PCR-RFLP at 18S rRNA gene for identification of Sarcocystis species and their prevalence in cattle of Andhra Pradesh

K MOUNIKA; SREEDEVI CHENNURU; R VENU; T SRINIVASA RAO; SUDHAKAR KROVVIDI

doi:10.56093/ijans.v89i4.89140

Authors

K MOUNIKA Scholar, Sri Venkateswara Veterinary University, Tirupati, Andhra Pradesh 517 502 India
SREEDEVI CHENNURU Professor, Sri Venkateswara Veterinary University, Tirupati, Andhra Pradesh 517 502 India
R VENU Associate Professor, Sri Venkateswara Veterinary University, Tirupati, Andhra Pradesh 517 502 India
T SRINIVASA RAO Associate Professor, Sri Venkateswara Veterinary University, Tirupati, Andhra Pradesh 517 502 India
SUDHAKAR KROVVIDI Assistant Professor, Sri Venkateswara Veterinary University, Tirupati, Andhra Pradesh 517 502 India

https://doi.org/10.56093/ijans.v89i4.89140

Keywords:

Andhra Pradesh, Cattle sarcocysts, PCR-RFLP, Sarcocystis cruzi, S. hirsuta, S. fusiformis

Abstract

In the present study, the species specific determination of Sarcocystis cysts isolated from 137 naturally infected and slaughtered cattle from different regions of Chittoor district, Andhra Pradesh, India was achieved by PCR-RFLP. Diagnosis of sarcocysts in cattle was based on macroscopic examination and pepsin digestion method. Genomic DNA was extracted individually from bradyzoites of all 137 sarcocystosis positive cattle and all the isolates were characterized at 18S rRNA. Digestion of PCR amplicons (900 bp) with restriction endonuclease revealed 3 different electromorphs which were referred to Sarcocystis cruzi (513 and 343 bp), S. hirsuta (525, 241 and 141 bp) and S. fusiformis (532 and 335 bp). Sarcocystis cruzi (93.43%) was significantly more prevalent in comparison with the S. hirsuta (4.38%) and S. fusiformis (2.19%). Infection of cattle with S. hominis was not observed in the study area. Occurrence of S. fusiformis in cattle supports that Sarcocystis species of buffaloes are not strictly intermediate host specific however can infect cattle.

Downloads

Download data is not yet available.

References

Badawy A I I, Abouzaid N Z and Ahmed H A. 2012. Sarcocystis hominis and other Sarcocystis species infecting cattle in Sharkia Province, Egypt. Journal of American Science 8: 271– 75.

Dafedar A, D’Souza P E and Mamatha G S. 2011. Prevalence and morphological studies on Sarcocystis species infecting cattle in Bangalore. Journal of Veterinary Parasitology 25: 183–84.

Daptardarkar M, Bagicha Singh B, Aulakh R S and Gill J P S. 2016. Prevalence and first molecular identification of Sarcocystis species in cattle and water buffaloes in India. Acta Parasitologica 61: 523–28. DOI: https://doi.org/10.1515/ap-2016-0069

Domenis L, Peletto S, Sacchi L, Clementi E, Genchi M, Felisari L, Felisari C, Mo P, Modesto P, Zuccon F, Campanella C, Maurella C, Guidetti C and Acutis P L. 2011. Detection of a morphogenetically novel Sarcocystis hominis-like in the context of a prevalence study in semi-intensively bred cattle in Italy. Parasitology Research 109: 1677–87. DOI: https://doi.org/10.1007/s00436-011-2441-1

Dubey J P, Speer C A and Fayer R. 1989. Sarcocystosis of Animals and Man. C R C Press, Boca Raton, Florida, pp 215.

