Microbiome analysis of bile from apparently healthy cats and cats with suspected hepatobiliary disease.

BACKGROUND: Bacterial infection of bile is a common cause of hepatobiliary disease in cats. Whether bile harbors a core microbiota in health or in cases of suspected hepatobiliary disease in cats is unknown. OBJECTIVES: Establish if gallbladder bile in apparently healthy cats harbors a core microbiota composed of bacterial taxa common to many individuals. Compare results of bile cytology, bile culture, and 16S rRNA gene amplicon sequencing in apparently healthy cats and cats with suspected hepatobiliary disease. ANIMALS: Forty-three client-owned cats with suspected hepatobiliary disease and 17 control cats. METHODS: Bile was collected by ultrasound guided cholecystocentesis (cats with suspected hepatobiliary disease) or laparotomy after euthanasia (controls). Bile samples underwent cytologic examination, aerobic and anaerobic culture, and DNA was extracted for 16S rRNA gene amplification and sequencing. RESULTS: Microbiome sequencing did not identify a core microbiota in control cats or cats having bile sampled because of clinical suspicion for hepatobiliary disease. Microbiome profiles from control cats were indistinguishable from profiles obtained from sampling instruments and reagents that were not exposed to bile (technical controls). Bacterial taxa that could not be explained by contamination or off-target amplification were identified only in samples from cats with bactibilia and positive bile culture results for Escherichia coli. In several E. coli positive samples, microbiome sequencing also identified a small number of potentially co-infecting bacterial genera not identified by culture. CONCLUSIONS AND CLINICAL IMPORTANCE: Cat bile does not harbor a core microbiota. Uncultured bacteria may contribute to pathogenesis of hepatobiliary disease in cats with bile E. coli infection.

Gallbladder microbiota in healthy dogs and dogs with mucocele formation.

To date studies have not investigated the culture-independent microbiome of bile from dogs, a species where aseptic collection of bile under ultrasound guidance is somewhat routine. Despite frequent collection of bile for culture-based diagnosis of bacterial cholecystitis, it is unknown whether bile from healthy dogs harbors uncultivable bacteria or a core microbiota. The answer to this question is critical to understanding the pathogenesis of biliary infection and as a baseline to exploration of other biliary diseases in dogs where uncultivable bacteria could play a pathogenic role. A pressing example of such a disease would be gallbladder mucocele formation in dogs. This prevalent and deadly condition is characterized by excessive secretion of abnormal mucus by the gallbladder epithelium that can eventually lead to rupture of the gallbladder or obstruction of bile flow. The cause of mucocele formation is unknown as is whether uncultivable, and therefore unrecognized, bacteria play any systematic role in pathogenesis. In this study we applied next-generation 16S rRNA gene sequencing to identify the culture-negative bacterial community of gallbladder bile from healthy dogs and gallbladder mucus from dogs with mucocele formation. Integral to our study was the use of 2 separate DNA isolations on each sample using different extraction methods and sequencing of negative control samples enabling recognition and curation of contaminating sequences. Microbiota findings were validated by simultaneous culture-based identification, cytological examination of bile, and fluorescence in-situ hybridization (FISH) performed on gallbladder mucosa. Using culture-dependent, cytological, FISH, and 16S rRNA sequencing approaches, results of our study do not support existence of a core microbiome in the bile of healthy dogs or gallbladder mucus from dogs with mucocele formation. Our findings further document how contaminating sequences can significantly contribute to the results of sequencing analysis when performed on samples with low bacterial biomass.

MRI features can help to confirm a diagnosis of progressive myelomalacia, but may not be accurate in dogs lacking characteristic clinical signs at the time of imaging.

