Estimation of percent body surface area in cats with use of computed tomography.

OBJECTIVE: To establish a baseline reference and create a user-friendly chart for normal body surface area (BSA) in the adult domestic cat similar to the "Rule of Nines" chart. DESIGN: Prospective cadaveric study. SETTING: University Teaching Hospital. ANIMALS: Seven adult domestic short-haired feline cadavers of normal body condition (4-6/9) and body weight. INTERVENTION: Individual whole-body computed tomography (CT) scan was performed on all cats. Images were analyzed with 3-dimensional technology to measure the BSA. A chart was created, illustrating the individual surface area of each body part. MEASUREMENTS AND MAIN RESULTS: Obtained measurements of the cats are different from surface areas of people and dogs. Percentage of total BSA for cats was as follows: head 13%, neck 5%, thorax 20%, abdomen 15%, pelvis and tail 9%, front legs 7% each, and pelvic legs 12% each. CONCLUSION: The distribution of BSA between body parts is different in cats compared to dogs and people. A species-specific chart was created to assure a more accurate estimation of BSA in cats.

Radiological characterization of gilthead seabream (Sparus aurata) by X-ray computed tomography.

In recent years, the increasing use of fish as new animal models in scientific research and the growth of fish farming (mainly for human consumption) have highlighted the need for advanced technology to deepen our knowledge of fish biology. Hence, the present study was carried out to radiologically analyse the whole body of gilthead seabream (Sparus aurata) specimens using X-ray computed tomography (CT). Images were acquired in an Albira SPECT/PET/CT tri-modal preclinical-scanner. Segmentation, measurements and three-dimensional reconstruction were made using the Carestream Molecular imaging Albira CT system in conjunction with Pmod, AMIDE and Amira software packages. The results showed that the density values of gilthead seabream are in the range -700 to +2500 HU for the whole body. We also determined the density ranges that topographically coincide with the swim bladder, soft tissues, fat, skin and skeleton. This work describes, validates and demonstrates the application of a fully automated image analysis technique to study and quantify fish body composition, whether segmented or as a whole. In addition, the basis for applying this image technique in other in vivo studies is established.

Utilization of whole body computed tomography in polytrauma patients.

OBJECTIVE: To determine the safety, feasibility, and utility of whole body computed tomography (WBCT) in polytrauma patients. A second objective was to describe the utilization of the VetMouse Trap for sedated WBCT in polytrauma patients. METHODS: A prospective, observational study in a high-volume private practice. Any cat or dog weighing <20 kg that presented to the emergency department following a polytrauma was eligible. Patients were given analgesia and sedation prior to placement in the VetMouse Trap. A WBCT was then performed. RESULTS: A total of 16 patients (8 dogs and 8 cats) met inclusion criteria. All patients presented with blunt trauma; 3 also had evidence of penetrating wounds. Five (31.25%) patients met inclusion criteria for WBCT based on their neurological evaluation. Five (31.5%) were non-ambulatory with suspicion of orthopedic injury, and 37.5% met additional criteria for WBCT. The most common areas of injury were head (43.7%), lungs (25%), and pelvis (25%). Four patients (25%) had evidence of cavitary effusion that was not seen on focused assessment using sonography for trauma (FAST) scan. No patient had any adverse events during the CT. CONCLUSION: This study demonstrated successful WBCT imaging of the sedated small animal polytrauma patient with the VetMouse Trap.

Survival analysis in dogs with urinary transitional cell carcinoma that underwent whole-body computed tomography at diagnosis.

