Sonographic features of the uncomplicated postoperative abdomen in dogs treated for pyometra by ovariohysterectomy.

Pyometra is a prevalent disease in intact bitches and the standard treatment is ovariohysterectomy (OHE). Published descriptions of normal sonographic findings after OHE are currently lacking. The aims of this prospective observational study were to describe and compare postoperative abdominal sonographic features for three timepoints following OHE in a group of dogs with pyometra and an uneventful recovery. A total of 22 dogs had sequential focused abdominal ultrasound examinations on days 1, 4-6, and 10-15 postsurgery. Recorded sonographic features for each examination time point and characteristics of the cervical stump and the mesovarium, size, and echogenicity of medial iliac lymph nodes (MILNs), presence of free peritoneal fluid, and pneumoperitoneum. The cervical stump appeared as a heterogenous area with a hypoechoic center surrounded by hyperechogenic tissue in all dogs. The cervical stump transverse-sectional area was larger on day 4-6 compared with day 1 and day 10-15 (P = .0009). Mesovarium ligature reactions were identified as heterogeneous and hyperechoic areas with central and/or edge shadowing in all dogs. The size and echogenicity of MILNs and the mesovarium reactions did not significantly differ among time points. Free peritoneal fluid was detected in 45%, 41%, and 9% and pneumoperitoneum in 95%, 82%, and 14% of dogs at sequential time points. Findings from this sample of dogs with an uneventful recovery following OHE due to pyometra can be used to assist veterinarians in interpreting postoperative abdominal ultrasonographic characteristics in future dogs treated surgically for pyometra.

A survey of testicular texture in canine ultrasound images.

Introduction: Computer-based texture analysis provides objective data that can be extracted from medical images, including ultrasound images. One popular methodology involves the generation of a gray-level co-occurrence matrix (GLCM) from the image, and from that matrix, texture fractures can be extracted. Methods: We performed texture analysis on 280 ultrasound testicular images obtained from 70 dogs and explored the resulting texture data, by means of principal component analysis (PCA). Results: Various abnormal lesions were identified subjectively in 35 of the 280 cropped images. In 16 images, pinpoint-to-small, well-defined, hyperechoic foci were identified without acoustic shadowing. These latter images were classified as having "microliths." The remaining 19 images with other lesions and areas of non-homogeneous testicular parenchyma were classified as "other." In the PCA scores plot, most of the images with lesions were clustered. These clustered images represented by those scores had higher values for the texture features entropy, dissimilarity, and contrast, and lower values for the angular second moment and energy in the first principal component. Other data relating to the dogs, including age and history of treatment for prostatomegaly or chemical castration, did not show clustering on the PCA. Discussion: This study illustrates that objective texture analysis in testicular ultrasound correlates to some of the visual features used in subjective interpretation and provides quantitative data for parameters that are highly subjective by human observer analysis. The study demonstrated a potential for texture analysis in prediction models in dogs with testicular abnormalities.

Collimation and Exposure Parameter Influence Image Quality and Potential Radiation Dose to the Eye Lens of Personnel in Computed Radiography of the Canine Pelvis.

Introduction: The purpose of this study was to evaluate the effect of collimation on image quality and radiation dose to the eye lenses of the personnel involved in computed radiography of the canine pelvis. Materials and Methods: A retrospective study of canine pelvic radiographs (N = 54) was undertaken to evaluate the relationship between image quality and the degree of field the collimation used. This was followed by a prospective cadaver study (N = 18) that assessed the effects on image quality and on scattered radiation dose of different collimation field areas and exposure parameters. All radiographs were analyzed for image quality using a Visual Grading Analysis (VGA) with three observers. Finally, the potential scattered radiation dose to the eye lens of personnel restraining a dog for pelvic radiographs was measured. Results: The retrospective study showed a slightly better (statistically non-significant) VGA score for the radiographs with optimal collimation. Spatial and contrast resolution and image sharpness showed the greatest improvement in response to minimizing the collimation field. The prospective study showed slightly better VGA scores (improved image quality) with the optimal collimation. Increasing the exposure factors especially the tube current and exposure time (mAs) resulted in improved low contrast resolution and less noise in the radiographs. The potential eye lens radiation dose increased by 14, 28, and 40% [default exposures, increased the tube peak potential (kVp), increased mAs, respectively] as a result of reduced collimation (increased beam size). Conclusion: The degree of collimation has no statistically significant on image quality in canine pelvic radiology for the range of collimation used but does have an impact on potential radiation dose to personnel in the x-ray room. With regard to radiation safety, increases in kVp are associated with less potential scatter radiation exposure compared to comparable increases in mAs.

