Corrigendum: Extracorporeal Shock Wave Therapy Enhances the In Vitro Metabolic Activity and Differentiation of Equine Umbilical Cord Blood Mesenchymal Stromal Cells.

[This corrects the article DOI: 10.3389/fvets.2020.554306.].

Extracorporeal Shock Wave Therapy Enhances the In Vitro Metabolic Activity and Differentiation of Equine Umbilical Cord Blood Mesenchymal Stromal Cells.

Extracorporeal shock wave therapy (ESWT) has been shown to induce different biological effects on a variety of cells, including regulation and stimulation of their function and metabolism. ESWT can promote different biological responses such as proliferation, migration, and regenerations of cells. Recent studies have shown that mesenchymal stromal cells (MSCs) secrete factors that enhance the regeneration of tissues, stimulate proliferation and differentiation of cells, and decrease inflammatory and immune reactions. Clinically, the combination of these two therapies has been used as a treatment for tendon and ligament lesions in horses; however, there is no scientific evidence supporting this combination of therapies in vivo. Therefore, the objectives of the study were to evaluate the effects of ESWT on equine umbilical cord blood mesenchymal stromal cells (CB-MSCs) proliferative, metabolic, migrative, differentiation, and immunomodulatory properties in vitro. Three equine CB-MSC cultures from independent donors were treated using an electrohydraulic shock wave generator attached to a water bath. All experiments were performed as triplicates. Proliferation, viability, migration and immunomodulatory properties of the cells were evaluated. Equine CB-MSCs were induced to evaluate their trilineage differentiation potential. ESWT treated cells had increased metabolic activity, showed positive adipogenic, osteogenic, and chondrogenic differentiation, and showed higher potential for differentiation toward the adipogenic and osteogenic cell fates. ESWT treated cells showed similar immunomodulatory properties to none-ESWT treated cells. Equine CB-MSCs are responsive to ESWT treatment and showed increased metabolic, adipogenic and osteogenic activity, but unaltered immunosuppressive properties. In vivo studies are warranted to determine if synergistic effects occur in the treatment of musculoskeletal injuries if ESWT and equine CB-MSC therapies are combined.

Case-Control Comparison of Cervical Spine Radiographs From Horses With a Clinical Diagnosis of Cervical Facet Disease With Normal Horses.

The accuracy of using radiographs to diagnose cervical facet osteoarthritis (CFA) in horses is undetermined. Further investigation is required to determine the clinical significance of radiographic evidence of CFA, the prevalence of radiographic changes in horses without clinical signs, and the long-term efficacy of intra-articular CFA treatment. The objectives of this study is to compare degenerative changes of the cervical facet joints of the cervical vertebrae on radiographs of horses with clinical signs of CFA with healthy cohort-matched horses, to compare clinical findings between groups, and to obtain follow-up information on the long-term outcome in treated horses. This is a retrospective case-control cohort-matched comparison study of horses treated for cervical facet disease versus horses with no clinical signs of cervical facet disease. Horses diagnosed with CFA and treated with intra-articular injection of corticosteroids were included. Follow-up information on recovery from treatment was obtained via telephone survey of owners/trainers. Healthy horses with no clinical signs of CFA were matched to treated horses by breed, sex, age, and sport as the control group. Two blinded radiologists reviewed cervical spine radiographs for each horse and recorded CFA score and intravertebral/intervertebral measurements. Clinical and radiographic parameters were compared between treatment and control groups. There was a significant difference in CFA grades for C5-6 and C6-7 between horses with presence of clinical signs and healthy horses. However, interobserver agreement between radiologists for grading CFA was moderate and only 56% of values were identical for both observers. Atrophy of the neck was present on clinical examination in most cases in the treatment group. Dressage horses were overrepresented. Overall, 64% of horses returned to their previous level of performance after treatment. Clinical examination data collected for the treatment group were retrospective and were obtained by different clinicians. Eight owners/trainers were not able to be reached for the survey. There was a significant difference in CFA grades for C5-6 and C6-7 between horses with presence of clinical signs and healthy horses. Despite these statistical differences, the clinical diagnosis of CFA based on radiographic grading alone is questionable because of the lack of agreement between the radiologists. To strengthen the diagnosis, clinical signs of facet disease, in particular atrophy of the neck muscles, need to be present to make this diagnosis. Intra-articular corticosteroid injection was effective at allowing most treated horses to return to athletic use..

