Ex vivo comparison of one-layer versus two-layer closures in end-to-end anastomoses in normal equine descending colon.

OBJECTIVE: The aim of the study was to evaluate time to closure, bursting pressures and luminal diameters of a single and double-layer end-to-end anastomoses in normal equine descending colon. STUDY DESIGN: Experimental study. SAMPLE POPULATION: Eight adult horses. METHODS: Four segments of descending colon from each horse were randomly assigned to a control (n = 8, CON), one-layer (n = 12, group 1; continuous Lembert pattern), or two-layer group (n = 12, group 2; simple continuous oversewn with Cushing pattern). Anastomoses were performed, and time to closure and luminal diameter were measured. Bursting pressures were determined, and location of failure was recorded. Mixed analysis of variance (ANOVA) was employed. RESULTS: Mean time to completion was decreased (p = .003) in group 1 (18.6 min ± 22.8 s) compared with group 2 (21.35 min ± 22.8 s). Luminal diameter was reduced (p < .0001) in group 2 (47 ± 2.46 mm) compared to group 1 (65 ± 2.77 mm). Location of failure was remote to the anastomosis in all but one segment; therefore, differences in anastomotic bursting pressures could not be determined. CONCLUSION: Two-layer closures resulted in significant luminal reduction and took longer to complete than one-layer anastomoses. CLINICAL SIGNIFICANCE: Use of one-layer closures may be advantageous primarily with respect to luminal diameter.

The influence of a tension band fixation as an adjunct for arthrodesis of the metacarpophalangeal joint in the horse.

OBJECTIVES: To determine the influence of a stainless-steel cable (SSC) tension band fixation as an adjunct to a locking compression plate (LCP) for arthrodesis of the equine metacarpophalangeal (MCP) joint. STUDY DESIGN: Experimental. An ex vivo biomechanical paired equine cadaver limb study. SAMPLE POPULATION: Five MCP joint pairs were collected from adult Thoroughbred horses, euthanized for reasons unrelated to orthopedic disease. METHODS: Each pair of MCP joints were randomly implanted with either a dorsally placed 5.5 mm LCP and a palmarly placed 2.0 mm SSC or a dorsally placed 5.5 mm LCP alone. Each construct was tested in cyclic loading followed by single cycle to failure in axial compression. Displacement at a target load of 1 kN over 3600 cycles at 1 Hz was recorded prior to single cycle to failure testing. RESULTS: In cyclic testing, displacement was not significantly different between the first and last 5% of testing cycles regardless of construct. Maximum displacement of each construct during cyclic testing was <1.1 mm. In single cycle testing, the observed yield point did not reveal any difference between LCP and LCP-SSC (p = .440). The maximum load at failure was significantly higher in LCP-SSC compared to constructs with the LCP alone (p = .046). CONCLUSION: The addition of the SSC to the LCP did not statistically affect construct displacement during cyclic loading or construct yield load during subsequent single cycle to failure. CLINICAL SIGNIFICANCE: This study provided much needed information regarding the necessity of a tension band SSC application in the arthrodesis of the MCP/MTP joint in horses.

Pharmacokinetics of intra-articular buprenorphine in horses with lipopolysaccharide-induced synovitis.

The objective of this study was to describe the pharmacokinetics of intra-articular (IA) administered buprenorphine in horses with lipopolysaccharide (LPS)-induced synovitis. Radiocarpal synovitis was induced in six healthy adult horses with the IA injection of LPS (0.5 ng/joint) on two occasions in a randomized cross-over design. Treatments (IA buprenorphine (IAB) at 5 µg/kg plus intravenous saline; and intravenous buprenorphine (IVB) at 5 µg/kg plus IA saline) were administered 4 h following LPS injection. Concentrations of buprenorphine were assessed in plasma and synovial fluid (SF) at 0.5, 2, 6, 12, and 24 h after administration. Pharmacokinetic parameters after IVB and IAB in plasma and synovial fluid were calculated using a nonlinear mixed effects model. IAB was detectable in SF of all horses at 24 h [median concentration of 6.2 (3.46-22.6) ng/mL]. IAB resulted in a median plasma concentration of 0.59 (0.42-1.68) ng/mL at 0.5 h and was detectable in all subjects for up to 6 h and in two horses for up to 12 h. IVB resulted in SF concentrations detected up to 6 h in all horses [median concentration of 0.12 (0.07-0.82) ng/mL]. Results suggest that IA buprenorphine remains present in the inflamed joint for at least 24 h and systemic absorption occurs.