Digital Analysis of Subtrochlear Sclerosis in Elbows Submitted for Dysplasia Screening.

Ulnar trochlear notch (UTN) subchondral bone sclerosis is observed in elbow dysplasia (ED) associated with the medial coronoid disease. However, its evaluation is based on a simple visual examiner assessment of bone radio-opacity level and is considered subjective. The purpose of this study was to objectively characterize the radiographic opacity of the ulnar trochlear notch (UTN) subchondral bone in mediolateral elbow projections classified, using the International Elbow Working Group guidelines. Records and mediolateral flexed elbow images from the Danish Kennel Club database for the ED screening scheme between 2012 and 2018 were available. Of the dogs in the database, those with an ED-negative status in the left limb were identified. From these, 20 dogs each having a status free from ED, or with Grade 1, 2, or 3 in the right limb, were randomly chosen. Joints with primary ununited anconeal process were excluded from the sample. A template was developed using the ImageJ software, for computer UTN sclerosis analysis. It was overlaid onto each image to define five regions of interest (ROIs): ROI-1, distal UTN; ROI-2, middle UTN; ROI-3, caudal UTN; ROI-4, cortical bone; and ROI-5, bone marrow. Mean pixel intensity for each UTN ROI was divided by the mean pixel intensity of ROI-4 to normalize the data. The mean ± standard deviation (SD) of the normalized pixel intensity in the disease joints (ED Grades 1, 2, and 3) was 1.18 ± 0.17, 1.03 ± 0.12, and 0.92 ± 0.09 for ROIs 1, 2, and 3, respectively. The corresponding values for the contralateral normal left joints were 1.16 ± 0.17, 1.01 ± 0.1, and 0.91 ± 0.08. There was a significant difference (P < 0.05) in the normalized mean pixel intensity in dysplastic vs. non-dysplastic elbow joints for ROIs 1 and 2. The raw mean pixel intensity from right and left cortical and marrow bone ROIs sometimes showed relatively large differences. Digital radiography is associated with exposure and post-processing variabilities. Differences in apparent radio-opacity (as indicated by pixel intensity) though statistically significant in dysplastic joints compared with contralateral normal joints are slight and are thus problematic for computer-aided assessments of UTN sclerosis.

Deep transfer learning can be used for the detection of hip joints in pelvis radiographs and the classification of their hip dysplasia status.

Reports of machine learning implementations in veterinary imaging are infrequent but changes in machine learning architecture and access to increased computing power will likely prompt increased interest. This diagnostic accuracy study describes a particular form of machine learning, a deep learning convolution neural network (ConvNet) for hip joint detection and classification of hip dysplasia from ventro-dorsal (VD) pelvis radiographs submitted for hip dysplasia screening. 11,759 pelvis images were available together with their Fédération Cynologique Internationale (FCI) scores. The dataset was dicotomized into images showing no signs of hip dysplasia (FCI grades "A" and "B", the "A-B" group) and hips showing signs of dysplasia (FCI grades "C", "D," and "E", the "C-E" group). In a transfer learning approach, an existing pretrained ConvNet was fine-tuned to provide models to recognize hip joints in VD pelvis images and to classify them according to their FCI score grouping. The results yielded two models. The first was successful in detecting hip joints in the VD pelvis images (intersection over union of 85%). The second yielded a sensitivity of 0.53, a specificity of 0.92, a positive predictive value of 0.91, and a negative predictive value of 0.81 for the classification of detected hip joints as being in the "C-E" group. ConvNets and transfer learning are applicable to veterinary imaging. The models obtained have potential to be a tool to aid in hip screening protocols if hip dysplasia classification performance was improved through access to more data and possibly by model optimization.

A De Novo Mutation in COL1A1 in a Holstein Calf with Osteogenesis Imperfecta Type II.