Clinical indications, complications, and long-term outcome of esophageal surgeries in 27 horses.

The main objective of this retrospective study was to describe clinical findings, management, and short- and long-term outcome in 27 horses that underwent various surgical techniques for esophageal disease. Surgical techniques (sometimes concurrently) performed were: esophagostomy (n = 14), esophagotomy with primary closure (n = 6), esophagomyotomy (n = 3), and esophagoplasty (n = 2). Esophageal perforation in 5 horses was treated by ventral drainage; 3 horses had the esophageal defect sutured (n = 3). Feeding tubes were placed in 15 horses. Postoperative complications occurred in 52% (14/27) with a median of 3 complications/horse (range: 1 to 7). Significantly more complications occurred in horses with a perforated esophagus. Eighteen horses (18/27; 67%) were discharged. Most horses (8/9; 89%) with a lesion located in the proximal esophagus were discharged. Horses with a higher number of postoperative complications, particularly postoperative infection, were more likely to be euthanized. One year after surgery, 41% of the horses were alive and free of complications.

Indications cliniques, complications et résultat à long terme de chirurgies œsophagiennes chez 27 chevaux. L'objectif principal de cette étude rétrospective a été de décrire les résultats cliniques, la gestion et les résultats à court et à long terme chez 27 chevaux qui ont subi diverses techniques de chirurgie pour le traitement de la maladie œsophagienne. Les techniques chirurgicales réalisées (parfois de manière concomitante) étaient : l'œsophagostomie (n = 14), l'œsophagotomie avec fermeture primaire (n = 6), l'œsophagomyotomie (n = 3) et l'œsophagoplastie (n = 2). La perforation œsophagienne chez 5 chevaux a été traitée par drainage ventral; 3 chevaux ont subi une suture du défaut œsophagien (n = 3). Des sondes d'alimentation ont été placées chez 15 chevaux. Les complications postopératoires se sont produites chez 52 % des chevaux (14/27) avec une médiane de 3 complications/cheval (fourchette : 1 à 7). Un nombre significativement supérieur de complications se sont produits chez les chevaux ayant une perforation de l'œsophage. Dix-huit chevaux (18/27; 67 %) ont reçu leur congé. La plupart des chevaux (8/9; 89 %) ayant une lésion situé dans l'œsophage proximal ont reçu leur congé. Il était plus probable que les chevaux avec un nombre supérieur de complications postopératoires, particulièrement une infection postopératoire, soient euthanasiés. Un an après la chirurgie, 41 % des chevaux étaient vivants et libres de complications.(Traduit par Isabelle Vallières).

Response to Intravenous Allogeneic Equine Cord Blood-Derived Mesenchymal Stromal Cells Administered from Chilled or Frozen State in Serum and Protein-Free Media.

Equine mesenchymal stromal cells (MSC) are commonly transported, chilled or frozen, to veterinary clinics. These MSC must remain viable and minimally affected by culture, transport, or injection processes. The safety of two carrier solutions developed for optimal viability and excipient use were evaluated in ponies, with and without allogeneic cord blood-derived (CB) MSC. We hypothesized that neither the carrier solutions nor CB-MSC would elicit measurable changes in clinical, hematological, or biochemical parameters. In nine ponies (study 1), a bolus of HypoThermosol(®) FRS (HTS-FRS), CryoStor(®) CS10 (CS10), or saline was injected IV (n = 3/treatment). Study 2, following a 1-week washout period, 5 × 10(7) pooled allogeneic CB-MSCs were administered IV in HTS-FRS following 24 h simulated chilled transport. Study 3, following another 1-week washout period 5 × 10(7) pooled allogeneic CB-MSCs were administered IV in CS10 immediately after thawing. Nine ponies received CB-MSCs in study 2 and 3, and three ponies received the cell carrier media without cells. CB-MSCs were pooled in equal numbers from five unrelated donors. In all studies, ponies were monitored with physical examination, and blood collection for 7 days following injection. CD4 and CD8 lymphocyte populations were also evaluated in each blood sample. In all three studies, physical exam, complete blood cell count, serum biochemistry, and coagulation panel did not deviate from established normal ranges. Proportions of CD4(+) and CD8(+) lymphocytes increased at 168 h postinjection in CB-MSC treatment groups regardless of the carrier solution. Decreases in CD4(+/)CD8(+) double positive populations were observed at 24 and 72 h in CB-MSC-treated animals. There was no difference in viability between CB-MSCs suspended in HTS-FRS and CS10. HTS-FRS and CS10 used for low volume excipient injection of MSC suspensions were not associated with short-term adverse reactions. HTS-FRS and CS10 both adequately maintain CB-MSC viability following hypothermic or frozen simulated transport, respectively. CB-MSCs do not elicit clinical abnormalities, but allogeneic stimulation of CD4(+) and CD8(+) lymphocyte populations may occur. Future studies should include in vitro or in vivo evaluation of cell-mediated or adaptive immunity to autologous, identical allogeneic, or MSC originating from additional unrelated individuals in order to better characterize this response.

