Comparison of indirect hemagglutination assay (IHA) and coprological diagnosis of schistosomosis in cows and buffaloes

Transcription

1 Indian J. Anim. Res., 49 (4) 2015 : Print ISSN: / Online ISSN: AGRICULTURAL RESEARCH COMMUNICATION CENTRE Comparison of indirect hemagglutination assay (IHA) and coprological diagnosis of schistosomosis in cows and buffaloes Sadaf Niaz 1, Tanweer Akhtar 2, Naser M. AbdEl-Salam 3, Sumaira Shams 1, Sultan Ayaz and Riaz Ullah 4 College of Veterinary Sciences and Animal Husbandry, Abdul Wali Khan University, Mardan, Pakistan. Received: Accepted: DOI: / ABSTRACT Schistosomosis is a disease of veterinary and medical importance caused by Schistosoma spp is characterized by frequent diarrhea with blood and mucous, weight loss and weakness in animals and causes significant morbidity, mortality and economic loss in cattle population. The study was designed to analyze/detect the schistosoma spp by coprological and serological examination (IHA) in cows and buffaloes of different areas of Punjab, Pakistan (Lahore, Sheikhupura, Gujranwala and Sialkot) from March February Both the techniques (coprological and serological examination (IHA) for Schistosomosis in cows and buffaloes were compared. Overall prevelance of Schistosomosis in cows and buffaloes was noted 14.0% and 13.6% by coprological examination and 17% and 16.3% by indirect hemagglutination assay (IHA), respectively. Statistical analysis showed non-significant difference in indirect hemagglutination assay (IHA) and fecal examination. Similarly month, area, age and sex wise seroprevalence of Schistosomosis was noted higher by IHA test than coprological examination in cows and buffaloes. Sensitivity (%) and specificity (%) of IHA test was noted 100% and >97.0%, respectively. The study reveals that Schistosomosis is prevalent in the study areas and needed proper preventive measure to minimize its prevalence. Besides that it is found that IHA is a good diagnostic tool for detection of Schistosomosis. Key words: Buffaloes, Cow, Indirect hemagglutination assay (IHA), Infectious disease, Schistosomosi. INTRODUCTION Schistosomosis caused by Schistosoma spp is characterized by frequent diarrhea with blood and mucous, weight loss and weakness in animals (Urquhart, et al. 2005). Diagnosis is a key step for the control of Schistosomosis. Determination of target population for chemotherapy in endemic communities, assessment of morbidity and the evaluation of control strategies all build on the results from diagnostic test (Van Dijk, et al. 2010, Makinson, et al. 2008). Schistosomosis diagnosis can be divided into direct parasitological techniques (parasite egg detection) and indirect approaches (detection of antibodies or circulating antigens in serum). Direct parasitological techniques include standard tests of fecal examination using the Kato-Katz thick smear (Katz, et al. 1972), which is currently the recommended method for diagnosis this disease and miracidium hatching test (WHO, 1994). Different bioassay used for immunodiagnostic are intradermal test (ID), circumoval precipitation test (COPT) Indirect hemagglutination assay (IHA), enzyme-linked immunosorbet assay (ELISA), dipstick dye immunoassay (DDIA) and so on. IHA is currently the most widely used immunodiagnostic assay in Schistosomosis endemic areas of China for the demonstration of antibodies against Schistosomosis, whereas ELISA has become more important over the past years (Sorgho H, et al. 2005, Xiang, et al. 2003). Immunodiagnostic methods based on the detection of antibodies continue to be most effective and practical method for diagnosis of imported Schistosomosis (Pardo, et al. 2004). Schistosoma bovis is a species whose final hosts are bovines. A commercial indirect hemagglutination (IHA) test using erythrocytes coated with Schistosoma mansoni adult worm antigen (SEA) were assessed for their use in serodiagnosis of imported Schistosomosis. The specificity of this combination in detecting Schistosomosis was 93.0%. Combination of SEA/ ELISA with WA/IHA (160) gave sensitivity of 98.7, 96.0, 98.0 and 80% with specificity of 97.2%. These findings suggest that WA/IHA and SEA/ELISA is each sensitive and *Corresponding author afridiriaz@yahoo.com. 1 Department of Zoology, Abdul Wali Khan University Mardar, KPK, Pakistan. 2 Department of Zoology, University of Punjab, Pakistan. 3 Riyadh Community College, King Saud University, Riyadh 11437, Saudi Arabia. 4 Department of Chemistry Government College Ara Khel FR Kohat KPK Pakistan.