Progressive myelomalacia (PMM) is a fatal sequela of acute thoracolumbar intervertebral disc extrusion in dogs, with unpredictable onset in the days after the inciting injury. No single reliable diagnostic test is currently available. Magnetic resonance imaging (MRI) features such as T2-weighted spinal cord hyperintensity and loss of subarachnoid signal in a half-Fourier single-shot turbo spin echo (HASTE) sequence have been associated with PMM, but are sometimes present in other dogs with severe deficits. Magnetic resonance imaging findings in 22 dogs with a clinical or histopathologic diagnosis of PMM and 38 deep pain-negative paraplegic dogs were compared in a retrospective case-control study. Length of T2-weighted hyperintense spinal cord change and HASTE signal loss were significantly associated with clinically evident PMM (P = .0019 and P = .0085), however, there were no significant differences between groups when analysis was restricted to dogs not yet showing clinical signs of PMM. The PMM group also had significantly shorter compressive lesions than the control group (P = 0.026), suggesting a possible role of more severe focal pressure at the extrusion site. A segment of total loss of contrast enhancement in the venous sinuses and meninges, a feature not previously described, was more common in the PMM group and the difference approached significance (P = 0.054). Findings show that MRI features can support the diagnosis in dogs with clinical evidence of PMM, and absence of these features supports absence of PMM at time of imaging. However, their absence does not reliably differentiate dogs with imminent progressive myelomalacia from other dogs with severe deficits following intervertebral disc extrusion.

Shear wave elastography evaluation of cats with chronic kidney disease.

Chronic kidney disease (CKD) is a major health condition in cats that can lead to poor quality of life and financial implications for therapy. Currently staging and identification of CKD is limited by diagnostic testing such as creatinine and urine-specific gravity, which do not change until late in the disease course. Other methods to evaluate CKD would be valuable in the clinical setting. Shear wave elastography is one novel ultrasound method, which has shown promise in identifying increases in tissue stiffness and identifying CKD in people. As CKD is often histologically characterized by tubulointerstitial fibrosis, shear wave elastography has the potential to identify CKD and differentiate between stages of CKD in cats. This prospective observational case-control study with 78 cats found no difference in shear wave velocities between groups (P = 0.33), a contradictory finding to one prior publication. There was no effect of weight (P = 0.65), nor the presence of mineralization (P = 0.31) or infarction (P = 0.52) on cortical shear wave velocities. There was a significant effect of age on shear wave velocity (P = 0.018) where velocities increased with age. The intraclass correlation coefficient was only moderate (0.62). Possible reasons for the difference in results between our work and that published prior, include differences in methodology and differences in instrumentation. Variability in measurements in our population may be due to the effects of respiratory motion or limitations in shear wave elastography software. As such, shear wave elastography is not currently recommended as a tool to evaluate CKD in cats and further work is necessary.

ACVR and ECVDI consensus statement for the standardization of the abdominal ultrasound examination.

This consensus statement is designed to provide a standard of care document and describes the ACVR and ECVDI definition for performing a standard abdominal ultrasound examination in dogs and cats. The ACVR and ECVDI define a standard abdominal ultrasonographic examination as a complete exam of the abdominal organs which is appropriately documented. The consensus statement intends to provide guidance to veterinary sonographers and veterinarians for the performance and documentation of high-quality diagnostic ultrasound examination. The document may also serve as a teaching aid for veterinary students, veterinarians, and residents in diagnostic imaging who seek proficiency in diagnostic ultrasound. Finally, it may serve an additional role in educating the public as to what a high-quality abdominal ultrasound examination should entail.

A Platelet-Rich Plasma-Derived Biologic Clears Staphylococcus aureus Biofilms While Mitigating Cartilage Degeneration and Joint Inflammation in a Clinically Relevant Large Animal Infectious Arthritis Model.

The leading cause of treatment failure in Staphylococcus aureus infections is the development of biofilms. Biofilms are highly tolerant to conventional antibiotics which were developed against planktonic cells. Consequently, there is a lack of antibiofilm agents in the antibiotic development pipeline. To address this problem, we developed a platelet-rich plasma (PRP)-derived biologic, termed BIO-PLY (for the BIOactive fraction of Platelet-rich plasma LYsate) which has potent in vitro bactericidal activity against S. aureus synovial fluid free-floating biofilm aggregates. Additional in vitro studies using equine synoviocytes and chondrocytes showed that BIO-PLY protected these cells of the joint from inflammation. The goal of this study was to test BIO-PLY for in vivo efficacy using an equine model of infectious arthritis. We found that horses experimentally infected with S. aureus and subsequently treated with BIO-PLY combined with the antibiotic amikacin (AMK) had decreased bacterial concentrations within both synovial fluid and synovial tissue and exhibited lower systemic and local inflammatory scores compared to horses treated with AMK alone. Most importantly, AMK+BIO-PLY treatment reduced the loss of infection-associated cartilage proteoglycan content in articular cartilage and decreased synovial tissue fibrosis and inflammation. Our results demonstrate the in vivo efficacy of AMK+BIO-PLY and represents a new approach to restore and potentiate antimicrobial activity against synovial fluid biofilms.

