Maria Lucia Manunta

Objective—To design and test a motion analysis protocol for the gait analysis of adult German Shepherd (GS) dogs with a focus in the analyses of their back movements. Animals—Eight clinically healthy adult large-sized GS dogs (age, 4 ± 1.3 years; weight, 38.8 ± 4.2 kg). Procedures—A six-camera stereo-photogrammetric system and two force platforms were used for data acquisition. Experimental acquisition sessions consisted of static and gait trials. During gait trials, each dog walked along a 6 m long walkway at self-selected speed and a total of six gait cycles were recorded. Results—Grand mean and standard deviation of ground reaction forces of fore and hind limbs are reported. Spatial-temporal parameters averaged over gait cycles and subjects, their mean, standard deviation and coefficient of variance are analyzed. Joint kinematics for the hip, stifle and tarsal joints and their average range of motion (ROM) values, and their 95% Confidence Interval (CI) values of kinematics curves are reported. Conclusions and Clinical Relevance—This study provides normative data of healthy GS dogs to form a preliminary basis in the analysis of the spatial-temporal parameters, kinematics and kinetics during quadrupedal stance posture and gait. Also, a new back movement protocol enabling a multi-segment back model is provided. Results show that the proposed gait analysis protocol may become a useful and objective tool for the evaluation of canine treatment with special focus on the back movement.

The objectives of this study were to test: (1) the repeatability of ultrasonographic examination of the optic nerve sheath diameter (ONSD) in the cat; (2) the association between the ONSD and age, sex and body weight in healthy cats; and (3) the difference in the ONSD between healthy cats and those suffering from presumed intracranial hypertension (ICH). This study had a prospective, blinded, observational cross-sectional study design. Two groups of animals were considered: healthy cats (group A) and cats with a diagnosis of presumed ICH (group B). The ONSD was evaluated, measured and compared between the two groups via an ultrasonographic transpalpebral approach. Repeatability of the procedure was evaluated through the intraclass correlation coefficient (ICC). Data were statistically compared using the Student’s t-test and linear regression analysis. A strong inter- and intraobserver ICC indicating good repeatability was observed. The interobserver ICC was 0.965 (P P P P  P <0.001). The transpalpebral ultrasonographic technique is a non-invasive, feasible and reproducible method to measure ONSD both in healthy cats and in cats suffering from suspected ICH.

Objective: To evaluate the effects of iso‐osmolar doses of 18% mannitol and 3% sodium chloride (NaCl) solutions in decreasing intracranial pressure (ICP) in animals with severe traumatic brain injury (TBI).Design: Prospective uncontrolled interventional study.Setting: Veterinary university teaching hospital.Animals: Two cats and 1 dog with TBI with a modified Glasgow Coma Scale score ≤8 after hemodynamic stabilization, and with brain magnetic resonance imaging changes suggestive of intracranial hypertension.Interventions: Animals were surgically instrumented for direct ICP measurement, then randomly treated with iso‐osmolar doses of 18% mannitol or 3% NaCl. Direct ICP and cerebral perfusion pressure (CPP) were recorded both before treatment and for 120 minutes following drug administration.Measurements and Main Results: Direct ICP and CPP were recorded both before treatment and at 5 additional time points following administration over the subsequent 120 minutes. Case 1 received 3% NaCl without any response to therapy; refractory posttraumatic hypertension was suspected. Case 2 was treated with 3% NaCl; ICP decreased by 40.7% and CPP increased by 15%; however, these effects were transient. Case 3 received 18% mannitol, and ICP decreased by 19% and CPP increased to normal. However, there was a rebound increase in ICP that was higher than pretreatment values, and CPP decreased slightly before it gradually increased to normal values towards the end of the study.Conclusions: Both mannitol and hypertonic saline decrease ICP and improve CPP, but the effect observed in this pilot study suggests that there might be differences in the duration of these effects. Appropriately designed studies in a larger and homogeneous population are warranted to further investigate these findings.