2 504 INDIAN JOURNAL OF ANIMAL RESEARCH specific serological test that is easy to use for the diagnosis of important schstosomiasis. The majority of workers found that commercially available IHA is a good diagnostic toll for detection of Schistosomosis (Azab and El-Zayat 1996, El- Ganayni and Youssef 1992). MATERIALS AND METHODS Study area and sample collection: 300 blood samples and 300 fecal samples were collected monthly from each cows and buffaloes (Total 600) of four different districts (Lahore, Sheikhupura, Gujranwala and Sialkot) of Punjab, from March Febrruary 2008.The fecal and blood samples were examined by carpological and IHA test, respectively. Eggs were identified on the basis of morphology (Yamaguti 1975). Prevalence of infection was recorded. Its correlation with age, sex, area and seasonal variations was given consideration. IHA test: All the samples were analyzed for specific IgG antischistosoma antibodies by using commercial test kit (Bilharziose Fumouze kits). The procedure for Indirect hemagglutination assay (IHA) was followed as mentioned by Fumouze laboratory (France). A positive and negative serum included in the kit were processed like test serum for the comparison of results. Titer equal or above 1:320 were considered positive for Schistosomosis. The sensitivity and specificity of the IHA test used for the detection of Schistos infection was calculated according to O Reilly (1995) ((Reilly 1995). Sensitivity (%) = True positive cases (True positive cases + False negative cases) Specificity (%) = True negative cases X 100 (True negative cases + False positive cases) RESULTS AND DISCUSSION Comparison of (IHA) with coprological diagnosis: Over all prevalence of Schistosomosis was noted 14.0% and 17.3% by faecal examination and IHA test respectively in cows while 13.66% and 16.3% in buffaloes respectively (Table 1 and 2). Statistical analysis by Chi-square (c2) test did not show any significant difference between infection rates noted by faecal TABLE 1: Comparison of coprological and serological (IHA) diagnosis of Schistosomosis in cows in relation to month, season, area, age and sex wise in 4 districts of Punjab from March 2007 to February Factors Time (Months) Season Areas Age Sex Chi 2 ( 2 ) test: P = N.S. (non significant) Total sample observed Coprological Examination Serodiagnosis (IHA test) No. of positive Prevalence (%) No. of positive Prevalence (%) March April May June July Aug Sep Oct Nov Dec Jan Feb Autumn Winter Summer Spring Lahore Sheikhupura Gujranwala Sialkot years > 2 years Male Female Total

3 Volume 49 Issue 4 (August 2015) 505 examination and IHA test in both species ((c2 = 0.290, d.f. = 1, P = 0.59 in cows and (c2 = 0.71, d.f. = 1, P = 0.71 in buffaloes). Month wise prevalence: In month wise data, both diagnostic methods showed same pattern of prevalence. The highest prevalence was noted in the month of September, 24.0% and 28.0% by faecal examination and (IHA), respectively, while lowest in May and June 0.0% and 4.0% by faecal examination and IHA, respectively in cows of Punjab (Table 1). In buffaloes highest infection rate was noted in the month of September by both diagnostic methods i.e., 28% by faecal examination and 32% by (IHA) while lowest, 0.0 (April to June) and 0.0% (June) by faecal examination and IHA respectively (Table 2). Season wise prevalence: Almost same pattern of overall season wise prevalence (%) was noted in cows by faecal examination and IHA test. Highest prevalence was