COMPARISON OF ENZYME-LINKED IMMUNOSORBENT ASSAY WITH ACID-FAST STAINING TO DETECT CRYPTOSPORIDIUM OF CATTLE FAECES
Keywords:Acid-fast staining, Bovine, Cryptosporidium, ELISA.
Cryptosporidiosis is a major cause of diarrhoea, weight loss and low productivity in various domestic animals. The acid-fast staining method used to detect the parasite in cattle may be responsible for the low prevalence rates recorded in previous studies in Nigeria. A comparison of the efficacy of an enzyme-linked immunosorbent assay (ELISA) and acid-fast staining techniques in detecting Cryptosporidium spp. in bovine faeces was carried out in this work. A total of 200 faecal samples were collected from cattle of different age groups by the use of a simple random sampling technique. The samples were analyzed microscopically using formalin-ethylacetate sedimentation method followed by modified Kinyounâ€™s acid-fast staining technique. All the samples were later tested for Cryptosporidium coproantigens by the use of a commercially available ELISA test kit. Microscopy detected Cryptosporidium oocysts in 24.0% while ELISA detected the antigens in 37.5% of the samples. The ELISA, with a sensitivity and specificity of 72.9% and 73.7% respectively, had a significantly higher (p<0.05) rate of detection of Cryptosporidium spp. than microscopy with a sensitivity and specificity of 46.7% and 89.6% respectively. The ELISA is therefore a preferable method than microscopy for detection of Cryptosporidium in faecal specimens and will be useful in routine diagnosis and screening of large number of samples in epidemiological surveys
Ayeni, A.O., Olubunmi, P.A., Ade, J.O. 1985. The occurrence of Cryptosporidium in faeces in livestock in Ile-Ife, Nigeria. Trop. Vet. 3: 96-100.
Ayinmode, A.B., Fagbemi, B.O. 2010. Prevalence of Cryptosporidium infection in cattle from southern Nigeria. Veterinarski Archiv. 80 (6): 723-731
Ayinmode, A.B., Fagbemi, B.O. 2011. Cross-Reactivity of Some Cryptosporidium Species with Cryptosporidium parvum Coproantigen in a Commercial ELISA Kit. Nigerian Veterinary Journal 32(1): 1 â€“ 4
El-Shazly, A. M., Gabr, A., Mahmoud, M.S., Aziz, S.S., Saleh, W.A. 2002. The use of Ziehl-Neelsen stain, enzyme-linked immunosorbent assay and nested Polymerase Chain Reaction in diagnosis of cryptosporidiosis in immunocompetent, -compromised patients. J. Egypt Soc. Parasitol. 32: 155-166.
Fayer, R., Santin, M., Trout, J.M., Greiner, E. 2006. Prevalence of species and genotypes of Cryptosporidium found in 1-2-year-old dairy cattle in the eastern United States. Veterinary Parasitology 135: 105-112.
Fayer, R., Santin, M., Trout J.M. 2008. Cryptosporidium ryanae n. sp. (Apicomplexa: Cryptosporidiidae) in cattle (Bos taurus). Vet Parasitol 156:191â€“198
Godekmerdan A., Kalkan, A., Erensoy, A., Kilic, S.S. 1999. Prevalence of Cryptosporidium spp. in children with diarrhoea. Acta Parasitol. Turcica. 23: 122-125. In: Yilmaz H., Zeynep, T., Mautalip, C. 2008. Investigation of cryptosporidiosis by enzyme-linked immunosorbent assay and microscopy in children with diarrhoea. Saudi Med. J. 29(4): 526-529.
Hamedi, Y., Safa, O., Haidari, M. 2005. Cryptosporidium infection in diarrhoeic children in southeastern Iran. Pediatr. Infect. Dis. J. 24: 86-88
Harrington B. J., Kama, H. 2002. A comparison of an immunoassay with acid-fast staining to detect Cryptosporidium. Laboratory Med. 33(6): 449-452.
Janoff, E. N., Reller, L.B. 1987. Cryptosporidium species, a protean protozoan. J Clinical Microbiol 25: 967â€“975
Johnston, S. P., Ballard, M.M., Beach, M.J., Causer, L., Wilkins, P.P. 2003. Evaluation of three commercial assays for detection of Giardia and Cryptosporidium organisms in fecal specimens. J. Clinical Microbiology 41: 623â€“626.
Kaur, R., Rawat, D., Kakkar, M., Uppal, B., Sharma, V.K. 2002. Intestinal parasites in children with diarrhoea in Delhi, India. Southeast Asian J. Trop. Med. Public Health 33: 725-729.
Mahdi, N. K., Ali, N.H. 2004. Cryptosporidiosis and other intestinal parasitic infections in patients with chronic diarrhoea. Saudi Med. J. 25: 1204-1207.
Morgan, U.M., Pallant, L., Dwyer, B.W., Forbes, D.A., Rich, G., Thompson R.C.A. 1998. Comparison of PCR and microscopy for detection of Cryptosporidium parvum in human faecal specimens: clinical trial. J Clin Microbiol. 36:995â€“998
Seyrafian S., Pestehchian, N., Kerdegari, M., Yousefi, H.A., Bastani, B. 2006. Prevalence rate of Cryptosporidium infection in hemodialysis patients in Iran. Haemodial. Int. 10: 375-379.
Thompson, R. C. A., Olson, M.E., Zhu, G., Enomoto, S., Abrahamsen, M.S., Hijjawi, N. S. 2005. Cryptosporidium and Cryptosporidiosis. Advances in Parasitology 59: 77-158.
Ungar, B. L. P. 1990. Enzyme-linked immunoassay for detection of Cryptosporidium antigens in fecal specimens. J. Clinical Microbiology 28: 2491-2495.
Xiao, L., Fayer, R. 2008. Molecular characterisation of species and genotypes of Cryptosporidium and Giardia and assessment of zoonotic transmission. Int J Parasitol. 38: 1239â€“1255.
Xiao, L., Fayer, R., Ryan, U., Upton, S.J. 2004. Cryptosporidium taxonomy: recent advances and implications for public health. Clin. Microbiol. Rev. 17:72â€“97.
Yilmaz H., Zeynep, T., Mautalip, C. 2008. Investigation of cryptosporidiosis by enzyme-linked immunosorbent assay and microscopy in children with diarrhoea. Saudi Med. J. 29(4): 526-529.
Yvonne K., Berit, B., Katja, D., Arwid, D., Ronald, S. 2012. Cryptosporidiosis: comparison of three diagnostic methods and effects of storage temperature on detectability of cryptosporidia in cattle faeces. Parasitol. Res. 111: 165-171.