Introduction: The genomes of nine H5 subtypes of low pathogenic avian influenza virus (LPAIV) strains identified in wild birds in Poland between 2010 and 2015 were sequenced, and their phylogenetic relationship was determined. Material and Methods: AIV genome segments were amplified by RT-PCR and the PCR products were sequenced using Sanger method. Phylogenetic trees were generated in MEGA6 software and digital genotyping approach was used to visualise the relationship between analysed strains and other AIVs. Results: High genetic diversity was found in the analysed strains as multiple subgroups were identified in phylogenetic trees. In the HA tree, Polish strains clustered in two distinct subclades. High diversity was found for PB2, PB1, PA and NP, since 5-8 sublineages could be distinguished. Each strain had a different gene constellation, although relationship of as much as six out of eight gene segments was observed between two isolates. A relationship with poultry isolates was found for at least one segment of each Polish strain. Conclusion: The genome configuration of tested strains indicates extensive reassortment, although the preference for specific gene constellation could be noticed. A significant relationship with isolates of poultry origin underlines the need for constant monitoring of the AIV gene pool circulating in the natural reservoir.

High-throughput sequencing (HTS) identifies random viral fragments in environmental samples metagenomically. High reliability gains it broad application in virus evolution, host-virus interaction, and pathogenicity studies. Deep sequencing of field samples with content of host genetic material and bacteria often produces insufficient data for metagenomics and must be preceded by target enrichment. The main goal of the study was the evaluation of HTS for complete genome sequencing of field-case rabies viruses (RABVs). The material was 23 RABVs isolated mainly from red foxes and one European bat lyssavirus-1 isolate propagated in neuroblastoma cells. Three methods of RNA isolation were tested for the direct metagenomics and RABV-enriched approaches. Deep sequencing was performed with a MiSeq sequencer (Illumina) and reagent v3 kit. Bioinformatics data were evaluated by Kraken and Centrifuge software and de novo assembly was done with metaSPAdes. Testing RNA extraction procedures revealed the deep sequencing scope superiority of the combined TRIzol/column method. This HTS methodology made it possible to obtain complete genomes of all the RABV isolates collected in the field. Significantly greater rates of RABV genome coverages (over 5,900) were obtained with RABV enrichment. Direct metagenomic studies sequenced the full length of 6 out of 16 RABV isolates with a medium coverage between 1 and 71. Direct metagenomics gives the most realistic illustration of the field sample microbiome, but with low coverage. For deep characterisation of viruses, e.g. for spatial and temporal phylogeography during outbreaks, target enrichment is recommended as it covers sequences much more completely.

Introduction: The objective of this study was to describe a pharmacokinetic–pharmacodynamic (PK/PD) approach for determination of a rational dosage of ampicillin (AMP) and depletion of the antibiotic residues in milk after intramammary administration to cows.Material and Methods: The cows came from different farms from the Lublin Province area. They (n = 9) received 5 g of the drug, containing 75 mg of AMP sodium in physiological solution, through a syringe tube by intramammary administration. Following single intramammary administration, the milk samples (5 mL) were collected after 2, 4, 6, 8, 10, 24, 36, 48, and 60 h. The liquid chromatography-mass spectrometry analysis was performed on the Agilent 1200 system connected to an AB Sciex API 4000™ mass spectrometer. The pharmacokinetic analysis of the concentrations of the antibiotic in milk was performed using software Phoenix® WinNonlin® 6.4. Calculations were made in non-compartmental (slopes, highest, amounts, and moments) and compartmental analysis.Results: The pharmacokinetic characteristics of AMP after intramammary administration indicate rapid elimination of the drug from milk. The mean residence time had a several-fold lower value than the designated elimination half-life and amounts to only 3.4 h. The concentration of the drug in the milk dropped relatively quickly and the process was very dynamic.Conclusion: The conducted research confirms the rationale of using the PK/PD model in order to verify the dosing regimen for other antibiotic groups and various indicators of the applied PK/PD model.

OBJECTIVE To describe methods to measure the 3-D orientation of the proximal, diaphyseal, and distal segments of the canine radius by use of computer-aided design software (CADS) and to compare the repeatability and reliability of measurements derived by those methods. SAMPLE 31 canine radii with biapical deformities and 24 clinically normal (control) canine radii. PROCEDURES Select CT scans of radii were imported into a CADS program. Cartesian coordinate systems for the humerus and proximal, diaphyseal, and distal radial segments were developed. The orientation of each radial segment in the frontal, sagittal, and transverse planes was measured in triplicate by 3 methods. The repeatability and reliability of those measurements were calculated and compared among the 3 measurement methods. RESULTS The mean ± SD within-subject repeatability of radial angular measurements for all 3 methods was 1.40 ± 0.67° in the frontal plane, 3.17 ± 2.21° in the sagittal plane, and 3.01 ± 1.11° in the transverse plane for control radii and 2.56 ± 1.95° in the frontal plane, 3.59 ± 2.39° in the sagittal plane, and 3.47 ± 1.19° in the transverse plane for abnormal radii. Mean ± SD bias between radial measurement methods was 1.88 ± 2.07° in the frontal plane, 6.44 ± 6.80° in the sagittal plane, and 2.27 ± 2.81° in the transverse plane. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that use of CADS to assess the 3-D orientation of the proximal, diaphyseal, and distal segments of normal and abnormal canine radii yielded highly repeatable and reliable measurements.

