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This case report describes the rescue of an eight-month-old male Italian wolf (Canis lupus italicus), the victim of a car accident that caused it a pulmonary contusion, a fracture of the shaft of right femur, and a metaphyseal fracture of the left stifle. A lateral surgical approach was performed to treat the animal’s multiple contusions and fractures. Afterwards the wolf was transferred to a wild animal recovery center for its rehabilitation, where it fully recovered. After 35 days in captivity the wolf was thus released into the supposed home-range of its original pack, and its movements were monitored by a GPS satellite collar. The collar worked correctly for 479 days. During that period the collar acquired a total of 1202 locations, indicating that the wolf had traveled at least 1590 km, with an average monthly distance (± SD) of 102 ± 40 km, exploring an overall area of about 270 km2. During the first 10 days after its release, the wolf remained in the area of its supposed native pack, whereas at about the age of 10 months the wolf began to make wide extraterritorial movements. The wolf’s last localization was acquired on 13th May 2018, about 17 months after its release, at a linear distance of about 65 km from the release site. These preliminary data showed that the wolf was alive and travelled long distances after its release, and demonstrates how a multidisciplinary management approach can support the recovery and successful release into nature of a rescued wild animal belonging to a flagship species with a notable ecological role, such as the Italian wolf., Ovaj prikaz slučaja opisuje spašavanje osmomjesečnog mužjaka talijanskog vuka (Canis lupus italicus) koji je bio žrtva naleta vozila s posljedičnim nagnječenjem pluća, prijelomom vrata desne bedrene kosti prijelomom u području metafize lijevoga koljena. Primijenjen je lateralni kirurški pristup kako bi se sanirala višestruka nagnječenja i prijelomi u životinje. Nakon toga je vuk prevezen u centar za oporavak divljih životinja gdje se potpuno oporavio. Nakon 35 dana u zatočeništvu vuk je pušten u okoliš nalik na njegov prirodni te je praćen preko GPS ogrlice. Ogrlica je ispravno radila 479 dana. Za to je vrijeme pokazala 1202 lokacije, upućujući na to da je vuk prešao 1590 km, u prosjeku mjesečno 102 ± 40 km (± SD), istražujući područje od 270 km2. Prvih 10 dana nakon oslobađanja vuk je ostao u području nalik na njegov prirodni okoliš, no u dobi od 10 mjeseci počeo je odlaziti na šire područje. Posljednja je njegova lokacija zabilježena 13. svibnja 2018., oko 17 mjeseci nakon puštanja, na zračnoj udaljenosti od oko 65 km od mjesta ispuštanja. Ovaj preliminarni podatak pokazuje da je vuk bio živ i prelazio velike udaljenosti nakon oslobađanja. Također pokazuje da multidisciplinarni pristup spašavanju može pospješiti oporavak i uspješno vraćanje u prirodu divljih životinja koje pripadaju vodećim vrstama s velikom ekološkom važnošću, kao što je talijanski vuk.

Large colon volvulus in horses is associated with a poor prognosis, especially when ischemic-reperfusion injury of the affected intestinal tract develops. Proteinase-activated receptor 2 (PAR2) plays an important role in the pathogenesis of inflammation in the gastrointestinal tract. The aim of this study was to evaluate the distribution and expression of PAR2in colonic pelvic flexure of horses spontaneously affected by large colon volvulus (CVH group). Eight horses admitted for severe abdominal colon volvolus and which underwent surgery were included. Colon samples were collected after enterotomy. Data previously obtained from healthy horses were used as a control group. Histologic evaluation was carried out to grade the severity of the colon lesions. Immunofluorescence, western blot and quantitative polymerase chain reaction (RT-qPCR) were carried out on colon samples to evaluate PAR2expression. In addition, the transcriptional profile of cytokines and chemokines was evaluated using RT2Profiler™ PCR Array Horse Cytokines & Chemokines. Three out of the eight patients were euthanised due to clinical deterioration. Immunostaining for PAR2was observed in the enterocytes, intestinal glands and neurons of the submucosal and myenteric plexi. In the CVH horses, the expression of PAR2mesenger RNA (mRNA) did not differ significantly from that of the healthy animals; western blots of the mucosa of the colon tracts showed a clear band of the expected molecular weight for PAR2(~44 kDa) and a band smaller than the expected molecular weight for PAR2(25kDa), suggesting its activation. The gene expressions for C-X-C motif ligand 1 (CXCL1); interleukin 8 (IL8), macrophage inflammatory protein 2 beta (MIP-2BETA) were upregulated in the colic horses as compared with the colons of the healthy horses. Therefore, in the present study, the expression and activation of PAR2in the colons of horses in the presence of an inflammatory reaction like that occurring in those with spontaneous colon volvulus was confirmed.

