Discrepancies between Genetic and Visual Coat Color Assignment in Sarcidano Horse.

This study aimed to evaluate the discrepancies between genetic and visual coat color assignment in the Sarcidano Horse and to elucidate potential reasons. Individual DNA from 90 Sarcidano Horses was used for genetic assignment of coat color to explore the correspondence with individual forms containing phenotypical traits. The MC1R exon 1 and ASIP exon 3 have been genotyped and sequenced to obtain a picture of the coat color distribution in this breed. Surprisingly, once we compared the genetic results with the individual forms reporting the phenotypic data for each subject, a certain degree of non-correspondence between the phenotypic and genetic data in relation to coat color emerged. From the genetic analysis, Chestnuts (n = 58) resulted the most common Sarcidano Horse (n = 58), followed by a quite large number of Blacks (n = 28) and a very small number of Bays (n = 4), whereas phenotypic distribution resulted in 38 Chestnuts, 40 Bays, only 2 Blacks, and 10 Grays (without the possibility of recognizing the true color they carried). Chestnut resulted a very representative coat color, while many horses that visually identified as Bays were genetically Blacks. This discrepancy, that could be due to a variety of individual and external factors, including age, time of year, living situation and dietary condition, suggesting the importance of accurate coat color identification to ensure adequate features registration and reliable prediction of offspring's coat color.

Student insights towards animal welfare science and law. Survey results from Sassari University, Italy.

In this paper, we describe the results of an online survey consisting of 23 questions created to evaluate the knowledge and interest on animal welfare by students attending 15 different scientific, medical, and biomedical courses at University of Sassari, Italy. The survey collected students' demographic data, level of knowledge both on animal welfare and 3Rs, as well as their opinions on animal experimentation. The majority of the cohort was female and over 24 years of age. About a third of the students responded that their graduate programme included subjects that taught science, ethics, and animal welfare legislation. Just 21.2% of respondents had heard about the concept of 3Rs. About a quarter of the students believed that animal models can be replaced by in vitro and in silico methods while half believed that both are needed. However, 70% of the participants did not know the existence of an Ethics and Animal Welfare Committee. The result showed the importance of an Animal Welfare Course for the professional future of a larger number of students and underlined the key role of veterinary medicine in promoting ethics and animal experimentation.

Characterization of the Sarcidano Horse Coat Color Genes.

The goal of this study was to contribute to the general knowledge of the Sarcidano Horse, both by the identification of the genetic basis of the coat color and by updating the exact locations of the genotyping sites, based on the current EquCab3.0 genome assembly version. One-hundred Sarcidano Horses, living in semi-feral condition, have been captured to perform health and biometric checks. From that total number, 70 individual samples of whole blood were used for DNA extraction, aimed to characterize the genetic basis of the coat color. By genotyping and sequencing analyses of the MC1R Exon 1 and ASIP Exon 3, a real image of the coat color distribution in the studied population has been obtained. Chestnut and Black resulted in the most representative coat colors both from a phenotypic and genotypic point of view, that is suggestive of no human domestication or crossbreeding with domestic breed. Due to its ancient origin and genetic isolation, an active regional plan for the conservation of this breed would be desirable, focused on maintenance of resident genotypes and genetic resources. Collection and management of DNA, sperm, embryos, with the involvement of research centers and Universities, could be a valid enhancing strategy.

Strain Elastography of Injured Equine Superficial Digital Flexor Tendons: A Reliability Study of Manual Measurements.

Early diagnosis of tendon injuries and accurate long-term monitoring of the healing process are key for equine veterinarians that use conventional ultrasonography. The development of strain elastography could improve the management of clinical cases. The aim of the study was to assess the intraobserver repeatability and interobserver reproducibility of manual measurements of the colored areas of the tendons within elastograms and to standardize this manual modality by comparing the analysis of the images with ImageJ. Twenty elastograms of the injured superficial digital flexor tendons (SDFTs) of horses were analyzed by two different operators after an acute injury was diagnosed with ultrasonography. Statistical analysis demonstrated excellent intraobserver repeatability (intraclass correlation coefficient, ICC = 0.949) and good interobserver reproducibility (ICC = 0.855) for manual measurements performed with tools available on the ultrasound unit. A good agreement between manual measurements and measurements performed with ImageJ (ICC = 0.849) was then demonstrated. Despite its subjectivity, the manual modality proved to be a valid method for analyzing images obtained with strain elastography.