The effects of bit chewing on borborygmi, duodenal motility, and gastrointestinal transit time in clinically normal horses.

OBJECTIVE: To determine the influence of bit chewing on gastrointestinal transit in clinically normal horses. STUDY DESIGN: Prospective crossover designed study. ANIMALS: Six healthy adult horses. METHODS: Horses were assigned randomly to treatment (apple flavored bit) and control (no-bit) groups and studied for 2 × 1-week trial periods with a 2-week washout period between trials. Horses were fasted for 24 h and slowly refed over 3 days. The bit was placed for 20 min every 6 h. Duodenal contractions and borborygmi auscultations were evaluated every 12 h, approximately 5 min following bit placement. Gastrointestinal total transit time (GI TTT) was measured by administering 200 colored beads via stomach tube and then collected in the manure until 50% and 80% were recovered. Measured variables were compared using an ANOVA or Wilcoxon signed-rank test and the P value was noted. RESULTS: The GI TTT was shortened in the bit chewing group (median: 106.37 h, range: 70-171 h) compared to the no-bit group (median: 170.1 h, range: 149-186 h) (P = .0156) at 80% bead passage (only 4/6 horses passed 80%). Borborygmi (P = .8193), duodenal contractions (P = .2605), and 50% bead passage (P = .0781) showed no differences. CONCLUSION: Bit chewing was safe, inexpensive, and well tolerated. Bit chewing shortened GI TTT and might be an adjunct therapy to augment GI TTT. Further clinical studies are warranted. CLINICAL SIGNIFICANCE: Ileus is a common complication following equine abdominal surgery with no current consistently successful treatment. Bit chewing may be a simple and inexpensive way to augment progressive GI motility.

A study of ultrasound-guided perineural injection of the caudal cervical spinal nerve roots in equine cadavers.

OBJECTIVES: To develop an ultrasound-guided cervical perineural injection technique for horses and to evaluate and compare the distribution of contrast agent among perineural, intra-articular and periarticular injections. STUDY DESIGN: Prospective, experimental cadaveric study. ANIMALS: A total of 14 equine cadaveric necks. METHODS: Bilateral ultrasound-guided perineural injection technique for the caudal cervical spinal nerve roots (CSNRs 5-7) was developed. Paramagnetic or iodinated contrast was injected and the distribution of contrast was evaluated using magnetic resonance (MR) or computed tomography (CT) imaging, respectively. The presence of contrast in the CSNR region was determined by an observer unaware of the technique used for each injection performed. The ability of the perineural injection technique to distribute contrast agent to the CSNR region was compared with intra-articular and periarticular injection techniques. RESULTS: Perineural injection delivered contrast agent to the CSNR region 100% of the time and was significantly different when compared with intra-articular injection (p = 0.008). There was no difference in ability to deliver contrast agent to the CSNR region between the perineural and periarticular injection techniques or between the intra-articular and periarticular injection techniques. CONCLUSION AND CLINICAL RELEVANCE: The ultrasound-guided perineural injection technique developed in this study accurately delivered contrast agent to the CSNR region in equine cadavers. This technique could potentially be used for the diagnosis and treatment of cervical pain in horses, particularly in cases where intra-articular cervical articular process joint injections have not been beneficial. Further studies are necessary to assess the effectiveness of the ultrasound-guided perineural injection technique in live horses.

Ovarian histopathology, pre- and post-operative endocrinological analysis and behavior alterations in 27 mares undergoing bilateral standing laparoscopic ovariectomy.

Determining if ovariectomy will abolish unwanted behaviors can be challenging in mares without ultrasonographic ovarian abnormalities and/or endocrine values not consistent with a granulosa cell tumor. The objective of the study was to identify mares that have a positive improvement in unwanted behaviors after elective standing laparoscopic ovariectomy and to determine if the change is associated with any pre- and post-operative endocrine profiles and/or with ovarian histological findings. During the study period 27 mares underwent a bilateral standing laparoscopic ovariectomy. Pre- and post-operative questionnaires and endocrine profiles, along with ovarian histopathology were performed and data were analyzed. Eighty-nine percent of clients were satisfied with the change in behavior after surgery. There were no significant associations between specific elevated hormones or absent luteal tissue with any specific unwanted behaviors. Pre-operative endocrine values were not predictive of post-operative owner satisfaction but bilateral ovariectomy for the treatment of unwanted behaviors in mares had a high likelihood of achieving owner satisfaction.