Osteogenesis imperfecta (OI) type II is a genetic connective tissue disorder characterized by bone fragility, severe skeletal deformities and shortened limbs. OI usually causes perinatal death of affected individuals. OI type II diagnosis in humans is established by the identification of heterozygous mutations in genes coding for collagens. The purpose of this study was to characterize the pathological phenotype of an OI type II-affected neonatal Holstein calf and to identify the causative genetic variant by whole-genome sequencing (WGS). The calf had acute as well as intrauterine fractures, abnormally shaped long bones and localized arthrogryposis. Genetic analysis revealed a private heterozygous missense variant in COL1A1 (c.3917T>A) located in the fibrillar collagen NC1 domain (p.Val1306Glu) that most likely occurred de novo. This confirmed the diagnosis of OI type II and represents the first report of a pathogenic variant in the fibrillar collagen NC domain of COL1A1 associated to OI type II in domestic animals. Furthermore, this study highlights the utility of WGS-based precise diagnostics for understanding congenital disorders in cattle and the need for continued surveillance for rare lethal genetic disorders in cattle.

Multimodal soft tissue markers for bridging high-resolution diagnostic imaging with therapeutic intervention.

Diagnostic imaging often outperforms the surgeon's ability to identify small structures during therapeutic procedures. Smart soft tissue markers that translate the sensitivity of diagnostic imaging into optimal therapeutic intervention are therefore highly warranted. This paper presents a unique adaptable liquid soft tissue marker system based on functionalized carbohydrates (Carbo-gel). The liquid state of these markers allows for high-precision placement under image guidance using thin needles. Based on step-by-step modifications, the image features and mechanical properties of markers can be optimized to bridge diagnostic imaging and specific therapeutic interventions. The performance of Carbo-gel is demonstrated for markers that (i) have radiographic, magnetic resonance, and ultrasound visibility; (ii) are palpable and visible; and (iii) are localizable by near-infrared fluorescence and radio guidance. The study demonstrates encouraging proof of concept for the liquid marker system as a well-tolerated multimodal imaging marker that can improve image-guided radiotherapy and surgical interventions, including robotic surgery.

Fingerprinting of Doppler audio signals from the common carotid artery.

Audio fingerprinting involves extraction of quantitative frequency descriptors that can be used for indexing, search and retrieval of audio signals in sound recognition software. We propose a similar approach with medical ultrasonographic Doppler audio signals. Power Doppler periodograms were generated from 84 ultrasonographic Doppler signals from the common carotid arteries in 22 dogs. Frequency features were extracted from each periodogram and included in a principal component analysis (PCA). From this 10 audio samples were pairwise classified as being either similar or dissimilar. These pairings were compared to a similar classification based on standard quantitative parameters used in medical ultrasound and to classification performed by a panel of listeners. The ranking of sound files according to degree of similarity differed between the frequency and conventional classification methods. The panel of listeners had an 88% agreement with the classification based on quantitative frequency features. These findings were significantly different from the score expected by chance (p < 0.001). The results indicate that the proposed frequency based classification has a perceptual relevance for human listeners and that the method is feasible. Audio fingerprinting of medical Doppler signals is potentially useful for indexing and search for similar and dissimilar audio samples in a dataset.

Pregabalin alleviates clinical signs of syringomyelia-related central neuropathic pain in Cavalier King Charles Spaniel dogs: a randomized controlled trial.

OBJECTIVE: We aimed to assess the efficacy and benefit-risk profile of pregabalin (PGN) to reduce the clinical signs of central neuropathic pain (CNeP) as reflected by scratching episodes in dogs with symptomatic syringomyelia (SM). STUDY DESIGN: Randomized, double-blind, placebo-controlled crossover study. ANIMALS: A total of 12 client-owned Cavalier King Charles Spaniels (age, 1.1-7.4 years, bodyweight, 8.2-10.8 kg) with magnetic resonance imaging-confirmed SM and clinical signs of CNeP. METHODS: Dogs were randomized to either PGN 150 mg or placebo for 25 days, followed by 48 hour washout period before crossover to the alternate phase of 25 days. The primary outcome was defined as number of scratching events during 10 minutes of video-recorded physical activity. Treatment effect was estimated using a generalized estimation equation model. Benefit-risk and quality of life assessments were obtained through owner interviews focusing on potential adverse events. RESULTS: The treatment effect estimate was an 84% (95% confidence interval = 75-89%) reduction in mean number of scratching events relative to baseline compared with placebo (p < 0.0001). Owner-assessed satisfactory quality of life was status quo and rated as 'good' or 'could not be better' in six/11 dogs and improved in four/11 dogs. The most prevalent adverse events were increased appetite in nine/12 dogs and transient ataxia in nine/12 dogs. There was one dog withdrawn by the owner 7 days after crossover to PGN owing to persistent ataxia. No dogs needed rescue analgesia during the trial. CONCLUSIONS AND CLINICAL RELEVANCE: PGN is superior to placebo in the reduction of clinical signs of SM-related CNeP in dogs. At a dose range of 13-19 mg kg-1 orally twice daily, the encountered adverse events were acceptable to all but one owner.