REAC regenerative treatment efficacy in experimental chondral lesions: a pilot study on ovine animal model.

Radioelectric asymmetric conveyor (REAC) technology is a platform designed to optimize cell polarity. Cell polarity is a universal biological phenomenon that is implicated in cell differentiation, proliferation, morphogenesis, aging, and rejuvenation. In this work, we investigate a timing and administration protocol for tissue optimization regenerative treatment type C, in order to treat aging-related chondral damage or injuries and gain insights into regenerative processes of articular cartilage in humans. The chondral lesion produced in this study in an animal model (6 knee joints of 4 adult sheep) was 6 mm in diameter and about 2 mm deep. These lesions, which did not involve subchondral bone, tend to increase in size and depth and are not completely repaired with normal hyaline articular cartilage since adult articular cartilage is avascular and has a very slow turnover at the cellular and molecular level. Moreover, the hydration of articular cartilage is reduced with aging and with decreased mitotic activity, synthesis, and population size of chondrocytes. Six months posttreatment, lesions appeared filled, though not completely, with newly generated tissue of the light opalescent color of healthy articular cartilage, which otherwise covered the underlying subchondral bone. The newly formed tissue surface appeared to be quite regular. Nearly complete regeneration of subchondral bone occurred, with little vascularization and ossification nuclei almost absent. The results of this study confirm previous data obtained in vitro on the regenerative effects of REAC technology on human normal and osteoarthritic chondrocytes exposed to IL-1ß. The present findings indicate that REAC tissue optimization-regenerative treatment type C is a promising therapeutic tool among the other REAC regenerative treatment protocols for the treatment of cartilage lesions.

Clinical evolution of cerebral coenurosis from invasive to chronic infection in sheep and a goat.

This survey describes the evolution in vivo of Coenurus cerebralis in small ruminants. At presentation, neurological signs and cerebrospinal fluid (CSF) features were suggestive of multifocal or diffuse inflammatory reaction. Magnetic resonance imaging (MRI) captured the transition between the invasive and quiescent phase of the infection, revealing the concurrent presence of meningitis and small cysts. During the quiescent phase, in all animals, neurological symptoms disappeared, and cerebrospinal fluid was unremarkable while cysts grew progressively. Subsequently, the onset of neurological symptoms coincided with MRI signs of diffuse or localized increase of intracranial pressure, as confirmed by direct intracranial pressure measuring. All the animals had an excellent post-surgical recovery. This is the first report describing the evolution of coenurosis in vivo. Sequential imaging allowed describing interesting such as the death of some coenuri and different parasite growth rate in the same host.

Physical reparative treatment in reptiles.