Histopathologie ovarienne, analyse endocrinienne pré- et post-opération et modifications du comportement chez 27 juments soumises à une ovariectomie bilatérale laparoscopique en position debout. Déterminer si une ovariectomie éliminera les comportements non-désirés peut représenter un défi chez les juments sans anomalie ovarienne détectable par échographie et/ou ayant des valeurs endocrines non cohérentes avec une tumeur des cellules de la granulosa. L'objectif de la présente étude était d'identifier les juments avec une amélioration positive des comportements non-désirés à la suite d'une ovariectomie élective par laparoscopie en position debout et de déterminer si le changement était associé avec un quelconque profil endocrinien pré- et post-opération et/ou avec des trouvailles histologiques dans les ovaires. Durant la période d'étude 27 juments ont été soumises à une ovariectomie laparoscopique bilatérale en position debout. Des questionnaires pré- et post-opération ainsi que des profils endocriniens et des analyses histopathologiques des ovaires furent réalisés et les données analysées. Quatre-vingt-neuf pourcents des clients étaient satisfaits avec le changement dans le comportement après la chirurgie. Il n'y avait pas d'association significative entre des niveaux élevés d'hormones spécifiques ou l'absence de tissu lutéal et des comportements non-désirés spécifiques. Les valeurs endocrines pré-opération n'étaient pas prédictives de la satisfaction post-opération des propriétaires mais l'ovariectomie bilatérale pour le traitement de comportements non-désirés chez les juments avait un grand potentiel d'obtenir la satisfaction du propriétaire.(Traduit par Dr Serge Messier).

Evaluation of post-operative pain after active desufflation at completion of laparoscopy in mares undergoing ovariectomy.

The cause of transient post-operative pain in a subset of horses undergoing laparoscopy is unclear. The objective of this study was to evaluate if residual pneumoperitoneum is associated with transient post-operative pain in mares undergoing ovariectomy. Thirty-eight mares undergoing routine standing laparoscopic ovariectomy were randomly allocated into 2 groups. At the completion of laparoscopy, either the abdominal cavity was actively desufflated or the cannulas were opened to achieve ambient pressure before incisional closure. Assessments were performed for 18 hours after surgery using a validated visceral/somatic pain scale for horses. Overall, pain was minimal in both groups (median score 2/39) post-surgery. Active desufflation of the pneumoperitoneum at the completion of laparoscopy approached statistical significance (P = 0.07) in decreasing pain at 12 hours after laparoscopy. However, effects of active desuffation were not significant throughout the monitored 18-hour post-surgery period. We conclude that the decision to actively desufflate at the completion of laparoscopy should be based on surgeon's preference.

Évaluation de la douleur postopératoire après un désoufflement actif à l'achèvement d'une laparoscopie chez les juments subissant une ovariectomie. La cause d'une douleur postopératoire transitoire dans un sous-groupe de chevaux subissant une laparoscopie est indéterminée. Cette étude avait pour objectif d'évaluer si le pneumopéritoine résiduel est associé à de la douleur postopératoire chez les juments subissant une ovariectomie. Trente-huit juments subissant une ovariectomie laparascopique debout de routine ont été réparties au hasard dans deux groupes. À l'achèvement de la laparoscopie, soit la cavité abdominale a été activement désoufflée ou les canules ont été ouvertes pour obtenir une pression ambiante avant la fermeture de l'incision. Des évaluations ont été réalisées pendant 18 heures après la chirurgie en utilisant une échelle de douleur viscérale/somatique validée pour les chevaux. Dans l'ensemble, la douleur a été minimale dans les deux groupes (note médiane de 2/39) après la chirurgie. La désoufflement actif du pneumopéritoine à l'achèvement de la laparoscopie s'approchait de l'importance statistique (valeur de P = 0,07) en réduisant la douleur à 12 heures après la laparoscopie. Cependant, les effets du désoufflement actif n'étaient pas significatifs pendant la période de surveillance 18 heures après la chirurgie. La décision de désouffler activement à l'achèvement de la laparoscopie est toujours sujette à la préférence du chirurgien.(Traduit par Isabelle Vallières).

