ULTRASOUND-GUIDED PERCUTANEOUS LIVER BIOPSY IN SNAKES

Percutaneous ultrasound-guided liver biopsies were conducted in 15 snakes using 18-gauge automatic biopsy devices. The liver samples collected ranged in size from 1.16 ± 0.40 mm to 8.3 ± 0.55 mm. Upon histologic examination, all biopsy samples were of diagnostic quality. Anesthesia with complete immobilization of the snake was necessary to allow proper selection of the target site and limit complications associated with biopsy collection. Percutaneous ultrasound-guided liver biopsy is a rapid, minimally invasive method for collecting liver tissue in snakes.     

Ultrasound-guided biopsy

Ultrasound-guided biopsy is a good and feasible technique in dogs. Although we have only used it for biopsy of liver and kidney, it can have further uses. In our use of ultrasound-guided biopsy of kidney and liver, we have found that we can achieve adequate biopsy samples using the Franklin modified Vim-Silverman and Tru Cut biopsy of kidney and liver. The fine-needle biopsies of the liver were adequate for examination of hepatocytes but inadequate to evaluate structure owing to small size of the sample. In contrast, the renal fine-needle samples were both poor in quality and quantity. This technique has the advantages of being noninvasive, quick, and easy, and it can be performed with the patient under local anesthesia. It has an advantage over blind percutaneous biopsy because the needle can be visualized in the organ and the organ scanned after biopsy for possible complications. Another advantage is that, unlike other radiographic biopsy procedures, ionizing radiation is not used for imaging.     

Ultrasound-guided biopsy

Ultrasound-guided biopsy is a good and feasible technique in dogs. Although we have only used it for biopsy of liver and kidney, it can have further uses. In our use of ultrasound-guided biopsy of kidney and liver, we have found that we can achieve adequate biopsy samples using the Franklin modified Vim-Silverman and Tru Cut biopsy of kidney and liver. The fine-needle biopsies of the liver were adequate for examination of hepatocytes but inadequate to evaluate structure owing to small size of the sample. In contrast, the renal fine-needle samples were both poor in quality and quantity. This technique has the advantages of being noninvasive, quick, and easy, and it can be performed with the patient under local anesthesia. It has an advantage over blind percutaneous biopsy because the needle can be visualized in the organ and the organ scanned after biopsy for possible complications. Another advantage is that, unlike other radiographic biopsy procedures, ionizing radiation is not used for imaging.     

Evaluation of hemorrhage, sample size, and collateral damage for five hepatic biopsy methods in dogs

OBJECTIVES: To compare the volume of hemorrhage, number of lobules, and portal triads available for histologic evaluation, and resultant collateral damage between 5 hepatic biopsy methods: biopsy punch, biopsy needle, ligature method, laparoscopic biopsy forceps, and ultrasonically activated scalpel (UAS). STUDY DESIGN: Experimental, repeated measures, block. ANIMALS: Twelve adult dogs. METHODS: Biopsies were obtained from the periphery and center of the left lateral liver lobe of each dog using each of 5 biopsy techniques. Hemorrhage was quantified and compared between methods and sites. Biopsy samples were evaluated histologically to characterize collateral damage and determine the number of lobules and portal triads sampled. RESULTS: Regardless of technique, liver biopsy resulted in minimal hemorrhage (<2 mL). For peripheral biopsies, UAS was comparable with the ligature method, but caused significantly less hemorrhage than all other methods, whereas for central biopsies, UAS caused significantly less hemorrhage than other methods. Except for the laparoscopic biopsy forceps, UAS caused significantly more collateral damage than other methods. UAS and ligature biopsy methods yielded specimens that had more portal triads per sample than other methods. Eight of 48 biopsy needle samples were inadequate for histologic evaluation, whereas other methods yielded adequate specimens. CONCLUSIONS: All biopsy methods produced minimal hemorrhage and except for needle biopsy yielded adequate tissue samples for histologic evaluation. CLINICAL RELEVANCE: Use of UAS is a reliable, safe alternative technique for liver biopsy and can be used laparoscopically to obtain large tissue samples.     

