Document Type : Short Communication
Department of Pathobiology, Faculty of Veterinary Medicine, Science and Research Branch, Islamic Azad University, Tehran, Iran
Department of Microbiology, Faculty of Basic Sciences, Saveh Branch, Islamic Azad University, Saveh, Iran
The present study was conducted to investigate the detection and identification of Cryptosporidium species via molecular techniques and evaluate the serum concentrations of inflammatory factors in Cryptosporidium species. The fecal samples (n = 256) were collected from pre-weaned (≤ 2.00 months) calves and the positive samples were identified utilizing Ziehl-Neelsen staining. Nested species-specific multiplex PCR (nssm-PCR) and restriction fragment length polymorphism (RFLP) were used to identify the species and sub-species. The serum concentrations of IL-1β, IL-6, IL-12, TNF-α, and IFN-γ were also assessed. The results revealed that 10.54% of samples were positive. The results of Nested-PCR showed that 92.59% of the samples were positive for C. parvum while 7.41% were positive for C. andersoni. The results of RFLP confirmed 92.59% of the samples for C. parvum, 3.70% for C. muris / C. andersoni, and 3.70% for C. muris. The serum concentrations of IL-1β, IL-6, IL-12, TNF-α, and IFN-γ were significantly higher in the infected calves compared to those in healthy calves. However, the serum concentration of IFN-γ was significantly higher in the calves infected with C. parvum while the serum concentrations of TNF-α and IL-6 were significantly higher in those infected with C. andersoni. In conclusion, C. parvum was prevalent in the region and the calves demonstrated inflammatory responses to Cryptosporidium species.
- Yildirim A, Adanir R, Inci A, et al. Prevalence and genotyping of bovine Cryptosporidium species in the Mediterranean and Central Anatolia Region of Turkey. Comp Immunol Microbiol Infect Dis 2020; 69:101425. doi: 10.1016/j.cimid.2020.101425.
- Xiao L. Molecular epidemiology of cryptosporidiosis: an update. Exp Parasitol 2010; 124(1): 80-89.
- Díaz P, Varcasia A, Pipia AP, et al. Molecular characterization and risk factor analysis of Cryptosporidium spp. in calves from Italy. Parasitol Res 2018; 117(10): 3081-3090.
- Imre K, Lobo LM, Matos O, et al. Molecular characterisation of Cryptosporidium isolates from pre-weaned calves in Romania: is there an actual risk of zoonotic infections? Vet Parasitol 2011; 181(2-4): 321-324.
- Leitch GJ, He Q. Cryptosporidiosis - an overview. J Biomed Res 2012; 25(1):1-16.
- Delirezh N, Norian R, Azadmehr A. Changes in some pro-and anti-inflammatory cytokines produced by bovine peripheral blood mononuclear cells following foot and mouth disease vaccination. Arch Razi Inst 2016; 71(3): 199-207.
- Kongara K, Dukkipati V, Tai HM, et al. Differential transcription of selected cytokine and neuroactive ligand-receptor genes in peripheral leukocytes from calves in response to cautery disbudding. Animals (Basel) 2020; 10(7): 1187. doi: 10.3390/ ani10071187.
- Aboelsoued D, Hendaw SHM, Abo-Aziza FAM, et al. Copro-microscopical and immunological diagnosis of cryptosporidiosis in Egyptian buffalo-calves with special reference to their cytokine profiles. J Parasit Dis 2020; 44(3): 654-660.
- Canals A, Pasquali P, Zarlenga DS, et al. Local ileal cytokine responses in cattle during a primary infection with Cryptosporidium parvum. J Parasitol 1998; 84(1):125-130.
- Clavel A, Arnal AC, Sánchez EC, et al. Evaluation of the optimal number of faecal specimens in the diagnosis of cryptosporidiosis in AIDS and immunocompetent patients. Eur J Clin Microbiol Infect Dis 1995; 14(1): 46-49.
- Thompson RCA, Ash A. Molecular epidemiology of Giardia and Cryptosporidium infections. Infect Genet Evol 2016; 40: 315-323.
- Feng Y, Ortega Y, He G, et al. Wide geographic distribution of Cryptosporidium bovis and the deer-like genotype in bovines. Vet Parasitol 2007; 144(1-2): 1-9.
- Bouzid M, Elwin K, Nader JL, et al. Novel real-time PCR assays for the specific detection of human infective Cryptosporidium Virulence 2016; 7(4): 395-399.
- Rekha KM, Puttalakshmamma GC, D’Souza E. Comparison of different diagnostic techniques for the detection of cryptosporidiosis in bovines. Vet World 2016; 9(2): 211-215.
- Thomson S, Innes EA, Nicholas N, et al. A Multiplex PCR test to identify four common cattle–adapted Cryptosporidium Parasitol Open 2019; 5, e1, 1. doi: 10.1017/pao.2018.16.
- Xiao L, Fayer R, Ryan U, et al. Cryptosporidium taxonomy: recent advances and implications for public health. Clin Microbiol Rev 2004; 17(1): 72-97.
- Fayer R, Santín M, Trout JM. Cryptosporidium ryanae sp. (Apicomplexa: Cryptosporidiidae) in cattle (Bos taurus). Vet Parasitol 2008; 156(3-4): 191-198.
- Xiao L, Morgan UM, Limor J, et al. Genetic diversity within Cryptosporidium parvum and related Cryptosporidium Appl Environ Microbiol 1999; 65(8): 3386-3391.
- Kasimanickam RK, Kasimanickam VR, Olsen JR, et al. Associations among serum pro- and anti-inflammatory cytokines, metabolic mediators, body condition, and uterine disease in postpartum dairy cows. Reprod Biol Endocrinol 2013; 11: 103. doi: 10.1186/1477-7827-11-103.
- Beheshtipour J, Raeeszadeh M. Evaluation of inter-leukin-10 and pro-inflammatory cytokine profile in calves naturally infected with neonatal calf diarrhea syndrome. Arch Razi Inst 2020; 75(2): 213-218.