Optimization of ultrasonographic ureteral jet detection and normal ureteral jet morphology in dogs.

Ureteral jets are visualized with ultrasound as echogenic streams extending from the ureterovesicular junction into the urinary bladder. In clinical practice, diuretics are sometimes administered to increase visibility of ureteral jets, however this has not been well described in the veterinary literature. The purpose of this prospective, crossover study was to describe the normal morphology of canine ureteral jets, determine an optimal protocol for diuretic administration to increase visibility of ureteral jets, and confirm in vitro the effect that differences in specific gravity and velocity have on visibility. Ultrasound of 10 normal dogs was performed at baseline and following 1 mg/kg furosemide administered intravenously or subcutaneously. Increased numbers of ureteral jets were seen post-furosemide administration compared to baseline, with an overall increased number of ureteral jets identified following intravenous administration when compared to subcutaneous administration. Time to first ureteral jet was significantly shorter with intravenous compared to subcutaneous administration. Urine specific gravity significantly decreased following furosemide administration. For the in vitro study, saline solutions of varying specific gravities were infused into a bath of hypertonic saline with specific gravity of 1.037. There was good visibility in vitro with infusion of solutions of specific gravities of 1.010-1.025. Infusion of saline solution with a specific gravity of 1.030 had reduced visibility, while infusion of saline with equivalent specific gravity (1.037) was not visible with B-mode ultrasonography. Both intravenous and subcutaneous furosemide administration significantly increase ureteral jet detection with ultrasound secondary to differences in specific gravity, confirming results of prior studies.

Investigation of the use of microwave ablation with and without cooling urethral perfusion for thermal ablation of the prostate gland in canine cadavers.

OBJECTIVE: To investigate the use of microwave ablation (MWA) with cooling urethral perfusion and with no perfusion (MWA-UP and MWA-NP, respectively) for prostate gland ablation in canine cadavers. ANIMALS: Cadavers of 18 sexually intact male dogs. PROCEDURES: After technique refinement in 2 cadavers, laparotomy with ultrasound-guided MWA-UP (n = 8) or MWA-NP (8) of the prostate gland was performed in 16 cadavers. Normograde cystourethroscopy was performed before and after treatment; recorded images were reviewed in a blinded manner for scoring of urethral mucosal discoloration and loss of integrity. Difficulty with cystoscope insertion was recorded if present. Excised prostate glands were fixed for serial sectioning, gross measurements, and calculation of percentage ablation. Percentages of prostate tissue necrosis from MWA, denuded urethral mucosa, and depth of epithelial surface loss in an adjacent section of the colon were estimated histologically. Variables of interest were statistically analyzed. RESULTS: Difficulty with cystoscope insertion after treatment was significantly more common and scores for urethral mucosal discoloration and loss of integrity were significantly higher (indicating more severe lesions) for the MWA-NP group than for the MWA-UP group. The histologically assessed percentage of denuded urethral mucosa was also greater for the MWA-NP group. Overall median percentage prostate gland ablation was 73%; this result was not associated with prostate gland volume or chronological order of treatment. CONCLUSIONS AND CLINICAL RELEVANCE: MWA-UP induced subtotal thermal necrosis of prostate glands in canine cadavers while limiting urethral mucosal injury. Further study is required to optimize the technique and evaluate its safety and efficacy in vivo as a future curative-intent treatment for prostatic tumors in dogs.

Proteinuria in dogs with gallbladder mucocele formation: A retrospective case control study.