A Comparison of Liver Sampling Techniques in Dogs

Background

The liver sampling technique in dogs that consistently provides samples adequate for accurate histopathologic interpretation is not known.

Hypothesis/Objectives

To compare histopathologic results of liver samples obtained by punch, cup, and 14 gauge needle to large wedge samples collected at necropsy.

Animals

Seventy dogs undergoing necropsy.

Methods

Prospective study. Liver specimens were obtained from the left lateral liver lobe with an 8 mm punch, a 5 mm cup, and a 14 gauge needle. After sample acquisition, two larger tissue samples were collected near the center of the left lateral lobe to be used as a histologic standard for comparison. Histopathologic features and numbers of portal triads in each sample were recorded.

Results

The mean number of portal triads obtained by each sampling method were 2.9 in needle samples, 3.4 in cup samples, 12 in punch samples, and 30.7 in the necropsy samples. The diagnoses in 66% of needle samples, 60% of cup samples, and 69% of punch samples were in agreement with the necropsy samples, and these proportions were not significantly different from each other. The corresponding kappa coefficients were 0.59 for needle biopsies, 0.52 for cup biopsies, and 0.62 for punch biopsies.

Conclusion and Clinical Importance

The histopathologic interpretation of a liver sample in the dog is unlikely to vary if the liver biopsy specimen contains at least 3–12 portal triads. However, in comparison large necropsy samples, the accuracy of all tested methods was relatively low.     

ULTRASOUND-GUIDED BIOPSY OF THE CANINE LIVER, KIDNEY, AND PROSTATE

Sixty-nine hepatic, 25 renal, and 16 prostatic biopsies were performed under ultrasound guidance using a biopsy guide. The majority (98 of 110) were tissue-core biopsies. Multiple attempts at obtaining a sample were required; however, in the kidney, the number of attempts was restricted to two. Adequate samples were obtained in 94% (65/69) hepatic, 88% (22/25) renal, and 94% (5/6) prostatic biopsies. Postbiopsy scanning did not demonstrate parenchymal hemorrhage. In three renal and one prostatic biopsy, gross hematuria, noted immediately following biopsy, resolved in 2–3 days. Animals with prostatic disease frequently had hematuria, making evaluation for this complication difficult. One animal died acutely 2 days following prostatic biopsy due to an unrelated problem, a ruptured aortic àneurysm. Complications were not encountered with the liver biopsy procedure. Animals biopsied under sedation tolerated the procedure well. The authors preferred to anesthetize uncooperative animals and those needing kidney biopsy to minimize the likelihood of complications. Problems encountered during the procedure, overlying bowel gas obscuring the target organ or poor visualization of the biopsy needle, were corrected by changing patient or transducer position or the procedure was postponed. The usefulness of the biopsy procedure is illustrated in four case reports.     

Effects of percutaneous needle liver biopsy on dairy cow behaviour

In cattle, percutaneous needle liver biopsy is used for scientific examination of liver metabolism. The impact of the biopsy procedure is, however, poorly investigated. Our aim was to examine the behaviour of dairy cows during and after liver biopsy. Data were collected from 18 dry cows. Percutaneous needle liver biopsies (after administration of local anaesthesia (2% Procaine)) and blood samples were taken during restraining. During the control treatment, animals were restrained and blood sampled. During the biopsy procedure, cows showed increased restlessness (P=0.008), frequency of head shaking (P=0.016), and decreased rumination (P=0.064). After biopsies, tail pressing (P=0.016) and time spent perching (P=0.058) increased. Time spent upright (P=0.10) and number of leg movements (P=0.033) increased during the night as compared to controls. Thus, liver biopsy induced behavioural changes for up to 19h - and particularly for behaviour previously associated with pain. Even though the exact welfare impact of percutaneous needle liver biopsies in cows is not known, and the magnitude of the behavioural changes was limited, pain always has negative effects on animal welfare. Therefore, if the present biopsy procedure - involving several biopsy passes - is to be used, improvement of the anaesthetic protocol as well as the inclusion of analgesics should be considered.     