Topical Wound Medications.

Topical therapies are used in equine wound healing to clean and decontaminate the wound environment after acute injury and to promote healing and decrease the risk of infection once the wound has initially been treated. Evolving antibiotic resistance has prompted judicious use of systemic antimicrobials, particularly in the treatment of local infections, such as wounds. The use of topical antiseptics to disinfect acute wounds and topical antimicrobials to manage chronic wounds is necessary to achieve successful healing. In addition, many topical medications can alter the wound environment to promote rapid and effective wound healing.

Apoptosis in epithelial cells and its correlation with leukocyte accumulation in lamellar tissue from horses subjected to experimental sepsis-associated laminitis.

Inflammation and apoptosis in the hoof lamellar interface both contribute to the early stages of sepsis-associated laminitis, but it is not clear whether apoptosis is occurring before the onset of inflammation or is being provoked by inflammation. Apoptosis and inflammation were therefore measured in lamellar tissues obtained at different time points throughout the early stages of experimentally induced laminitis. Apoptotic cells and leukocyte were enumerated in archived paraffin embedded lamellar tissue samples from previous experiments in which acute laminitis was induced using Black Walnut Extract (BWE) or starch (CHO). BWE-derived samples from 20 horses were allocated into four groups: Control (CON = 5); Early Time Point (ETP, 1.5 h after induction, n = 5); Developmental Time Point (DTP, 3-4 h after induction, n = 5); Obel Grade 1 (OG1, Onset of Lameness, n = 5). CHO-derived samples from 25 horses were allocated into four groups: CON (n = 8); DTP (10-12 h after induction, n = 6); OG 1 (n = 6); Obel 3 (OG3, lameness progression, n = 5). Apoptotic cells were enumerated using a horse validated TUNEL technique. Compared to controls, significant increases in apoptotic cell counts were not detected in lamellar epithelial cells during the developmental phase or at the onset of lameness during laminitis induction. A negative correlation between apoptosis and leukocyte infiltration was detected in the BWE model (P < 0.05). In conclusion, apoptosis does not play an important role in the initial stages of sepsis-related laminitis.

Pharmacokinetics and pharmacodynamics of clodronate disodium evaluated in plasma, synovial fluid and urine.

BACKGROUND: Clodronate is a non-nitrogenated bisphosphonate approved for use in horses. There are no peer-reviewed published reports describing the pharmacokinetics or evaluating renal health indices and urinary excretion patterns in conjunction with plasma and synovial fluid concentration following the systemic administration of clodronate to horses. OBJECTIVES: Describe clodronate concentrations in plasma, urine and synovial fluid and evaluate the effects on renal indices after intramuscular administration to healthy horses. STUDY DESIGN: Experimental study with repeated measures. METHODS: Six healthy adult horses received a single intramuscular dose of clodronate (1.8 mg/kg). Blood, synovial fluid and urine were collected prior to and after administration of clodronate up to 72, 48 and 168 hours respectively. Drug concentrations were measured using LC-MS/MS and noncompartmental pharmacokinetic analysis was performed. Renal function indices were also evaluated. RESULTS: Clodronate was quantifiable for up to 24 hours in plasma and 48 hours in synovial fluid and detected at all time points in urine. Maximum plasma concentration of clodronate 210 ± 68.2 ng/mL occurred at approximately 34.8 ± 0.2 minutes after administration, while peak synovial concentration (57.7 ± 32.8 ng/mL) occurred at 2.67 ± 2.32 hours after administration and peak urine concentration (88 358.2 ± 79 521.4 ng/mL) occurred at 2.67 ± 2.58 hours post administration. Terminal half-life in plasma was 3.32 ± 1.25 and was 4.8 ± 3.05 hours in synovial fluid. Creatinine concentrations rose significantly after treatment but remained within normal adult reference ranges at all times. MAIN LIMITATIONS: Limited number of animals and sampling times and the absence of urine collection for determination of concentration beyond 7 days. CONCLUSIONS: Clodronate is rapidly cleared from the blood and synovial fluid. It has variable and biphasic urinary excretion. While significant increase in blood creatinine concentrations was present after a single intramuscular dose of clodronate, values were never above the normal reference range. Further studies are warranted in horses undergoing exercise and those undergoing multiple dosing schemes.