BACKGROUND: The tissue growth necessary to achieve a complete or partial restitution ad integrum as a result of injury to soft tissue and/or hard times in reptiles is variable and often needs long time in relation to the species, to the habitat and to their intrinsic physiological characteristics. The purpose of this work was to see if the tissue optimization (TO) treatment with radio electric asymmetric conveyer (REAC) provided good results in these animals and whether its use translates into reduced time of tissue repair. This paper describes preliminary results with in promoting the tissue repair in reptiles. CASES PRESENTATION: A 5 year old male Testudo graeca (Leo) and Trachemys scripta scripta (Mir) and a 15 year old female Testudo hermanni (Juta) were evaluated because of soft tissue injuries. A female 25 year old Trachemys scripta elegans (Ice), a female 2.5 year old Trachemys scripta scripta (Penelope) as well as a 50 year old male Testudo graeca (Margherito) were evaluated because of wounds of the carapace. Following debridement and traditional therapies, Leo, Penelope and Margherito were exposed to the radio electric asymmetric conveyer (REAC) device, with a specific treatment protocol, named tissue optimization-basic (TO-B). Also Ice and Mir were subjected to REAC treatment after wounds debridement. Juta was treated only with REAC treatment. Complete wound healing was evident after 17 days for Leo, 7 days for Penelope, 27 days for Mir, 78 days for Ice and after 14 days for Margherito. Juta showed a considerable tissue activation in 2 days and complete wound healing in 5 days. CONCLUSION: Our findings suggest that REAC TO-B treatment may provide advantages over other traditional methods after complete wound healing in Leo, and also suitable healing in the other patients. Then REAC device with its specific treatment TO-B protocol, which induces tissue repair without causing severe stress to the patient, could be a potential therapy for tissue damage healing in reptiles. Further studies still need to be conducted to support our observations.

Magnetic resonance imaging of the brain and skull of sheep with cerebral coenurosis.

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.

Repeated platelet concentrate injections enhance reparative response of microfractures in the treatment of chondral defects of the knee: an experimental study in an animal model.

PURPOSE: To assess the histology and biomechanics of repair cartilage after microfractures with and without repeated local injections of platelet concentrate for the treatment of full-thickness focal chondral defects of the knee. METHODS: A full-thickness chondral lesion on the medial femoral condyle was created in 30 sheep and treated with microfractures. Animals were divided into 2 groups, according to postoperative treatment: in group 1 we performed 5 weekly injections of autologous conditioned plasma, whereas group 2 did not undergo further treatments. Animals were killed at 3, 6, and 12 months after treatment. Macroscopic, histologic, and biomechanical evaluations were performed. Differences between groups at each time interval and differences over time within groups were analyzed for each outcome. Significance was set at P < .05. RESULTS: Group 1 showed significantly better macroscopic, histologic, and biomechanical results than group 2 at each time interval. Analysis of time effect within groups showed that in group 1, quality of repair tissue significantly improved from 3 to 6 months after treatment and remained stable over time for all the outcomes; in group 2 a significant histologic and mechanical deterioration was observed between 6 and 12 months' follow-up. CONCLUSIONS: Five repeated local injections of autologous conditioned plasma after microfractures in the treatment of full-thickness cartilage injuries promoted a better and more durable reparative response than isolated microfractures, although they did not produce hyaline cartilage. CLINICAL RELEVANCE: Periodical intra-articular injections of platelet concentrate after microfractures may improve cartilage repair and prevent further degenerative changes.

Experimental use of labial mucosa free graft urinary conduit in the Mitrofanoff principle.

PURPOSE: One method of treating urinary incontinence is to create a catheterizable and continent vesicostomy by the Mitrofanoff principle, based on the use of a conduit, which in the original method is the appendix. The authors present an experimental technique in which a continent vesicostomy is created by the Mitrofanoff principle, using a conduit made of a labial mucosa free graft. METHODS: Six 30-day-old pigs underwent surgery to create a continent vesicostomy by the Mitrofanoff principle. A tube, made of labial mucosa free graft from the lower lip, was used. The labial mucosal conduit was anastomosed to the bladder mucosa. The tunnel was then prepared, part of which passed through the fibers of the right rectus muscle. The conduit was taken through the tunnel and anastomosed to the skin in the right iliac fossa. RESULTS: Fifty days after the operation, the vesicostomy was continent, pervious, and easily catheterizable in 5 animals. In the sixth pig, the cystostomic tube did not graft successfully because it was infected. CONCLUSIONS: A number of investigators have extrapolated Mitrofanoff method using the ileum, the ureter, or others. The use of a tube made by labial mucosa free graft is an easily performed technique, and as with the Mitrofanoff method, continence and easy catheterization are successfully achieved.

Mapping a sex hormone-sensitive gene determining female resistance to liver carcinogenesis in a congenic F344.BN-Hcs4 rat.

