Use of high-frequency ultrasonography for evaluation of skin thickness in relation to hydration status and fluid distribution at various cutaneous sites in dogs

OBJECTIVE: To assess the usefulness of high-frequency diagnostic ultrasonography for evaluation of changes of skin thickness in relation to hydration status and fluid distribution at various cutaneous sites in dogs. ANIMALS: 10 clinically normal adult dogs (6 males and 4 females) of various breeds. PROCEDURES: Ultrasonographic examination of the skin was performed before and after hydration via IV administration of an isotonic crystalloid solution (30 mL/kg/h for 30 minutes). A 13-MHz linear-array transducer was used to obtain series of ultrasonographic images at 4 different cutaneous sites (the frontal, sacral, flank, and metatarsal regions). Weight and various clinicopathologic variables (PCV; serum osmolality; and serum total protein, albumin, and sodium concentrations) were determined before and after the infusion. These variables and ultrasonographic measurements of skin thickness before and after hydration were compared. RESULTS: Among the 10 dogs, mean preinfusion skin thickness ranged from 2,211 microm (metatarsal region) to 3,249 microm (sacral region). Compared with preinfusion values, weight was significantly increased, whereas PCV; serum osmolality; and serum total protein, albumin, and sodium concentrations were significantly decreased after infusion. After infusion, dermal echogenicity decreased and skin thickness increased significantly by 21%, 14%, 15%, and 13% in the frontal, sacral, flank, and metatarsal regions, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Cutaneous site and hydration were correlated with cutaneous characteristics and skin thickness determined by use of high-frequency ultrasonography in dogs. Thus, diagnostic ultrasonography may be a useful tool for the noninvasive evaluation of skin hydration in healthy dogs and in dogs with skin edema.     

Bacterial population and histologic changes in dogs with perianal fistula

Ages of 44 dogs with perianal fistula, ranged from 6 months to 13 years (mean, 5.2 years). German Shepherd Dogs and Irish Setters were statistically (P less than 0.01) over-represented compared with those breeds in a canine hospital population (n = 22,047) for the same period. There was a 2:1 male-to-female ratio, with 38 (86.4%) of dogs sexually intact and 6 (13.6%) of dogs neutered. Eleven types of bacterial organisms were recovered from deep perianal tissues of which Escherichia coli, Staphylococcus aureus, beta-hemolytic streptococci, and Proteus mirabilus were most common. Organisms were not recovered from 7 dogs. Of 93 isolates, 88.3% were susceptible to gentamicin, 80.5% to cephalothin, 79.2% to chloramphenicol, and 74% to trimethoprim-sulfamethoxazole. Fifty-one biopsy specimens from 44 dogs were classified as having early, intermediate-, and late-stage lesions based on the amount of fibrosis, severity of the inflammatory response, and, if present, depth of sinus tracts. In most biopsy specimens, all 3 stages were represented in the same histologic section. In 45 specimens, most inflamed lesions were in the dermis of the zona cutanea. Hidradenitis was present in 22 biopsy specimens and was associated with the formation of epithelial-lined sinus tracts.     

Histologic comparison of canine skin biopsies collected using monopolar electrosurgery, CO2 laser, radiowave radiosurgery, skin biopsy punch, and scalpel

OBJECTIVE: To compare the histologic appearance of canine skin biopsies collected by use of a scalpel, skin biopsy punch, monopolar electrosurgery, CO2 laser, and radio wave radiosurgery in fully rectified wave form (RWRS). STUDY DESIGN: Experimental, randomized design. ANIMALS: Healthy adult grayhounds (n=4). METHODS: Skin biopsies were collected using 5 techniques. Cut margins of biopsy specimens and adjacent peripheral skin were evaluated using light microscopy to compare penetration of the dermis by tissue carbonization (char). RESULTS: No char occurred in skin specimens collected by biopsy punch and scalpel. Char penetration occurred in all specimens collected by electrosurgery, CO2 laser, and RWRS. Mean char penetration in skin biopsies collected by RWRS (0.158 mm) was significantly less than for monopolar electrosurgery (0.223 mm) and CO2 laser (0.215 mm). Mean char penetration in adjacent peripheral skin surrounding biopsies collected by RWRS (0.171 mm) was significantly less than monopolar electrosurgery (0.255 mm) but not less than CO2 laser (0.215 mm, P<.07). CONCLUSIONS: RWRS (blended waves in cut-coagulate mode) caused less lateral thermal damage to canine skin biopsies than monopolar electrosurgery and CO2 laser and less lateral thermal injury to peripheral skin than monopolar electrosurgery. CLINICAL RELEVANCE: Excision of canine skin biopsies with heat-generating devices may not allow reliable histologic interpretation, particularly when assessing margins of small biopsy specimens. RWRS may be less traumatic to canine skin than monopolar electrosurgery and CO2 laser when used to make incisions.     