BACKGROUND: Proteinuria is an independent risk factor for morbidity and mortality in dogs. An association between proteinuria and gallbladder mucocele formation in dogs is unknown. OBJECTIVE: Determine if gallbladder mucocele formation or clinicopathologic comorbidities are associated with proteinuria. ANIMALS: Twenty-five dogs with mucocele formation and 25 breed and age-matched control dogs from a prior study. METHODS: Retrospective case control study. Proteinuria defined by calculated urine dipstick protein concentration (mg/mL) to urine specific gravity (USG) ratio. Clinicopathologic findings, postcosyntropin cortisol concentration, thyroid function profile, and illness severity score were recorded. RESULTS: Median urine dipstick protein concentration to USG ratio and number of dogs having a ratio ≥1.5 were significantly higher for dogs with mucocele formation compared to control dogs. Proteinuria was not significantly associated with CBC or serum biochemistry profile abnormalities but increased in relation to severity of illness. CONCLUSIONS AND CLINICAL IMPORTANCE: Gallbladder mucocele formation is significantly associated with proteinuria in dogs. Diagnosis and treatment of proteinuria in dogs with mucocele formation might minimize long term kidney morbidity in these patients.

Treatment of feline gastrointestinal intermediate- or large-cell lymphoma with lomustine chemotherapy and 8 Gy abdominal cavity radiation therapy.

OBJECTIVES: The goal of this study was to document the outcomes and toxicity of a novel multimodality treatment protocol for feline gastrointestinal intermediate- or large-cell lymphoma (FGL) in which cats were treated at 21-day intervals. METHODS: This was a prospective, single-arm study. Twelve client-owned cats with cytologically diagnosed FGL were treated with a combination of abdominal cavity radiation therapy (RT; 8 Gy total dose administered in two 4 Gy fractions, 21 days apart), lomustine chemotherapy (approximately 40 mg/m2, administered orally at 21-day intervals for four treatments), prednisolone (5 mg PO q24h) and cobalamin (250 µg/week SC). RESULTS: Three cats were euthanized prior to the second treatment and it was difficult to discern treatment-associated toxicity from progressive disease. Four of the remaining cats developed cytopenias, resulting in 7-14-day lomustine treatment delays and/or dose reductions. Six cats had a partial response to treatment and three had stable disease based on ultrasound at day 21 (50% overall response rate). Three of these six cats completed the study and lived >240 days; one died of refractory diabetes mellitus with no clinical evidence of FGL, and the other two died as a result of FGL. The median overall survival time was 101 days (95% confidence interval [CI] 9-240). The median progression-free survival time was 77 days (95% CI 8-212). Necropsies were performed in eight cats, which revealed multifocal lymphoma throughout the gastrointestinal tract and other organs. CONCLUSIONS AND RELEVANCE: Oncological outcomes reported herein are comparable to those achieved with multiagent injectable chemotherapy (eg, CHOP). Treatment was seemingly well tolerated in most cats and was relatively cost-effective. It is therefore plausible that improved disease control may be achievable through continued optimization and intensification of the combinatorial chemoradiotherapy protocol.

Ability of positive and negative contrast computed tomographic peritoneography to delineate canine liver lobe fissures.

OBJECTIVE: To determine the efficacy of contrast peritoneography in delineating liver lobe fissures by using computed tomography (CT). STUDY DESIGN: Prospective cross-sectional study. ANIMALS: Canine cadavers (n = 10). METHODS: An abdominal CT image of each cadaver was obtained under three conditions: without peritoneal contrast (precontrast), after insufflation of the peritoneal cavity with CO2 (negative contrast), and after intraperitoneal administration of iohexol (positive contrast). Images were reconstructed in three planes. Three reviewers independently scored each plane/contrast combination. Observers measured the length of each fissure in each plane. The measurable length of fissures was compared between techniques but was not compared with the actual length on cadavers. RESULTS: When assessment of all fissures was combined, measurements were obtained in 96 to 108 of 120 (80%-90%) positive contrast studies, 56 to 96 of 120 (47%-80%) negative contrast studies, and 12 to 32 of 120 (10%-27%) precontrast CT images. All observers measured fissures in all planes that were approximately two times longer on positive compared with negative contrast studies and two times longer on negative contrast studies compared with CT images without contrast. Measurable fissure lengths were greater on positive contrast CT images (P < .0001) than on other CT images. CONCLUSION: Positive contrast CT peritoneography improved the delineation of hepatic interlobar fissures in normal canine cadavers. CLINICAL SIGNIFICANCE: These results justify further studies to determine the ability to locate large hepatic masses in dogs with positive CT. Positive contrast CT peritoneography may assist treatment planning in dogs with large hepatic tumors.