Gastroesophageal reflux (GER) is a common event during general anaesthesia but is often underdiagnosed in veterinary medicine. The oesophageal pH in anaesthetised dogs undergoing endoscopic evaluation of the upper gastrointestinal tract (END group, n = 12) or orthopaedic surgery (ORT group, n = 12) was measured using an oesophageal probe. The dogs were sedated with acepromazine or with methadone or butorphanol, and anaesthesia was induced with propofol and maintained with isoflurane. Of the 24 dogs in this study, 21 (87.5%) had an episode of GER during anaesthesia. The incidence of GER, as well as the first, the minimum, and the maximum pH values, did not differ significantly between the groups. The mean maximum difference versus the first pH value was higher for dogs in the END group (&minus, 2.6 &plusmn, 3.5) as compared with those in the ORT group (&minus, 0.7 &plusmn, 2.5), although they were not statistically significant (p = 0.25). The administration of methadone or butorphanol had no significant effect on the development of acidic reflux or biliary reflux. In the acepromazine-sedated dogs, the incidence of GER did not differ significantly between patients undergoing an endoscopic procedure and those undergoing orthopaedic surgery, however, during endoscopy, fluctuations in the oesophageal pH can be expected, even without any clinical signs of GER.

High dose medetomidine 0.13 mg/kg can be used for semen collection in cats with variable results in terms of quantity and quality. Therefore, a variation in terms of distribution and elimination among patients has been hypothesised. The aim of the study was to characterise the pharmacokinetics of medetomidine (0.13 mg/kg) administered intramuscularly (IM) in healthy male cats. Eighteen male cats undergoing castration were included, and medetomidine (0.13 mg/kg) was administered IM. Venous blood samples were collected at 20, 30, 40, 50, 60, 75 and 90 minutes after medetomidine administration. Before orchiectomy, at T20, sperm collection was attempted. Plasma medetomidine concentrations were determined by liquid chromatography/mass spectrometry analysis. Semen collection was successful in 15/18 cats. The medetomidine plasma concentration following the IM administration of a bolus was best described using a non-compartment model. Time of maximum concentration was observed at 40 minutes (range 20−90); maximum concentration was 32.8 ng/mL (range 26.8−51.2). The median apparent clearance was 11.9 mL/kg/minute (range 0.7−43.8). In conclusion, medetomidine administered IM at 0.13 mg/kg reached its peak plasma concentration slowly and with variability among patients. In addition, it was characterised by low total body clearance probably due to the cardiovascular alterations associated with medetomidine administration.

Proteinase activated receptor 4 (PAR4) in the gastrointestinal tract is involved in the regulation of inflammation and pain pathways. The aim of the present study was to evaluate the distribution and expression of PAR4 in the jejunum of healthy horses and in the pathologic tracts from horses undergoing surgery for herniation of the small intestine through the epiploic foramen. Eight healthy horses (Group H) and eight horses with epiploic hernia (Group EH) were included; the jejunum samples were collected at the slaughter or intraoperatively after enterectomy, respectively. To evaluate PAR4 expression in sections of the jejunum, immunofluorescence, western blot and quantitative polymerase chain reaction (qRT-PCR) were performed. Immunohistochemistry of PAR4 in the jejunum of the healthy horses showed that receptors are predominantly expressed in the immune cell population scattered throughout the lamina propria of the mucosa and in the submucosa. Quantitative PCR data demonstrated that PAR4 mRNA was detectable in all of the samples analyzed without any difference between the H and the EH groups, however the PAR4 protein level was significantly lower in the jejunums of the EH horses. In the Group EH horses, PAR4 immunoreactivity was mainly expressed in the mast cells and was extensively distributed in the sierosa. In the lamina propria of mucosa of Group EH, leukocytes were less abundant than in Group H. In this study, the distribution and expression of PAR4 in the jejunums of the healthy horses and in those with spontaneous occurring epiploic hernia was demonstrated.

