PJ BS Bosxiczs so Pakistan Journal of Biological Sciences ANSIez n Network for Scientific Information 308 Lasani Towns Sargodha Road, Faisalabad - PakistanPakistan Joumal of Biological Seienees 10(11): 1938-1940, 2007 ISSN 1028-8880 (© 2007 Asian Network For Selentifie Information. Evaluation of Different Methods for Diagnosis of Dirofitaria immitis ‘Shahrokh Ranjbar-Bahadori “Ali Eslami and ?Seid Bokaic ‘Department of Parasitology, College of Veterinary Medicine, Islamic Azad University, Garmsar Branch, Garmsar, Iran Department of Parasitology. "Department of Bpidemiology, College of Veterinary Medicine, Tehran University, Tehran, Iran Abstract: Divefilaria immitis, « common parasite of the cardiovascular system of the carniveres all over the world, is reported from dogs, cals and, humans in Iran, Knolt method is the most common test fer its diagnosis Jnmany regions including Iran_In the present study, during ene year period from 2002 to 2003, blood samples of 110 dogs were examinied using modified Knott method and commercial antigen detet test kit (WITNESS. (CHW IL kit). All dogs were subjected to necropry to evaluate the specificity and sensitivity of both tests. In Knott method, 22 harbored dogs (20%) were microfilaracmic of which (of the firs instance}, 20 dogs (18.18%) D. immits and 21-82%) D. reconditum. Accordingly, by using commercial antigen detecticn test kit 16 dogs (14546) were found to be infected with D. immitis. At necropsy 14 out of 110 dogs (12.73%) harbored D. inomitis in the right ventricle of the heart, Thus, it was concluded that 6 out of 20 positive dogs with _D. mits were mieroflaraemie and these micrfilaria belonged to other filarial except D. mnmitis. To compare the findings with goldon test (necropsy) the sensitivity and specificity of Knott and antigen deteeton test were 85.71 and 91.66%, 92:85 and 9687 was higher, differences were not significant (p>0.05), Key words: Diroflaria inmmitis, diagnosis, dog, Iran INTRODUCTION Canine dirofilariosis caused by Divofifara iments is usually diagnosed by specific antigen testing and/or idantifcation of microfilariae. However, D. inomits and at least sx ote ilariae can produce caine microfilaremias ‘with negative heartworm antigen tests (Rishnite ef al, 2006), Accurate diagnosis of canine filarosis is essential for choosing correct therapeutis approach, Therefore, reliable methods for diseriminating among the different filaral infections in dogs are needed (Casiraghi et a, 2006). A dot enzyme-linked immunosorbent assay (Doc ELISA) wed for detecting eisculating antigens inthe sera of dogs infected with D. immitis (Ripoll etal, 1991; Matsumura et al, 1988). Also to estimate the seroprevalence of D. inenitis infection in domestic dogs in Taiwan, they uulized a commercial ELISA kit (Sump, IDEXX, USA) for detecting circulating antigens released by female adult worms (Fan e¢ af, 2001), In the present sty, the attempts were made to compare modified Knott ‘method with that of antigen detection test in order 10 present more accurate method and to determine the prevalence of the infection in stay dogs of Golestan provinee, nerth of Tran 5, Teapectively. MeNemar test shoved that although sensitivity ofthe later MATERIALS AND METHODS ‘Blood samples: Ina period of one year, from 2003-2004, 110 blood samples taken from saplien or cephalic vein of stray dogs in Golestan provinee (north of Tran) were ‘examisted using modified Knott method and an antigen detection test kit (WITNESS CHW II kit) and then necropsy vas caried out to confirm the accuracy ofthese ‘mothods ‘Modified Knott method: In Krott method, the sediments ‘were searched for mierofilarine and identification of which was performed using the key presented by Etinger and Felden (2000) key. ‘Heartworm antigen test kit (WITNESS CHW IT kit: In ‘antigen detection test, examination procedures were followed according to manufacturer's recommendation. ‘The appearance of pink ot purple lines in window 2 and 3, respectively were talcen as positive with D. inmate ig. D. Necropsy: All dozs were subjected to necropsy and then pulmonary arteries and hearts were searched for the (Corresponding Author: ‘Shalokih Ranjr-Babadon, Department of Veterinary Parasitology, Veterinary Collage of Islamic Azad University, Garmsar Branch, Gatmsar, Semman, fran ‘Te: #98-232-4229962 Fan: +98-232-4220700-1 1938)Pak. J. Biol. Se., 1O(41): 19881940, 2007 B Fig. 1: Results of WITNESS CHW II kit. A: Negative result (The presence of line in win. 3 and its absence in win, 2). B: Positive reslt (The presence of both lines in win, 2 and3) presence of adil D, ‘mnitis. The collected worms were counted in preserved in 70% alcohol containing $%@ alyeerin Stacstess MeNemsr test has been used for evaluation of one findings. RESULTS ‘The reslts ofboth tests are summatized in Table 1. Necropsy as a golden test showed that 14 out of 110, examined dogs (12.73%) were found to be infected with ‘Drums whereas in Knott method20 dogs (18.18%) and boy antigen detection tet 16 dogs (14.54%) showed ‘microlaria of D. erupitis in that onde, Thus it can be postulated thot in Knott method, 6 dogs were rmictoflaraemic andthese microfilaia were belong to other Silarial except. ivmsiis (5.45% foe positive results) and ‘wo out of 90 non infected dogs shown by Knott method (1.8290) were postive at necropsy (false negative = occult infection). Meanwhile in antigen detection test out of 16 dogs infected with heartworm antigen (1.8288) were not positive at necropsy ifalse postive) and 1 (.91%%) out of non infected dog was positive at necropsy (false negative = oceult infection). although sensitivity of tanfigen detection test was higher, no difference was shown using MeNemar test (p>0.05). At necropsy, 14 out of 110 dogs were found to be infected with ini ‘Thenumber of worms found ranged fom 1-7 of which fo dogs harbored only one worm in the right ventricle that proves two cases of occult infection, DISCUSSION “The results of this investigation show that in Knott method and antigen detection test kit, the sensitivity and specificity were high (85.71 and 91.66%, 92.85 and 96.87%), respectively. Present findings in the case of antigen detection tes were similar with many other workers using different commercial kits (Atkins, 2003; Courtney and Zeng, 2001). This shows that antigen detection test regardless of the type of kit is a reliable method andits accuracy isnot microfilaraemia dependent, thus can be used to detect amicrotilaraemic or occult, infection or for rapid screening of heartworm infection Knott method was also accurate enough for diagnosis of a high percentage of infected dogs with D. immu in the present study, Some workers believe that this is not as good as antigen detection test (Atkins, 2003), although Ieir advantages make of the latter a choice method. It seems likely that occult heartworm die to treatment of infection dogs with Ivermectine of Levamisole is the main source of high occult infection in many country (Rawlings et ai, 1982), thus low oceult infection reported in this survey seems to be reasonable because anf D. smmiis treatment is not practiced at all in Iran, except in experimental study (Esfami etal, 2004), although tccording to clinical signs and postive response to treetment with Melarsomine dichlothydrate 3 ont of 30 infected (109%) harbored occult infection, 2 Snding Similar to present investigation. Accordingly, 18.18 and “ble Rem of did Kn mrad ecient heare stn tet CWTTMESS CHW Ge doe se enon hen Mo. of eagle Toe Toe Hie Pe Smstoly— Spmibety Accuacy edicee Receute ee gostve sense poasre ease) Sa Ga mune) vale Noiieimanedid 1 ¢ 2 oa ‘ca 0s mon at wmnesscwwns 4 1 ous ong) sess 1939)