Articular cartilage lacks a regenerative response. Embryonic stem cells (ESCs) are a source of pluripotent cells for cartilage regeneration. Their use, however, is associated with a risk of teratoma development, which depends on multiple factors including the number of engrafted cells and their degree of histocompatibility with recipients, the immunosuppression of the host and the site of transplantation. Colonies of sheep embryonic stem-like (ES-like) cells from in vitro-produced embryos, positive for stage-specific embryonic antigens (SSEAs), alkaline phosphatase (ALP), Oct 4, Nanog, Sox 2 and Stat 3 gene expression, and forming embryoid bodies, were pooled in groups of two-three, embedded in fibrin glue and engrafted into osteochondral defects in the left medial femoral condyles of 3 allogeneic ewes (ES). Empty defects (ED) and defects filled with cell-free glue (G) in the condyles of the controlateral stifle joint served as controls. After euthanasia at 4 years post-engraftment, the regenerated tissue was evaluated by macroscopic, histological and immunohistochemical (collagen type II) examinations and fluorescent in situ hybridization (FISH) assay to prove the ES-like cells origin of the regenerated tissue. No teratoma occurred in any of the ES samples. No statistically significant macroscopic or histological differences were observed among the 3 treatment groups. FISH was positive in all the 3 ES samples. This in vivo preclinical study allowed a long-term evaluation of the occurrence of teratoma in non-immunosuppressed allogeneic adult sheep engrafted with allogeneic ES-like cells, supporting the safe and reliable application of ES cells in the clinic.

Diagnosing high intracranial pressure by clinical and diagnostic imaging is particularly challenging for chronic or slow‐growing lesions. The aim of this prospective case‐control study is to determine whether the neuroscore and brain magnetic resonance imaging (MRI) are related to the direct measurement of intracranial pressure in sheep affected by intracranial slow‐growing lesions due to chronic cerebral coenurosis (Coenurus cerebralis). Seventeen affected and 10 control sheep were included. All animals underwent a neurological examination, MRI of the brain, and direct measurement of intracranial pressure. The severity of clinical signs and MRI findings were scored. Data were statistically analyzed. The invasive intracranial pressure value was higher in affected animals. A severely altered neuroscore is related to an increased intracranial pressure beyond the normal threshold (P < 0.05). The volume of the calvarium was larger in affected animals than in control animals (P = 0.0001) and was positively influenced by the presence and volume of the parasitic cyst (r = 0.7881, P < 0.01). Several degrees of deviation and deformation of both the ventricular system and brain parenchyma were detected by MRI. Subjective MRI findings were not associated with intracranial hypertension. In conclusion, this study shows that in sheep affected by slow‐growing lesions, severe alterations in the neuroscore and the results of objective MRI are related to an increased intracranial pressure beyond the normal threshold.

Shepherd and stray dogs are thought to represent the primary definitive hosts of Coenurosis by Taenia multiceps, due to their feeding habits which translate into high chances of coming into contact with infected intermediate hosts. Nonetheless, little attention has been paid to the role of the red fox (Vulpes vulpes) in the epidemiology of coenurosis. In fact a knowledge gap exists on the role played by red foxes in the epidemiology of Taenia multiceps and the capability of this parasite to produce fertile and viable eggs in this wild canid, i.e. on the occurrence of a sylvatic cycle. This study investigates the role of the red fox (Vulpes vulpes) in the epidemiology of T. multiceps and related metacestodoses. The small intestine of 63 red foxes was macroscopically examined for the presence of cestodes. Adult parasites were identified morphologically as being T. multiceps. Tapeworm eggs were counted and stored at 4 °C in physiological saline solution prior to experimental infection of four sheep and one goat. Sheep were inoculated orally on Day 0 with 3000 (sheep 1), 5000 (sheep 2 and 3) or 7000 eggs (sheep 4), while the goat was infected with 5000 eggs of T. multiceps. The animals were followed-up regularly by MRI and underwent surgical treatment between days 180 to day 240 post infection. Collected coenuri were identified using morphological and molecular methods. A total of 6.3 % of red foxes were found infected with T. multiceps and the eggs obtained from the worms were determined to have a viability of 45.4 %. Two of the challenged sheep and the goat developed disease compatible with T. multiceps. Morphometrical features of the cysts were consistent with those of T. multiceps; nucleotide amplification and sequencing of mitochondrial genes (i.e., cox1 and Nd1) from the metacestode material confirmed the identification. The present study is the first to provide evidence of the role of the red fox as a competent definitive host for T. multiceps, thus changing the epidemiological scenarios of infections by this cestode.