Aspiration, but not injection, decreases cultured equine mesenchymal stromal cell viability.

BACKGROUND: Recently, equine multipotent mesenchymal stromal cells (MSC) have received significant attention as therapy for various conditions due to their proposed regenerative and immune-modulating capacity. MSC are commonly administered to the patient through a hypodermic needle. Currently, little information is available on the effect of such injection has on equine MSC immediate and delayed viability. We hypothesize that viability of equine MSC is not correlated with needle diameter during aspiration and injection. RESULTS: Using a 3 mL syringe, manual injection of equine cord blood (CB) or bone marrow-derived (BM) MSC with no needle and needles ranging in size from 18 to 30 Ga did not affect immediate MSC viability. Similarly, 24 h post-injection, MSC delayed viability was not different between any of the tested needles as determined by a resazurin-based proliferation assay. Using a 3 mL syringe, aspiration of MSC through 20, 25, and 30 Ga needles resulted in significant decreases in immediate viability with no change in delayed viability when compared to aspiration without a needle. BM- and CB-MSC were observed to be of similar size with a diameter ± SD of 19.8 ± 2.7 and 20.4 ± 2.2 µm, respectively. In comparison, the smallest needles, (30 Ga) have an internal diameter of 160 µm. CONCLUSIONS: Following injection, needle diameter did not affect immediate or delayed viability of equine MSC. Following aspiration through needles sizes 20 Ga and smaller, immediate viability, but not delayed viability, decreased. As a result, an 18 Ga or larger needle should be utilized for aspiration of cell suspensions. In contrast, needle selection for MSC injection should be based on clinical preference and experience rather than concerns over decreasing MSC viability.

Post-thaw non-cultured and post-thaw cultured equine cord blood mesenchymal stromal cells equally suppress lymphocyte proliferation in vitro.

Multipotent mesenchymal stromal cells (MSC) are receiving increased attention for their non-progenitor immunomodulatory potential. Cryopreservation is commonly used for long-term storage of MSC. Post-thaw MSC proliferation is associated with a lag-phase in vitro. How this lag-phase affect MSC immunomodulatory properties is unknown. We hypothesized that in vitro there is no difference in lymphocyte suppression potential between quick-thawed cryopreserved equine cord blood (CB) MSC immediately included in mixed lymphocyte reaction (MLR) and same MSC allowed post-thaw culture time prior to inclusion in MLR. Cryopreserved CB-MSC from five unrelated foals were compared using two-way MLR. For each of the five unrelated MSC cultures, paired MLR assays of MSC allowed five days of post-thaw culture and MSC included in MLR assay immediately post-thawing were evaluated. We report no difference in the suppression of lymphocyte proliferation by CB-MSC that had undergone post-thaw culture and MSC not cultured post-thaw (p<0.0001). Also, there was no inter-donor variability between the lymphocyte suppressive properties of MSC harvested from the five different donors (p = 0.13). These findings suggest that cryopreserved CB-MSC may have clinical utility immediately upon thawing. One implication hereof is the possibility of using cryopreserved CB-MSC at third party locations without the need for cell culture equipment or competencies.

Evaluation of ultrasmall superparamagnetic iron oxide contrast agent labeling of equine cord blood and bone marrow mesenchymal stromal cells.