Cerebrospinal fluid administration of recombinant adeno-associated viral (rAAV) vectors has been demonstrated to be effective in delivering therapeutic genes to the central nervous system (CNS) in different disease animal models. However, a quantitative and qualitative analysis of transduction patterns of the most promising rAAV serotypes for brain targeting in large animal models is missing. Here, we characterize distribution, transduction efficiency, and cellular targeting of rAAV serotypes 1, 2, 5, 7, 9, rh.10, rh.39, and rh.43 delivered into the cisterna magna of wild-type pigs. rAAV9 showed the highest transduction efficiency and the widest distribution capability among the vectors tested. Moreover, rAAV9 robustly transduced both glia and neurons, including the motor neurons of the spinal cord. Relevant cell transduction specificity of the glia was observed after rAAV1 and rAAV7 delivery. rAAV7 also displayed a specific tropism to Purkinje cells. Evaluation of biochemical and hematological markers suggested that all rAAV serotypes tested were well tolerated. This study provides a comprehensive CNS transduction map in a useful preclinical large animal model enabling the selection of potentially clinically transferable rAAV serotypes based on disease specificity. Therefore, our data are instrumental for the clinical evaluation of these rAAV vectors in human neurodegenerative diseases.

Objective To measure plasma methadone concentrations in bitches and the umbilical cords of their puppies after systemic or epidural administration. Study design Prospective, randomized, clinical study. Animals A total of 27 healthy pregnant female dogs undergoing caesarean section, 4.3 ± 2.3 years of age and weighing 19.9 ± 13.2 kg. Methods The dogs were randomly divided into three groups: 1) intramuscular methadone (0.3 mg kg–1) (group MET; n = 9); 2) epidural methadone (0.1 mg kg–1) (group METEPI; n = 9); and 3) epidural lidocaine (4.4 mg kg–1) [group CON (control group); n = 9]. Ten minutes before induction, methadone was administered intramuscularly to the group MET dogs. Anaesthesia was induced with propofol and maintained with isoflurane. Cardiovascular and respiratory parameters were monitored throughout the anaesthesia. After induction, epidural anaesthesia was administered to dogs in groups METEPI and CON. Before any treatment (T0) and, as soon as the last foetus was removed from the uterus (T1), venous blood samples were collected from each dog into heparinized tubes; the umbilical cords were collected and stored at –80 °C until pharmacological analysis was carried out. The samples were analysed using ultra performance liquid chromatography. Results The cardiorespiratory parameters of the bitches and of the puppies at birth, and the Apgar scores did not differ significantly between groups. At T1 both the median maternal methadone plasma concentration and the median methadone umbilical cord concentration were higher in group MET compared to group METEPI (p = 0.0018 and p = 0.004, respectively). The maternal plasma concentration was higher than the concentration in the umbilical cords (p = 0.05) in group METEPI but not in group MET (p = 0.25). Conclusions and clinical relevance Epidural methadone (0.1 mg kg–1) administered to bitches undergoing caesarean section is associated with lower umbilical cord methadone concentrations as compared with intramuscularly administered methadone at higher dosages (0.3 mg kg–1).

Objective To investigate the pharmacokinetics of buprenorphine and its main active metabolite, norbuprenorphine, after administration of an intravenous loading dose followed by constant rate infusion (CRI) in dogs. Study design Prospective, clinical study. Animals A total of seven healthy dogs undergoing elective ovariectomy. Methods Buprenorphine was administered as a loading dose (intravenous bolus of 15 μg kg−1) followed by CRI (2.5 μg kg−1 hour−1 for 6 hours). Moreover, intraoperative analgesia was supplemented by an intramuscular carprofen (4 mg kg−1) injection, administered prior to surgery, and by lidocaine, administrated through subcutaneous infiltration and through a splash on the ovarian vascular pedicle during surgery. Pain and sedation were scored for all animals throughout the 24-hour study period and rescue analgesia was administered when a visual analogue scale score was > 40 mm. Blood samples were collected from a jugular catheter at regular intervals, and plasma concentrations of buprenorphine and norbuprenorphine were determined by a validated liquid chromatography–tandem mass spectrometry method. Results Buprenorphine showed a two-compartment kinetic profile. Maximum concentration was 23.92 ± 8.64 ng mL−1 at 1 minute (maximum time); elimination half-life was 41.87 ± 17.35 minutes; area under the curve was 486.68 ± 125.66 minutes ng−1 mL−1; clearance was 33.61 ± 13.01 mL minute−1 kg−1, and volume of distribution at steady state was 1.77 ± 0.50 L kg−1. In no case was rescue analgesia required. Norbuprenorphine resulted below the lower limit of quantification in almost all samples. Conclusions and clinical relevance The results suggest that a buprenorphine CRI can be a useful tool for providing analgesia in postoperative patients, considering its minor side effects and the advantages of a CRI compared to frequent boluses. The negligible contribution of norbuprenorphine to the therapeutic effect was confirmed.