Hepatocellular carcinoma (HCC) is prevalent in human and rodent males. Hepatocarcinogenesis is controlled by various genes in susceptible F344 and resistant Brown Norway (BN) rats. B alleles at Hcs4 locus, on RNO16, control neoplastic nodule volume. We constructed the F344.BN-Hcs4 recombinant congenic strain (RCS) by introgressing a 4.41-cM portion of Hcs4 from BN strain in an isogenic F344 background. Preneoplastic and neoplastic lesions were induced by the "resistant hepatocyte" protocol. Eight weeks after initiation, lesion volume and positivity for proliferating cell nuclear antigen (PCNA) were much higher in lesions of F344 than BN rats of both sexes. These variables were lower in females than in males. Lesion volume and PCNA values of male RCS were similar to those of F344 rats, but in females corresponded to those of BN females. Carcinomatous nodules and HCC developed at 32 and 60 weeks, respectively, in male F344 and congenics and, rarely, in F344 females. BN and congenic females developed only eosinophilic/clear cells nodules. Gonadectomy of congenic males, followed by beta-estradiol administration, caused a decrease in Ar expression, an increase in Er-alpha expression, and development of preneoplastic lesions comparable to those from BN females. Administration of testosterone to gonadectomized females led to Ar increase and development of preneoplastic lesions as in F344 males. This indicates a role of homozygous B alleles at Hcs4 in the determination of phenotypic patterns of female RCS and presence at Hcs4 locus of a high penetrance gene(s), activated by estrogens and inhibited/unaffected by testosterone, conferring resistance to females in which the B alleles provide higher resistance.

Substitution of thoracic oesophagus by interposition of a pedicled gastric tube, preserving LES function: clinical and histological follow-up.

Assessment of clinical evolution and histological findings in a group of animals experimentally operated on to substitute the thoracic oesophagus with a gastric tube. Six piglets underwent oesophageal replacement with a gastric tube, constructed from the greater curvature of stomach and pedicled on the gastroepiploic vessels, which was interposed between the oesophageal stumps. At follow-up, all animals were found to be growing and eating normally, apart from case no 1 (stenosis of the lower oesophageal anastomosis). Ph-metry showed a neutral pH on the gastric tube. Postmortem histological analysis of the gastric tube and native oesophagus samples did not show any significant lesions, except in case no 1 (inflammation of the gastric tube and upper oesophagus due to food stasis). The technique of substitution of the oesophagus with an interposed pedicled gastric tube can be a breakthrough in existing surgical methods of oesophageal replacement.

Evaluation of bone plug and soft tissue anterior cruciate ligament graft fixation over time using transverse femoral fixation in a sheep model.

PURPOSE: To evaluate the mechanical role of bone-anterior cruciate ligament (ACL) graft junction in comparison with primary fixation of the graft. TYPE OF STUDY: Ex vivo controlled biomechanical study. METHODS: An ACL reconstruction was performed on 2 groups of 40 sheep each. The ACL graft was patellar tendon in group 1, and free tendon in group 2. Load-to-failure tests were performed at 1, 2, 3, and 6 months. Each group was divided into 2 subgroups: In subgroup A the femoral fixation devices were removed before testing and in subgroup B they were left in place. RESULTS: At 1 month, structural properties of subgroup 2A were significantly lower than other subgroups. Until the third month, structural properties of each subgroup were significantly lower than normal ACL. Grafts always failed at midsubstance, except for subgroup 2A at 1 month, which pulled out of the femoral tunnel. CONCLUSIONS: Bone plug incorporation was stronger than graft strength at 1 month whereas soft tissue tendon incorporation was not stronger than graft until 2 months. At 3 months, both groups failed at approximately 30% of the native ACL strength, and at 6 months, both groups were nearly equal in terms of load to failure and stiffness. CLINICAL RELEVANCE: High fixation strength and stiffness of ACL graft does not allow earlier return to sports activities.