Articular cartilage has poor intrinsic capacity for regeneration because of its avascularity and very slow cellular turnover. Defects deriving from trauma or joint disease tend to be repaired with fibrocartilage rather than hyaline cartilage. Consequent degenerative processes are related to the width and depth of the defect. Since mesenchymal stem cells (MSCs) deriving from patients affected by osteoarthritis have a lower proliferative and chondrogenic activity, the systemic or local delivery of heterologous cells may enhance regeneration or inhibit the progressive loss of joint tissue. Embryonic stem cells (ESCs) are very promising, since they can self-renew for prolonged periods without differentiation and can differentiate into tissues from all the 3 germ layers. To date only a few experiments have used ESCs for the study of the cartilage regeneration in animal models and most of them used laboratory animals. Sheep, due to their anatomical, physiological and immunological similarity to humans, represent a valid model for translational studies. This experiment aimed to evaluate if the local delivery of male sheep embryonic stem-like (ES-like) cells into osteochondral defects in the femoral condyles of adult sheep can enhance the regeneration of articular cartilage. Twenty-two ewes were divided into 5 groups (1, 2, 6, 12 and 24 months after surgery). Newly formed tissue was evaluated by macroscopic, histological, immunohistochemical (collagen type II) and fluorescent in situ hybridization (FISH) assays. Regenerated tissue was ultimately evaluated on 17 sheep. Samples engrafted with ES-like cells had significantly better histologic evidence of regeneration with respect to empty defects, used as controls, at all time periods. Histological assessments demonstrated that the local delivery of ES-like cells into osteochondral defects in sheep femoral condyles enhances the regeneration of the articular hyaline cartilage, without signs of immune rejection or teratoma for 24 months after engraftment.

Objective—To determine MRI characteristics of the skulls and brains of sheep with chronic cerebral coenurosis (CC) caused by naturally acquired Taenia multiceps infection. Animals—33 sheep with CC and 10 healthy control sheep. Procedures—Sheep underwent MRI of the head. Volumes of the cranial cavity and rostral and caudal fossas of the cranial cavity were determined. For CC-affected sheep, the number, location, and volume of T multiceps cysts were determined and the percentage volumes of cysts in the cranial cavity and rostral and caudal fossas of the cranial cavity were calculated. Focal and diffuse abnormalities of cranial bones in CC-affected sheep were identified. Brain edema and hemorrhage and signs of increased cranial pressure (ICP) in MRI images were determined. Results—Volumes of the cranial cavity and rostral and caudal fossas of the cranial cavity were significantly larger for CC-affected sheep versus healthy control sheep. Total volumes of cysts ranged from 4.40% to 46.93% in cranial cavities of sheep, 4.12% to 51.53% in rostral fossas of cranial cavities of sheep, and 15.24% to 68.30% in caudal fossas of cranial cavities of sheep. Moderate to severe diffuse cranial bone abnormalities and signs of increased ICP in MRI images were detected in 21 and 24 sheep, respectively, and were positively correlated with cyst volumes. Conclusions and Clinical Relevance—Results suggested that cranial cavity volume and morphological abnormalities can be detected in sheep with CC. These changes may reflect abnormalities in ossification of the cranial bones secondary to chronically increased ICP caused by development of T multiceps cysts.