The aim of this retrospective study was to describe the use of simple interrupted suture pattern for the closure of the abdomen after ventral midline celiotomy in horses and to evaluate the prevalence of resulting incisional complications. Seventy horses undergoing abdominal surgery (2006–2015) and surviving at least 30 days were included in the study. Closure technique of the abdominal wall was performed on three layers. Linea alba was closed with a simple interrupted pattern using number two coated braided absorbable polyglycolic acid suture. Follow-up information was recovered at the 90th day postoperatively. Incisional complications considered included drainage, infection, dehiscence, and herniation. The complications observed in the present study using simple interrupted suture pattern were 4.29% (3 of 70), lower than reported in literature with different suture pattern. Simple interrupted suture pattern could be a valid option for the closure of linea alba after ventral midline celiotomy in horses.

The purpose of this study was to provide data on the disposition of stanozolol after bilateral intra-articular injection in the tarsal joints, to discover the length of time for which the drug can be detected in plasma after administration. Fourteen horses were included in the study. After aseptically preparing the injection site, 1 mL of an aqueous suspension containing 5 mg of stanozolol was injected into both the right and left tarsal joints of 12 horses; the two remaining animals were not treated and were used as a control group. Five milliliters of blood was collected immediately before stanozolol administration (t0) and at 1, 2, 4, 6, 12, 18, 24, 36, 48, 60, 72, 84, 96, 108, 120, 132, 144, 156, and 168 hours after injection. The plasma concentration was determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS) after solid phase extraction. The maximum plasma concentration was 1.7 ng/mL (range, 0.5–3.0 ng/mL), measured at 6 hours (range, 4–12 hours). The plasma elimination half life varied between 4 and 12 hours, whereas the plasma clearance per fraction of dose absorbed was in the 257.85–820.88 L/h range. The results of the present study make a preliminary contribution toward understanding the elimination profile of intra-articularly administered stanozolol in the horse. The drug passes rapidly into the systemic circulation, is eliminated rapidly, and is detected in plasma for no more than 36 hours after local administration.

Porcine models of ophthalmological diseases are often used in pre-clinical translational studies due to pigs’ similarities to humans. In particular, the iodoacetic acid (IAA) model of photoreceptor degeneration seems to mimic well the endstage phenotype of human pathologies as retinitis pigmentosa and age-related macular degeneration, with high potential for prosthesis/retinal devices testing. IAA is capable of inducing photoreceptor death by blockage of glycolysis, and its effects on the retina have been described. Nonetheless, up to date, literature lacks of a comprehensive morpho-functional characterization of the entire visual system of this model. This gap is particularly critical for prosthesis testing as inner retinal structures and optic pathways must be preserved to elicit cortical responses and restore vision. In this study, we investigated the functional and anatomical features of the visual system of IAA-treated pigs and compared them to control animals. IAA was administered intravenously at 12 mg/kg; control animals received saline solution (NaCl 0.9% w/v). Electrophysiological analyses included full-field (ffERGs) and pattern (PERGs) electroretinograms and flash visually evoked potentials (fVEPs). Histological evaluations were performed on the retina and the optic pathways and included thickness of the different retinal layers, ganglion cells count, and immunohistochemistry for microglial cells, macroglial cells, and oligodendrocytes. The histological results indicate that IAA treatment does not affect the morphology of the inner retina and optic pathways. Electrophysiology confirms the selective rod and partial cone degeneration, but is ambiguous as to the functionality of the optic pathways, seemingly preserved as indicated by the still detectable fVEPs. Overall, the work ameliorates the characterization of such rapid and cost-effective model, providing more strength and reliability for future pre-clinical translational trials.

This study aims to validate a tool, the Bologna healing stifle injury index (BHSII), for the evaluation of the clinical picture and the healing after surgical treatment for cranial cruciate ligament (CCL) rupture. The study included 158 client-owned dogs with CCL rupture and 20 healthy dogs. The BHSII is a questionnaire made up of 34 multiple-choice questions, divided into a part directed to the clinician and a part for the dog's owners. It was applied twice in the healthy dogs in order to test and retest the device. It was evaluated for reliability, validity, and responsiveness to clinical changes involving the dogs treated at the time of surgery, and 1, 3, and 6 months postoperatively. Statistical analyses were performed and the intraclass correlation coefficient test was ≥0.9 and the Cronbach-α was 0.84 suggesting good stability and good internal consistency of the tool. The area under the curve of the receiver operating characteristic curve was >0.9, indicative of the high accuracy of this tool. The clinician survey correlated with the owner questionnaire. In dogs with CCL rupture, the scores of the BHSII increased significantly postoperatively as compared with baseline. In conclusion, this clinical study proved the reliability, validity, and responsiveness of the BHSII. The results achieved from the BHSII provided an instantaneous, collective complete vision of the healing process of the stifle joints treated. It can be considered a valid tool for collecting data and for assessing successful surgical treatment in clinical practice.