This retrospective study aimed to evaluate factors associated with survival and to compare characteristics between tumour localizations in dogs with urinary transitional cell carcinoma (TCC) that underwent whole-body computed tomography (CT) at diagnosis. Dogs with histologically confirmed TCC that received medical therapy between 2010 and 2017 were included; dogs that underwent surgery or radiotherapy for the primary tumour were excluded. According to the CT findings, primary tumour localization (classified into the Bladder, Urethra and Bladder and Urethra groups), prostate involvement, iliosacral lymphadenomegaly, sternal lymphadenomegaly and metastasis to the bone and lung were evaluated for survival analysis. CT at diagnosis revealed iliosacral lymphadenomegaly, sternal lymphadenomegaly, bone metastasis and lung metastasis in 47.7%, 18.5%, 24.6% and 35.4% of the 65 included dogs, respectively. The overall median survival time was 196 days. On multivariable analysis, TCC localization (hazard ratio [HR], 1.90; P = .037), bone metastasis (HR, 2.76; P = .013) and sternal lymphadenomegaly (HR, 3.56; P = .004) were significantly associated with survival. Compared to the Bladder group (n = 16), the Urethra group (n = 26) had higher metastasis rates to the bone (6.3% vs 42.3%; P = .045) and lung (6.3% vs 46.2%; P = .022). The survival time was shorter in the Urethra group than in the Bladder group (121.5 vs 420 days; P < .001), and it was similar only in female dogs (247 vs 420 days; P = .031). These findings suggest that whole-body CT could be valuable for predicting the prognosis in urinary TCC.

Validation of whole-body radiographs for examining uterine disorders in sexually intact female rabbits.

OBJECTIVE: The present study evaluated whole-body radiographs of female rabbits regarding uterine disorders. MATERIAL AND METHODS: Based on radiographs of 143 rabbits, visibility, size, shape and density of the uterus, nutritional status of the animal and possible metastasis in other organs were examined. A suspected diagnosis was deduced. The radiological findings were compared with the results of the histopathological examination. RESULTS: Especially the nutritional status was important regarding the visibility of the genital tract. In 99 of the 130 visible uteri (76.2 %), a pathological alteration was detected. Thereof, endometrial hyperplasia was most frequently seen (63.6 %), followed by neoplasia (altogether 42.4 %, 39.4 % of these with adenocarcinoma), endometritis (35.4 %) and other diseases (18.2 %). Forty-six rabbits showed only one disease, 42 rabbits had two, 10 rabbits three and one rabbit four different diseases. Among 13 of the 143 uteri that were not visible in the radiograph four were altered pathologically. In 31 of the 130 rabbits with a visible uterine region (21.7 %) the uterus was unaltered. Visualising the uterus proved impossible in two of four skinny rabbits (2.8 %), whereas in all 39 obese rabbits the uterus was visible. Only in one of the four skinny rabbits was the uterus without pathological findings and six of the 39 obese rabbits showed no pathological alteration, visibility notwithstanding. Calcification indicated a tumourous occurrence in 23 of 53 cases. Except for one rabbit, the mineralised neoplasia was an adenocarcinoma (n = 22). Five animals with calcification in the uterine region showed no abnormalities. Of these, the calcification of four animals was no longer existent in the radiograph after ovariohysterectomy. Neoplastic changes in the uterus did not always show calcification. In 20 of the 44 patients with uterine neoplasia no calcification was seen. Changes in uterine size occurred in 118 of the 143 rabbits (82.5 %) and alteration of the uterine shape in 109 of the 143 rabbits (67.1 %). CONCLUSION AND CLINICAL RELEVANCE: Thus, the above presented parameters - especially size, shape and mineralisation - are very important regarding uterine diseases.

Evaluation of Two Potent and Selective PET Radioligands to Image COX-1 and COX-2 in Rhesus Monkeys.

This study assessed whether the newly developed PET radioligands 11C-PS13 and 11C-MC1 could image constitutive levels of cyclooxygenase (COX)-1 and COX-2, respectively, in rhesus monkeys. Methods: After intravenous injection of either radioligand, 24 whole-body PET scans were performed. To measure enzyme-specific uptake, scans of the 2 radioligands were also performed after administration of a nonradioactive drug preferential for either COX-1 or COX-2. Concurrent venous samples were obtained to measure parent radioligand concentrations. SUVs were calculated from 10 to 90 min. Results:11C-PS13 showed specific uptake in most organs, including spleen, gastrointestinal tract, kidneys, and brain, which was blocked by COX-1, but not COX-2, preferential inhibitors. Specific uptake of 11C-MC1 was not observed in any organ except the ovaries and possibly kidneys. Conclusion: The findings suggest that 11C-PS13 has adequate signal in monkeys to justify its extension to human subjects. In contrast, 11C-MC1 is unlikely to show significant signal in healthy humans, though it may be able to do so in inflammatory conditions.

Use of iDXA spine scans to evaluate total and visceral abdominal fat.