OBJECTIVE: To evaluate the efficacy and effects of labeling equine umbilical cord blood (UCB)- and bone marrow (BM)-derived multipotent mesenchymal stromal cells (MSCs) with an ultrasmall superparamagnetic iron oxide (SPIO) contrast agent and the detection of labeled MSCs by use of MRI. SAMPLE: UCB MSCs from placental tissues of 5 foals and BM MSCs from 5 horses. PROCEDURES: UCB and BM MSC cultures were seeded in duplicate (5,000 cells/cm(2)). One duplicate was incubated with SPIO (50 µg/mL); the other was processed identically, but without SPIO. Mesenchymal stromal cells were expanded in triplicates for 5 passages and assessed for viability and proliferative capacity, labeling efficacy, and labeled cell proportion. For MRI detection, 5 × 10(6) labeled BM MSCs from passage 1 or 2 were injected into a collagenase-induced superficial digital flexor tendon defect of an equine cadaveric forelimb from 2 horses. RESULTS: For passages 1, 2, and 3, labeling efficacy and cell proportion for UCB MSCs (99.6% [range, 98.8% to 99.9%], 16.6% [range, 6.5% to 36.1%], and 1.0% [range, 0.4% to 2.8%], respectively) were significantly higher than for BM MSCs (99.2% [range, 97.8% to 99.7%], 4.5% [range, 1.6% to 11.8%], and 0.2% [range, 0.1% to 0.6%], respectively). Labeling was not detectable after passage 3. Viability of MSCs was not affected, but cell doubling time increased in labeled MSCs, compared with that of unlabeled MSCs. On MRI 3-D T2*-weighted fast gradient echo sequences, decreased signal intensity was observed for BM passage 1 MSCs. CONCLUSIONS AND CLINICAL RELEVANCE: Equine UCB and BM MSCs were labeled with SPIO at high efficiencies.

Evaluation of a broad range real-time polymerase chain reaction (RT-PCR) assay for the diagnosis of septic synovitis in horses.

Septic synovitis is a potentially debilitating and life-threatening disorder in horses. We hypothesized that a universal bacterial real-time PCR (RT-PCR) assay would have improved sensitivity and decreased turn-around time for detection of bacteria in synovial fluid (SF) samples. Forty-eight SF samples were collected from 36 horses that presented to two referral institutions with suspected septic synovitis. Universal RT-PCR, bacterial culture and SF analysis were performed on all samples, and an interpretation on the sample being septic or not was derived by three board certified specialists from the history, clinical assessment and SF characteristics. RT-PCR results were compared to a composite standard comprised of positive culture and interpretation by all three specialists of samples as "septic". For 41 of 48 samples (85%), culture and RT-PCR results were concordant. Compared to the composite standard, 83% of samples were correctly classified by RT-PCR (turn-around time of approximately 4 hours). Relative sensitivity and specificity of RT-PCR were 87% and 72% respectively, and 56% and 86% for culture. Hence, universal RT-PCR was a rapid and highly sensitive test, which may accelerate diagnosis and improve outcome for horses with septic synovitis.

La synovite septique est une condition potentiellement débilitante et mortelle chez les chevaux. Nous avons émis l'hypothèse qu'une épreuve universelle d'amplification en chaîne par la polymérase en temps réel (RT-PCR) pourrait avoir une sensibilité augmentée et une diminution du délai d'obtention des résultats pour la détection de bactérie dans des échantillons de liquide synovial (SF). Quarante-huit échantillons de SF ont été prélevés à partir de 36 chevaux présentés à deux centres de référence avec une synovite septique suspectée. L'épreuve RT-PCR universelle, la culture bactérienne et l'analyse du SF ont été effectuées sur tous les échantillons et une interprétation à savoir si l'échantillon était septique ou non était obtenue de trois spécialistes certifiés à partir de l'histoire du cas, de l'évaluation clinique et des caractéristiques du SF. Les résultats du RT-PCR ont été comparés à un composite standard consistant en une culture et une interprétation positives par les trois spécialistes d'échantillons comme étant «septique¼. Pour 41 des 48 échantillons (85 %), les résultats de la culture et du RT-PCR concordaient. Comparativement au composite standard, 83 % des échantillons ont été classés correctement par le RT-PCR (délai d'obtention des résultats approximatif de 4 heures). La sensibilité et la spécificité relatives du RT-PCR étaient de 87 % et 72 %, respectivement, 56 % et 86 % pour la culture. Ainsi, l'épreuve RT-PCR universelle était un test rapide et hautement sensible, ce qui pourrait accélérer le diagnostic et améliorer le sort des chevaux avec une synovite septique.(Traduit par Docteur Serge Messier).

Effect of unfocused extracorporeal shock wave therapy on growth factor gene expression in wounds and intact skin of horses.