Ultrasound-guided serial transabdominal cardiac biopsies in cats

A percutaneous/transdiaphragmatic core needle biopsy technique was developed in cats to obtain serial biopsies from different locations of the left ventricle, through which morphological and molecular changes within the same individual can be studied to unravel the mechanisms of feline cardiomyopathies. Transmural left ventricular myocardial samples were obtained from 29 anesthetized, healthy, adult cats with ultrasound guidance. An 18G automatic biopsy needle was inserted between the last left rib and the sternum through the diaphragm into the thorax. Biopsies were obtained from the left ventricular wall. In five cats, three single biopsies were taken with 4-week intervals. Autopsy was performed on six cats, of which three cats had serial biopsies. In total, 87 biopsies were obtained without long-term effects on cardiac function or structure. The biopsies caused transient single ventricular premature complexes and mild pericardial effusion without tamponade. Necropsy revealed a minimal amount of fibrous connective tissue in the diaphragm and the heart without any significant microscopic lesions in the adjacent muscle tissue. The high quality biopsy material was suitable for morphological and molecular studies. This minimally invasive, ultrasound-guided cardiac biopsy technique thus allows for the safe collection of serial biopsies to study feline cardiomyopathies in an experimental setting.     

COMPARISON OF GLOMERULAR NUMBER AND SPECIMEN LENGTH OBTAINED FROM 100 DOGS VIA PERCUTANEOUS ECHO-ASSISTED RENAL BIOPSY USING TWO DIFFERENT NEEDLES

Our objective was to evaluate possible differences in the number of glomeruli and length of renal biopsies collected in canine subjects by two different types of biopsy needles: a semiautomatic 18-gauge Trucut and an automated 18-gauge Jamshidi modified (Biopince). One hundred biopsy samples obtained from dogs of different ages and gender affected by different nephropathies were evaluated retrospectively. All animals were biopsied using one of the two different needles. Biopsies were performed under ultrasound guidance and evaluated by a single pathologist. Statistical analysis was performed to evaluate possible differences in the number of glomeruli and length of renal biopsies collected comparisons were determined between subgroups of dogs with or without the identification of renal interstitial infiltrates and/or fibrosis. Neither the mean difference of the number of glomeruli nor the length of tissue sample collected with the different needles was significantly different. Likewise, the average biopsy length did not differ in dogs with or without renal interstitial infiltrate in animals biopsied with either biopsy needle. Both the Biopince and the Trucut devices provide diagnostically adequate biopsy renal specimens using ultrasound-guidance.     

Needle biopsy for hepatic vitamin A levels in lions (Panthera leo)

Hypovitaminosis A (HA)-related skull malformations resulting in neurologic abnormalities and death have been, and still are, reported in captive lions (Panthera leo) worldwide. Liver vitamin A (VA) concentration is the most reliable indicator of animals' VA status, and its assessment is essential in prevention and treatment of HA in lions. A percutaneous needle liver biopsy using high-performance liquid chromatography ultraviolet retinoid analysis for VA concentration measurement was validated. It was first assessed in vitro using chicken liver. Later, the safety and feasibility of ultrasound-guided percutaneous needle liver biopsy was assessed in living lions. Hepatic VA concentrations in lion liver were measured using the above laboratory method. Mean chicken hepatic VA concentration in needle biopsy (NB) and wedge biopsy (WB) of the same liver lobes were 108.66 and 60.89 microg/g wet tissue, respectively, and were significantly (P = 0.03) correlated (r = 0.74). The calculated linear regression for predicting VA concentration in WB using NB VA for chicken liver was 25.194 + 0.3234x NB (microg/g). Four ultrasound-guided percutaneous needle liver biopsies were obtained from each of the four lions under general anesthesia. Mean hepatic VA concentration was 8.25 microg/g wet tissue (range 1.43-25.29 microg/g). Mean serum VA concentration, measured in these four lions was 1,011.1 nmol/L with a standard deviation of 337.91 nmol/L (range 590.26-1,077.2 nmol/L). The lions recovered uneventfully, and no complications were observed during a 4-yr follow-up period. In conclusion, the percutaneous needle liver biopsy technique is a reliable, practical, safe tool for obtaining liver tissue samples antemortem for assessment of the VA status in lions and can be used in future studies.     