observed in autumn i.e., 22% and 24% followed by winter (18% and 22%) summer (9% and 12%) and lowest in spring season (8% and 10%) by coprological examination and (IHA), respectively (Table 1). In buffaloes season wise overall highest prevalence (%) was observed in autumn (22% and 24%) followed by winter (19% and 23%), summer (8% and 10%) and lowest in spring (6% and 8%) by faecal examination and (IHA), respectively (Table 2). Area wise prevalence: In area wise data of cows highest prevalence (%) was observed in Lahore (17.3% and 20.0%) followed by Sheikhupura (16% and 18.5%), Gujranwala (12.0% and 16.0%) and lowest in Sialkot (10.6% and 13.3%) by coprological examination and (IHA), respectively (Table 1). In area wise data of buffaloes highest prevalence (%) was observed in Lahore (18.6% and 21.3%) followed by Sheikhupura (17.3% and 20%), Gujranwala (10.6% and 13.3%) and lowest in Sialkot (8.0% and 10.6%) by coprological examination and IHA test, respectively (Table2). Age wise prevalence: Adult cows showed higher prevalence (%) i.e., 16.1% and 19.0% than young ones (11.0% and 14.1%) in faecal and blood serum examination, respectively (Table1). In age wise data it was noted that adult buffaloes were found more (16.9% and 19.2%) susceptible than young ones (9.3% and 12.4%) (Table 2). TABLE 2: Comparison of coprological and serological (IHA) diagnosis of Schistosomosis in buffaloes in relation to month, season, area, age and sex wise in 4 districts of Punjab from March 2007 to February Time (Months) Season Areas Age Sex Factors Total sample observed Coprological Examination Serodiagnosis (IHA test) No. of positive Prevalence (%) No. of positive Prevalence (%) March April May June July Aug Sep Oct Nov Dec Jan Feb Autumn Winter Summer Spring Lahore Sheikhupura Gujranwala Sialkot years > 2 years Male Female Total N.S Chi 2 ( 2 ) test: P = N.S. (non significant)

4 506 INDIAN JOURNAL OF ANIMAL RESEARCH Gender wise: In sex wise coprological and blood examination (IHA test) male cows (15.6% and 19.0% respectively) were found more susceptible than females (12.4% and 15.0% respectively) for Schistosomosis (Table 1). In gender wise faecal and blood examination male buffaloes (15.9% and 18.8%) were found more susceptible than female 11.7% and 14.1%, respectively (Table 2). Comparison of diagnostic test: Although area, month, season, age and sex wise prevalence were found higher by IHA test as compared to faecal examination, but statistical analysis by Chi-square (c2) showed non significant (P>0.05) difference (Table 1 and 2). Sensitivity and specificity and (IHA) test: Sensitivity and specificity of IHA test were calculated 100% and >97.0 respectively for cows as well as buffaloes (Table 3). Considering the importance of diagnosis in Schistosomosis, we compared the role of coprological and serological investigation. In present study, 17% cows and 16.3% buffaloes were found positive for Schistosomosis when tested by IHA test, cows were 4.1% more infected than TABLE 3: Sensitivity and specificity of (IHA) test Factors Cows Buffaloes Sensitivity Specificity buffaloes. Serological diagnosis by IHA test showed highest prevalence of schitosomiasis in cows as well as in buffaloes, it showed100% sensitivity and >97.0% specificity. Similar study was conducted by (Zhou et al 2007). (Al-Mofarreh et al 2000) also found higher values in serological diagnosis by IHA. The reason may be that coprological methods are positive only after 2-3 months while immunodiagnostic