Investigation of adrenal and thyroid gland dysfunction in dogs with ultrasonographic diagnosis of gallbladder mucocele formation.

Gallbladder mucocele formation is an emerging disease in dogs characterized by increased secretion of condensed granules of gel-forming mucin by the gallbladder epithelium and formation of an abnormally thick mucus that can culminate in obstruction of the bile duct or rupture of the gallbladder. The disease is associated with a high morbidity and mortality and its pathogenesis is unknown. Affected dogs have a significantly increased likelihood of concurrent diagnosis of hyperadrenocorticism, hypothyroidism, and hyperlipidemia. Whether these endocrinopathies represent coincidental primary disease processes that exacerbate gallbladder mucocele formation in predisposed dogs or reflect a concurrent disruption of endocrine and lipid metabolism is unclear. In this study, we investigated a hypothesis that dogs with gallbladder mucocele formation would have a high prevalence of occult and atypical abnormalities in adrenal cortical and thyroid gland function that would suggest the presence of endocrine disruption and provide deeper insight into disease pathogenesis. We performed a case-control study of dogs with and without ultrasonographic diagnosis of gallbladder mucocele formation and profiled adrenal cortical function using a quantitative mass spectrometry-based assay of serum adrenal-origin steroids before and after administration of synthetic cosyntropin. We simultaneously profiled serum thyroid hormone concentrations and evaluated iodine sufficiency by measurement of urine iodine:creatinine ratios (UICR). The studies were complemented by histological examination of archival thyroid tissue and measurements of thyroid gland organic iodine from dogs with gallbladder mucocele formation and control dogs. Dogs with gallbladder mucocele formation demonstrated an exaggerated cortisol response to adrenal stimulation with cosyntropin. A prevalence of 10% of dogs with gallbladder mucocele formation met laboratory-based criteria for suspect or definitive diagnosis of hyperadrenocorticism. A significantly greater number of dogs with gallbladder mucocele formation had basal serum dehydroepiandrosterone (DHEAS) increases compared to control dogs. A high percentage of dogs with gallbladder mucocele formation (26%) met laboratory-based criteria for diagnosis of hypothyroidism, but lacked detection of anti-thyroglobulin antibodies. Dogs with gallbladder mucocele formation had significantly higher UICRs than control dogs. Examination of thyroid tissue from an unrelated group of dogs with gallbladder mucocele formation did not demonstrate histological evidence of thyroiditis or significant differences in content of organic iodine. These findings suggest that dogs with gallbladder mucocele formation have a greater capacity for cortisol synthesis and pinpoint DHEAS elevations as a potential clue to the underlying pathogenesis of the disease. A high prevalence of thyroid dysfunction with absent evidence for autoimmune thyroiditis suggest a disrupted thyroid hormone metabolism in dogs with gallbladder mucocele formation although an influence of non-thyroidal illness cannot be excluded. High UICR in dogs with gallbladder mucocele formation is of undetermined significance, but of interest for further study.

Qualitative metabolomics profiling of serum and bile from dogs with gallbladder mucocele formation.