Fine-needle aspirate cytology suggesting hepatic lipidosis in four cats with infiltrative hepatic disease

Four cats are reported in which cytology smears obtained by ultrasound-guided fine needle aspiration of the liver were interpreted as indicative of hepatic lipidosis. However, histopathology of hepatic tissue samples obtained with Tru-Cut-like needles or wedge biopsy revealed that the cats had inflammatory or neoplastic hepatic disease causing their clinical signs. Fine needle aspiration and cytology may not detect infiltrative lesions, particularly those that are nodular, multifocal, or localised around the portal regions. Fine needle aspirate cytology is a useful diagnostic procedure with many advantages, but care must be taken to avoid diagnosing hepatic lipidosis as the cause of illness when an infiltrative lesion is responsible.     

Pilot evaluation of a vacuum-assisted biopsy instrument for percutaneous renal biopsy in dogs

Kidney biopsies in dogs are commonly obtained using automated spring-loaded biopsy instruments. Interpretation of biopsies from dogs with glomerular disease requires examination of at least 5-10 glomeruli, with at least two biopsies usually required for full evaluation. The purpose of this study was to compare quality and interpretability of renal biopsies obtained from healthy dogs with a large-gauge, vacuum-assisted biopsy instrument versus two biopsies obtained with a spring-loaded biopsy needle. Twenty dogs were randomized into two groups, and percutaneous, ultrasound-guided renal biopsies were evaluated using standard criteria. There were no significant differences in the number of biopsies that contained renal tissue, cortex, or medulla. Biopsies obtained with either instrument contained an adequate number of glomeruli and an equivalent number of arterioles and severity of tissue compression. Differences included easier penetration of the renal capsule and collection of sufficient tissue for interpretation with only one instrument pass when using the vacuum-assisted device (vs two passes required with the spring-loaded instrument). Before use in client-owned dogs, future studies should evaluate whether these differences are clinically relevant advantages in the diagnostic evaluation of dogs with kidney disease, and determine the prevalence and severity of complications when using this larger gauge device.     

Correlation of ultrasonographic appearance of lesions and cytologic and histologic diagnoses in splenic aspirates from dogs and cats: 32 cases (2002-2005)

OBJECTIVE: To determine the accuracy of cytologic diagnosis, compared with histologic diagnosis, in determination of disease in ultrasound-guided fine-needle aspirates of splenic lesions. DESIGN: Retrospective study. SAMPLE POPULATION: Splenic specimens from 29 dogs and 3 cats. PROCEDURES: Records were searched for dogs and cats that had undergone ultrasound-guided splenic aspiration. Criteria for inclusion were ultrasonographic identification of splenic lesions and cytologic and histologic evaluation of tissue from the same lesion. Cytologic samples were obtained by fine-needle aspiration, and histologic specimens were obtained via surgical biopsy, ultrasound-guided biopsy, or necropsy. RESULTS: Cytologic diagnoses corresponded with histologic diagnoses in 19 of 31 (61.3%) cases and differed in 5 of 31(16.1%) cases, and 1 aspirate was inadequate for evaluation. In 7 of 31 (22.6%) cases, histologic evaluation of tissue architecture was required to distinguish between reactive and neoplastic conditions. On the basis of histologic diagnosis in 14 animals with nonneoplastic conditions, the cytologic diagnosis was correct in 11 cases, not definitive in 2 cases, and incorrect in 1 case. In 17 animals with malignant neoplastic diseases, the cytologic diagnosis was correct in 8 cases, not definitive but consistent with possible neoplasia in 5 cases, and incorrect in 4 cases. Multiple similar-appearing nodules were significantly associated with malignancy, whereas single lesions were more often benign. CONCLUSIONS AND CLINICAL RELEVANCE: Ultrasound-guided aspiration of splenic lesions is a minimally invasive tool for obtaining specimens for cytologic evaluation. Although cytologic diagnoses often reflect histologic results, if missampling or incomplete sampling occurs or tissue architecture is required to distinguish between reactive and neoplastic conditions, accurate diagnosis with fine-needle aspiration may not be possible.     