Stifle Joint Rotational Variation in Extended Stifles in Healthy Dogs Undergoing Computed Tomography Examinations.

OBJECTIVES: The aim of this study was to evaluate stifle joint rotational stability in response to body position and repositioning in dogs undergoing computed tomography (CT). MATERIALS AND METHODS: Nine dogs (18 stifles) with no history or clinical signs of stifle injury undergoing CT examinations for other reasons were included in the study. Dogs were positioned in alternating dorsal and sternal recumbency with the pelvic limbs extended caudally and unrestrained, for a total of four examinations. Scans included the entire tibia and distal femur. Using defined landmarks on the tibia and femur, stifle joint rotational angles were measured from multiplanar reconstructions made by two independent observers, and repositioning effects and intra- and interobserver agreement evaluated. RESULTS: Repositioning produced repeatable stifle joint rotational angles (pooled within-subject standard deviations of 0.9° and 1.4°) and intraobserver repeatability was good (within-subject standard deviations 1.4°), but interobserver agreement was poor. Dorsal and sternal positioning produced equivalent results. CLINICAL SIGNIFICANCE: The results indicate that little rotation occurs in the healthy extended canine stifle joint during positioning for CT and support CT as a method for assessing limb alignment around the stifle joint, provided that the limb distal to the femorotibial joint is unrestrained. Clinically, it is likely that measured changes in rotational angles across (and distant to) the stifle joint are real, rather than being due to rotation artefact during imaging.

Assessment of Image Quality in Digital Radiographs Submitted for Hip Dysplasia Screening.

Digital radiography is widely seen to be forgiving of poor exposure technique and to provide consistent high quality diagnostic images. Optimal quality images are however not universal; sub-optimal images are encountered. Evaluators on hip dysplasia schemes encounter images from multiple practices produced on equipment from multiple manufacturers. For images submitted to the Danish Kennel Club for hip dysplasia screening, a range of quality is seen and the evaluators are of the impression that variations in image quality area associated with particular equipment. This study was undertaken to test the hypothesis that there is an association between image quality in digital radiography and the manufacturer of the detector equipment, and to demonstrate the applicability of visual grading analysis (VGA) for image quality evaluation in veterinary practice. Data from 16,360 digital images submitted to the Danish Kennel Club were used to generate the hypothesis that there is an association between detector manufacturer and image quality and to create groups for VGA. Image quality in a subset of 90 images randomly chosen from 6 manufacturers to represent high and low quality images, was characterized using VGA and the results used to test for an association between image quality and system manufacturer. The range of possible scores in the VGA was -2 to +2 (higher scores are better). The range of the VGA scores for the images in the low image quality group (n = 45) was -1.73 to +0.67, (median -1.2). Images in the high image quality group (n = 44) ranged from -1.52 to +0.53, (median -0.53). This difference was statistically significant (p < 0.001). The study shows an association between VGA scores of image quality and detector manufacturer. Possible causes may be that imaging hardware and/or software are not equal in terms of quality, that the level of support sought and given differs between systems, or a combination of the two. Clinicians purchasing equipment should be mindful that image quality can differ across systems. VGA is practical for veterinarians to compare image quality between systems or within a system over time.

A de novo missense mutation of FGFR2 causes facial dysplasia syndrome in Holstein cattle.