Comparison of B-mode and Doppler ultrasonographic findings with histologic features of benign and malignant mammary tumors in dogs

OBJECTIVE: To compare and correlate B-mode and color Doppler ultrasonographic characteristics with the histologic findings of benign and malignant mammary tumors in dogs. STUDY POPULATION: 49 mammary tumors in 26 dogs. PROCEDURES: Before excision, tumors were evaluated via B-mode and color Doppler ultrasonography to assess size, echogenicity, echopattern, acoustic transmission, invasiveness, and vascularity. Paraffin-embedded microsections of the tumors were stained with H&E and examined for presence of necrosis, cysts, cartilage, bone, mineralization, invasion of surrounding tissue, and tissue heterogeneity. To assess vascularity, the number and distribution of vessels that were stained by the Verhoeff van Gieson technique were recorded. RESULTS: Tumor echogenicity and echopattern on ultrasonographic images correlated with tissue heterogeneity detected histologically. Acoustic enhancement was correlated with the presence of necrotic or cystic areas. Tumor invasion into surrounding tissues as determined ultrasonographically did not correlate with the histologic findings. There was a significant correlation between the number of detected vessels and distribution of flow within the tumors determined via ultrasonographic and histologic examinations. CONCLUSIONS AND CLINICAL RELEVANCE: In canine mammary tumors, ultrasonographic characteristics appear to be correlated with histopathologic changes. Data suggest that ultrasonography may have an important role in the evaluation of mammary tumors in dogs, particularly in the evaluation of tissue composition and tumor vascularity.     

Biochemical evaluation of mitochondrial respiratory chain enzymes in canine skeletal muscle

OBJECTIVE: To perform respiratory chain enzymatic activity assays on canine skeletal muscle biopsy specimens and establish reference range values of skeletal muscle enzyme activities for dogs. SAMPLE POPULATION: Biopsy specimens from the vastus lateralis muscle were obtained from 24 dogs (8 sexually intact males and 14 sexually intact females) ranging from 15 months to 6 years of age. PROCEDURE: Mean values of citrate synthase, cytochrome-c oxidase, succinate dehydrogenase, succinate dehydrogenase-cytochrome-c reductase, nicotinamide adenine dinucleotide (NADH) dehydrogenase, and NADH dehydrogenase-cytochrome-c reductase activities were established by use of 6 standard spectrophotometric assays for respiratory chain enzyme analysis. RESULTS: Compared with published data for skeletal muscle enzyme activities in humans, skeletal muscle enzyme activities in dogs were 2- to 4-fold higher. Additionally, citrate synthase activity, a marker for mitochondrial volume, was positively correlated with age in dogs, suggesting that mitochondrial volume increases with age, although no apparent change in respiratory chain enzymatic activity with an increase in age was found. CONCLUSIONS AND CLINICAL RELEVANCE: Reference range values for skeletal muscle enzyme activities of dogs are needed to accurately interpret results of respiratory chain enzymatic activity assays. During investigation of metabolic myopathies, if skeletal muscle biopsy specimens are evaluated for respiratory chain enzyme kinetics, they should be performed and evaluated in concert with skeletal muscle biopsy specimens from clinically normal animals of the same species.     