Mucocele formation is characterized by secretion of abnormally thick mucus by the gallbladder epithelium of dogs that may cause obstruction of the bile duct or rupture of the gallbladder. The disease is increasingly recognized and is associated with a high morbidity and mortality. The cause of gallbladder mucocele formation in dogs is unknown. There is a strong breed predisposition and affected dogs have a high incidence of concurrent endocrinopathy or hyperlipidemia. These observations suggest a significant influence of both genetic and metabolic factors on disease pathogenesis. In this study, we investigated a theory that mucocele formation is associated with a syndrome of metabolic disruption. We surmised that a global, untargeted metabolomics approach could provide unique insight into the systemic pathogenesis of gallbladder mucocele formation and identify specific compounds as candidate biomarkers or treatment targets. Moreover, concurrent examination of the serum and hepatic duct bile metabolome would enable the construction of mechanism-based theories or identification of specific compounds responsible for altered function of the gallbladder epithelium. Abnormalities observed in dogs with gallbladder mucocele formation, including a 33-fold decrease in serum adenosine 5'-monophosphate (AMP), lower quantities of precursors required for synthesis of energy transporting nucleotides, and increases in citric acid cycle intermediates, suggest excess metabolic energy and a carbon surplus. Altered quantities of compounds involved in protein translation and RNA turnover, together with accumulation of gamma-glutamylated and N-acetylated amino acids in serum suggest abnormal regulation of protein and amino acid metabolism. Increases in lathosterol and 7α-hydroxycholesterol suggest a primary increase in cholesterol synthesis and diversion to bile acid formation. A number of specific biomarker compounds were identified for their ability to distinguish between control dogs and those that formed a gallbladder mucocele. Particularly noteworthy was a significant decrease in quantity of biologically active compounds that stimulate biliary ductal fluid secretion including adenosine, cAMP, taurolithocholic acid, and taurocholic acid. These findings support the presence of significant metabolic disruption in dogs with mucocele formation. A targeted, quantitative analysis of the identified serum biomarkers is warranted to determine their utility for diagnosis of this disease. Finally, repletion of compounds whose biological activity normally promotes biliary ductal secretion should be examined for any therapeutic impact for resolution or prevention of mucocele formation.

Comparisons between elastographic stiffness scores for benign versus malignant lymph nodes in dogs and cats.

Lymph node evaluation is an important component of oncology staging in dogs and cats, however diagnosis of malignancy currently requires cytology or histopathology. Elastography is an ultrasound technique that allows estimation of tissue stiffness. In people, it has been shown to increase the sensitivity of sonography in detection of metastatic lymph nodes. The objective of this prospective, cross-sectional study was to determine if elastographic stiffness differs for malignant versus benign canine and feline lymph nodes that were considered abnormal in gray-scale ultrasound imaging. Animals scheduled for ultrasound-guided fine needle aspirates of a lymph node at a single center were prospectively enrolled. Elastography was performed by a board-certified veterinary radiologist (G.S.) prior to tissue sampling. Softness was scored qualitatively as 1-4 (1 = hard; 4 = soft) on an image depicted by a color scale (blue = hard, red = soft). Quantitative analysis was performed using custom-made software. Fifty-one lymph nodes were included in the evaluation, 21 had benign and 30 had neoplastic cytology. Benign lymph nodes were softer (median score of 2.5, range 1-4) than malignant lymph nodes (median = 2, range 1-3), and differences were significant in qualitative and quantitative analyses (P < 0.01). Of the lymph nodes with a score of 2 or below (hard), 19/25 (76%) were malignant, of the lymph nodes with a score over 2 (soft), 24/36 (67%) were benign. Findings indicated that elastographic stiffness scores differed between benign and malignant lymph node groups; however overlapping scores for the two groups limited the use of this technique for routine clinical diagnosis of malignancy.

The computed tomographic "tree-in-bud" pattern: Characterization and comparison with radiographic and clinical findings in 36 cats.

In humans, a CT "tree-in-bud" pattern has been described as a characteristic of centrilobular bronchiolar dilation, with bronchiolar plugging by mucus, pus, or fluid. Aims of this retrospective, descriptive, multi-center study were to characterize the CT appearance of a "tree-in-bud" pattern in a group of cats, and compare this pattern with radiographic and clinical findings. Databases from four hospitals were searched during the period of January 2012 to September 2015 and cats with thoracic radiographs, thoracic CT scans and CT reports describing findings consistent with a "tree-in-bud" pattern were included. Images were reviewed by two veterinary radiologists and characteristics were recorded based on consensus. Clinical findings were recorded by one observer from each center. Thirty-six cats met inclusion criteria. Six cats were asymptomatic, 12 were diagnosed with bronchial disease and 23 were suspected to have bronchial disease. Right cranial and right caudal lung lobes were most commonly affected on both imaging modalities. Localization of the "tree-in-bud" pattern was most often peripheral. On radiographs, the CT "tree-in-bud" pattern often appeared as soft-tissue opacity nodules; their number and affected pulmonary segments were often underestimated compared with CT. In conclusion, the "tree-in-bud" pattern should be considered as a differential diagnosis for radiographic soft tissue opaque nodules in feline lungs. Based on lesion localization and presence or suspicion of a concomitant bronchial disease for cats in this sample, authors propose that the CT "tree-in-bud" pattern described in humans is also a characteristic of bronchial or bronchiolar plugging and bronchial disease in cats.