OBJECTIVE To evaluate the feasibility of contrast-enhanced CT for assessment of pancreatic perfusion in healthy dogs. ANIMALS 6 healthy purpose-bred female Treeing Walker Coonhounds. PROCEDURES Contrast-enhanced CT of the cranial part of the abdomen was performed with 3-mm slice thickness. Postprocessing computer software designed for evaluation of human patients was used to calculate perfusion data for the pancreas and liver by use of 3-mm and reformatted 6-mm slices. Differences in perfusion variables between the pancreas and liver and differences in liver-specific data of interest were evaluated with the Friedman test. RESULTS Multiple pancreatic perfusion variables were determined, including perfusion, peak enhancement index, time to peak enhancement, and blood volume. The same variables as well as arterial, portal, and total perfusion and hepatic perfusion index were determined for the liver. Values for 6-mm slices appeared similar to those for 3-mm slices. The liver had significantly greater median perfusion and peak enhancement index, compared with the pancreas. CONCLUSIONS AND CLINICAL RELEVANCE Measurement of pancreatic perfusion with contrast-enhanced CT was feasible in this group of dogs. Hepatic arterial and pancreatic perfusion values were similar to previously published findings for dogs, but hepatic portal and hepatic total perfusion measurements were not. These discrepancies might have been attributable to physiologic differences between dogs and people and related limitations of the CT software intended for evaluation of human patients. Further research is warranted to assess reliability of perfusion variables and applicability of the method for assessment of canine patients with pancreatic abnormalities.

OBJECTIVE To determine the feasibility of radiographic measurement of liver area in small-breed dogs and to assess correlations between CT liver volume measurements (reference standard) and radiographic liver size measurements. ANIMALS 107 small-breed dogs (body weight, ≤ 10 kg) that had previously undergone orthogonal thoracic and abdominal radiography and abdominal CT. PROCEDURES In a retrospective study design, dogs were allocated to groups (normal liver [n = 36], microhepatia [34], and hepatomegaly [37]) on the basis of radiographic liver size and clinicopathologic findings. Radiographic liver area (RLA) was automatically calculated from archived radiographic images by free-hand outlining of the liver margins by use of DICOM viewer software, and other standard radiographic measurements were performed. Liver volume was measured on CT images. Intraoperator repeatability of RLA and CT measurements was assessed (duplicate measurements 2 weeks apart). To control for various breed conformations, radiographic values were normalized to body weight and T11 area. RESULTS Mean ± SD ratios of RLA to T11 area and RLA to body weight for dogs with normal livers were 32.7 ± 6.2 and 7.0 ± 1.4, respectively. Excellent intraobserver agreement was observed in RLA measurements within groups (intraclass correlation coefficients, 0.861 to 0.989), and RLA measurements had the highest correlation with CT liver volume measurements (r = 0.94) of all radiographic measurements. CONCLUSIONS AND CLINICAL RELEVANCE Findings indicated that RLA measurement in small-breed dogs with or without liver disease was useful and accurate for estimation of liver size, compared with CT measurement, and might be particularly useful for monitoring of changes in liver size.

OBJECTIVE To evaluate the diagnostic yield of dental radiography (Rad method) and cone-beam CT (CBCT) methods for the identification of 31 predefined dental disorders in brachycephalic dogs. ANIMALS 19 client-owned brachycephalic dogs admitted for evaluation and treatment of dental disease. PROCEDURES 31 predefined dental disorders were evaluated separately and scored by use of dental radiography and 3 CBCT software modules (serial CBCT slices and custom cross sections, tridimensional rendering, and reconstructed panoramic views). A qualitative scoring system was used. Dental disorders were grouped into 10 categories for statistical analysis. Point of reference for presence or absence of a dental disorder was determined as the method that could be used to clearly identify the disorder as being present. Accuracy, sensitivity, specificity, and positive and negative predictive values were calculated with the McNemar χ2 test of marginal homogeneity of paired data. RESULTS When all 3 CBCT methods were used in combination, the diagnostic yield of CBCT was significantly higher than that of dental radiography for 4 of 10 categories (abnormal eruption, abnormally shaped roots, periodontitis, and tooth resorption) and higher, although not significantly so, for all categories, except for 1 (loss of tooth integrity). CONCLUSIONS AND CLINICAL RELEVANCE CBCT provided more detailed information than did dental radiography. Therefore, CBCT would be better suited for use in diagnosing dental disorders in brachycephalic dogs.