OBJECTIVE: To compare the effect of extracorporeal shock wave therapy (ESWT) on expression of fibroblast growth factor-7 (FGF-7), transforming growth factor-ß1 (TGF-ß1), insulin-like growth factor-1 (IGF-1), platelet-derived growth factor-A (PDGF), and vascular endothelial growth factor-A (VEGF) in skin with surgically created skin wounds and intact skin in horses. ANIMALS: 14 healthy horses. PROCEDURE: 8 horses were treated with ESWT at 6 locations along the neck at 36, 24, 12, 6, 2, or 1 hour prior to collection of full-thickness biopsy specimens from each location; a control specimen was collected from a sham-treated location. In 6 horses, 5 full-thickness wounds were created in each forelimb. Wounds in 1 forelimb/horse received ESWT immediately after creation and subsequently on days 7, 14, and 21; wounds in the contralateral forelimb remained untreated. Biopsy specimens were collected from 1 wound on each forelimb on days 7, 14, 21, 28, and 35. Expression levels of FGF-7, TGF-ß1, IGF-1, PDGF, and VEGF were assessed in tissue samples from the horses' necks and forelimbs. RESULTS: In surgically created wounds, ESWT treatment was associated with reduced TGF-ß1 expression, compared with expression in control wounds, during the entire study period. At 28 days following wound creation, IGF-1 expression was significantly increased for treated and untreated wounds, compared with findings on days 7, 14, 21, and 35. There was no significant effect of treatment on FGF-7, TGF-ß1, IGF-1, PDGF, or VEGF expression in intact skin. CONCLUSIONS AND CLINICAL RELEVANCE: Intervention with ESWT to suppress TGF-ß1 may decrease granulation tissue production, resulting in improved wound healing on the distal portion of horses' limbs.

Effects of unfocused extracorporeal shock wave therapy on healing of wounds of the distal portion of the forelimb in horses.

OBJECTIVE: To determine effects of extracorporeal shock wave therapy (ESWT) on healing of wounds in the distal portion of the forelimb in horses. ANIMALS: 6 horses. PROCEDURES: Five 6.25-cm2 superficial wounds were created over both third metacarpi of 6 horses. Forelimbs were randomly assigned to treatment (ESWT and bandage) or control (bandage only) groups. In treated limbs, each wound was treated with 625 shock wave pulses from an unfocused electrohydraulic shock wave generator. In control limbs, each wound received sham treatment. Wound appearance was recorded weekly as inflamed or healthy and scored for the amount of protruding granulation tissue. Standardized digital photographs were used to determine the area of neoepithelialization and absolute wound area. Biopsy was performed on 1 wound on each limb every week for 6 weeks to evaluate epithelialization, fibroplasia, neovascularization, and inflammation. Immunohistochemical staining for A smooth muscle actin was used to label myofibroblasts. RESULTS: Control wounds were 1.9 times as likely to appear inflamed, compared with treated wounds. Control wounds had significantly higher scores for exuberant granulation tissue. Treatment did not affect wound size or area of neoepithelialization. No significant difference was found for any of the histologic or immunohistochemical variables between groups. CONCLUSIONS AND CLINICAL RELEVANCE: Treatment with ESWT did not accelerate healing of equine distal limb wounds, but treated wounds had less exuberant granulation tissue and appeared healthier than controls. Therefore, ESWT may be useful to prevent exuberant granulation tissue formation and chronic inflammation of such wounds, but further studies are necessary before recommending ESWT for clinical application.

Evaluation of endotoxin activity in blood measured via neutrophil chemiluminescence in healthy horses and horses with colic.

OBJECTIVE: To evaluate the performance of a chemiluminescent endotoxin activity assay in horses with colic and healthy horses. ANIMALS: 20 horses with colic and systemic inflammatory response syndrome (SIRS group), 8 horses with colic with no SIRS (NSIRS group), and 20 healthy horses. PROCEDURES: Venous blood was collected into EDTA blood collection tubes after completion of a physical examination, and a chemiluminescent endotoxin activity assay was performed within 60 minutes of collection. Medical or surgical interventions and outcome were recorded for each horse. RESULTS: Mean + or - SE endotoxin activity was 0.16 + or - 0.05 for healthy horses, 0.18 + or - 0.07 for the NSIRS group, and 0.53 + or - 0.05 for the SIRS group and was significantly different among the groups. Mean endotoxin activity was significantly higher in the SIRS group than in the NSIRS group and the healthy group. No significant difference between the healthy and NSIRS groups was present. The higher the measured endotoxin activity, the more likely it was for horses to be euthanized. CONCLUSIONS AND CLINICAL RELEVANCE: The chemiluminescent endotoxin assay was easy to use, required a short time to perform, could be completed at the patient's side, and with some modifications, may be a useful component in the clinical assessment and prognostication of horses with colic.

