with low tick infestations (P < 0.001). These data support A. americanum as the most common tick species infesting horses in central Oklahoma from May through July and suggest horses are also commonly exposed to an Ehrlichia sp.

Neutrophil and cytokine dysregulation in hyperinsulinemic obese horses.

Equine metabolic syndrome is characterized by obesity and regional adiposity coupled with evidence of recurrent laminitis. Although inflammation has been well characterized in several experimental models of acute laminitis, the inflammatory events associated with endocrinopathic laminitis are not well documented. The aim of this study was to characterize selected markers of inflammation in horses with clinical evidence of equine metabolic syndrome (EMS). Neutrophil phagocytosis and oxidative burst, as well as endogenous and stimulated cytokine expression were evaluated. A marked increase in neutrophil reactive oxygen species production upon phagocytosis was observed in horses with EMS that was strongly correlated to the blood insulin concentration. Increased oxidative burst activity of neutrophils in hyperinsulinemic horses may predispose horses with metabolic syndrome to laminitis. In contrast, peripheral blood cells of obese hyperinsulinemic horses showed decreased endogenous proinflammatory cytokine gene expression (IL-1 and IL-6) and similar cytokine response following immune stimulation compared to that of control horses. This may suggest that, unlike in people, cytokine-mediated inflammation does not increase in direct response to obesity or insulin resistance in horses. This species-specific disparity may explain the difference in clinical outcomes observed in obese horses compared to obese people.

Gastric and enteric phytobezoars caused by ingestion of persimmon in equids.

CASE DESCRIPTION-13 equids (10 horses, 2 donkeys, and 1 pony) were examined for signs of colic (n = 7), weight loss (6), anorexia (3), and diarrhea (2). Ten equids were evaluated in the fall (September to November). Seven equids had a history of persimmon ingestion. CLINICAL FINDINGS-A diagnosis of phytobezoar caused by persimmon ingestion was made for all equids. Eight equids had gastric persimmon phytobezoars; 5 had enteric persimmon phytobezoars. Gastroscopy or gastroduodenoscopy revealed evidence of persimmon ingestion in 8 of 10 equids in which these procedures were performed. TREATMENT AND OUTCOME-2 of 13 equids were euthanatized prior to treatment. Supportive care was instituted in 11 of 13 equids, including IV administration of fluids (n = 8) and treatment with antimicrobials (5), NSAIDs (5), and gastric acid suppressants (4). Persimmon phytobezoar-specific treatments included dietary modification to a pelleted feed (n = 8); oral or nasogastric administration of cola or diet cola (4), cellulase (2), or mineral oil (2); surgery (4); and intrapersimmon phytobezoar injections with acetylcysteine (1). Medical treatment in 5 of 7 equids resulted in resolution of gastric persimmon phytobezoars. Seven of 8 equids with gastric persimmon phytobezoars and 1 of 5 equids with enteric persimmon phytobezoars survived > 1 year after hospital discharge. CLINICAL RELEVANCE-Historical knowledge of persimmon ingestion in equids with gastrointestinal disease warrants gastroduodenoscopy for evaluation of the presence of persimmon phytobezoars. In equids with gastric persimmon phytobezoars, medical management (including administration of cola or diet cola and dietary modification to a pelleted feed) may allow for persimmon phytobezoar dissolution.