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Evaluation of SomeImmunodiagnostic Tests forHydatidosis in BuffaloesS. K. Binjola a & S. N.S. Gaur aa Department of Parasitology , College of VeterinarySciences G.B. Pant University of Agriculture andTechnology , Pantnagar, 263145, IndiaPublished online: 11 Nov 2011.

To cite this article: S. K. Binjola & S. N.S. Gaur (1994) Evaluation of SomeImmunodiagnostic Tests for Hydatidosis in Buffaloes, Journal of Applied AnimalResearch, 6:2, 183-186

To link to this article: http://dx.doi.org/10.1080/09712119.1994.9706041

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J. Appl. h i m . Res. 6 (1994) 183-186

Evaluation of Some Immunodiagnostic Tests for Hydatidosis in Buffaloes

S.K. Binjola*, S.N.S. Gaur** Department of Parasitology

College of Veterinary Sciences G.B. Pant University of Agriculture and Technology

Pantnagar-263145 India (Received December 7, 1993, accepted September 15, 1994)

Abstract Binjola, S.K. and Gaur, S.N.S. 1994. Evaluation of some immuno- diagnostic tests for hydatidosis in buffaloes. J. Appl. h i m . Res., 6 : 183-186.

Indirect haemagglutination test (IHA) was compared with immunoelectmphoresis test (IEP) and counter immunoelectmphoresis test (CIEP) by using two antigens for diagnosis of hydatidosis in buffaloes. Total 48 positive and 30 uninfected normal sera were collected from slaughtered buffaloes. Fractionated first peak antigen (F,) prepared by column chromatography gave more sensitive and specific reactions as compared to the antigen prepared by salt precipitation of hydatid fluid. The IHA was found to be the most sensitive test while the specificity of CIEP was the highest.

Key words : Hydatidosis, buffaloes, immuno diagnosis

Introduction The immunodiagnosis of hydatidosis has undergone considerable technical advancement since the introduction of Ghedini's compliment fixation and Casoni's intradermal tests. The major problem in its immunodiagnosis is cross 1918 Hospital Road, Vikasnagar-248 198, Dehradun, India

'*Professor Parasitology, School of Veterinary Medicine, Shiraz Univ. Shiraz, Iran.

183 J. Appl. h i m . Res. 0971-2119/93/$05.00 0 GSP, India

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184 S. K. Binjolu and S. N. S. Gaur

reactivity with other taenid metacostodos and other parasitic infections (Kagan, 1968) and a positive reaction may indicate only a previous exposure to Echimcoccus antigens.

Indirect haemagglutination, immunoeledrophoresis and counter immunoelectrophoresis tests extensively evaluated and compared by various workers including Garabedian et al. (1957), Yarzabal et al. (1974), Kelkar and Kotwal(l975) with variable results, have again been evaluated in order to asses their field application by using salt precipitated and fractionated antigens of hydatid fluid.

Materials and Methods Ammonium sulphate hydatid fluid antigen designated as partially purified antigen (€'PA) and fractionated first peak antigen (F,) obtained by fractionation of PPA were prepared according to Deka and Gaur (1991). The concentration of protein was found to be 0.3 mgml and 0.2mgrnl in F, and PPA respectively. Hyperimmune sera were raised in rabbits against each antigen by conventional method. Serum samples were collected from 48 buffaloes (age above 10 years) naturally infected with hydatid cysts of Echimcoccus grunulosus. Sera from 30 other healthy buffaloes were also collected to serve as control. The sera were subjected to the following tests : Indirect haemagglutination test (Garabedian et al., 1957) was observed for five hours. Immunoelectrophoresis test @eka and Gaur, 1991) was run at room temperature in barbitone buffer (PH 8.2) at 6 volts per cm and 10 mA per slide. A small drop of bromophenol blue was placed in the well near the cathode end of slide. Current was continuously passed till the dye migrated upto 3.5 cm distance. Counter immunoelectrophoresis test Valwar et al., 1986) was carried out at room temperature for 2 h at lOmA per slide.