Validation of ultrasonographic muscle thickness measurements as compared to the gold standard of computed tomography in dogs.

OBJECTIVE: The objective was to quantitatively evaluate the validity of ultrasonographic (US) muscle measurements as compared to the gold standard of computed tomography (CT) in the canine. DESIGN: This was a prospective study. POPULATION: Twenty-five, client-owned dogs scheduled for CT as part of a diagnostic work-up for the management of their primary disease process were included. MATERIALS AND METHODS: Specific appendicular (cubital flexors and extensors, coxofemoral flexors and extensors) and axial (temporalis, supraspinatus, infraspinatus, lumbar epaxials) muscle groups were selected for quantitative measure based on CT planning and patient position. Prior to CT scan, the skin over the muscle sites was shaved and marked with a permanent marker. Patient body position was determined based on the patient's CT plan; positioning was consistent between CT and US imaging. To ensure identical imaging position for both CT and US measurements, radio-opaque fiducial markers were placed directly over the skin marks once the dog was positioned. Quantitative measurements (cm) for both lean muscle mass (LMM) and subcutaneous adipose (SQA) were recorded. Statistical comparisons between CT and US values were done separately for each site and type. RESULTS: Muscle groups and associated SQA measured by US and CT were not statistically different based on an adjusted p-value using Bonferroni's correction (p < 0.0031). In addition, all LMM and SQA sites had good reliability and agreement (Cronbach's α = 0.8 - 1.0) between the two metrics, excluding the coxofemoral extensor muscle group (Cronbach's α = 0.73232). Linear regression analysis of muscle measures indicated close agreement (slope range 0.93-1.09) and minimal bias of variation (intercept range 0.05-0.11) between CT versus US modalities, with the exception of the coxofemoral extensor muscle. Similarly, SQA CT and US measures indicated close agreement with the slope range of 0.88-1.02 and minimal bias of variation with an intercept range of 0.021-0.098, excluding the cubital flexor and extensor groups. Additionally, the R2 values for these remaining LMM and SQA sites are reported as >0.897 for LLM and >0.8289 for SQA. CONCLUSIONS: Ultrasound imaging of selected appendicular and axial muscle groups in dogs can provide comparable assessment of muscle thickness to the current gold standard, CT. In consideration of both statistical reliability to CT and cage-side accessibility, the temporalis, supraspinatus, infraspinatus, and lumbar epaxial LMM sites are considered the most useful targets for US LMM assessment in the canine. Our findings support the potential utility of US as a clinical tool in veterinary medicine to assess LMM status in patients. Additional studies are indicated to develop standardized protocols of its use in a cage-side setting and to elucidate the benefit of this modality, in conjunction with nutritional interventions, to manage body LLM stores in compromised patients.

Eosinophilic meningomyelitis associated with T-cell lymphoma in a cat.

A 12-year-old cat was presented for evaluation of progressive tetraparesis. Magnetic resonance imaging of the cervical spine demonstrated T2-hyperintensity, and contrast enhancement within the C4-C7 spinal cord, with marked meningeal contrast enhancement and segmental nerve root thickening. Lumbar cerebrospinal fluid contained 407 total nucleated cells/µL, with 99% eosinophils. The cat transiently improved with prednisolone, clindamycin, and ivermectin therapy, but subsequently worsened and was euthanized. Necropsy revealed an asymmetric infiltration predominantly of the white matter, meninges, and nerve roots of the C4-C6 spinal cord segments by an unencapsulated, poorly demarcated neoplasm composed of atypical lymphocytes admixed with eosinophils, causing perivascular hemorrhage and lytic necrosis. The neoplastic cells were immunoreactive for CD3, ultimately confirming T-cell lymphoma.

FEASIBILITY AND SAFETY OF CONTRAST-ENHANCED ULTRASOUND IN THE DISTAL LIMB OF SIX HORSES.