The Raphe Pallidus (RPa) is a region of the brainstem that was shown to modulate the sympathetic outflow to many tissues and organs involved in thermoregulation and energy expenditure. In rodents, the pharmacological activation of RPa neurons was shown to increase the activity of the brown adipose tissue, heart rate, and expired CO2, whereas their inhibition was shown to induce cutaneous vasodilation and a state of hypothermia that, when prolonged, leads to a state resembling torpor referred to as synthetic torpor. If translatable to humans, this synthetic torpor-inducing procedure would be advantageous in many clinical settings. A first step to explore such translatability, has been to verify whether the neurons within the RPa play the same role described for rodents in a larger mammal such as the pig. In the present study, we show that the physiological responses inducible by the pharmacological stimulation of RPa neurons are very similar to those observed in rodents. Injection of the GABAA agonist GABAzine in the RPa induced an increase in heart rate (from 99 to 174bpm), systolic (from 87 to 170mmHg) and diastolic (from 51 to 98mmHg) arterial pressure, and end-tidal CO2 (from 49 to 62 mmHg). All these changes were reversed by the injection in the same area of the GABAA agonist muscimol. These results support the possibility for RPa neurons to be a key target in the research for a safe and effective procedure for the induction of synthetic torpor in humans.

Proteinase-activated receptor 2 (PAR2) is a G-protein-coupled receptor for trypsin and mast cell tryptase; it is highly expressed at the intestinal level with multiple functions, such as epithelial permeability and intestinal motility. The aim of the study was to evaluate the distribution and expression of proteinase-activated receptor 2 in the small intestine during herniation through epiploic foramen. In this prospective clinical study, eight horses admitted for colic and which underwent exploratory laparotomy were considered. During surgery, the jejunum or the ileum was sampled by enterectomy. Morphological examination (histology, PAR2 immunohistochemistry) and molecular biology analysis (western blot and quantitative polymerase chain reaction) were carried out on the resected intestinal samples. The Marginal Injured Tracts (MITs) and Central Injury Tracts (CITs) were defined as the oral and caudal marginal segments of the resected bowel tract and as the geometric centre of the intestinal ischaemic lesion length, respectively. The PAR2 immunoreactivity was particularly evident in the epithelial cells, with higher immunoreactivity in the MIT rather than in the CIT. Moreover, a different immune localisation was observed in the MITs at the cell membrane level and in the CITs in the cytoplasm. No statistical difference was observed in PAR2 mRNA and protein (44 kDa) expression between the MIT and the CIT. The PAR2 protein content in the intestinal tracts which were removed from horses with herniation was lower when compared with the control animals. This study provided data concerning the PAR2 presence and distribution in horses with intestinal herniation through the epiploic foramen.

Poor data could be found in the literature on the effects of intra-articular (IA) administration of stanozolol in horses affected by osteoarthritis (OA). To verify the clinical effects of IA stanozolol in acute and chronic cases of OA in horses, a clinical double-blinded trial involving 60 client-owned horses was performed. Veterinary practitioners selected horses, all showing clinical signs of acute or chronic OA, diagnosed in a single joint. After lameness evaluation and synovial fluid collection, the clinicians administered one to four (acute cases) or one to six (chronic cases) weekly IA doses of 5 mg of stanozolol or placebo. Positive result was considered a grade 1 improvement in lameness scored according to the American Association of Equine Practitioner scale. The characteristics of the synovial fluid were also evaluated. Of the 60 horses included in the study, 31 were affected by acute OA and 29 by chronic OA. Horses were treated blindly with placebo. The overall outcomes were positive in 82.50% of cases. Lameness disappeared completely in 15 of 21 (acute) and 11 of 19 (chronic) animals, and there was a significant reduction after two and four treatments, in the acute and chronic cases, respectively. Improvement in the physical characteristics of the synovial fluid sampled was evident after the third treatment. Stanozolol injected IA at a dose of 5 mg significantly resolved lameness in affected joints. Stanozolol could be considered a safe option for the treatment of horses suffering from OA.