Use of a locking compression plate for equine proximal interphalangeal joint arthrodesis: 29 cases (2008-2014).

OBJECTIVE To describe clinical use of a locking compression plate (LCP) for proximal interphalangeal joint (PIPJ) arthrodesis in horses and compare outcomes for horses that underwent the procedure as treatment for fracture of the middle phalanx (P2) versus other causes. DESIGN Retrospective case series. ANIMALS 29 client-owned horses. PROCEDURES Medical records of 2 veterinary teaching hospitals from 2008 through 2014 were reviewed to identify horses that underwent PIPJ arthrodesis of 1 limb. Signalment, surgical, and outcome-related variables were recorded. Owners were contacted from 1 to 6 years after surgery to determine rehabilitation time, current use of the horse, and overall owner satisfaction with the procedure. Success was determined on the basis of owner satisfaction and outcome for intended use. Variables of interest were compared statistically between horses that underwent surgery for P2 fracture versus other reasons. RESULTS 14 horses underwent surgery for treatment of P2 fracture, and 15 had surgery because of osteoarthritis, subluxation, or osteochondrosis. Median convalescent time after surgery (with no riding or unrestricted exercise) was 7 months. Four horses were euthanized; of 23 known alive at follow-up, 22 were not lame, and 18 had returned to their intended use (8 and 10 at higher and lower owner-reported levels of work, respectively). Horses undergoing arthrodesis for reasons other than fracture were significantly more likely to return to their previous level of work. Twenty-two of 24 owners contacted indicated satisfaction with the procedure. CONCLUSIONS AND CLINICAL RELEVANCE Surgical arthrodesis of the PIPJ was successful in most horses of the study population. Various nuances of the system for fracture repair need to be understood prior to its use.

Effects of intramuscular administration of acepromazine on palmar digital blood flow, palmar digital arterial pressure, transverse facial arterial pressure, and packed cell volume in clinically healthy, conscious horses.

OBJECTIVE: To determine the magnitude and duration of effects of acepromazine administered intramuscularly (IM) on digital and systemic hemodynamic variables in clinically healthy horses. STUDY DESIGN: Experimental study. ANIMALS: Healthy adult horses (n=12). Methods- An ultrasonic Doppler flow probe was surgically implanted around the medial palmar digital artery before the study. Catheters were inserted in the transverse facial artery, lateral palmar digital artery, and jugular vein. A treatment group (n=6) was administered 0.04 mg/kg body weight of acepromazine IM; control horses (n=6) were administered an equivalent volume of saline IM. Palmar digital blood flow, and digital and facial arterial pressures were measured at baseline and for 6 hours after administration. Venous blood was collected for measurement of packed cell volume (PCV). RESULTS: Horses administered acepromazine had significantly lower facial arterial pressure compared with control horses administered saline. Palmar digital arterial blood flow in acepromazine-treated horses was not significantly different from that in control horses but increased significantly post-administration, compared with the respective baseline values for acepromazine-treated horses. PCV significantly decreased in horses administered acepromazine compared with their respective baseline value. CONCLUSION: IM acepromazine causes hypotension and increases palmar digital blood flow over time but the magnitude of the effect on digital blood flow was not sufficient to yield differences compared with saline-treated horses. CLINICAL RELEVANCE: IM acepromazine has a modest effect on palmar digital blood flow, facial arterial pressures and PCV in healthy horses with minimal sedation.

Evaluation of two methods for topical application of contrast medium to the pharyngeal and laryngeal region of horses.

