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Clinicopathologic Findings in an Experimental Spirocerca Lupi Infection in Dogs

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Clinicopathologic Findings in an Experimental Spirocerca Lupi Infection in Dogs ISRAEL JOURNAL OF VETERINARY MEDICINE CLINICOPATHOLOGIC FINDINGS IN AN EXPERIMENTAL SPIROCERCA LUPI INFECTION IN DOGS Aroch, I.,1* Harrus, S.,1 Amit T.,1 Bark, H.,1 Markovics, A.,1 Hagag, A.,1 Aizenberg, Z.,1 and Lavy, E.1 1 Koret School of Veterinary Medicine The Robert H. Smith Faculty of Agriculture, Food and Environment The Hebrew University of Jerusalem, Israel. 2 Department of Parasitology, Kimron Veterinary Institute, PO Box 12, Beit Dagan, Israel. Short title: clinocopathologic findings in spirocercosis *Corresponding author Prof. Itamar Aroch Koret School of Veterinary Medicine, The Hebrew University of Jerusalem, PO Box 12, 76100 Rehovot, Israel Tel: 972-3-9688556, Fax: 972-3-9604079, Email: [email protected] ABSTRACT Spirocerca lupi is primarily a parasite of dogs, which typically causes esophageal nodules, aortic aneurysms and spondylitis. In this study, the hematology and serum biochemistry parameters in experimental canine spirocercosis were investigated. Ten beagle dogs were orally challenged with 40 infectious S. lupi larvae. One dog did not become infected. Two dogs died within 2 months post infection (PI) due to rupture of an aortic aneurism. Seven dogs became infected, presented with esophageal nodules and worm eggs in their feces. All were then treated with doramectin, survived and were followed for 21 months PI. Blood and fecal samples and esophagoscopy were performed periodically. Significant differences in the eosinophil counts were noticed in repeated measures between different time points during the study course. Eosinophil counts increased significantly (P<0.018) two weeks PI and their relative and absolute counts were above the reference interval (RI) (18.8%; RI: 0-10%; 2.19x109/L; RI: 0.1-1.0x109/L, respectively). Both relative and absolute counts decreased significantly (P<0.018) at 4 weeks PI (8.9% and 0.75x109/L, respectively) and remained within RI throughout the study course. Appearance of esophageal nodules, number of eggs shed in the feces and doramectin treatment had no association with eosinophil counts. There were no other significant differences in any hematologic and serum biochemistry analytes between different time points in repeated measures during the study course. In conclusion, with exception of a transient moderate eosinophilia, 14 days PI, hematologic and serum biochemistry abnormalities were absent in the early disease stages of experimental canine spirocercosis. Key Words: Spirocercosis; Canine; Hematology; Eosinophil; Serum biochemistry; Nematode. INTRODUCTION The pathogenesis of spirocercosis results from the The nematode Spirocerca lupi is mostly a parasite of migration and prolonged presence of infective larvae and adults dogs, although other animals, particularly carnivores might in the host’s tissues. Typical esophageal nodular masses, that be affected (1, 2). It has a worldwide distribution, but is most might undergo neoplastic transformation, and aortic scars and prevalent in warm climates (1-4). It is transmitted to dogs aneurysms are the most frequent lesions, and are considered through ingestion of coprophagous beetles or paratenic hosts, pathognomonic (5). Spondylitis and spondylosis of the caudal including birds, lizards and rodents (5). In Israel, it is most thoracic vertebrae (T6 -T12) are also typical lesions (3-8). commonly transmitted through ingestion of the dung beetle Hypertrophic osteopathy is observed, mostly in the neoplastic Ontophagus sellatus (3), although in other countries, paratenic form of the disease (9). hosts seem to play a major role in disease transmission (5). A The clinical signs might related to the esophageal lesions previous study indicated an emerging outbreak of spirocercosis that often interfere with swallowing and cause persistent in Israel. In spite of increased awareness, the disease prevalence regurgitation and/or vomiting (1, 5, 6, 10). Sudden death and its geographic distribution have been increasing (3). might be caused due to aortic aneurysm rupture resulting in Volume 66 (1) 2011 website: www.isrvma.org 19 ARTICLES hemothorax (5, 11, 12). Abberant migration to different body Infectious larvae were harvested, divided into 10 aliquots of 40 organs has been reported in canine spirocercosis, causing a larvae each and kept in saline at 4 °C pending inoculation (up to variety of clinical signs (5, 13, 14). Pyothorax and aortic 3 days post harvesting). Dogs were randomly allocated to two thromboembolism have been reported in canine spirocercosis 5-dog groups. In one group (study group), the dogs were treated (5, 15, 16). with doramectin (Dectomax, Pfizer, Amboise, France, 400 μg/ Spirocercosis can be suspected