methods can make early detection of disease. (Van et al 2002) stated that IHA gave the most reliable outcome because its sensitivity increased considerably in all stages of infection. ACKNOWLEDGMENT: The authors are thankful to the Deanship of Scientific Research, King Saud University, Riyadh, Saudi Arabia, for funding the work through the research Group project No. RGP REFERENCES Al-Mofarreh, M., Ahmad, A., Hasan, S. and Al-Mofleh, I. (2000). Formalether concentration method in the diagnosis of active schistosoma in patient with detectable IHA. Saudi J. Gastroenterol., 6: Azab, M.E. and El-Zayat, E.A. (1996). Evaluation of purified antigens in haemagglutination test (IHA) for determination of cross reactivities in diagnosis of fasciolosis and Schistosomosis. J. Egypt Soc. Parasitol., 26: El-Ganayni, G.A. and Youssef, M.E. (1992). Evaluation of adult schistosoma and cercarial antigens in serodiagnosis of Schistosomosis using IHAT and ELISA. J. Egypt Soc. Parasitol., 22: Katz, N., Chaves, A. and Pellegrino, J. (1972). A simple device for quantitative stool thicksmear technique in Schistosomosis. Rev. Inst. Med. Trop. Sao. Paulo., 14: Makinson, A., Morales, R. J., Basset, D., Bouchaud, O., Verdon, R., Hosseini, H., Moing, V. L., Delaporte, E. and Reynes, J. (2008). Diagnostic approaches to imported schistosomal myeloradiculopathy in travelers. Neurology 71: Pardo, J., Carranza, C., Turrients, M.C., Arellano, J.L.P., Velez, R.L., Ramajo, V. and Muro, A. (2004). Utility of S. bovis adult worm antigen for diagnosis of human Schistosomosis by ELISA and Electroimmunotransfer blot techniques. Clin. Diag. Lab. Immunol., 11: Reilly, O.M.M. (1995). Tuberculin shin tests sensitivity and specificity in Mycobacterium bovis infection in animals and humans, 1st ed. Iowa State University Press/Ames, pp Sorgho, H., Bahgat, M., Poda, J.N., Song, W., Kirsten, C., Doenhoff, M.J. and Ruppel, A. (2005). Serodiagnosis of Schistosomosis in an endemic area of Burkina Faso. Performance of several immunological test with parasite antigens. Acta Trop., 93: Urquhart, G.M., Armour, J., Duncan, J.L., Dunn, A.M. and Jennings, F.W. (2005). Veterinary Parasitology, ed. ELBS Langman, U.K. Van, Dijk, K., Starink, M.V., Bart, A., Nijhuis, E.W.P., van der Wal, A.C., van Thiel, P. P. A. M., de Vries, H. J. C. and van Gool, T. (2010). The Potential of Molecular Diagnosis of Cutaneous Ectopic Schistosomosis. Am J Trop Med Hyg 83:

5 Volume 49 Issue 4 (August 2015) 507 Van, G.T., Vetter, H., Vervoort, T., Doenhoff, M.J., Wetsteyn, J. and Overbosch, D. (2002). Serodiagnosis of imported Schistosomosis by a combination of commercial indirect hemagglutination test with adult worm antigens and an enzyme-linked immunosorbent assay with schistosoma egg antigen. J. Clin. Microbiol., 40: WHO (1994). The control of Schistosomosis report of a WHO expert communities. World Health Organization Technical Report Series. Xiang, X., Tianping, W. and Zhigang, T. (2003). Development of a rapid sensitive dye immunoassay for Schistosomosis diagnosis. J. Immunol. Method, 280: Yamaguti, S. (1975). A synoptical review of life historics of digenetic trematode of vertebrate with special reference to the morphology of their larvae. Keiguka Publ. Co. Tokyo, pp Zhou, Y.B., Yang, M.X., Wang, Q.Z., Zhao, G.M., Wei, J.G. and Jiang, Q.W. (2007). Field comparison of immunodiagnostic and parasitological techniques for the detection of Schistosoma japonicum in the people s Republic of China. Am. J. Trop. Med. Hyg., 76: