Author(s): Dina Aboelsoued, Seham H. M. Hendawy, Faten A. M. Abo-Aziza, Kadria N. Abdel Megeed
Keywords: Cryptosporidiosis, Buffalo-calves, Diagnosis, Coprological examination, ELISA, Cytokines
Cryptosporidiosis is considered to be one of the most devasting gastrointestinal diseases in calves. The aim of this study was to investigate Cryptosporidium parvum infection (C. parvum) in buffalo-calves with both copromicroscopic examination and enzyme linked immunosorbent assay (ELISA) using two C. parvum prepared antigens with regards to their cytokines profile; interferon- c (IFNc), interleukin (IL)-12 and IL-14 to achieve a proper diagnosis. All collected buffalo- calves’ fecal samples were examined by modified Ziehl–Neelsen staining technique. ELISA was performed to evaluate the diagnostic accuracy of the two C. parvum prepared antigens; crude whole oocyst (CWO) and crude sonicated oocyst (CSO) in detection of anti-C. parvum IgG in buffalo-calves’ sera. As well, concentrations of INF-c, IL-12 and IL-14 in the buffalo-calves’ serum samples were estimated. The results revealed that the overall parasitological incidence of cryptosporidiosis was 40%. However, the serological diagnosis by ELISA assay showed 53.75% and 27.5% when using CWO and CSO antigen, respectively. Also, the diagnostic efficacy parameters of both antigens; CWO and CSO showed a significant high specificity (83.3%) achieved by CSO antigen and a high sensitivity (71.8%) by CWO antigen. The levels of INF-c, IL-12 and IL-14 were significantly increased in positive Cryptosporidium infected group by both coprological and serological assays followed by the group which was positive for cryptosporidiosis by copro-microscopic examination only. The present study concluded that a combination of coprological and serological examination with reference to the cytokines profile is needed for proper diagnosis of cryptosporidiosis in buffalo-calves.
Aboelsoued D, Toaleb NI, Abdel Megeed KN, Hassan SE, Ibrahim S (2019) Cellular immune response and scanning electron microscopy in the evaluation of Moringa leaves aqueous extract effect on Cryptosporidium parvum in buffalo intestinal tissue explants. J Parasit Dis 43(3):393–401. https://doi.org/10.1007/s12639-019-01103-9
Akdis M, Aab A, Altunbulakli C, Azkur K, Costa RA, Crameri R et al (2016) Interleukins (from IL-1 to IL-38), interferons, transforming growth factor b, and TNF-a: receptors, functions, and roles in diseases. J Allergy Clin Immunol 138(4):984–1010. https://doi.org/10.1016/j.jaci.2016.06.033
Auray G, Facci MR, van Kessel J, Buchanan R, Babiuk LA, Gerdts V (2013) Porcine neonatal blood dendritic cells, but not monocytes, are more responsive to TLRs stimulation than their adult counterparts. PLoS One 8(5):e59629. https://doi.org/10.1371/journal.pone.0059629
Bauer C, Steng G, Prevot F, Dorchies P (2002) Seroprevalence of Oestrus ovis infection in sheep in southwestern Germany. Vet Parasitol 110(1–2):137–143 Bones AJ, Josse´ L, More C, Miller CN, Michaelis M, Tsaousis AD (2019) Past and future trends of Cryptosporidium in vitro research. Exp Parasitol 196:28–37. https://doi.org/10.1016/j.exppara.2018.12.001
Canals A, Pasquali P, Zarlenga DS, Fayer R, Almeria S, Gasbarre LC (1998) Local ileal cytokine responses in cattle during a primary infection with Cryptosporidium parvum. J Parasitol 84:125–130 Checkley W, White AC, Jaganath D, Arrowood MJ, Chalmers RM, Chen XM et al (2015) A review of the global burden, novel diagnostics, therapeutics, and vaccine targets for Cryptosporidium. Lancet Infect Dis 15(1):85–94. https://doi.org/10.1016/s1473-3099(14)70772-8
Current WL, Reese NC (1986) A comparison of endogenous development of three isolates of Cryptosporidium in suckling mice. J Protozool 33:98–108 Danisova´ O, Hala´nova´b M, Valenca´kova´ A, Lupta´kova´ L (2018) Sensitivity, specificity and comparison of three commercially available immunological tests in the diagnosis of Cryptosporidium species in animals. Braz J Microbiol 49(1):177–183. https://doi.org/10.1016/j.bjm.2017.03.016