OBJECTIVES: Abdominal fat may be a better predictor than body mass index (BMI) for risk of metabolically-related diseases, such as diabetes, cardiovascular disease, and some cancers. We sought to validate the percent fat reported on dual energy X-ray absorptiometry (DXA) regional spine scans (spine fat fraction, SFF) against abdominal fat obtained from total body scans using the iDXA machine (General Electric, Madison, WI), as previously done on the Prodigy model. METHODS: Total body scans and regional spine scans were completed on the same day (N = 50). In alignment with the Prodigy-based study, the following regions of interest (ROI) were assessed from total body scans and compared to the SFF from regional spine scans: total abdominal fat at (1) lumbar vertebrae L2-L4 and (2) L2-Iliac Crest (L2-IC); (3) total trunk fat; and (4) visceral fat in the android region. Separate linear regression models were used to predict each total body scan ROI from SFF; models were validated by bootstrapping. RESULTS: The sample was 84% female, a mean age of 38.5 ± 17.4 years, and mean BMI of 23.0 ± 3.8 kg/m2 . The SFF, adjusted for BMI, predicted L2-L4 and L2-IC total abdominal fat (%; Adj. R2 : 0.90) and total trunk fat (%; Adj. R2 : 0.88) well; visceral fat (%) adjusted R2 was 0.83. Linear regression models adjusted for additional participant characteristics resulted in similar adjusted R2 values. CONCLUSIONS: This replication of the strong correlation between SFF and abdominal fat measures on the iDXA in a new population confirms the previous Prodigy model findings and improves generalizability.

A prototype PET/SPECT/X-rays scanner dedicated for whole body small animal studies.

OBJECTIVE: To present a prototype tri-modal imaging system, consisting of a single photon emission computed tomography (SPET), a positron emission tomography (PET), and a computed tomography (CT) subsystem, evaluated in planar mode. MATERIALS AND METHODS: The subsystems are mounted on a rotating gantry, so as to be able to allow tomographic imaging in the future. The system, designed and constructed by our group, allows whole body mouse imaging of competent performance and is currently, to the best of our knowledge, unequaled in a national and regional level. The SPET camera is based on two Position Sensitive Photomultiplier Tubes (PSPMT), coupled to a pixilated Sodium Iodide activated with Thallium (NaI(Tl)) scintillator, having an active area of 5x10cm2. The dual head PET camera is also based on two pairs of PSPMT, coupled to pixelated berillium germanium oxide (BGO) scintillators, having an active area of 5x10cm2. The X-rays system consists of a micro focus X-rays tube and a complementary metal-oxide-semiconductor (CMOS) detector, having an active area of 12x12cm2. RESULTS: The scintigraphic mode has a spatial resolution of 1.88mm full width at half maximum (FWHM) and a sensitivity of 107.5cpm/0.037MBq at the collimator surface. The coincidence PET mode has an average spatial resolution of 3.5mm (FWHM) and a peak sensitivity of 29.9cpm/0.037MBq. The X-rays spatial resolution is 3.5lp/mm and the contrast discrimination function value is lower than 2%. CONCLUSION: A compact tri-modal system was successfully built and evaluated for planar mode operation. The system has an efficient performance, allowing accurate and informative anatomical and functional imaging, as well as semi-quantitative results. Compared to other available systems, it provides a moderate but comparable performance, at a fraction of the cost and complexity. It is fully open, scalable and its main purpose is to support groups on a national and regional level and provide an open technological platform to study different detector components and acquisition strategies.

Establishing a reliable gait evaluation method for rodent studies.

BACKGROUND: CatWalk is one of the most popular tools for evaluating gait recovery in preclinical research, however, there is currently no consensus on which of the many gait parameters captured by CatWalk can reliably model recovery. There are conflicting interpretations of results, along with many common but seldom reported problems such as heel walking and poor compliance. NEW METHOD: We developed a systematic manual classification method that overcomes common problems such as heel walking and poor compliance. By correcting automation errors and removing inconsistent gait cycles, we isolated stretches of recordings that are more reliable for analysis. Recovery outcome was also assessed by quantitative histomorphometric analysis of myelinated axons. RESULTS: While 40-60% of runs were erroneously classified without manual intervention, we corrected all errors with our new method, and showed that Stand Time, Duty Cycle, and Swing Speed are able to track significant differences over time and between experimental groups (all p<0.05). The usability of print area and intensity parameters requires further validation beyond the capabilities of CatWalk. COMPARISON WITH EXISTING METHOD(S): There is currently no strategy that addresses problems such as heel walking and poor compliance, and therefore no standard set of parameters that researchers can rely on to report their findings. CONCLUSION: Manual classification is a crucial step to generate reliable CatWalk data, and Stand Time, Duty Cycle, and Swing Speed are suitable parameters for evaluating gait recovery. Static parameters such as print area and intensity should be used with extreme caution.