Glomerular lesions in dogs infected with Leishmania organisms

OBJECTIVE: To histologically identify glomerular lesions in dogs infected with Leishmania organisms. ANIMALS: 41 dogs (17 sexually intact males and 14 sexually intact and 10 ovariohysterectomized females) that had positive results when tested for leishmaniosis as determined by use of serologic evaluation (indirect fluorescent antibody test, titers of 1:80 to 1:640) and direct microscopic identification of the protozoal organisms. PROCEDURE: Urine samples were collected by use of cystocentesis and examined by qualitative SDS-agarose gel electrophoresis (AGE). All dogs had non-selective (glomerular) or mixed (glomerular and tubular) proteinemia. Specimens were obtained from each dog during ultrasound-assisted renal biopsy and used for histologic examination. Each specimen was stained with H&E, periodic acid-Schiff, Goldner's trichrome, methenamine silver, and Congo Red stains. Specimens were adequate for evaluation when they contained at least 5 glomeruli/section, except for specimens stained with Congo Red in which 1 glomerulus/section was adequate. RESULTS: Examination of renal biopsy specimens revealed various glomerular lesions in all dogs and interstitial or tubular (or both) lesions in 23 of 41 (55%) dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Glomerular lesions that develop in dogs during infection with Leishmania organisms can be classified histologically as mesangial glomerulonephritis, membranous glomerulonephritis, membranoproliferative glomerulonephritis, and focal segmental glomerulonephritis. Tubulointerstitial histopathologic conditions were not observed as the primary lesion, despite being evident in 23 of 41 (55%) dogs. Use of SDS-AGE for qualitative evaluation of proteinuria and successive collection of specimens during renal biopsies following diagnosis of nonselective glomerular proteinuria provides the possibility for early identification of renal lesions.     

Ultrasonographic findings in dogs with hyperammonemia: 90 cases (2000-2002)

OBJECTIVE: To determine ultrasonographic abnormalities in dogs with hyperammonemia. DESIGN: Retrospective study. ANIMALS: 90 client-owned dogs with hyperammonemia. PROCEDURE: Ultrasonography of the abdominal vessels and organs was performed in a systematic way. Dogs in which the ultrasonographic diagnosis was a congenital portosystemic shunt were included only if they underwent laparotomy or necropsy. Dogs in which the abdominal vasculature appeared normal and dogs in which the ultrasonographic diagnosis was acquired portosystemic shunts and portal hypertension were included only if liver biopsy specimens were submitted for histologic examination. RESULTS: Ultrasonography excluded portosystemic shunting in 11 dogs. Acquired portosystemic shunts were found in 17 dogs, of which 3 had arterioportal fistulae and 14 had other hepatic abnormalities. Congenital portosystemic shunts were found in 61 dogs, of which 19 had intrahepatic shunts and 42 had extrahepatic shunts. Intrahepatic shunts originated from the left portal branch in 14 dogs and the right portal branch in 5. Extrahepatic shunts originated from the splenic vein, the right gastric vein, or both and entered the caudal vena cava or the thorax. Ultrasonography revealed splenic-caval shunts in 24 dogs, right gastric-caval shunts in 9 dogs, splenic-azygos shunts in 8 dogs, and a right gastric-azygos shunt in 1 dog. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that ultrasonography is a reliable diagnostic method to noninvasively characterize the underlying disease in dogs with hyperammonemia. A dilated left testicular or ovarian vein was a reliable indicator of acquired portosystemic shunts.     

Assessment of proliferative activity of canine dermal mast cells by bromodeoxyuridine and proliferating cell nuclear antigen labelling

Cutaneous mast cells from skin biopsies of three healthy dogs and three dogs with atopic dermatitis were assessed for their proliferative potential using bromodeoxyuridine and proliferating cell nuclear antigen labelling. Mast cells isolated from the skin of two healthy dogs were also studied using bromodeoxyuridine labelling. Mast cells in skin biopsy specimens and mast cells isolated from the skin of healthy dogs did not incorporate bromodeoxyuridine. Two mast cells expressing proliferating cell nuclear antigen were seen around two superficial vessels in the dermis of one atopic dog. Epidermal cells, glandular epithelial cells, fibroblasts and endothelial cells incorporated bromodeoxyuridine and showed positive staining for proliferating cell nuclear antigen. These results suggest that canine mature mast cells do not proliferate in the dermis.     