Effect of sample quality on the sensitivity of endoscopic biopsy for detecting gastric and duodenal lesions in dogs and cats

Background: The quality of histopathology slides of endoscopic biopsies from different laboratories varies, but the effect of biopsy quality on outcome is unknown.
Hypothesis: The ability to demonstrate a histologic lesion in the stomach or duodenum of a dog or cat is affected by the quality of endoscopic biopsy samples submitted. More endoscopic samples are needed to find a lesion in poor-quality tissue specimens.
Animals: Tissues from 99 dogs and 51 cats were examined as clinical cases at 8 veterinary institutions or practices in 5 countries.
Methods: Histopathology slides from sequential cases that underwent endoscopic biopsy were submitted by participating institutions. Quality of the histologic section of tissue (inadequate, marginal, adequate), type of lesion (lymphangiectasia, crypt lesion, villus blunting, cellular infiltrate), and severity of lesion (normal, mild, moderate, severe) were determined. Sensitivity of different quality tissue samples for finding different lesions was determined.
Results: Fewer samples were required from dogs for diagnosis as the quality of the sample improved from inadequate to marginal to adequate. Duodenal lesions in cats displayed the same trend except for moderate duodenal infiltrates for which quality of tissue sample made no difference. Gastric lesions in dogs and mild gastric lesions in cats had the same trend, whereas the number of tissue samples needed to diagnose moderately severe gastric lesions in cats was not affected by the quality of tissue sample.
Conclusions and Clinical Importance: The quality of endoscopically obtained tissue samples has a profound effect on their sensitivity for identifying certain lesions, and there are differences between biopsies of canine and feline tissues.     

A COMPARISON OF MANUAL AND AUTOMATIC ULTRASOUND-GUIDED BIOPSY TECHNIQUES

A manual, ultrasound-guided biopsy technique was compared to a new automatic method using a new biopsy device. The liver and the left kidney of 10 dogs were biopsied using the two methods. The specimens from the biopsies were compared for size and quality. Results demonstrate that larger and higher quality samples were obtained with the new automatic method.     

Diagnostic accuracy of pre‐treatment biopsy for grading soft tissue sarcomas in dogs

Histologic grade is an important prognostic factor for both local recurrence and metastatic potential with canine soft tissue sarcoma (STS). Pre‐treatment biopsy with identification of tumour grade may aid in prognostication and determination of surgical margins necessary for local control. The purpose of this study was to evaluate the grading accuracy of various pre‐treatment biopsy techniques (wedge, punch, needle‐core) for STS in dogs. Medical records of 68 dogs diagnosed with a STS via pre‐treatment biopsy and confirmed by excisional biopsy were evaluated. The concordance in grade between excisional and pre‐treatment biopsies was 59%. Of the 41% that lacked concordance, 29% of pre‐treatment biopsies underestimated and 12% overestimated grade. The method of pre‐treatment biopsy did not significantly effect grade concordance. Based on these data, needle‐core biopsy appears to be similar in accuracy compared to open biopsy, however, grading determined by pre‐treatment biopsy in general should be interpreted with caution.     

Effects of serial ultrasound-guided renal biopsies on kidneys of healthy adolescent dogs

Ten healthy mixed-breed dogs were used to evaluate the functional and structural effects of serial ultrasound-guided renal biopsies obtained with an automated biopsy needle. In each dog, one lateral renal cortex was biopsied at 2, 4, and 6 months of age; the other kidney was the control. Five dogs had two tissue cores and five dogs had four tissue cores taken on each biopsy occasion, and one core was examined microscopically. One week before each biopsy and a month after the final biopsy, the glomerular filtration rate (GFR) was determined by renal scintigraphy. Dogs were then euthanized for evaluation of gross and microscopic lesions attributable to the biopsies. There was no difference between GFR values for biopsied kidneys and those of control kidneys ( P >0.05). Microscopic lesions were not identified in biopsies taken at 2 and 4 months, but focal lesions were found in three of 10 specimens taken at 6 months of age. At necropsy, six of 10 biopsied kidneys had small visible capsular scars, and linear tracts <2 mm wide were observed on cut surfaces in six of 10 biopsied kidneys cut transversely into slices 5 mm thick. Discrete light microscopic lesions were observed in 25 of 452 (5.5%) of randomly selected 6-mm-diameter sections of renal cortex from biopsied kidneys. We conclude that serial renal cortical biopsies can be obtained by our method from healthy adolescent dogs with minimal risk of inducing changes that might be confused with those of a progressive renal disease.     