OBJECTIVE To determine the pharyngeal and laryngeal distribution of radiopaque contrast medium administered orally or via nasopharyngeal catheter to standing horses. ANIMALS 5 healthy adult horses. PROCEDURES A crossover study was conducted. Radiopaque contrast medium (12 mL) was administered orally and via nasopharyngeal catheter to each horse. Pharyngeal and laryngeal distribution of contrast medium was determined by examination of radiographs obtained immediately after administration of contrast medium, compared with those obtained before administration. Regional distribution of contrast medium was graded. Endoscopic examination of the nasopharynx, laryngopharynx, and larynx was performed to confirm radiographic results. RESULTS Examination of radiographs obtained after nasopharyngeal administration revealed contrast medium in the nasopharynx (n = 5), oropharynx (2), laryngopharynx (3), and larynx (5) of the 5 horses. Examination of radiographs obtained after oral administration revealed contrast medium in the oropharynx (n = 4) and larynx (1) of the 5 horses. Endoscopic examination confirmed radiographic findings and was found to be sensitive for detection of contrast medium in the laryngopharynx, whereby detection rates were higher for both administration methods. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that medication administered by use of a nasopharyngeal catheter will result in topical distribution within the nasopharynx, including the dorsal surface of the soft palate, and larynx, although distribution should be evaluated in horses with clinical airway disease to confirm these findings. Oral administration did not result in consistently detectable topical laryngeal distribution but could be used for selected conditions (eg, palatitis).

Concentrations of serum amyloid A and plasma fibrinogen in horses undergoing emergency abdominal surgery.

OBJECTIVE: To compare the perioperative response of serum amyloid A (SAA) to fibrinogen in horses requiring exploratory celiotomy for colic and to determine if SAA could be used to predict complications and outcome. DESIGN: Prospective observational clinical study. SETTING: University teaching hospital. ANIMALS: Eighteen horses undergoing exploratory celiotomy for colic. Inclusion criteria for the study included survival and anesthetic recovery from exploratory celiotomy, no history of surgery within the past year. INTERVENTIONS: Blood was obtained via jugular venipuncture before surgery (time 0) and at 24, 48, 72, and 96 hours after recovery from anesthesia. MEASUREMENTS AND MAIN RESULTS: Quantitative and semiquantitative fibrinogen, SAA, total nucleated cell counts, and total protein were evaluated at each time point. Multivariable linear regression was used to assess differences at each time point and after grouping horses according to duration of colic prior to surgery, strangulating surgical lesion or not, presence of systemic inflammatory response syndrome (SIRS) on admission, and postsurgical complications. Significant (P < 0.05) increases in SAA concentrations occurred in all cases after surgery compared to fibrinogen concentration, which only demonstrated a mild, clinically insignificant increase postsurgery. SAA concentrations were also significantly increased (P < 0.05) in cases identified with SIRS prior to surgery and postoperatively at 48 (P = 0.05) and 72 hours (P = 0.02) in horses that developed complications. CONCLUSIONS: Measurement of SAA is a more sensitive indicator of inflammation than fibrinogen in the perioperative period of horses requiring exploratory celiotomy for colic. Serial measurement of SAA at 48, 72, and 96 hours after surgery may be helpful to determine risk of complications and guide postoperative management. Measurement of SAA on admission also allows for quantification of SIRS when it is detected clinically.

The effect of hypothermia on influx of leukocytes in the digital lamellae of horses with oligofructose-induced laminitis.

Sepsis-related laminitis (SRL) is a common complication in the septic/endotoxemic critically-ill equine patient, in which lamellar injury and failure commonly lead to crippling distal displacement of the distal phalanx. Similar to organ injury in human sepsis, lamellar injury in SRL has been associated with inflammatory events, including the influx of leukocytes into the lamellar tissue and markedly increased expression of a wide array of inflammatory mediators at the onset of Obel grade 1 (OG1) laminitis. The only treatment reported both clinically and experimentally to protect the lamellae in SRL, local hypothermia ("cryotherapy"), has been demonstrated to effectively inhibit lamellar expression of multiple inflammatory mediators when initiated at the time of administration of a carbohydrate overload in experimental models of SRL. However, the effect of hypothermia on leukocyte influx into affected tissue has not been assessed. We hypothesized that cryotherapy inhibits leukocyte emigration into the digital lamellae in SRL. Immunohistochemical staining using leukocyte markers MAC387 (marker of neutrophils, activated monocytes) and CD163 (monocyte/macrophage-specific marker) was performed on archived lamellar tissue samples from an experimental model of SRL in which one forelimb was maintained at ambient temperature (AMB) and one forelimb was immersed in ice water (ICE) immediately following enteral oligofructose administration (10g/kg, n=14 horses). Lamellae were harvested at 24h post-oligofructose administration (DEV, n=7) or at the onset of OG1 laminitis (OG1, n=7). Both MAC387-positive and CD163-positive cells were counted by a single blinded investigator on images [n=10 (40× fields/digit for MAC387 and 20x fields/digit for CD163)] obtained using Aperio microscopy imaging analysis software. Data were assessed for normality and analyzed with a paired t-test and one-way ANOVA with significance set at p<0.05. MAC387-positive cells were present in low numbers in the lamellar tissue and were decreased in the hypothermic limbs (vs. AMB limbs, p<0.05) in the OG1 group; no change in CD163-positive cell numbers was noted across the conditions of the model. This study demonstrated that hypothermia of the distal limbs instituted early in the disease process in the horse at risk of SRL significantly attenuates the increase of MAC387-positive leukocytes in the digital lamellae, but has minimal effect on increases in lamellar concentrations of the major leukocyte cell type present in that tissue, CD163-positive mononuclear cells.