FIM$

In vivo whole-body imaging of small animals plays an important role for biomedical studies. In particular, animals like the fruit fly Drosophila melanogaster or the nematode Caenorhabditis elegans are popular model organisms for preclinical research since they offer sophisticated genetic tool-kits. Recording these translucent animals with high contrast in a large arena is however not trivial. Furthermore, fluorescent proteins are widely used to mark cells in vivo and report their functions. This paper introduces a novel optical imaging technique called FIM2c enabling simultaneous detection of the animals posture and movement as well as fluorescent markers like green fluorescent protein (GFP). FIM2c utilizes frustrated total internal reflection of two distinct wavelengths and captures both, reflected and emitted light. The resultant two-color high-contrast images are superb compared to other imaging systems for larvae or worms. This multipurpose method enables a large variety of different experimental approaches. For example, FIM2c can be used to image GFP positive cells/tissues/animals and supports the integration of fluorescent tracers into multitarget tracking paradigms. Moreover, optogenetic tools can be applied in large-scale behavioral analysis to manipulate and study neuronal functions. To demonstrate the benefit of our system, we use FIM2c to resolve colliding larvae in a high-throughput approach, which was impossible given the existing tools. Finally, we present a comprehensive database including images and locomotion features of more than 1300 resolved collisions available for the community. In conclusion, FIM2c is a versatile tool for advanced imaging and locomotion analysis for a variety of different model organisms.

An FIM-Based Long-Term In-Vial Monitoring System for Drosophila Larvae.

Drosophila larvae are an insightful model and the automated analysis of their behavior is an integral readout in behavioral biology. Current tracking systems, however, entail a disturbance of the animals, are labor-intensive, and cannot be easily used for long-term monitoring purposes. Here, we present a novel monitoring system for Drosophila larvae, which allows us to analyze the animals in cylindrical culture vials. By utilizing the frustrated total internal reflection in combination with a multi-camera/microcomputer setup, we image the complete housing vial surface and, thus, the larvae for days. We introduce a calibration scheme to stitch the images from the multi-camera system and unfold arbitrary cylindrical surfaces to support different vials. As a result, imaging and analysis of a whole population can be done implicitly. For the first time, this allows us to extract long-term activity quantities of larvae without disturbing the animals. We demonstrate the capabilities of this new setup by automatically quantifying the activity of multiple larvae moving in a vial. The accuracy of the system and the spatio-temporal resolution are sufficient to obtain motion trajectories and higher level features, such as body bending. This new setup can be used for in-vial activity monitoring and behavioral analysis and is capable of gathering millions of data points without both disturbing the animals and increasing labor time. In total, we have analyzed 107 671 frames resulting in 8650 trajectories, which are longer than 30 s, and obtained more than 4.2 × 106 measurements.

Retrospective Evaluation of Whole Body Computed Tomography for Tumor Staging in Dogs with Primary Appendicular Osteosarcoma.

OBJECTIVE: To evaluate whole body computed tomography (CT) for staging canine appendicular osteosarcoma. STUDY DESIGN: Retrospective case series. ANIMALS: Client-owned dogs diagnosed with appendicular osteosarcoma (n=39). METHODS: Medical records for client-owned dogs diagnosed with appendicular osteosarcoma from August 2008 to July 2014 were reviewed. Dogs were included if they had a confirmed diagnosis of appendicular osteosarcoma and were staged using whole body CT. Data collected included signalment, body weight, primary tumor location, serum alkaline phosphatase (ALP) activity, findings on 3-view thoracic radiographs, cytologic or histologic results, and findings on CT. RESULTS: Thirty-nine dogs (median age 8.5 years; median body weight 37 kg) had osteosarcoma of the distal radius (n=17), proximal humerus (11) and other sites. Serum ALP activity was elevated in 14 dogs. Bone metastasis was not detected in any dog on whole body CT. Pulmonary metastasis was considered definitive on CT based on board certified radiologist assessment in 2/39 dogs (5%). Two additional dogs (2/39, 5%) had soft tissue masses diagnosed on CT, consistent with concurrent, non-metastatic malignancies. CONCLUSION: Bone metastases were not identified in any dog with whole body CT. Thoracic and abdominal CT detected lung lesions and concurrent neoplasia in dogs with primary appendicular osteosarcoma. Whole body CT may be a useful adjunct to other screening tests for disseminated malignancy.