The study clinically evaluated the effectiveness of autologous platelet-rich plasma (PRP), obtained using a double transfusion bag after an ultrasound-guided intralesion injection into an acute lesion of proximal portion of suspensory ligament (SL). Twenty horses with acute proximal suspensory ligament desmitis were included in the study. An ultrasound evaluation of the proximal region of the SL was carried out, and the cross-sectional area (CSA) percentage and fiber score alignment were determined. Four hundred fifty milliliters of blood was collected, and PRP was obtained by means of the buffy coat method, using a double transfusion bag. The PRP was injected into the lesion using an ultrasound guide. A long-term follow-up was carried out by means of a telephonic interview with the owner. The median CSA of the SL lesion at the first examination was 30%. The median platelet count in the PRP was 1,045 × 10 3 platelets/μL; the final concentration ranged from six to 20 times higher than the platelet blood count. Sixteen horses returned to activity within a period ranging from 12 to 24 weeks from the last PRP injection, and four animals recidivated in different regions of suspensory ligament: midbody desmitis and medial branch. Clinical results confirm that the use of autologous PRP supports healing and reduces recurrence in acute SL lesions.

Equine ileocolonic aganglionosis, which is also called lethal white foal syndrome (LWFS), is a severe congenital condition characterized by the unsuccessful colonization of neural crest progenitors in the caudal part of the small intestine and the entire large intestine. LWFS, which is attributable to a mutation in the endothelin receptor B gene, is the horse equivalent of Hirschsprung's disease in humans. Affected foals suffer from aganglionosis or hypoganglionosis of the enteric ganglia resulting in intestinal akinesia and colic. In other species with aganglionosis, fibers of extrinsic origin show an abnormal distribution pattern within the gut wall, but we have no information to date regarding this occurrence in horses. Our present aim is to investigate the distribution of extrinsic sympathetic and sensory neural fibers in LWFS, focusing on ileum and the pelvic flexure of the colon of two LWFS foals compared with a control subject. The sympathetic fibers were immunohistochemically identified with the markers tyrosine hydroxylase and dopamine beta-hydroxylase. The extrinsic sensory fibers were identified with the markers Substance P (SP) and calcitonin gene-related peptide (CGRP). Since SP and CGRP are also synthesized by subclasses of horse intramural neurons, LWFS represents a good model for the selective study of extrinsic fiber distribution. Affected foals showed large bundles of extrinsic fibers, compared with the control, as observed in Hirschsprung's disease. Furthermore, altered adrenergic pathways were observed, prominently in the pelvic flexure. The numbers of SP- and CGRP-immunoreactive fibers in the muscle, a target of enteric neurons, were dramatically reduced, whereas fibers deduced to be extrinsic sensory axons persisted around submucosal blood vessels. Fiber numbers in the mucosa were reduced. Thus, extrinsic innervation, contributing to modulate enteric functions, might also be affected during LWFS.

Objective: To assess agreement and trending ability of transpulmonary thermodilution (TPTD), calibrated pulse contour (PiCCO), and pulse power (PulseCO) methods compared to pulmonary artery thermodilution (PATD) for determination of cardiac output (CO) in anesthetized dogs. Design: Experimental, prospective study. Setting: University teaching hospital. Animals: Six adult Beagle dogs. Interventions: Dogs were anesthetized with sevoflurane and instrumented with pulmonary and femoral artery thermodilution catheters. CO was measured at baseline and at 5, 15, 30, 45, 60, 120, 180, and 240 minutes after IV administration of ketamine or s-ketamine. Baseline PATD and TPTD calibrated PulseCO and PiCCO, respectively. Agreement and trending ability was analyzed with Bland–Altman, concordance, and polar plot methodology. Measurements and Main Results: Median (range) CO values of 2.27 (0.98–3.4) L/min were measured with PATD, and 2.8 (1.9–4.04) L/min with TPTD, which resulted in a mean bias (± standard deviation) of −0.66 (± 0.36) L/min. Concordance rate was 91% and radial limits of agreement (RLOA) were ±35°. PATD against PiCCO resulted in a mean bias of −0.71 (± 0.62) L/min and PATD against PulseCO in a mean bias of 0.13 (± 0.46) L/min. The continuous techniques resulted in concordance rates of 77% for PATD-PiCCO and 74% for PATD-PulseCO and RLOA of ±57° and ±60°, respectively. Conclusions: Intermittent TPTD showed marginal trending ability, while continuous pulse contour and pulse power methods showed poor trending ability over a 4-hour period. The poor performance and possible side effects of the methods tested in this study suggest that they should not be recommended for use in critical patients.

