USE OF CONTRAST HARMONIC ULTRASOUND FOR THE DIAGNOSIS OF CONGENITAL PORTOSYSTEMIC SHUNTS IN THREE DOGS

Contrast harmonic ultrasound was used to determine macrovascular and perfusion patterns in three dogs with congenital extrahepatic solitary portosystemic shunts (PSS). With coded harmonic angiographic ultrasound, the size and tortuosity of the hepatic arteries were subjectively increased. Single pulse intermittent low-amplitude harmonic perfusion imaging provided contrast enhancement time-intensity curves from regions of interest in the liver. Mean (+/- standard deviation) peak perfusion times of dogs with PSS were significantly shorter (p = 0.01; 7.0 +/- 2.0 s) than reported in normal dogs (22.8 +/- 6.8 s). The contrast inflow slope for the dogs with PSS (14.6 +/- 3.7 pixel intensity units [PIU] was significantly (p = 0.05) larger than reported for normal dogs (3.6 +/- 1.4 PIU/s). These results indicate that combined coded harmonic angiographic and contrast harmonic perfusion sonography can be used to detect increased hepatic arterial blood flow as an indicator of PSS in dogs.     

CONTRAST HARMONIC ULTRASOUND OF SPONTANEOUS LIVER NODULES IN 32 DOGS

Thirty-two dogs with spontaneous hepatic nodules were given intravenous ultrasound contrast medium (Definity or Sonovue) and imaged with contrast harmonic software on a conventional ultrasound machine system. Digital video images were initially reviewed to describe the perfusion pattern of malignant nodules. The images were reviewed again to test this pattern against all individual nodules. Subjectively, there was improved conspicuity of malignant nodules after contrast enhancement compared with conventional imaging and increased numbers of malignant nodules were often noted. There was decreased conspicuity of benign nodules and no additional nodules were seen after contrast enhancement. There was a highly significant (P < 0.0001) association of malignancy with a hypoechoic nodule at surrounding normal liver peak contrast enhancement. Benign nodules were isoechoic to the surrounding normal liver at peak contrast enhancement. Only one benign nodule (hepatoma) had regions of hypoechogenicity compared with the surrounding normal liver at peak liver contrast enhancement. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were highly significant (P < 0.0001) (100%, 94.1%, 93.8%, 100%, and 96.9%, respectively). No complications or morbidity was noted throughout the course of the study. Contrast harmonic ultrasound appears to be accurate at discriminating between naturally occurring benign and malignant nodules in the liver of dogs.     

COMPARISON OF SONOGRAPHIC FEATURES OF BENIGN AND NEOPLASTIC DEEP LYMPH NODES IN DOGS

The differentiation of benign vs. neoplastic lymph nodes impacts patient management. Specific sonographic features are typically considered when assessing lymph nodes in dogs. However, the usefulness of these criteria in distinguishing benign vs. malignant lymph nodes remains largely unknown, especially for deep lymph nodes. Our aim was to compare sonographic features in benign and neoplastic deep lymph nodes with the hope of identifying predictive criteria. Thirty‐one deep lymph nodes (16 mesenteric, 10 medial iliac, three hepatic, one sternal, and one cranial mediastinal) in 31 dogs were examined prospectively with B‐mode and Color flow Doppler. Lymph nodes were aspirated using ultrasound‐guidance and final diagnosis were established based on cytologic and/or histopathologic interpretation. Prevalence of each sonographic feature and combinations of two features was calculated for each group and compared using a χ²‐test or Student's t‐test for unequal variances. Ten lymph nodes were benign (hyperplastic and/or inflammatory) and 21 were neoplastic. All were hypoechoic, except for one neoplastic lymph node. Maximal short‐axis diameter (P=0.0006) and long‐axis diameter (P=0.01), and SA/LA ratio (P=0.008) were increased significantly for neoplastic (2.8, 5.5 cm, and 0.50, respectively) vs. benign (1.2, 3.8 cm, and 0.34, respectively) lymph nodes. The prevalence of other features was similar between groups. Doppler evaluation was possible in 77% of lymph nodes, but there was no significant difference between groups. When any two ultrasound features were combined, the only difference between benign and neoplastic lymph nodes was for the combination of contour regularity and appearance of the perinodal fat (P=0.03).     