Use of multichannel electrogastrography for noninvasive assessment of gastric myoelectrical activity in dogs.

OBJECTIVE: To evaluate whether changes in gastric myoelectrical activity in healthy, awake dogs can be detected via multichannel electrogastrography (EGG). ANIMALS: 6 healthy hound-breed dogs. PROCEDURES: For each dog, 8-channel EGG was performed after food had been withheld for 12 hours and at 30 minutes after subsequent feeding; 60 minutes after feeding, atropine (0.04 mg/kg) was administered IM to induce ileus, and 30 minutes later, EGG was again performed. Mean cycles per minute (cpm) values of the dominant frequency (a measure of the rhythmicity of gastric electrical activity) and mean power ratios (ie, power measured after treatment divided by the power measured when food was withheld) were calculated. Motility of the gastric antrum was assessed via B-mode ultrasonography during the same phases; contractions determined ultrasonographically were correlated with EGG power for each channel in each phase. RESULTS: The criterion for stability (SD of the dominant frequency < 15% of the cpm value in at least 3 of the 8 EGG channels) was met in 4 of the 6 dogs (only in long-distance channels). The mean power ratios were significantly higher in the postprandial phase than in the ileus phase. Compared with the postprandial phase, significantly fewer contractions per minute were evident ultrasonographically in the ileus and food-withholding phases. There was a significant and good correlation between EGG power and ultrasonographic findings in all 8 channels. CONCLUSIONS AND CLINICAL RELEVANCE: Electrogastrography may be useful in assessing gastric myoelectrical activities in awake dogs with naturally occurring gastrointestinal disease, including gastric dilatation-volvulus.

Use of multichannel electrointestinography for noninvasive assessment of myoelectrical activity in the cecum and large colon of horses.

OBJECTIVE: To evaluate whether changes in myoelectrical activity in the cecum and large colon of horses can be detected via multichannel electrointestinography (EIG). ANIMALS: 6 healthy mature horses. PROCEDURES: Each horse underwent 3 EIG procedures. Intestinal myoelectrical activity (cecum and large colon) was recorded during a 20-minute period following i.v. administration of physiologic saline (0.9% NaCl) solution (20 mL; baseline), erythromycin lactobionate (0.5 mg/kg), or detomidine (0.015 mg/kg); intestinal contractions were concurrently viewed via B-mode ultrasonography. By use of computer software, 8-channel EIG recordings were analyzed and the mean of the dominant frequency (a measure of the rhythmicity of gastric electrical activity) expressed in cycles per minute (cpm) was obtained. Total power (muV(2)) was calculated, and treatment effect was expressed as the power ratio (ie, treatment-associated power divided by the baseline power). RESULTS: The dominant frequency cpm values were not stable, and no significant differences between treatments were detected. Compared with the effects of saline solution treatment, detomidine significantly reduced the mean cecal and colonic power ratios. Erythromycin significantly reduced the cecal power ratio and increased the colonic power ratio, although the increase was significant in only 1 channel. Ultrasonographic findings and total power (predominantly from the long-distance electrode pairs) were significantly correlated. CONCLUSIONS AND CLINICAL RELEVANCE: In horses, EIG was useful for assessment of changes in myoelectrical activity in the cecum and large colon. Multiple electrodes should be used to cover a larger area of the intestine, and agreement between multiple channels is needed to make the analysis meaningful.

Congenital colonic malformation ("short colon") in a 4-month-old standardbred foal.

During exploratory laparotomy of a foal with colic, a congenital abnormally developed large colon was identified incidentally. Long-term follow-up showed that the colt was more prone to gas-colic with diet and exercise changes than were other horses, due possibly to the short colon.

Use of a circular external skeletal fixator for stabilization of a comminuted diaphyseal metatarsal fracture in an alpaca.