Three-dimension (3D) scaffolds for bone tissue regeneration were produced combining three different phases: nanometric hydroxyapatite (HA) was synthesized by precipitation method and the crystals nucleation took place directly within collagen fibrils following a biologically inspired mineralization process; polycaprolactone was employed to give the material a 3D structure. The chemico-physical analysis carried out to test the material's properties and composition revealed a high similarity in composition and morphology with biologically mineralized collagen fibrils and a scaffold degradation pattern suitable for physiological processes. The micro- computerized tomography (micro-CT) showed 53.53% porosity and a 97.86% mean interconnected pores. Computer-aided design and computer-aided manufacturing (CAD-CAM) technology was used for molding the scaffold's volume (design/shape) and for guiding the surgical procedure (cutting guides). The custom made scaffolds were implanted in sheep mandible using prototyped surgical guides and customized bone plates. After three months healing, scanning electron microscopy (SEM) analysis of the explanted scaffold revealed a massive cell seeding of the scaffold, with cell infiltration within the scaffold's interconnected pores. The micro-CT of the explanted construct showed a good match between the scaffold and the adjacent host's bone, to shield the implant primary stability. Histology confirmed cell penetration and widely documented neoangiogenesis within the entire scaffold's volume. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 723-734, 2017.

Objective To assess the differences in the pharmacokinetic profiles of S-ketamine, R-ketamine and their metabolites, S-norketamine and R-norketamine, and to measure relevant physiologic variables after intravenous administration of racemic (RS) ketamine or S-ketamine alone in Beagle dogs sedated with medetomidine. Study design Experimental, blinded and randomized crossover study. Animals A total of six (three female and three male) adult Beagle dogs. Methods Medetomidine (450 μg m–2) was administered intramuscularly, followed by either S-ketamine (2 mg kg–1) or RS-ketamine (4 mg kg–1) 20 minutes later, both administered intravenously. Blood samples were collected before medetomidine administration and at multiple time points 1–900 minutes following the ketamine administration. Plasma samples were analysed using liquid chromatography–tandem mass spectrometry. Heart rate, respiratory rate, noninvasive blood pressure, haemoglobin saturation with oxygen and body temperature were measured at baseline, before ketamine administration, and 1, 2, 5, 10, 15, 20 and 30 minutes after ketamine administration. All cardiovascular variables, blood glucose, haemoglobin and lactate concentrations were analysed using different linear mixed effects models; the significance was set at p Results S-ketamine showed a two-compartment kinetic profile; no statistically significant differences were observed between its concentrations or in the calculated pharmacokinetic parameters following S- or RS-ketamine. When the racemic mixture was administered, no differences were detected between R- and S-ketamine concentrations, but the area under the curve (AUC) for R-norketamine was significantly lower than that for S-norketamine. Clinically relevant physiologic variables did not show statistically significant differences following the administration of the racemic mixture or of S-ketamine alone. Conclusions and clinical relevance This study performed in dogs showed that RS-ketamine and S-ketamine combined with medetomidine showed enantioselective pharmacokinetics as S- and R-norketamine AUCs were different, but S-ketamine levels were identical.

The objective of the study is to evaluate the anesthetic effect produced by the tiletamine/zolazepam association in horses sedated with medetomidine undergoing orchiectomy under field conditions. Ten uncastrated male horses, American Society of Anesthesiologists category I, undergoing orchiectomy, were enrolled in the study. The horses received acepromazine (0.03 mg kg−1 intramuscularly) 30 minutes before sedation with medetomidine (8 μg kg−1 intravenously). Anesthesia was induced with tiletamine/zolazepam (0.7 mg kg−1 intravenously). During anesthesia, heart rate (HR), pulse quality, respiratory rate, temperature, SpO2, movement, and reflexes were measured every 5 minutes. Arterial blood gases were evaluated before the premedication and during the surgical procedure. The quality of induction and recovery was recorded. The sedation was appropriate, and the quality of recovery was evaluated as good. A statistically significant difference was revealed between the basal and intra-anesthesia HR. No differences were observed in blood gas parameters during the procedure. The results suggested that general anesthesia produced with tiletamine/zolazepam for induction in horses sedated with medetomidine is suitable during orchiectomy under field conditions.