based on the clinical signs, kg SC, 3 times, q30d). The 5 other dogs served as untreated and the diagnosis might be aided by radiography and computed controls. All dogs presented at least a single esophageal nodule tomography, demonstrating spondylosis and spondylitis of the of at least 1 cm in diameter and had positive fecal flotation for thoracic vertebrae, aortic aneurisms and deformations and a S. lupi eggs. One month after the last doramectin injection to caudal mediastinal thoracic mass (3-8). Radiography can lead the study group dogs, all dogs, including the control dogs, were to definite diagnosis of spirocercosis in those cases where orally challenged with 40 infectious S. lupi larvae as described esophageal nodule is present. However, it cannot definitely previously (20, 21). A dog was considered to be infected when differentiate neoplastic from non-neoplastic cases or identify typical esophageal nodules were observed during endoscopy early spirocercosis cases, when thoracic involvement is and S. lupi eggs were detected in its feces, or when S. lupi absent (5). A definite diagnosis of spirocercosis is made by larvae or typical histopathological changes were found in the detecting the characteristic eggs in the feces of infected dogs aorta on post mortem examination. At days 228 and 250 post through coproscopy or polymerase-chain reaction (PCR) inoculation (PI), for the doramectin-untreated and -treated using the mitochondrial cytochrome-C oxidase, subunit 1 dogs, respectively, a treatment protocol (doramectin, 400 (cox1) gene (17-19). The latter has been proven to be more µgr/kg SC every 14 days, 6 times) was implemented for all sensitive compared to routine coproscopical methods (19), but the surviving infected dogs. The dogs were followed for 21 it still relies on the presence of fertile female worms in the months post-inoculation (PI). esophagus. Typical esophageal nodules might be demonstrated by esophagoscopy or post mortem (3, 5, 6, 20). Diagnostic Fecal examination methods that allow a definite diagnosis of spirocercosis are Fecal samples (at least 30 g/sample, twice weekly) were limited to the advanced stages of the disease, when adult worms collected from day 128 PI, homogenized, and three 5-gram are present in the esophagus (5). The ante mortem diagnosis of aliquots per dog were analyzed per collection, using a sugar the disease in its early latent stages is almost impossible. flotation method (17). The number of S. lupi eggs/gram feces The aim of the present study was to evaluated and characterize for each sample was calculated as the mean egg number in the the clinicopathologic changes that occured during an three aliquots. experimental S. lupi infection in dogs, in order to assess their diagnostic usefulness in this disease, mostly in its early Hematology and serum biochemistry stages. The data for the study was collected during a study Samples for CBC and serum biochemistry analysis that assessed the prophylactic and therapeutic efficacy of were collected from all dogs every 10-30 days by jugular doramectin in experimental canine spirocercosis. The results venipuncture in potassium-EDTA and plain tubes. A CBC relating to the efficacy of doramectin have been published was performed within 30 minutes from collection using an elsewhere (21, 22). automatic impedance hematology analyzer (Abacus, Diatron, Wien, Austria). The CBC included the white blood cell (WBC) MATERIALS AND METHODS count, red blood cell (RBC) count, hemoglobin concentration, Animals hematocrit, (MCV), mean corpuscular hemoglobin, mean Ten beagle dogs (6 males, 4 females), aged 6-10 years corpuscular hemoglobin concentration and platelet count. The participated in this study. They were deemed healthy based on differential WBC and the morphologic assessment of blood normal physical examinations, complete blood count (CBC), cells were performed manually by examination of modified serum biochemistry and negative fecal flotations. These CBC Wright’s-stained blood smears (Hema-Tek 2000 Slide Stainer, and serum biochemistry results were used as base-line values model 4488B, Bayer Corporation, Elkhart, IN, USA, Stain: for each dog in further analyses. Dogs were housed in separate Hematek stain pack; Modified Wright's Stain) by a single cages in an enclosed kennel and fed a dry commercial canine veterinary clinician. Samples for serum biochemistry analysis diet. Feces were cleared and collected daily. This study was were allowed to clot at room temperature for 30 minutes, approved by the Joint Instititional Animal Use And Care centrifuged, and sera were separated and stored at 4 °C Committee of the Hebrew University Veterinary Teaching pending analysis, generlly within 24 hours from collection Hospital (HUVTH) and the Kimron Veterinary Institute (KVI). using a wet chemistry autoanalyzer (Kone lab, Kone, Espoo, Finland, at 37 °C). Serum
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