Fluorescence and Cerenkov luminescence imaging. Applications in small animal research.

This review addresses small animal optical imaging (OI) applications in diverse fields of basic research. In the past, OI has proven to be cost- and time-effective, allows real-time imaging as well as high-throughput analysis and does not imply the usage of ionizing radiation (with the exception of Cerenkov imaging applications). Therefore, this technique is widely spread - not only geographically, but also among very different fields of basic research - and is represented by a large body of publications. Originally used in oncology research, OI is nowadays emerging in further areas like inflammation and infectious disease as well as neurology. Besides fluorescent probe-based contrast, the feasibility of Cerenkov luminescence imaging (CLI) has been recently shown in small animals and thus represents a new route for future applications. Thus, this review will focus on examples for OI applications in inflammation, infectious disease, cell tracking as well as neurology, and provides an overview over CLI.

IMAGING DIAGNOSIS--USE OF RADIOGRAPHY AND COMPUTED TOMOGRAPHY IN THE DIAGNOSIS OF A MINERALIZED YOLK SAC IN A BROWN KIWI (APTERYX MANTELLI).

A 12-day-old Brown Kiwi (Apteryx mantelli) was presented with anorexia, torticollis, head-tilt, and coelomic distension. Radiographs showed an ill-defined, fat-opaque, coelomic mass displacing viscera craniodorsally. Curvilinear mineral opacities were superimposed over the ventral aspect of the mass. Computed tomography demonstrated the presence of mineral within the periphery of a fat attenuating mass consistent with a retained yolk sac. A deutectomy (yolk sac excision) was performed. Histopathology of the excised tissue confirmed the diagnosis of a retained yolk sac with multifocal mineralization.

MRI compatible small animal monitoring and trigger system for whole body scanners.

Performing magnetic resonance imaging (MRI) experiments with small animals requires continuous monitoring of vital parameters, especially the respiration rate. Clinical whole-body MR scanners represent an attractive option for preclinical imaging as dedicated animal scanners are cost-intensive in both investment and maintenance, thus limiting their availability. Even though impressive image quality is achievable with clinical MR systems in combination with special coils, their built-in physiologic monitoring and triggering units are often not suited for small animal imaging. In this work, we present a simple, MRI compatible low cost solution to monitor the respiration and heart rate of small animals in a clinical whole-body MR scanner. The recording and processing of the biosignals as well as the optimisation of the respiratory trigger generation is decribed. Additionally rat and mouse in-vivo MRI experiments are presented to illustrate the effectiveness of the monitoring and respiratory trigger system in suppressing motion artifacts.

A throughput-optimized array system for multiple-mouse MRI.

MRI is a versatile tool for the systematic assessment of anatomical and functional changes in small-animal models of human disease. Its noninvasive nature makes it an ideal candidate for longitudinal evaluations of disease progression, but relatively long scan times limit the number of observations that can be made in a given interval of time, imposing restrictions on experimental design and potentially compromising statistical power. Methods that reduce the overall time required to scan multiple cohorts of animals in distinct experimental groups are therefore highly desirable. Multiple-mouse MRI, in which several animals are simultaneously scanned in a common MRI system, has been successfully used to improve study throughput. However, to best utilize the next generation of small-animal MRI systems that will be equipped with an increased number of receive channels, a paradigm shift from the simultaneous scanning of as many animals as possible to the scanning of a more manageable number, at a faster rate, must be considered. This work explores the tradeoffs between the number of animals to scan at once and the number of array elements dedicated to each animal, to maximize throughput in systems with 16 receive channels. An array system consisting of 15 receive and five transmit coils allows acceleration by a combination of multi-animal and parallel imaging techniques. The array system was designed and fabricated for use on a 7.0-T/30-cm Bruker Biospec MRI system, and tested for high-throughput imaging performance in phantoms and live mice. Results indicate that up to a nine-fold throughput improvement of a single sequence is possible compared with an unaccelerated single-animal acquisition. True data throughput of a contrast-enhanced anatomical study is estimated to be improved by just over six-fold.