Results and Discussion The results of the immunodiagnostic tests are summarized in Table-1. In all the three tests carried out F, antigen was found to be better in terms of both sensitivity and specificity which was in agreement with Varela Diaz et al. (1975), who reported purified hydatid fluid antigen to be more sensitive than whole hydatid fluid of sheep origin. IHA test with F, antigen and homologous hyperimmune rabbit sera gave a titre of 1 : 2048. However, with PPA with its homologous rabbit sera, a titre of 1 : 512 was obtained. When the test was performed with antisera obtained from buffaloes naturally infected with E. granulosus the titres ranged between 1 : 256 to 1:1024 with F, antigen and 1:64 to 1512 with PPA. All the naturally infected buffaloes showed a titre higher than 1:64. The variation in titres in the sera of infected buffaloes was mostly influenced by the intensity of infection and the antigen used, as high titre was observed in the heavily infected buffaloes. The sensitivity observed in the test

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Immunodiagnostic tests for Hydatidosis 185

Table 1

Comparison of immunodiagnostic tests for diagnosis of hydatidosis in buffaloes

Test No. of No. of Positive False Sensiti- Specificity

Partially purified antigen ( P A )

IHA 48 30 28 6 70.58 83.33

IEP 48 30 24 12 66.66 71.40

CIEP 48 30 22 3 64.86 90.90

Fractionated antigen (F1)

IHA 48 30 36 4 80.00 88.23

IEP 48 30 28 8 70.58 78.94

CIEP 48 30 28 0 70.58 100.00

positive sera normal sera reaction positive vity (Yo) (Yo)

was comparable to Vanna and Malviya (1989) who reported this test to be 81.70% sensitive in naturally infected buffaloes. CIEPtest was found to be rapid and highly specific for the serodiagnosis of hydatidosis in buffaloes with PPA and F, antigens. The reactions were observed within 2 hours with F, antigen and 3 hours with PPA after keeping the slide in refrigerator. These findings are in agreement with those of Mahajan et al. (1976) who reported this test to be 98% specific. IEPtest was found to be least specific (78.94% and 71.42%respectively, with F, and PPA antigen) but its sensitivity (70.58% and 66.66Y0, respectively with F, and PPA antigen) was almost similar to CIEP.

The variation in the sensitivity might be due to the age of animals, location of the cysts, nature of the infection and application of the test (Kagan, 1968). The slaughtered buffaloes in the present study were above ten years of age and most of the cysts were infertile (cysts without scolices) which might explain the reduced sensitivity. Some of the false positive reactions were probably due to non specific components of the antigens.

Acknowledgements The authors are grateful to the Dean, College of Veterinary Sciences and Head

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186 S. K . Binjokz and S. N. S. Gaur

Department of Parasitology for providing facilities.

References Deka, D.K. and Gaur, S.N.S. 1991. Immunoelodrophoretic patterns in hydatidosis. J.

Vet. Parasit., 5 : 19-23.

Garabedian, G.A., Matosian, M. and Djanian, A.Y. 1957. An indirect haemagglutination test for hydatid disease. J. Immunol., 78 : 269-272.

Kagan, I.G. 1968. A review of serological tests for the diagnosis of hydatid disease. Bull. Wld. Hlth. Org., 39 : 25-37.

Kelkar, S.S. and Kotwal, S.E. 1975. Counter immunoelectrophoresis in diagnosis of hydatid disease. Lancet, 1 : 755-756.

Mahajan, R.C., Ganguli, N.K., Sharma, S., Chandani, R.E. and Chitara, N.L. 1976. Counter immunoeledrophoresis for a rapid serodiagnosis of human hydatid disease. Indian J. Med. Res., 64 : 1173-1176.

Talwar, V., Iqbal Kaur and Gupta, H.C. 1986. CIEP for serological diagnosis of typhoid

Varela-Diaz, V.M., Guisantes, J.A., Ricardes, M.I., Yarzabal, L.A. and Coltorti, E.A. 1975. Evaluation of whole and purified hydatid fluid antigens in the diagnosis of human hydatidosis by immunoeletrophoresis test. Am. J. Trop. Med. Hyg., 24 :

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Yarzabal, L.A., Leiton, J. and Lopez-Lemes, M.H. 1974. The diagnosis of pulmonary human hydatidosis by immunoeledrophoretic test. Am. J. Trop. Med. Hyg., 23 : 662-666.

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