Histologic comparison of skin biopsy specimens collected by use of carbon dioxide or 810-nm diode lasers from dogs

OBJECTIVE: To compare histologic artifacts caused by carbon dioxide (CO2) or 810-nm diode surgical lasers used to obtain small biopsy specimens of skin from healthy dogs. DESIGN: Prospective study. ANIMALS: 4 dogs. PROCEDURE: 21 skin biopsy specimens were collected from each dog. Three biopsy specimens were obtained with a CO2 or an 810-nm diode laser at 3 operating settings each, and 3 biopsy specimens were obtained with a 6-mm biopsy punch instrument (controls). After processing, biopsy specimens were examined for artifacts related to laser-tissue interactions. Microscopically visible char was measured from the lateral edge of each specimen obtained with a laser. RESULTS: There were no significant differences among mean char distances in biopsy specimens obtained with the CO2 laser at various settings. Mean char distance was significantly greater in all skin biopsy specimens obtained with the diode laser, compared with those obtained with the CO2 laser. Mean char distance was significantly greater in biopsy specimens obtained with the 810-nm diode laser at high power, compared with biopsy specimens obtained with the 810-nm diode laser at low power. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that the CO2 laser caused less thermal injury at margins of skin biopsy specimens; therefore, if a surgical laser is used for removal of cutaneous masses or to obtain skin biopsy specimens, use of the CO2 laser is recommended. Veterinarians performing a biopsy by using a surgical laser should be aware that laser-induced artifacts may render small biopsy specimens useless for providing accurate histologic diagnosis.     

Evaluation of papillomaviruses associated with cyclosporine-induced hyperplastic verrucous lesions in dogs

OBJECTIVE: To determine whether cyclosporine A-induced hyperplastic skin lesions of dogs were associated with papillomavirus infections. ANIMALS: 9 dogs that were treated with cyclosporine A and developed hyperplastic skin lesions. PROCEDURE: History and clinical and histopathologic data were collected. Paraffin-embedded skin biopsy specimens from hyperplastic skin lesions were immunostained for common papillomavirus genus-specific structural antigens by use of a polyclonal rabbit anti-bovine papillomavirus type 1 antiserum. Sections from each tissue block underwent DNA extraction, and polymerase chain reaction (PCR) assays were performed with several sets of primers to amplify a wide range of papillomavirus DNA from humans and other animals. RESULTS: In 7 of 9 dogs, there were more than 10 hyperplastic skin lesions that microscopically resembled those of psoriasiform lichenoid dermatosis. In those dogs, results of testing for papillomavirus via immunohistochemical analyses and PCR assays were negative. In the other 2 dogs, there were only 1 and 3 verrucous lesions, and in those dogs, histologic evaluation revealed koilocytes and nuclear viral inclusions that were immunoreactive for papillomavirus antigens. Papillomavirus DNA was amplified from both dogs. One of the sequences was characteristic for the canine oral papillomavirus, whereas the other had similarities with the recently described canine papillomavirus 2. CONCLUSIONS AND CLINICAL RELEVANCE: In dogs, hyperplastic skin lesions occasionally develop during treatment with cyclosporine A. Most of the lesions resemble those of psoriasiform lichenoid dermatosis, although papillomavirus can be detected in some instances.     

Microscopic anatomy of the skin of the woodchuck (Marmota monax): comparison of woodchuck hepatitis virus-infected and non-infected animals.

Thirty-three woodchucks were used in this study. Seventeen animals were healthy adults, not infected with woodchuck hepatitis virus (WHV); 10 were healthy adults infected with WHV; 4 were noninfected neonates; 2 were infected neonates. Within the 4 groups of woodchucks, no histologic differences were detected on the basis of sex or age. Neither were histologic findings different between infected and noninfected woodchucks of similar ages. The average thickness of skin (as measured from the skin surface to the inner limit of the dermis) from the general haired body area was 2394 microns. The skin was thickest on dorsal body areas, and gradually became thinner on ventral body and medial limb areas. The epidermis consisted of 4 layers: stratum basale, stratum spinosum, stratum granulosum, and stratum corneum. A stratum lucidum was present only in the epidermis of the footpads. There was no clear distinction between the superficial dermis and the deep dermis, except for the subtle differences in arrangement and size of collagen fibers. Elastic fibers were seen throughout the dermis, being more prominent in the superficial portion. Both compound and simple hair follicle arrangements were seen, with compound being more common. The arrectores pilorum muscles were largest in the skin over the dorsal body areas. Sebaceous glands were present either within the outer root sheath of hair follicles or in the dense connective tissue surrounding hair follicles. No apocrine sweat glands were found. However, there were abundant eccrine sweat glands in the subcutaneous fat of the footpads.     