Vascular alterations play important roles in many orthopedic diseases such as osteoarthritis, tendonitis, and synovitis in both human and equine athletes. Understanding these alterations could enhance diagnosis, prognosis, and treatment. Contrast-enhanced ultrasound (CEUS) could be a valuable method for evaluation of blood flow and perfusion of these processes in the equine distal limb, however no reports were found describing feasibility or safety of the technique. The goal of this prospective, experimental study was to describe the feasibility and safety of distal limb CEUS in a sample of six horses. For each horse, CEUS of the distal limb was performed after intravenous injections of 5 and 10 ml, as well as intra-arterial injections of 0.5 and 1 ml contrast medium. Vital parameters were monitored and CEUS images were assessed qualitatively and quantitatively for degree of contrast enhancement. None of the horses had clinically significant changes in their vital parameters after contrast medium injection. One horse had a transient increase in respiratory rate, and several horses had mild increases of systolic blood pressure of short duration after intravenous, but not after intra-arterial injections. Intra-arterial injection was possible in all horses and resulted in significantly improved contrast enhancement both quantitatively (P = 0.027) and qualitatively (P = 0.019). Findings from this study indicated that CEUS is a feasible and safe diagnostic test for evaluation of the equine distal limb. Future studies are needed to assess the clinical utility of this test for horses with musculoskeletal diseases.

Excess Secretion of Gel-Forming Mucins and Associated Innate Defense Proteins with Defective Mucin Un-Packaging Underpin Gallbladder Mucocele Formation in Dogs.

Mucosal protection of the gallbladder is vital yet we know very little about the mechanisms involved. In domestic dogs, an emergent syndrome referred to as gallbladder mucocele formation is characterized by excessive secretion of abnormal mucus that results in obstruction and rupture of the gallbladder. The cause of gallbladder mucocele formation is unknown. In these first mechanistic studies of this disease, we investigated normal and mucocele-forming dog gallbladders to determine the source, identity, biophysical properties, and protein associates of the culprit mucins with aim to identify causes for abnormal mucus behavior. We established that mucocele formation involves an adoptive excess secretion of gel forming mucins with abnormal properties by the gallbladder epithelium. The mucus is characterized by a disproportionally significant increase in Muc5ac relative to Muc5b, defective mucin un-packaging, and mucin-interacting innate defense proteins that are capable of dramatically altering the physical and functional properties of mucus. These findings provide an explanation for abnormal mucus behavior and based on similarity to mucus observed in the airways of people with cystic fibrosis, suggest that abnormal mechanisms for maintenance of gallbladder epithelial hydration may be an instigating factor for mucocele formation in dogs.

ELASTOGRAPHIC EVALUATION OF NATURALLY OCCURING TENDON AND LIGAMENT INJURIES OF THE EQUINE DISTAL LIMB.

Compression elastography is an ultrasonographic technique that estimates tissue strain and may have utility in diagnosing and monitoring soft tissue injuries in the equine athlete. Recently, elastography has been proven to be a feasible and repeatable imaging modality for evaluating normal tendons and ligaments of the equine distal forelimb. The purposes of this prospective study were to investigate the ability of elastography to detect spontaneously occurring lesions of equine tendons and ligaments diagnosed with magnetic resonance imaging (MRI) and gray-scale ultrasound (US) and to characterize the differences in the elastographic appearance of acute vs. chronic injuries. Fifty seven horses with a total of 65 lesions were evaluated. Images were assessed quantitatively and qualitatively. Acute lesions were found to be significantly softer (P < 0.0001) than chronic lesions (P < 0.0001) and the stiffness of lesions increased with progression of healing (P = 0.0138). A negative correlation between lesion hypoechogenicity and softness was appreciated with more hypoechoic lesions appearing softer (P = 0.0087) and more hyperechoic regions harder (P = 0.0002). A similar finding occurred with increased signal intensity on short tau inversion recovery (STIR) and proton density (PD) MRI sequences correlating with increased softness on elastography (P = 0.0164). Using US and MRI as references, commonly encountered soft tissue injuries of the equine distal limb could be detected with elastography. However, elastography was limited for detecting small, proximal injuries of the hindlimb proximal suspensory ligament. Elastographic evaluation of equine tendons and ligaments may allow better characterization of lesion chronicity and severity, and sequential examinations may optimize lesion management, rehabilitation, and return to training.