Prevalence and Diagnostic Methods for Cryptosporidium in Different Dairy Farm Environments

Kamran Ullah; Iftikhar Ali Shah; Hina Khan; Walayat  Ullah; Zulfiqar Ahmad; Aneela Ali; Muhammad Abbas; Saddam Hussain; Shahzada Khurram Adrian Shah

Prevalence and Diagnostic Methods for Cryptosporidium in Different Dairy Farm Environments

Authors

Kamran Ullah College of Veterinary Sciences and Animal Husbandry, Abdul Wali Khan University Mardan, Khyber Pakhtunkhwa, Pakistan.
Iftikhar Ali Shah Institute of Biological Science, Gomal University, Dera Ismail Khan, Khyber Pakhtunkhwa, Pakistan.
Hina Khan Institute of Biological Science, Gomal University, Dera Ismail Khan, Khyber Pakhtunkhwa, Pakistan.
Walayat Ullah Department of Basic and Applied Zoology, Shaheed Benazir Bhutto University, Sheringal Dir Upper, Khyber Pakhtunkhwa, Pakistan.
Zulfiqar Ahmad Department of Parasitology, University of Veterinary and Animal Sciences Lahore, Pakistan.
Aneela Ali Department of Microbiology, University of Veterinary and Animal Sciences Lahore, Pakistan.
Muhammad Abbas Department of Basic and Applied Zoology, Shaheed Benazir Bhutto University, Sheringal Dir Upper, Khyber Pakhtunkhwa, Pakistan.
Saddam Hussain Department of Basic and Applied Zoology, Shaheed Benazir Bhutto University, Sheringal Dir Upper, Khyber Pakhtunkhwa, Pakistan.
Shahzada Khurram Adrian Shah Department of Animal Health, The University of Agriculture Peshawar, Pakistan.

Keywords:

Cryptosporidium, Calves, Infection, Microscopic Analysis

Abstract

The prevalence of Cryptosporidium infection varied among different dairy farms, with Iqbal Dairy Farm recording the lowest prevalence (12.5%), while Nawan Kalay Farm (36.66%) and Toru Dairy Farm (33.33%) had the highest. Sex-based prevalence analysis indicated a slightly higher infection rate in female calves (25.12%) compared to males (22.75%). Among breeds, Holstein Friesians had the highest prevalence (30.53%), whereas Jersey (22.58%) and crossbreeds (21.98%) exhibited lower infection rates. A total of 384 samples were examined using modified Ziehl–Neelsen (MZN) staining, of which 92 were identified as positive for Cryptosporidium. PCR analysis was performed for confirmation, revealing that 60 of the 92 samples (53.57%) were true positives, while 52 (46.42%) tested negative. The molecular detection method (PCR) demonstrated a statistically significant difference in sensitivity compared to microscopy, confirming Cryptosporidium infection in 60 out of 112 samples (53%). These findings highlight the variability in Cryptosporidium prevalence across farms, breeds, and sex, as well as the importance of molecular techniques for accurate diagnosis.

References

. Thompson, R. C. A., Ash, A., & Lymbery, A. J. (2016). The epidemiology of zoonotic cryptosporidiosis in humans, animals and the environment. International Journal for Parasitology, 46(3), 197-206.

. Ryan, U., Paparini, A., & Monis, P. (2021). Cryptosporidium in humans and animals: A one health perspective. International Journal for Parasitology, 51(3-4), 235-246.

. Fayer, R., Santín, M., & Trout, J. M. (2017). Cryptosporidium in dairy cattle: From observation to intervention. Trends in Parasitology, 33(12), 958-970.

. Santín, M. (2020). Clinical and subclinical cryptosporidiosis in animals. Veterinary Parasitology, 276, 109-119.

. Khan, N.U., Usman, T., Sarwar, M.S., Ali, H., Gohar, A., Asif, M., Rabbani, F., Khan, R.U., Sultana, N., Khan, N.A. and Mobashar, M., 2022. The prevalence, risk factors analysis and evaluation of two diagnostic techniques for the detection of Cryptosporidium infection in diarrheic sheep from Pakistan. Plos one, 17(7), p.e0269859.

. Ullah, K., Ali, U., Ahsan, M. and Khan, N.U., 2024. Risk Factors Analysis of Bovine Cryptosporidiosis in Diarrheic Calves. Journal of Animal and Plant Research, 1(4), pp.1-9.

. Urie, N.J., Lombard, J.E., Shivley, C.B., Kopral, C.A., Adams, A.E., Earleywine, T.J., Olson, J.D. and Garry, F.B., 2018. Preweaned heifer management on US dairy operations: Part V. Factors associated with morbidity and mortality in preweaned dairy heifer calves. Journal of dairy science, 101(10), pp.9229-9244.

. Causapé, A.C., Quılez, J., Sánchez-Acedo, C., Del Cacho, E. and López-Bernad, F., 2002. Prevalence and analysis of potential risk factors for Cryptosporidium parvum infection in lambs in Zaragoza (northeastern Spain). Veterinary parasitology, 104(4), pp.287-298.

. Lechevallier, M.W. And Moser, R.H., 1995. Occurrence of Giardia and Cryptosporidium in Raw and Finished Drinking Water. J. Am. Water Works Assoc, 87, Pp.54-68.

. Lefay, D., Naciri, M., Poirier, P. and Chermette, R., 2000. Prevalence of Cryptosporidium infection in calves in France. Veterinary Parasitology, 89(1-2), pp.1-9.

. Maurya, P.S., Rakesh, R.L., Pradeep, B., Kumar, S., Kundu, K., Garg, R., Ram, H., Kumar, A. and Banerjee, P.S., 2013. Prevalence and risk factors associated with Cryptosporidium spp. infection in young domestic livestock in India. Tropical Animal Health and Production, 45(4), pp.941-946.

. Wang, Y., Zhang, B., Li, J., Yu, S., Zhang, N., Liu, S., Zhang, Y., Li, J., Ma, N., Cai, Y. and Zhao, Q., 2021. Development of a quantitative real-time PCR assay for detection of Cryptosporidium spp. infection and threatening caused by Cryptosporidium parvum subtype IIdA19G1 in diarrhea calves from Northeastern China. Vector-Borne and Zoonotic Diseases, 21(3), pp.179-190.

. Wegayehu, T., Adamu, H. and Petros, B., 2013. Prevalence of Giardia duodenalis and Cryptosporidium species infections among children and cattle in North Shewa Zone, Ethiopia. BMC infectious diseases, 13(1), pp.1-7.

. Uppal, B., Singh, O., Chadha, S. and Jha, A.K., 2014. A comparison of nested PCR assay with conventional techniques for diagnosis of intestinal cryptosporidiosis in AIDS cases from northern India. Journal of parasitology research, 2014(1), p.706105.

. Morris, A.J., Noble, S., Lindo, J.F. and Ong, C.S., 2022.First report of Cryptosporidium spp. In: Cattle from Jamaica. https://doi.org/10.21203/rs.3.rs-1503219/v

. Helmy, Y.A., Krücken, J., Nöckler, K., von Samson-Himmelstjerna, G. and Zessin, K.H., 2013. Molecular epidemiology of Cryptosporidium in livestock animals and humans in the Ismailia province of Egypt. Veterinary parasitology, 193(1-3), pp.15-24.