ULTRASONOGRAPHY OF CANINE GASTRIC EPITHELIAL NEOPLASIA

Signalment, clinical and ultrasonographic findings from 16 dogs with histologically confirmed gastric epithelial neoplasia were reviewed. The most common clinical findings were vomiting, anorexia and weight loss. Hematemesis and melena were uncommon findings. Ten (10/16) dogs were female and there were four Chows. The most common ultrasonographic findings were transmural thickening of the gastric wall associated with altered wall layering. A poorly echogenic lining often was noted on the innermost and/or the outermost portions of the gastric wall, separated by a more echogenic central zone. The distribution of these changes was variable. This ultrasonographic feature, called pseudolayering, was present in 14 dogs. It was believed to most likely correlate to the unevenly layered tumor distribution noted histopathologically. The maximum wall thickening ranged from 1 cm to 2.7 cm. Regional lymphadenopathy was identified ultrasonographically in 15 dogs. Ultrasound-guided fineneedle aspiration biopsy or automated microcore biopsy, endoscopic biopsy, surgical biopsy or necropsy resulted in the diagnosis of carcinoma in 15 dogs and of carcinoid tumor in one dog. The results of this study suggested that ultrasonography was a useful tool for the detection and diagnosis of canine gastric epithelial neoplasia. Furthermore, ultrasonography can assist in obtaining diagnostic samples and in clinical staging of the tumor.     

ULTRASONOGRAPHIC FINDINGS IN CANINE GASTRIC NEOPLASIA IN 13 PATIENTS

The ultrasonographic findings in 13 canine patients with histopathologically proven gastric tumors were reviewed. The echogenicity of gastric wall lesions was variable and independent of the type of tumor. All dogs had an increase in gastric wall thickness and in only one dog was the wall layering intact. Regional lymph node involvement was a common finding, but abnormalities within the liver were seen in only one patient. Gastric neoplasia was observed most commonly in middle-aged and old, medium- to large-sized dogs. Carcinoma was the most common tumor found. In this study, with a limited number of animals and limited tumor types, it appeared that thickening of the gastric wall, accompanied by loss of gastric wall layering and enlargement of regional lymph nodes were ultrasonographic signs likely to be found with gastric neoplasia. In this limited study the histologic type of gastric tumor could not be predicted on the basis of the ultrasound examination.     

Comparison of B-mode and Doppler ultrasonographic findings with histologic features of benign and malignant superficial lymph nodes in dogs

OBJECTIVE: To compare and correlate B-mode and color Doppler ultrasonographic characteristics with histopathologic findings of benign and malignant superficial lymph nodes in dogs. STUDY POPULATION: 50 superficial lymph nodes that were normal, abnormally large on physical examination, or represented regional lymph nodes draining an area of suspected primary malignancy in 30 dogs. PROCEDURES: Before excision, lymph nodes were evaluated via B-mode and color Doppler ultrasonography to assess size, echogenicity, presence of a hilus, acoustic transmission, and vascular flow. Formalin-fixed, paraffin-embedded tissue sections of excised lymph nodes were stained with H&E and examined for the presence and extent of necrosis, fibrosis, fat, metastases, and tissue heterogeneity. To assess vascularity, the number and distribution of vessels stained by the Verhoeff van Gieson technique were recorded. RESULTS: In superficial lymph nodes, a varied echogenicity corresponded to tissue heterogeneity. The ultrasonographic detection of a hilus was associated with the presence of fibrous tissue, fat, or both in the hilar region. Acoustic enhancement corresponded to presence of areas of intranodal necrosis. There was significant correlation between both the distribution and the number of vessels detected via ultrasonography and that detected by histopathology. The amount of flow estimated via ultrasonography was typically higher than that estimated via histologic examination. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that histopathologic changes in canine lymph nodes have associated ultrasonographic changes and suggest that lymph node ultrasonography has an important role in the evaluation of lymph nodes in dogs in general and in dogs with neoplastic disease in particular.     