QUANTITATIVE CONTRAST HARMONIC ULTRASOUND IMAGING OF NORMAL CANINE LIVER

Eight adult dogs with no evidence of liver disease, weighing between 8 and 25 kg were imaged after injection of a microbubble contrast medium using harmonic ultrasound imaging. All dogs received three separate bolus contrast injections, and six dogs also received three separate constant rate infusions each. Time/Mean Pixel Value curves were generated for selected regions of the liver. Upslope, downslope, baseline, peak, change, and time to peak were calculated. For bolus injection (averaging all subjects), upslope was 3.85 +/- 1.50 Mean Pixel Values/s, downslope was -0.71 +/- 0.30 Mean Pixel Values/s, baseline was 72.38 +/- 17.82 Mean Pixel Values, peak was 120.26 +/- 17.44 Mean Pixel Value, change from baseline to peak was 47.88 +/- 6.92 Mean Pixel Values, time to peak (from injection) was 22.88 +/- 6.79 s, and time to peak (from first upslope) was 13.88 +/- 1.55 s. Data acquisition and analysis from constant rate infusions was more cumbersome than for bolus, and results were less repeatable. There were significant differences (p < .005) in upslope, downslope, peak values, and time to peak between the two methods. These baseline data may prove useful in the evaluation of dogs with diffuse hepatic disease.     

CONTRAST HARMONIC ULTRASOUND APPEARANCE OF CONSECUTIVE PERCUTANEOUS RENAL BIOPSIES IN DOGS

Ultrasound‐guided percutaneous renal biopsy may be associated with complications, especially when using larger needles. Contrast harmonic ultrasound increases blood pool echo intensity, enhancing parenchymal lesions. Therefore, contrast harmonic ultrasound is a potential alternative screening method for postbiopsy renal lesions. Renal biopsies were performed using 14 G needles in 11 healthy Beagles, at three occasions: 0 (“Baseline Biopsy”; BB), 4 (“Biopsy 2”; B2), and 6 months (“Biopsy 3”; B3). Ultrasound and contrast harmonic ultrasound of biopsied kidneys were performed approximately 30 min after biopsy (week 0) at BB and B2, and repeated once every week (weeks 1–3) until normal appearance. At B3, only contrast harmonic ultrasound was performed, both immediately and 30‐min postbiopsy. Contrast harmonic ultrasound images were reviewed using subjective and semiquantitative methods to describe lesions including number, shape, size, sharpness, echogenicity, and evolution. More renal lesions were detected with contrast harmonic ultrasound (22/22) compared with conventional ultrasound (14/22). The majority appeared at week 0 as hypoechoic tract(s) (27/33), the other (6/33) as ill‐defined areas or area/tract combination, all having variable size, shape, and echogenicity. Seven tracts had a small subcapsular hematoma. In most kidneys, similar or gradual decrease of size and sharpness, and increased echogenicity was observed until normal appearance occurred at week 1 (1/22), week 2 (18/22), or week 3 (22/22). Two Beagles developed complications. At B3, immediately postbiopsy, tracts were hyperechoic in 9/11 kidneys, becoming hypoechoic again 30 min later. Contrast harmonic ultrasound is a valuable method to evaluate postbiopsy renal lesions in dogs.     

CHARACTERIZATION OF NORMAL AND ABNORMAL CANINE SUPERFICIAL LYMPH NODES USING GRAY-SCALE B-MODE, COLOR FLOW MAPPING, POWER, AND SPECTRAL DOPPLER ULTRASONOGRAPHY: A MULTIVARIATE STUDY

The objective of this study was to characterize the ultrasonographic patterns of normal superficial lymph nodes and to evaluate whether ultrasonography can help discriminate between different lymphadenopathies (reactive, lymphoma, and metastases) in dogs. Three hundred and eighteen superficial lymph nodes in 142 dogs were studied by B-mode, color flow mapping, power, and spectral Doppler ultrasonography. Size, echogenicity, nodal border definition, presence of a nodal hilus, acoustic enhancement and distribution of vascular flow, as well as perfusion indices were measured. Multivariate statistics using discriminant analysis was used to determine which parameters can be used to predict the diagnosis of the lymph node. The size of the lymph node, distribution of vascular flow within the lymph node, and pulsatility index (PI) in combination gave a classification error of 23% for the four groups of lymph nodes. This was improved to 11% if the nodes were divided into two groups: benign and malignant. There was a significant difference in resistive index (RI) and PI between benign and malignant nodes. Cut-off values were determined using receiver operator curves, 0.68 RI and 1.49 PI.     