Usefulness of Ki-67 proliferation marker in the cytologic identification of liver tumors in dogs

BACKGROUND: Performing a biopsy is currently the best method of diagnosing liver disease. To reduce possible risk factors resulting from a biopsy, liver cytology can provide an alternative technique. The diagnostic accuracy of cytology for identifying liver tumors is, however, limited. The results of cytology might be improved by using immunochemistry for Ki-67, a proliferation marker, on liver cytology specimens. OBJECTIVES: The purpose of this study was to investigate the value of Ki-67 immunochemistry on liver cytologic specimens from dogs for identifying neoplastic diseases of the liver, by comparing the results to histologic findings. METHODS: Liver biopsy and cytology samples were obtained from 30 dogs with hepatic disease. All samples were evaluated by an anatomic pathologist and a cytopathologist. Parallel Ki-67 immunochemistry of histologic and cytologic samples was performed. The gradation of Ki-67 expression in histologic and cytologic samples was assessed. RESULTS: Cytologic specimens of liver tumors (n = 9) showed <50% Ki-67-positive cells. Twenty of 21 cases of non-neoplastic liver disease had no or few single Ki-67-positive cells. Using Ki-67, the diagnostic accuracy of cytologic evaluation was increased from 78% to 100% for malignant neoplasia. CONCLUSIONS: Based on the results of this study, the cytologic evaluation of liver together with Ki-67 immunochemistry can improve the diagnostic accuracy of cytology for liver neoplasia.     

ULTRASOUND LOCALIZATION AND GUIDANCE FOR RENAL BIOPSY IN THE HORSE

Real-time ultrasonography was used to assist percutaneous renal biopsies in seven normal horses. Ultrasound (US) guidance and blind biopsy following US localization were compared. Ten biopsies (seven right kidney and three left kidney) were performed. A transverse approach to the lateral border of the right kidney through the 17th intercostal space was favored, with the needle angle being approximately perpendicular to the body wall. Both biopsy techniques were effective, but blind biopsy following US localization was favored as it was easily performed by one operator without the need for specialized biopsy equipment. The spleen had a variable location and its penetration with the biopsy needle was necessary to obtain two left kidney biopsies. Macroscopic and microscopic hematuria was observed postbiopsy in one and four horses, respectively, out of six horses examined. Perirenal hemorrhage was a prominent finding on necropsy in five of six horses euthanized.     

Measurement of the status of trace elements in cattle using liver biopsy samples

Serum, plasma, or urine samples are usually used for the measurement of the trace elements copper; zinc, iron, selenium, because these samples are easy to obtain; however; these samples are not always appropriate. For example, it is not possible to measure molybdenum, the major antagonist of copper; in blood or urine. Therefore measurement of trace elements in liver tissue is considered the gold standard. For the assessment of selenium the method of choice remains determination of glutathion peroxidase in erythrocytes and for the assessment of magnesium determination of magnesium in urine. We determined the accuracy and repeatability of measuring trace elements in liver biopsies and whole liver homogenates. The levels of trace elements measured were similar in both preparations (92% agreement). Liver biopsy in live animals is a relatively simple procedure but not common in The Netherlands. Reference levels of trace elements, classified as too low, low, adequate, high, and too high, were established on the basis of our research and information in the literature. In a second study we investigated the practical aspects of obtaining liver tissue samples and their use. Samples were collected from cattle on a commercial dairy farm. Liver biopsy provided additional information to that obtained from serum and urine samples. We prepared a biopsy protocol and a test package, which we tested on 14 farms where an imbalance of trace minerals was suspected. Biopsy samples taken from 4 to 6 animals revealed extreme levels of trace elements.