Recovery of chemical estimates by field inhomogeneity neighborhood error detection (REFINED): fat/water separation at 7 tesla.

PURPOSE: To reduce swaps in fat-water separation methods, a particular issue on 7 Tesla (T) small animal scanners due to field inhomogeneity, using image postprocessing innovations that detect and correct errors in the B0 field map. MATERIALS AND METHODS: Fat-water decompositions and B0 field maps were computed for images of mice acquired on a 7T Bruker BioSpec scanner, using a computationally efficient method for solving the Markov Random Field formulation of the multi-point Dixon model. The B0 field maps were processed with a novel hole-filling method, based on edge strength between regions, and a novel k-means method, based on field-map intensities, which were iteratively applied to automatically detect and reinitialize error regions in the B0 field maps. Errors were manually assessed in the B0 field maps and chemical parameter maps both before and after error correction. RESULTS: Partial swaps were found in 6% of images when processed with FLAWLESS. After REFINED correction, only 0.7% of images contained partial swaps, resulting in an 88% decrease in error rate. Complete swaps were not problematic. CONCLUSION: Ex post facto error correction is a viable supplement to a priori techniques for producing globally smooth B0 field maps, without partial swaps. With our processing pipeline, it is possible to process image volumes rapidly, robustly, and almost automatically.

Imaging diagnosis-multicentric lymphoma of granular lymphocytes imaged with FDG PET/CT in a dog.

A 5-year-old female spayed bulldog was referred for mild dyspnea, decreased activity and appetite, occasional nonproductive cough, polydipsia, and polyuria. A 2-deoxy-2-[(18) F]fluoro-D-glucose (FDG) positron emission tomography/computed tomography (PET/CT) scan revealed unexpected activity in the heart, lungs, and mild generalized lymphadenopathy that led to the diagnosis of lymphoma of granular lymphocytes after nonspecific findings on imaging with standard modalities of echocardiography, thoracic radiography, and abdominal ultrasound. PET/CT scanning is a useful whole body imaging modality with high sensitivity for changes associated with canine lymphoma.

Assessment of blood pool, soft tissue, and skeletal uptake of sodium fluoride F 18 with positron emission tomography-computed tomography in four clinically normal dogs.

OBJECTIVE: To determine the ideal interval to image acquisition after IV injection of sodium fluoride F 18 ((18)F-NaF) and evaluate biodistribution of the radiopharmaceutical in clinically normal skeletally immature dogs. ANIMALS: 4 female dogs. PROCEDURES: Each dog was anesthetized for evaluation with a commercial hybrid positron emission tomography (PET)-CT instrument. A low-radiation dose, whole-body CT scan was acquired first. An IV injection of (18)F-NaF (0.14 mCi/kg) was administered, and a dynamic PET scan centered over the heart and liver was acquired during a period of 120 minutes. Uptake of (18)F-NaF in the blood pool, soft tissues, and skeletal structures was evaluated via region of interest analysis to derive standardized uptake values and time-activity curves, which were used to determine the optimal postinjection time for skeletal image acquisition. Biodistribution was also assessed from a final whole-body PET-CT scan acquired after the dynamic scan. RESULTS: Time-activity curves revealed a rapid decrease in the amount of radiopharmaceutical in the blood pool and soft tissues and a rapid increase in the amount of radiopharmaceutical in bones soon after injection. At 50 minutes after injection, the greatest difference in uptake between soft tissues and bones was detected, with continued subtle increase in uptake in the bones. Uptake of (18)F-NaF was slightly increased at growth plates and open ossification centers, compared with that at other parts of the bone. CONCLUSIONS AND CLINICAL RELEVANCE: At 50 minutes after IV injection of (18)F-NaF at the dose evaluated, PET-CT yielded excellent bone-to-background ratio images for evaluation of the skeletal system in dogs.