QUANTITATIVE CONTRAST ULTRASOUND ANALYSIS OF RENAL PERFUSION IN NORMAL DOGS

Eight normal dogs with no evidence of renal disease, weighing between 8 and 25 kg were imaged using contrast harmonic ultrasound after injection of a microbubble contrast medium. All dogs received three separate bolus injections of 0.05 ml of commercial contrast medium (Definity). Time/mean pixel value (MPV) curves were generated for selected regions in the cortex and medulla of the left kidney in each dog. Upslope, downslope, baseline, peak intensity, and time to peak were calculated for each zone. For a bolus injection, within the renal cortex (averaging all subjects) the upslope was 7.4 +/- 1.5 MPV/s, downslope was -0.4 +/- .2 MPV/s, baseline was 66.8 +/- 9.3 MPV, peak was 103.6 +/- 8.2 MPV, time to peak (from injection) was 12.8 +/- 5.3 s and from time of contrast medium reaching the kidney was 5.1 +/- 2.0 s. Within the renal medulla (averaging all subjects), upslope was 2.8 +/- 1.7 MPV/s, downslope was -0.3 +/- .2 MPV/s, baseline was 39.3 +/- 6.0 MPV, peak was 65.2 +/- 14.3 MPV, time to peak from injection was 20.9 +/- 6.4 s and from time of contrast reaching the kidney was 11.6 +/- 4.1 s. These baseline data may prove useful in the evaluation of dogs with diffuse disease or vascular compromise.     

QUANTITATIVE EVALUATION OF IMAGENT® AS AN ABDOMINAL ULTRASOUND CONTRAST MEDIUM IN DOGS

Diagnostic ultrasound currently provides a noninvasive, economical means to evaluate internal organ structure. Methods to increase the amount of diagnostic information obtained from the sonographic examination are being explored. A new area of research has recently focused on the ability of microbubble contrast agents to help provide additional information. A prospective study was undertaken to quantitatively evaluate the amount of gray scale enhancement of ultrasound images of six canine livers, spleens, and left kidneys that occurs after intravenous injection of Imagent. Computerized analysis was used to assess the percent change in brightness of these images. A transient decrease in the percent change in brightness was seen to occur after injection of higher doses of Imagent. This effect was more pronounced in the deeper portions of the organs of interest. Imagent provides a measurable change in the brightness of the organs of interest and would be potentially useful in a clinical setting.     

CONTRAST‐ENHANCED HARMONIC ULTRASONOGRAPHY OF MEDIAL ILIAC LYMPH NODES IN HEALTHY DOGS

Herein, we describe the normal contrast‐enhanced harmonic, color, and power Doppler ultrasonographic characteristics of the medial iliac lymph nodes in healthy dogs. Contrast‐enhanced harmonic ultrasonography of the medial iliac lymph nodes was performed on 14 healthy dogs after intravenous administration of the lipoprotein‐bound inert gas‐filled microbubble contrast media Definity^®. Time–pixel intensity curves were generated for 1‐min postinjection. Quantification of these curves was performed using Philips QLab software. Non‐contrast‐enhanced power and color Doppler examinations were performed in each node to assess vascular patterns subjectively. Normal lymph nodes exhibited a mean contrast wash‐in phase beginning at 6.3 s from the time of injection with mean peak pixel intensity at 12.1 s. Angioarchitecture was best visualized with contrast‐enhanced harmonic ultrasound compared with power and color Doppler. Normal lymph nodes in dogs have a central artery with a centrifugal and uniform branching pattern. Contrast‐enhanced harmonic ultrasonography is a noninvasive examination that demonstrates improved visibility of the intranodal architecture of healthy medial iliac lymph nodes in dogs compared with conventional, non‐contrast‐enhanced Doppler methods that may have future clinical applications.     

FEASIBILITY OF CONTRAST‐ENHANCED ULTRASOUND‐GUIDED BIOPSY OF SENTINEL LYMPH NODES IN DOGS

Our goal was to develop and validate a technique to identify the sentinel lymph nodes of the mammary glands of healthy dogs with contrast‐enhanced ultrasound, and evaluate the feasibility of obtaining representative samples of a sentinel lymph node under ultrasound guidance using a new biopsy device. Three healthy intact female adult hounds were anesthetized and each received an injection of octafluoropropane‐filled lipid microspheres and a separate subcutaneous injection of methylene blue dye around a mammary gland. Ultrasound was then used to follow the contrast agent through the lymphatic channel to the sentinel lymph node. Lymph node biopsy was performed under ultrasound guidance, followed by an excisional biopsy of the lymph nodes and a regional mastectomy procedure. Excised tissues were submitted for histopathologic examination and evaluated as to whether they were representative of the node. The ultrasound contrast agent was easily visualized with ultrasound leading up to the sentinel lymph nodes. Eight normal lymph nodes (two inguinal, one axillary in two dogs; two inguinal in one dog) were identified and biopsied. Lymphoid tissue was obtained from all biopsy specimens. Samples from four of eight lymph nodes contained both cortical and medullary lymphoid tissue. Contrast‐enhanced ultrasound can be successfully used to image and guide minimally invasive biopsy of the normal sentinel lymph nodes draining the mammary glands in healthy dogs. Further work is needed to evaluate whether this technique may be applicable in patients with breast cancer or other conditions warranting evaluation of sentinel lymph nodes in animals.     