OBJECTIVE To evaluate accuracy of quantification of right ventricle volume (RVV) by use of 3-D echocardiography (3DE) and ECG-gated multidetector CT (MDCT). ANIMALS 6 healthy hound-cross dogs. PROCEDURES ECG-gated MDCT and complete 3DE examinations were performed on each dog. Right ventricular end-diastolic volumes (EDVs), end-systolic volumes (ESVs), stroke volume (SV), and ejection fraction (EF) were measured for 3DE and MDCT data sets by use of software specific for RVV quantification. Correlation and level of agreement between methods were determined. Intraobserver and interobserver variability were assessed for 3DE. RESULTS No significant differences were detected between SV and EF obtained with MDCT and 3DE. Significant differences were detected between right ventricular EDV and ESV obtained with MDCT and 3DE. No significant difference in heart rate was detected between methods. The correlation between MDCT and 3DE was very good (r = 0.87) for EDV and ESV, moderate (r = 0.60) for EF, and poor (r = 0.31) for SV. Bland-Altman analysis revealed a systematic underestimation of RVV derived by use of 3DE, compared with the RVV derived by use of MDCT (mean bias, 15 and 10.3 mL for EDV and ESV, respectively). Intraobserver (EDV, 12%; ESV, 18%) and interobserver (EDV, 14%; ESV, 11%) variability were acceptable for 3DE. CONCLUSIONS AND CLINICAL RELEVANCE There was substantial variance for RVV measured by use of 3DE in healthy dogs and a significant underestimation of volumes, compared with results for MDCT, despite the fact there were no significant differences in SV and EF.

OBJECTIVE To evaluate the 3-D geometry of canine pelves and to characterize the long-term effects of juvenile pubic symphysiodesis (JPS) on pelvic geometry by comparing the pelvic configuration between littermates that did and did not undergo the procedure. ANIMALS 24 Labrador Retriever, Golden Retriever, or Labrador Retriever–Golden Retriever crossbred service dogs from 13 litters. PROCEDURES At 16 weeks old, puppies with a hip joint distraction index ≥ 0.5 were randomly assigned to undergo thermal JPS (n = 9), mechanical JPS (7), or a sham (control) surgical procedure (8). Ten years later, each dog underwent a CT scan of the pelvic region. Modeling software was used to create 3-D reconstructions from the CT scans, and various pelvic measurements were made and compared among the 3 treatments. RESULTS Compared with the control treatment, thermal and mechanical JPS increased the hemipelvis acetabular angle by 4°, the acetabular angle of lateral opening by 5°, and the orientation of the medial acetabular wall in a transverse plane by 6°, which indicated that JPS increased dorsal femoral head coverage by the acetabulum. Both JPS procedures decreased the pelvic canal area by approximately 20% and acetabular inclination by 6° but did not alter acetabular retroversion. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that thermal and mechanical JPS were equally effective in altering the 3-D pelvic geometry of dogs. These findings may help guide future studies of alternatives for optimizing canine pelvic anatomy to minimize the risk of hip dysplasia and associated osteoarthritis.

OBJECTIVE To predict histologic grade of meningiomas in dogs via texture analysis (TA) of MRI scans of the brain and spinal cord. SAMPLE 58 sets of MRI scans of the brain and spinal cord of dogs with histologically diagnosed meningioma. PROCEDURES MRI sequences were divided into a training set and a test set, and results of histologic assessment were obtained. Tumors were histologically grouped as benign (stage I) or atypical-anaplastic (stage II or III). Texture analysis was performed by use of specialized software on T2-weighted (T2W) and pre- and postcontrast T1-weighted (T1W) images. A set of 30 texture features that provided the highest discriminating power between the 2 histologic classes in the training set was automatically selected by the TA software. Linear discriminant analysis was performed, and the most discriminant factor (MDF) was calculated. The previously selected texture features were then used for linear discriminant analysis of the test set data, and the MDF was calculated. RESULTS For the training set, TA of precontrast T1W images provided the best diagnostic accuracy; a cutoff MDF of < 0.0057 resulted in a sensitivity of 97.4% and specificity of 95.0% for discriminating benign from atypical-anaplastic meningiomas. Use of postcontrast T1W and T2W images yielded poorer diagnostic performances. Application of the MDF cutoff calculated with the training set to the MDF calculated with the test set provided a correct classification rate of 96.8% for precontrast T1W images, 92.0% for postcontrast T1W images, and 78.9% for T2W images. CONCLUSIONS AND CLINICAL RELEVANCE Findings supported the potential clinical usefulness of TA of MRI scans for the grading of meningiomas in dogs.