CASE DESCRIPTION: A 3-year-old male alpaca was evaluated because of non-weight-bearing lameness (grade 5/5) in the left hind limb. CLINICAL FINDINGS: Clinical and radiographic examination revealed a closed, comminuted, nonarticular, displaced diaphyseal fracture of the left third and fourth metatarsal bones. TREATMENT AND OUTCOME: Initial attempts at treatment via reduction of the fracture under traction and subsequent application of a cast were unsuccessful, and more stable fracture fixation was pursued. The alpaca underwent closed reduction of the fracture, which was stabilized by the application of a 3-ring circular external skeletal fixator (CESF). Improved weight bearing on the affected limb was evident soon after surgery and gradually increased; full weight bearing was evident by the seventh day after discharge from the hospital (day 20 after application of the CESF). Lameness was hardly noticeable during walking at that time. After 3 months, complete fracture healing was evident and the CESF was removed; mild outward rotation of the distal fragment and metatarsophalangeal joint was present. A Robert Jones bandage was applied to the limb, and the alpaca was kept in a stall for another 4 weeks. Eleven months after CESF application, the owners and referring veterinarian reported that the alpaca was healthy, not lame, and serving as a stallion without apparent impediment. CLINICAL RELEVANCE: Although mostly restricted to small animals, application of a CESF can be a viable alternative for management of long bone fractures in South American camelids.

Effect of intraluminal distension or ischemic strangulation obstruction of the equine jejunum on jejunal motilin receptors and binding of erythromycin lactobionate.

OBJECTIVE: To determine whether inflammation of the jejunum of horses decreases the number of motilin receptors and amounts of motilin receptor mRNA and alters erythromycin lactobionate binding affinity to the motilin receptor in jejunal tissues. SAMPLE POPULATION: Jejunal segments in 6 adult horses. PROCEDURE: Each horse was anesthetized, and a ventral median celiotomy was performed; 2 segments of jejunum underwent a sham operation, 2 segments underwent ischemic strangulation obstruction (ISO), and 2 segments underwent intraluminal distension (ILD). Treatments were maintained for 120 minutes. From each segment, full-thickness biopsy samples were collected and smooth-muscle homogenates were prepared. Affinity and distribution of motilin binding to these preparations were determined by use of iodine 125 (125I)-labeled synthetic porcine motilin. Via displacement experiments, competition between 125I-labeled motilin and erythromycin lactobionate for binding to motilin receptors in the different segments was investigated. A quantitative real-time PCR technique was used to assess motilin receptor mRNA content in the muscle preparations. RESULTS: Compared with the ISO or ILD segments, the number of motilin receptors was significantly higher in the sham-operated segments; ILD segments contained the lowest number of motilin receptors. The expression of motilin receptor mRNA was significantly decreased in ILD segments but not in ISO segments. Erythromycin lactobionate displacement of 125I-labeled motilin from motilin receptors did not differ significantly among the jejunal segments. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that downregulation and decreased production of motilin receptors in inflamed jejunal tissue contribute to the altered prokinetic response to erythromycin in horses with gastrointestinal disease.

Binding of radiolabeled porcine motilin and erythromycin lactobionate to smooth muscle membranes in various segments of the equine gastrointestinal tract.

OBJECTIVE: To identify and characterize motilin receptors in equine duodenum, jejunum, cecum, and large colon and to determine whether erythromycin lactobionate competes with porcine motilin for binding to these receptors. SAMPLE POPULATION: Specimens of various segments of the intestinal tracts of 4 adult horses euthanatized for reasons unrelated to gastrointestinal tract disease. PROCEDURE: Cellular membranes were prepared from smooth muscle tissues of the duodenum, jejunum, pelvic flexure, and cecum. Affinity and distribution of motilin binding on membrane preparations were determined by use of 125I-labeled synthetic porcine motilin. Displacement studies were used to investigate competition between 125I-labeled synthetic porcine motilin and erythromycin lactobionate for binding to motilin receptors in various segments of bowel. RESULTS: Affinity of 125I-labeled synthetic porcine motilin for the equine motilin receptor was estimated to be 6.1nM. A significantly higher number of motilin receptors was found in the duodenum than in the pelvic flexure and cecum. The jejunum had a significantly higher number of motilin receptors than the cecum. Erythromycin lactobionate displacement of 125I-labeled porcine motilin from the equine motilin receptor did not differ significantly among various segments of bowel. CONCLUSIONS AND CLINICAL RELEVANCE: Motilin receptors were found in the duodenum, jejunum, pelvic flexure, and cecum of horses. The highest number of motilin receptors was in the duodenum, and it decreased in more distal segments of bowel. Erythromycin lactobionate competed with motilin binding in the equine gastrointestinal tract. This suggests that 1 of the prokinetic actions of erythromycin in horses is likely to be secondary to binding on motilin receptors.