DOPPLER ULTRASOUND OF THE PROSTATE IN NORMAL DOGS AND IN DOGS WITH CHRONIC LYMPHOCYTIC-LYMPHOPLASMOCYTIC PROSTATITIS

The prostate gland of 11 normal dogs and five dogs with histological confirmed chronic lymphocytic or lymphoplasmocytic prostatitis were imaged with grey-scale and Doppler ultrasound. Three vessel types (prostatic artery, capsular artery and parenchymal artery) were identified with color Doppler and the resistive index and maximum and minimum velocities were measured with pulsed wave Doppler. No differences between normal dogs and dogs with prostatitis was identified in either grey-scale ultrasound or in any Doppler parameters measured. Regardless of histologic diagnosis, acepromazine (0.05 mg/kg IV) caused a significant decrease in capsular artery maximum and minimum velocities and prostatic artery resistive index. This study establishes normal Doppler ultrasound parameters for the intact male canine prostate gland. Additional studies are necessary to further evaluate the clinical utility of Doppler ultrasound in canine prostatic diseases.     

IMPROVED DETECTION OF METASTATIC HEPATIC HEMANGIOSARCOMA NODULES WITH CONTRAST ULTRASOUND IN THREE DOGS

Three dogs with a splenic hemangiosarcoma were imaged with conventional gray-scale ultrasound and no lesions were identified in the liver. After administration of intravenous ultrasound contrast medium (Definity) small, poorly enhanced, hypoechoic nodules were identified in the liver in each dog. The spleen and liver lesions were identified at surgery and the dogs underwent splenectomy and nodule biopsy. All lesions were identified histologically as hemangiosarcoma. These preliminary results suggest that contrast ultrasound may result in improved detectability of metastatic hepatic hemangiosarcoma.     

CONTRAST HARMONIC IMAGING OF THE NORMAL CANINE SPLEEN

The purpose of this study was to assess the perfusion pattern and perfusion dynamics in the normal canine spleen using contrast harmonic imaging. Twenty-five dogs without clinical or ultrasonographic evidence of splenic disease were studied. Twenty-three dogs were scanned with only manual restraint; two dogs were sedated with buprenorphin. All dogs received an intravenous bolus of a microbubble contrast medium (SonoVue). The perfusion pattern during the blood pool phase represented a skewed bell-shaped curve. A tissue-specific late phase, similar to humans, was not observed. Time/intensity curves were generated for a selected region. Mean average-derived peak intensity (PI) was 6.6dB, mean time to peak intensity calculated from the initial rise (TTP) was 25.6 s and mean area under the curve (AUC) was 523.6 dBs. If dogs were divided into two body weight groups (< or =15 and >15 kg body weight), average derived peak intensity area, time to peak intensity, and area under the curve were lower for the smaller dogs than for the larger animals. However, differences were not statistically significant (P = 0.2, 0.05, and 0.08, respectively). No significant association was found between hematocrit, hemoglobin concentration, red blood cell count, blood pressure, heart rate, age, gender, and the perfusion variables. In conclusion, these baseline data may prove useful in the evaluation of dogs with diffuse or focal splenic disease.     

TISSUE HARMONIC ULTRASOUND FOR IMAGING NORMAL ABDOMINAL ORGANS IN DOGS AND CATS

Normal cats (n = 38) and dogs (n = 40) were imaged with fundamental ultrasound and tissue harmonic ultrasound. Images of the liver, gall bladder, spleen, left kidney, urinary bladder, and jejunum were collected in all animals. Images of the left adrenal gland were collected in all dogs. All normal cats and dogs had improved imaging with tissue harmonic ultrasound. The number of organs with improved conspicuity ranged from one to all organs imaged. The most common organ to have improved conspicuity was the jejunum (100% of dogs and 89% of cats). Significant improvement by tissue harmonic ultrasound was seen in images of gall bladder (p = .05) and left adrenal gland (p = .02) in dogs, and spleen, urinary bladder, and intestinal images (p = .01) in cats. Significant improvement was seen in tissue harmonic ultrasound images of the gall bladder in dogs weighing greater than 16 kilograms (p = .03) and in the images of the urinary bladder of dogs weighing less than 16 kilograms (p = .02). These data suggest that image quality improvement of normal organs using tissue harmonic ultrasound is consistent but not predictable. The exception was the jejunum, where improvement was seen in all dogs. Sonographers should be cognizant of the potential benefits of tissue harmonic ultrasound.     