ULTRASONOGRAPHIC EVALUATION OF THE CANINE SHOULDER

The aim of this study was to determine the normal ultrasonographic anatomy of the canine shoulder. Fourteen shoulders from 7 clinically normal mid-sized dogs were radiographed and imaged using high frequency ultrasound. Each shoulder was isolated postmortem, and the ultrasonographic and gross anatomy was studied during dissection. The ultrasonographic appearance of the shoulder specimens was similar to that found in the live dogs. Twenty-four shoulders isolated postmortem from 12 variably sized dogs were also used to characterize the normal ultrasound anatomy over a range of sizes. Important anatomic structures that could be consistently evaluated were the biceps tendon and bursa, the bicipital groove surface, the supraspinatous tendon, the infraspinatous tendon, the teres minor tendon, and the caudal aspect of the humeral head. Results of ultrasonographic examination of 4 dogs with shoulder lameness are described to illustrate some applications of canine shoulder ultrasonography in the evaluation of the canine shoulder. In these dogs, ultrasound was a valuable tool to evaluate effusion and synovial proliferation within the bicipital bursa, supraspinatous and biceps tendinitis, biceps tendon strain, and dystrophic calcification.     

Evaluation of rhinoscopy and rhinoscopy-assisted mucosal biopsy in diagnosis of nasal disease in dogs: 119 cases (1985-1989).

The case records of 149 dogs examined from 1985 to 1989 with clinical signs of nasal disease were reviewed. Gross rhinoscopy was performed in 119 dogs, and rhinoscopy-assisted pinch biopsy was performed in 109. Rhinoscopy was performed by use of a 2.7-mm rigid fiberoptic endoscope. Mucosal biopsy specimens were obtained with rhinoscopic guidance by use of a 2 x 3-mm biopsy forcep. Gross, cytologic, and histologic findings are summarized. Ninety-four of 119 cases could be evaluated on the basis of diagnostic and follow-up criteria established by the authors. The diagnostic success rate of gross rhinoscopy with rhinoscopy-assisted biopsy was 83% (78 of 94 evaluated cases). Protracted hemorrhage was a complication in 2 of 109 cases in which rhinoscopy-assisted biopsy was performed. It was concluded that rhinoscopy with rhinoscopy-assisted biopsy contributes important diagnostic information in dogs with nasal disease without the relative invasiveness, expense, and risk of surgery.     

Detection of free radicals in ischemic and reperfused canine gracilis muscle flaps by use of spin-trapping electron paramagnetic resonance spectroscopy

OBJECTIVE: To determine whether free radicals are produced in ischemic and reperfused canine skeletal muscle, whether free radicals can be detected from effluent blood by use of spin-trapping electron paramagnetic resonance (EPR) spectroscopy, and whether free radical-induced skeletal muscle damage is detectable by use of light microscopy. ANIMALS: 6 healthy mixed-breed dogs. PROCEDURES: Dogs were anesthetized and both gracilis muscles were isolated, leaving only the major vascular pedicle intact. Ischemia was induced in 1 flap for 4 hours; the contralateral flap served as the control. Ischemic flaps were then reperfused for 15 minutes. alpha-Phenyl-N-tert-butylnitrone, a spin-trapping agent, was administered intravenously to each dog 1 hour prior to reperfusion. Following reperfusion, effluent blood samples from muscle flaps were obtained and processed for EPR spectroscopy. Muscle biopsy specimens were obtained for histologic evaluation, and dogs were euthanatized. RESULTS: Spin adducts were not detected in blood from control flaps. However, spin adducts were detected in all ischemic-reperfused muscle flaps. Principal signals identified were characteristic of oxygen- and carbon-centered radicals. Significantly more muscle damage was detected in ischemic-reperfused flaps, compared with control flaps. CONCLUSIONS AND CLINICAL RELEVANCE: Free radicals may be an important component of injury induced by ischemia and reperfusion of canine skeletal muscle. Spin-trap adducts of free radicals can be detected in effluent blood of canine muscle flaps by use of spin-trapping EPR spectroscopy. Spin-trapping EPR spectroscopy may be useful for the study of antioxidants and free radical scavengers in attenuating ischemia and reperfusion-mediated skeletal muscle damage.