Fayer R. 2004. Sarcocystis spp. in human infections. Clinical Microbiology Reviews 17: 894–902. DOI: https://doi.org/10.1128/CMR.17.4.894-902.2004

Gjerde B, Hilali M and Abbas I E. 2016. Molecular differentiation of Sarcocystis buffalonis and Sarcocystis levinei in water buffaloes (Bubalus bubalis) from Sarcocystis hirsuta and Sarcocystis cruzi in cattle (Bos taurus). Parasitology Research 115: 2549–71. DOI: https://doi.org/10.1007/s00436-016-4998-1

Hornok S, Mester A, Takacs N, Baska F, Majoros G, Fok E, Biksi I, Nemet Z, Hornyak A, Janosi S and Farkas R. 2015. Sarcocystis-infection of cattle in Hungary. Parasites and Vectors 8: 69. DOI: https://doi.org/10.1186/s13071-015-0685-9

Huong L T T, Dubey J P, Nikkila T and Uggla A. 1997. Sarcocystis buffalonis n.sp. (Protozoa: Sarcocystidae) from the water buffalo (Bubalus bubalis) in Vietnam. Journal of Parasitology 83: 471–74. DOI: https://doi.org/10.2307/3284413

Jehle C, Dinkel A, Sander A, Morent M, Romig T, Luc P V, De T V, Thai V V and Mackenstedt U. 2009. Diagnosis of Sarcocystis spp. in cattle (Bos taurus) and water buffalo(Bubalus bubalis) in northern Vietnam. Veterinary Parasitology 166: 314–20. DOI: https://doi.org/10.1016/j.vetpar.2009.08.024

Li Q Q, Yang Z Q, Zuo Y X, Attwood S W, Chen X W and Zhang Y P. 2002. A PCR-based RFLP analysis of Sarcocystis cruzi in Yunnan Province PR China reveals the water buffalo (Bubalus bubalis) as a natural intermediate host. Journal of Parasitology 88: 1259–61. DOI: https://doi.org/10.1645/0022-3395(2002)088[1259:APBRAO]2.0.CO;2

Mounika K, Chennuru S, Ravipati V, Tumati S R and Krovvidi S. 2018. Studies on prevalence and histomorphology of Sarcocystis species infecting cattle in Andhra Pradesh, India. Journal of Parasitic Disease 42: 77–80. DOI: https://doi.org/10.1007/s12639-017-0968-5

Sambrook J, Fritsch E F and Maniatis T. 1989. Molecular Cloning: A Laboratory Manual. 2nd edn. Cold Spring Harbor Laboratory Press, NY, USA.

Vangeel L, Houf K, Chier K, Vercruysse J, D’Herde K and Ducatelle R. 2007. Molecular based identification of Sarcocystis hominis in Belgian minced beef. Journal of Food Protection 70: 1523–26. DOI: https://doi.org/10.4315/0362-028X-70.6.1523

Velasquez J N, Di Risio C, Etchart C B, Chertcoff A V, Mendez N, Cabrera M G, Labbe J H and Carnevale S. 2008. Systemic sarcocystosis in a patient with acquired immune deficiency syndrome. Human Pathology 39: 1263–67. DOI: https://doi.org/10.1016/j.humpath.2008.01.016

Wouda W, Snoep J J and Dubey J P. 2006. Eosinophilic myositis due to Sarcocystis hominis in a beef cow. Journal of Comparative Pathology 135: 249–53. DOI: https://doi.org/10.1016/j.jcpa.2006.07.004

World Health Organization. 1981. Intestinal protozoan and helminthic infections. Vol. 666. World Health Organization, Geneva.

Xiang Z, He Y, Zhao H, Rosenthal B M, Dunams D B, Li X, Zuo Y, Feng G, Cui L and Yang Z. 2011. Sarcocystis cruzi: Comparitive studies confirm natural infections of buffaloes. Experimental Parasitology 127: 460–66. DOI: https://doi.org/10.1016/j.exppara.2010.10.012

Yang Z Q, Li Q Q, Zuo Y X, Chen X W, Chen Y J, Nie L, Wei C G, Zen J S X, Attwood S W, Zhang X Z and Zhang Y P. 2002. Characterization of Sarcocystis species in domestic animals using a PCR-RFLP analysis of variation in the 18S rRNA gene, a cost-effective and simple technique for routine species identification. Experimental Parasitology 102: 212–17. DOI: https://doi.org/10.1016/S0014-4894(03)00033-X