PATHOLOGIC CORRELATION OF RESISTIVE AND PULSATILITY INDICES IN CANINE ABDOMINAL LYMPH NODES

We assessed the ability of the resistive index (RI) and pulsatility index (PI) to allow differentiation between normal, reactive, and neoplastic lymph nodes. Forty-seven medial iliac and 54 mesenteric lymph nodes from 83 dogs were evaluated sonographically. A cytologic sample was obtained in each dog that allowed categorization into one of the categories defined above. We found a significant difference in the RI and PI in nonneoplastic vs. neoplastic medial iliac and mesenteric lymph nodes. Values higher than 0.67 for the RI and 1.02 for the PI in medial iliac lymph nodes and higher than 0.76 for the RI and 1.23 for the PI in mesenteric lymph nodes had a high sensitivity and specificity for differentiating benign from neoplastic lymph nodes.     

HARMONIC ULTRASOUND: A REVIEW

Harmonic ultrasound is a technique based on the principle of transmitting at frequency f and receiving at frequency 2f (or 1/2f). This technology has become available through the development of wide-bandwidth transducers. Microbubble contrast media produce a large amount of harmonic signal. Contrast harmonic ultrasound provides the opportunity to image patterns of high flow vasculature and overall perfusion. Regions of poor perfusion, including necrosis or infarction, can be identified with contrast harmonic ultrasound. While proportionately lower, tissues also produce harmonic signals. Tissue harmonic ultrasound sequences often improve subjective image quality compared to fundamental ultrasound in echocardiographic and abdominal examinations. This review will discuss the physical principles of harmonic ultrasound signal generation, medical and animal research applications, and an overview of current veterinary experiences.     

ULTRASONOGRAPHIC EXAMINATION OF ABDOMINAL LYMPH NODES IN THE DOG

Ultrasonography is a valuable diagnostic tool for examination of the abdomen. Normal lymph nodes are often indistinct during routine abdominal ultrasonographic examination. Lymph nodes altered by infiltrative or inflammatory processes may become sonographically visible due to enlargement and/or altered echogeńicity. A working knowledge of the anatomic distribution of lymph nodes and the scheme of lymphatic drainage is important for increasing the sensitivity and specificity of abdominal lymph node assessment during ultrasonographic examination.     

EVALUATION OF EXPERIMENTALLY INDUCED CANINE HEPATIC CIRRHOSIS USING DUPLEX DOPPLER ULTRASOUND

Portal blood flow was measured with duplex Doppler ultrasound in ten normal dogs and in ten dogs with hepatic cirrhosis induced by common bile duct ligation 4 weeks previously. Mean portal blood flow velocity in the 10 dogs with experimentally induced hepatic cirrhosis was markedly reduced (9.2 ± 1.70 cm/sec vs. normal 18.1 ± 7.6 cm/sec, p < 0.01). Mean portal blood flow was also significantly decreased compared to normal (17.2 ± 4.9 cc/min/kg versus normal 31.06 ± 9.1 cc/min/kg, p < 0.01) while portal vein diameter remained unchanged. The dogs with induced hepatic cirrhosis developed extensive extrahepatic portosystemic shunting that was confirmed at necropsy. It was concluded that decreased portal velocity and portal flow which resulted from hepatic cirrhosis was detectable noninvasively with Doppler ultrasound.     

ASSOCIATION BETWEEN MALIGNANCY AND SONOGRAPHIC HETEROGENEITY IN CANINE AND FELINE ABDOMINAL LYMPH NODES

Abdominal lymph node enlargement is frequently noted sonographically. Certain sonographic features can be used to suggest whether lymphadenopathy is more likely benign or malignant. Specific changes in size, shape, echogenicity, and Doppler flow patterns have an association with malignancy. In this retrospective case-control study, the association between abdominal lymph node heterogeneity and malignancy was evaluated. Twenty-three canine and 18 feline patients with ultrasonographically heterogeneous abdominal nodes were evaluated for presence of benign or malignant lymphadenopathy. Controls were animals with lymph node enlargement of uniform echogeneity. Twenty-one (91%) of heterogeneous canine lymph nodes were malignant, and there was a significant association between heterogeneity and malignancy in canine abdominal lymph nodes (P= 0.024). Seven (63%) heterogeneous feline lymph nodes were malignant, with no significant association between heterogeneity and malignancy (P = 0.537).