BACKGROUND: Surveillance for bovine genetic diseases in Denmark identified a hitherto unreported congenital syndrome occurring among progeny of a Holstein sire used for artificial breeding. A genetic aetiology due to a dominant inheritance with incomplete penetrance or a mosaic germline mutation was suspected as all recorded cases were progeny of the same sire. Detailed investigations were performed to characterize the syndrome and to reveal its cause. RESULTS: Seven malformed calves were submitted examination. All cases shared a common morphology with the most striking lesions being severe facial dysplasia and complete prolapse of the eyes. Consequently the syndrome was named facial dysplasia syndrome (FDS). Furthermore, extensive brain malformations, including microencephaly, hydrocephalus, lobation of the cerebral hemispheres and compression of the brain were present. Subsequent data analysis of progeny of the sire revealed that around 0.5% of his offspring suffered from FDS. High density single nucleotide polymorphism (SNP) genotyping data of the seven cases and their parents were used to map the defect in the bovine genome. Significant genetic linkage was obtained for three regions, including chromosome 26 where whole genome sequencing of a case-parent trio revealed two de novo variants perfectly associated with the disease: an intronic SNP in the DMBT1 gene and a single non-synonymous variant in the FGFR2 gene. This FGFR2 missense variant (c.927G>T) affects a gene encoding a member of the fibroblast growth factor receptor family, where amino acid sequence is highly conserved between members and across species. It is predicted to change an evolutionary conserved tryptophan into a cysteine residue (p.Trp309Cys). Both variant alleles were proven to result from de novo mutation events in the germline of the sire. CONCLUSIONS: FDS is a novel genetic disorder of Holstein cattle. Mutations in the human FGFR2 gene are associated with various dominant inherited craniofacial dysostosis syndromes. Given the phenotypic similarities in FDS affected calves, the genetic mapping and absence of further high impact variants in the critical genome regions, it is highly likely that the missense mutation in the FGFR2 gene caused the FDS phenotype in a dominant mode of inheritance.

Pulmonary vasculature in dogs assessed by three-dimensional fractal analysis and chemometrics.

Fractal analysis of canine pulmonary vessels could allow quantification of their space-filling properties. Aims of this prospective, analytical, cross-sectional study were to describe methods for reconstructing three dimensional pulmonary arterial vascular trees from computed tomographic pulmonary angiogram, applying fractal analyses of these vascular trees in dogs with and without diseases that are known to predispose to thromboembolism, and testing the hypothesis that diseased dogs would have a different fractal dimension than healthy dogs. A total of 34 dogs were sampled. Based on computed tomographic pulmonary angiograms findings, dogs were divided in three groups: diseased with pulmonary thromboembolism (n = 7), diseased but without pulmonary thromboembolism (n = 21), and healthy (n = 6). An observer who was aware of group status created three-dimensional pulmonary artery vascular trees for each dog using a semiautomated segmentation technique. Vascular three-dimensional reconstructions were then evaluated using fractal analysis. Fractal dimensions were analyzed, by group, using analysis of variance and principal component analysis. Fractal dimensions were significantly different among the three groups taken together (P = 0.001), but not between the diseased dogs alone (P = 0.203). The principal component analysis showed a tendency of separation between healthy control and diseased groups, but not between groups of dogs with and without pulmonary thromboembolism. Findings indicated that computed tomographic pulmonary angiogram images can be used to reconstruct three-dimensional pulmonary arterial vascular trees in dogs and that fractal analysis of these three-dimensional vascular trees is a feasible method for quantifying the spatial relationships of pulmonary arteries. These methods could be applied in further research studies on pulmonary and vascular diseases in dogs.

Effect of Skull Type on the Relative Size of Cerebral Cortex and Lateral Ventricles in Dogs.

Volume measurements of the brain are of interest in the diagnosis of brain pathology. This is particularly so in the investigation hydrocephalus and canine cognitive dysfunction (CCD), both of which result in thinning of the cerebral cortex and enlarged ventricles. Volume assessment can be made using computed tomography or more usually magnetic resonance imaging (MRI). There is, however, some uncertainty in the interpretation of such volume data due to the great variation in skull size and shape seen in dog. In this retrospective study, we examined normal MRI images from 63 dogs <6 years of age. We used a continuous variable, the cranial index (CrI) to indicate skull shape and compared it with MRI volume measurements derived using Cavalieri's principle. We found a negative correlation between CrI and the ratio of cortical to ventricular volume. Breeds with a high CrI (large laterolateral compared to rostrocaudal cranial cavity dimension) had a smaller ratio of cortical to ventricular volume (low C:V ratio) than breeds with lower CrI skull types. It is important to consider this effect of skull shape on the relative volume estimates of the cerebral cortex and ventricles when trying to establish if pathology is present.