Development and evaluation of an enzyme-linked immunosorbent assay for the serodiagnosis of pythiosis in dogs.

Pythiosis (caused by the aquatic oomycete Pythium insidiosum) is a devastating and often fatal cause of either severe transmural gastroenteritis or locally invasive subcutaneous disease in dogs living in the southeastern United States. Although early diagnosis is essential for successful treatment, tools available for this task are limited. Therefore, we developed and evaluated an enzyme-linked immunosorbent assay (ELISA) for the detection of anti-P insidiosum antibodies in canine serum. A soluble mycelial extract of P insidiosum was utilized as antigen in the ELISA, which was used to evaluate serum from 43 dogs with pythiosis, 8 dogs with lagenidiosis (another canine oomycosis), 16 dogs with nonoomycotic fungal or algal infections, 22 dogs with nonfungal gastrointestinal or skin disease, and 55 healthy dogs. Results were expressed as percent positivity (PP) relative to a strong positive control serum run on each plate. Medians and ranges for each of the 5 groups were as follows: pythiosis (81.7%, 50.6-98.5%), lagenidiosis (17.3%, 11.3-29.2%), other fungal or algal infections (8.2%, 4.7-15.4%), nonfungal gastrointestinal or skin disease (6.2%, 3.9-20.7%), and healthy dogs (6.7%, 3.0-15.2%). When using a cutoff value of 40% PP, the sensitivity and specificity of the ELISA both were 100%. In addition, ELISA values measured after successful surgical therapy in 2 dogs showed a decrease of anti-P insidiosum antibody concentrations into the normal range as early as 2 months after treatment. We conclude that the ELISA is a sensitive and specific test for the diagnosis of canine pythiosis, and may be a useful tool for monitoring response to medical or surgical therapy.

Laminar chemokine mRNA concentrations in horses with carbohydrate overload-induced laminitis.

Chemokines play a vital role in leukocyte activation and emigration that reportedly plays a central role in laminar injury in equine laminitis. The purpose of this study was to evaluate the pattern of laminar chemokine expression in horses in the classical carbohydrate overload (CHO)-model of laminitis. Laminar samples were obtained 24h following water administration in the control group (CON, n=8), and at the onset of fever (≥ 102°F, 12-22 h post CHO, DEV group, n=8) and at the onset of lameness (20-48 h post CHO, LAM group, n=8) in induced horses. Real time quantitative PCR was performed on all samples in order to determine laminar mRNA concentrations of both CXC chemokines (CXCL1, CXCL6, CXCL8) and CC chemokines (CCL2 [MCP-1], CCL3 [MIP-1α], and CCL8 [MCP-2]). Data were subjected to ANOVA followed by Student-Newman-Keuls (P<0.05). Laminar mRNA concentrations for all CXC chemokines were increased (P<0.05) at both the DEV and LAM horses when compared to the control horses, whereas mRNA concentrations of CCL2 and CCL8 were only increased in the LAM horses when compared to controls and the DEV horses. When taken in context with our previous studies, CXCL1, CXCL6 and CXCL8 increases precede peak laminar leukocyte accumulation. Additionally, CCL2 and CCL8 expression corroborate previous reports of monocyte/macrophage accumulation in affected laminae. Compared with previous studies, our findings demonstrate that increased laminar CXC chemokine expression consistently precedes peak leukocyte accumulation and onset of lameness in CHO laminitis models. Chemokine antagonists may be considered as possible therapeutic targets to decrease the influx of leukocytes that occurs during the development of equine laminitis.