The aim of the present work was to isolate, cultivate, differentiate, and conduct cellular characterization of mesenchymal stem cells (MSCs) derived from equine adipose tissue (eAT) and bone marrow (eBM). Isolated and characterized cells were used in racehorses suffering from a superficial flexor tendon injury. Equine adipose tissue collection was performed at the base of the horse tail, whereas eBM was aspirated from iliac crest. Mononuclear cell fraction was isolated and cultured. In vitro differentiation and molecular characterization at P3 of culture were performed. No statistically significant differences in the number of cell doublings were found among different culture passages (P > .05). Doubling time was greater for eBM than eAT (3.2 1.5 vs. 1.3 0.7; P < .05). Positive von Kossa and Alizarin Red staining confirmed osteogenesis. Alcian Blue and Oil Red O staining illustrated chondrogenesis and adipogenesis, respectively. Isolated cells resulted positive for CD90, CD44, and CD105, whereas negative for hematopoietic markers, CD14, CD45, and CD34. Using isolated cells for injured tendon therapy, no adverse reactions were observed, and all inoculated horses returned to race competitions. In vitro results revealed the immunophenotypic characterization of isolated cells similar to that observed in human MSCs from the same sources; furthermore, in the present study, their clinical use proves the safety of eBM-derived and eAT-derived MSCs and a successful outcome for the treated animals that returned to their previous level of sport activity.

The aim of this pilot study was to test a new customized biomimetic composite scaffolds to restore bone defects using CAD–CAM technology. The new biomaterial was synthesized using biomimetic mineralized collagen, and poly (ɛ-caprolactone) (PCL). The mold used in scaffolding, the surgical guide, and the bony plate used during the surgery, were rapid prototyped (CAD–CAM technology). Our in vitro results showed that sheep mesenchymal stem cells (MSC) seeding efficiency was very high (>94%). Furthermore, cells can adhere, grow and maintain cell viability up to 7 days of in vitro cultivation on the scaffold as revealed by SEM and confocal laser scanning microscope analysis. These results are indicative of good scaffold biocompatibility. The use of the sheep experimental model allowed exploring the efficacy of the customized bony plate used to give extra mechanical support to the scaffold, the potential of CAD–CAM technology in customization of the scaffold, and the in vivo biomimetic properties of the composite biomaterial.

The aims of this study were to measure plasma levels of R- and S-ketamine and their major metabolites R- and S-norketamine following single intravenous bolus administration of racemic or S-ketamine in sevoflurane anaesthetised dogs and to calculate the relevant pharmacokinetic profiles. Six adult healthy beagle dogs were used in the study. An intravenous bolus of 4 mg/kg racemic ketamine (RS-KET) or 2 mg/kg S-ketamine (S-KET) was administered, with a three-weeks washout period between treatments. Venous blood samples were collected at fixed times until 900 min and R- and S-ketamine as well as R- and S-norketamine plasma levels determined by liquid chromatography coupled with tandem mass spectrometry. Cardiovascular parameters were recorded during the anaesthesia until 240 min. All dogs recovered well from anaesthesia. No statistical differences between groups were detected in any cardiovascular parameter. The pharmacokinetics of S-ketamine did not differ when injected intravenously alone or as part of the racemic mixture in dogs anaesthetised with sevoflurane. Following racemic ketamine, the area under the curve of R-norketamine was statistically higher than the one of S-norketamine.

Background Serotonin plays a pivotal role in regulating gut motility, visceral sensitivity, and fluid secretion via specific receptors. Among these receptors, 5-HT4 exerts a prominent control on gut motor function. Although the prokinetic effect exerted by 5-HT4 agonists is well known, the cellular sites of 5-HT4 expression remain poorly understood in large mammals, e.g., horses. In this study, we evaluated the distribution of 5-HT4 in the horse intestine and in foals with enteric aganglionosis, reminiscent of human Hirschsprung's disease. Methods The intestine and spinal ganglia were obtained from three healthy horses and two foals with hereditary ileocolonic aganglionosis. Tissues were processed for immunohistochemistry using a specific antibody to 5-HT4 and a variety of neuronal markers. Myenteric and submucosal plexus 5-HT4-immunoreactive (IR) neurons were quantified as relative percentage (mean±SD) to the total number of neurons counted. Furthermore, the density of 5-HT4-IR nerve fibers was evaluated in the mucosa and tunica muscularis. Key Results The 5-HT4 immunoreactivity was localized to large percentages of myenteric neurons ranging from 28±9% (descending colon) to 63±19% (ileum), and submucosal neurons ranging from 54±6% (ileum) to 68±14% (duodenum). The 5-HT4-immunoreactivity was co-expressed by some substance P-IR (SP-IR) spinal ganglion neurons and extrinsic sensory fibers of aganglionic foals. Conclusions & Inferences The presence of 5-HT4 in many enteric and extrinsic sensory neurons and nerve fibers provides solid morphological evidence of the cellular sites of 5-HT4 expression in horses. The evidence of SP-IR sensory neurons positive for 5-HT4 suggests its role in visceral sensitivity.