Online Radiology Reporting with Peer Review as a Learning and Feedback Tool in Radiology; Implementation, Validity, and Student Impressions.

Communicating radiological reports to peers has pedagogical value. Students may be uneasy with the process due to a lack of communication and peer review skills or to their failure to see value in the process. We describe a communication exercise with peer review in an undergraduate veterinary radiology course. The computer code used to manage the course and deliver images online is reported, and we provide links to the executable files. We tested to see if undergraduate peer review of radiological reports has validity and describe student impressions of the learning process. Peer review scores for student-generated radiological reports were compared to scores obtained in the summative multiple choice (MCQ) examination for the course. Student satisfaction was measured using a bespoke questionnaire. There was a weak positive correlation (Pearson correlation coefficient = 0.32, p < 0.01) between peer review scores students received and the student scores obtained in the MCQ examination. The difference in peer review scores received by students grouped according to their level of course performance (high vs. low) was statistically significant (p < 0.05). No correlation was found between peer review scores awarded by the students and the scores they obtained in the MCQ examination (Pearson correlation coefficient = 0.17, p = 0.14). In conclusion, we have created a realistic radiology imaging exercise with readily available software. The peer review scores are valid in that to a limited degree they reflect student future performance in an examination. Students valued the process of learning to communicate radiological findings but do not fully appreciated the value of peer review.

Characterization of hemorrhages in the tenderloins of slaughter pigs.

Muscle hemorrhages are regularly observed in especially the tip of the tenderloin muscles of slaughter pigs. In order to characterize the hemorrhages, a macro- and microscopic examination of tenderloins with (n=5) and without (n=4) hemorrhages and the associated vertebral column was carried out. Furthermore, all columns were radiographed and two were CT scanned. Histologically, the muscle hemorrhages contained cells from bone marrow and growth line cartilage. Ventral epiphysiolysis in either the cranial or caudal epiphysis of the first lumbar vertebrae (L1) in 8 out of the 9 vertebral columns was present. In the 5 cases with tenderloin hemorrhage, similar hemorrhage with growth line cartilage was found within the fracture of the epiphysis. The hemorrhages develop secondarily to epiphysiolysis in the lumbar vertebrae, where the tenderloin attaches to the spine. The lesions probably develop around the time of stunning.

A CHRNB1 frameshift mutation is associated with familial arthrogryposis multiplex congenita in Red dairy cattle.

BACKGROUND: Bovine arthrogryposis multiplex congenita (AMC) is a syndromic term for a congenital condition characterized by multiple joint contractures. Rare inherited forms of bovine AMC have been reported in different breeds. For AMC in Angus cattle a causative genomic deletion encompassing the agrin (AGRN) gene, encoding an essential neural regulator that induces the aggregation of acetylcholine receptors (AChRs), is known. In 2015, three genetically related cases of generalized AMC affecting Red dairy calves were diagnosed in Denmark. RESULTS: The family history of three affected calves suggested an autosomal recessive inheritance. Single nucleotide polymorphism (SNP) genotyping showed a single genomic region of extended homozygosity of 21.5 Mb on chromosome 19. Linkage analysis revealed a maximal parametric LOD score of 1.8 at this region. By whole genome re-sequencing of the three cases, two private homozygous non-synonymous variants were detected in the critical interval. Both variants, located in the myosin phosphatase Rho interacting protein (MPRIP) and the cholinergic receptor nicotinic beta 1 subunit gene (CHRNB1), were perfectly associated with the AMC phenotype. Previously described CHRNB1 variants in humans lead to a congenital myasthenic syndrome with impaired neuromuscular transmission. The cattle variant represents a single base deletion in the first exon of CHRNB1 (c.55delG) introducing a premature stop codon (p.Ala19Profs47*) in the second exon, truncating 96 % of the protein. CONCLUSIONS: This study provides the first phenotypically and genetically characterized example of a bovine AMC phenotype that represents an inherited neuromuscular disorder corresponding to human congenital myasthenic syndrome. The identified CHRNB1 loss of function variant is predicted to have a deleterious effect on fetal AChR function, which could explain the lethal phenotype reported in this study. The identification of this candidate causative mutation thus widens the known phenotypic spectrum of CHRNB1 mutations and enables selection against this pathogenic variant in Red dairy cattle.

Lethal chondrodysplasia in a family of Holstein cattle is associated with a de novo splice site variant of COL2A1.

BACKGROUND: Lethal chondrodysplasia (bulldog syndrome) is a well-known congenital syndrome in cattle and occurs sporadically in many breeds. In 2015, it was noticed that about 12% of the offspring of the phenotypically normal Danish Holstein sire VH Cadiz Captivo showed chondrodysplasia resembling previously reported bulldog calves. Pedigree analysis of affected calves did not display obvious inbreeding to a common ancestor, suggesting the causative allele was not a rare recessive. The normal phenotype of the sire suggested a dominant inheritance with incomplete penetrance or a mosaic mutation. RESULTS: Three malformed calves were examined by necropsy, histopathology, radiology, and computed tomography scanning. These calves were morphologically similar and displayed severe disproportionate dwarfism and reduced body weight. The syndrome was characterized by shortening and compression of the body due to reduced length of the spine and the long bones of the limbs. The vicerocranium had severe dysplasia and palatoschisis. The bones had small irregular diaphyses and enlarged epiphyses consisting only of chondroid tissue. The sire and a total of four affected half-sib offspring and their dams were genotyped with the BovineHD SNP array to map the defect in the genome. Significant genetic linkage was obtained for several regions of the bovine genome including chromosome 5 where whole genome sequencing of an affected calf revealed a COL2A1 point mutation (g.32473300 G > A). This private sequence variant was predicted to affect splicing as it altered the conserved splice donor sequence GT at the 5'-end of COL2A1 intron 36, which was changed to AT. All five available cases carried the mutant allele in heterozygous state and all five dams were homozygous wild type. The sire VH Cadiz Captivo was shown to be a gonadal and somatic mosaic as assessed by the presence of the mutant allele at levels of about 5% in peripheral blood and 15% in semen. CONCLUSIONS: The phenotypic and genetic findings are comparable to a previously reported COL2A1 missense mutation underlying lethal chondrodysplasia in the offspring of a mosaic French Holstein sire (Igale Masc). The identified independent spontaneous splice site variant in COL2A1 most likely caused chondrodysplasia and must have occurred during the early foetal development of the sire. This study provides a first example of a dominant COL2A1 splice site variant as candidate causal mutation of a severe lethal chondrodysplasia phenotype. Germline mosaicism is a relatively frequent mechanism in the origin of genetic disorders and explains the prevalence of a certain fraction of affected offspring. Paternal dominant de novo mutations are a risk in cattle breeding, especially because the ratio of defective offspring may be very high and be associated with significant animal welfare problems.

Ultrasonographic predictors of response of European eels (Anguilla anguilla) to hormonal treatment for induction of ovarian development.

OBJECTIVE To examine ultrasonographic predictors of ovarian development in European eels (Anguilla anguilla) undergoing hormonal treatment for assisted reproduction. ANIMALS 83 female European eels. PROCEDURES Eels received weekly IM injections of salmon pituitary extract (first injection = week 1). Ultrasonography of the ovaries was performed twice during hormonal treatment (weeks 7 and 11). Eels were identified on the basis of body weight as having an adequate response by weeks 14 to 20 or an inadequate response after injections for 21 weeks. Eels were euthanized at the end of the experiment and classified by use of ovarian histologic examination. Ovarian cross-sectional area and size of eel (ie, length (3) ) were used to classify eels (fast responder, slow responder, or nonresponder) and to calculate an ultrasonographic-derived gonadosomatic index. Gray-level co-occurrence matrices were calculated from ovarian images, and 22 texture features were calculated from these matrices. RESULTS The ultrasonographic-derived gonadosomatic index differed significantly between fast responders and slow responders or nonresponders at both weeks 7 and 11. Principal component analysis revealed a pattern of separation between the groups, and partial least squares discriminant analysis revealed signals in the ovarian texture that discriminated females that responded to treatment from those that did not. CONCLUSIONS AND CLINICAL RELEVANCE Ovarian texture information in addition to morphometric variables can enhance ultrasonographic applications for assisted reproduction of eels and potentially other fish species. This was a novel, nonlethal method for classifying reproductive response of eels and the first objective texture analysis performed on ultrasonographic images of the gonads of fish.