Duranti A, Caccio SM, Pozio E, di Egidio A, de Curtis M, Battisti A, Scaramozzino P (2009) Risk factors associated with Cryptosporidium parvum infection in cattle. Zoonoses Public Health 56(4):176–182. https://doi.org/10.1111/j.1863-2378.2008.01173.x
Essa SH, Galila EM, Abdelwahab MG, Moustafa AM, Hamouda FK, El-Akabawy L (2014) Compare microscopy staining and polymerase chain reaction for diagnosis of cryptosporidium infection among Frisian calves in Minufiya governorate. BVMG 26(1):205–212 Garcia LS (2001) Diagnostic Medical Parasitology, 4th edn. ASM, Washington, DC Ghaffari S, Kalantari N (2014) Recognition of Cryptosporidium oocysts in fresh and old stool samples: comparison of four techniques. Asian Pac J Trop Biomed 4(2):S570–S574. https://doi.org/10.12980/APJTB.4.2014APJTB-2014-0067
Gharpure R, Perez A, Miller AD, Wikswo ME, Silver R, Hlavsa MC (2019) Cryptosporidiosis Outbreaks-United States, 2009–2017. Morb Mortal Wkly Rep 68:568–572. https://doi.org/10.15585/mmwr.mm6825a3
Gunasekera S, Zahedi A, O’Dea M, King B, Monis P, Thierry B et al (2020) Organoids and bioengineered intestinal models: potential solutions to the Cryptosporidium culturing dilemma. Microorganisms 8(5):E715. https://doi.org/10.3390/microorganisms8050715
Henriksen SA, Pohlenz JF (1981) Staining of cryptosporidia by a modified Ziehl–Neelsen technique. Acta Vet Scand 22:594–596 Kaushik K, Khurana S, Wanchu A, Malla N (2009) Serum immunoglobulin G, M and A response to Cryptosporidium parvum in Cryptosporidium-HIV co-infected patients. BMC Infect Dis 9:179. https://doi.org/10.1186/1471-2334-9-179
Lang C, Groß U, Lu¨der GGK (2007) Subversion of innate and adaptive immune responses by Toxoplasma gondii. Parasitol Res 100(2):191–203. https://doi.org/10.1007/s00436-006-0306-9
Laurent F, Lacroix-Lamande (2017) Innate immune responses play a key role in controlling infection of the intestinal epithelium by Cryptosporidium. Int J Parasitol 47:711–721. https://doi.org/10.1016/j.ijpara.2017.08.001
Leitch GJ, He Q (2012) Cryptosporidiosis- an overview. J Biomed Res 25(1):1–16. https://doi.org/10.1016/s1674-8301(11)60001-8
Lowry OH, Rosebrough NJ, Farr AB, Randall RJ (1951) Protein measurement with the folin-phenol reagent. J Biol Chem 193:265–275 McNair NN, Mead JR (2013) CD4? Effector and memory cell populations protect against Cryptosporidium Parvum infection. Microbes Infect 15:599–606 Morsy GH, Abdel Megeed KN, Hammam AM, Seliem MME, Khalil FAM, Aboelsoued D (2014) Prevalence of Cryptosporidium infection in buffalo calves with special reference to urea and creatinine levels. Global Vet 13:662–667 Nydam DV, Wade SE, Schaaf SL, Mohammed HO (2001) Number of Cryptosporidium parvum oocysts or Giardia spp cysts shed by dairy calves after natural infection. Am J Vet Res 62(10):1612–1615. https://doi.org/10.2460/ajvr.2001.62.1612
Nydam DV, Lindergard G, Guard CL, Schaaf SL, Wade SE, Mohammed HO (2002) Serological detection of exposure to Cryptosporidium parvum in cattle by ELISA and its evaluation in relation to coprological tests. Parasitol Res 88(9):797–803. https://doi.org/10.1007/s00436-002-0665-9
Handley RM, Cockwill C, McAllister TA, Jelinski M, Morck DW, Olson ME (1999) Duration of naturally acquired giardiosis and cryptosporidiosis in dairy calves and their association with diarrhea. J Am Vet Med Assoc 214:391–396 O’Donoghue PJ (1995) Cryptosporidium and cryptosporidiosis in man and animals. Int J Parasitol 25:139–195 Ouakli N, Belkhiri A, de Lucio A, Ko¨ster PC, Djoudi M et al (2018) Cryptosporidium-associated diarrhoea in neonatal calves in Algeria. Vet Parasitol Reg Stud Reports 12:78–84. https://doi.org/10.1016/j.vprsr.2018.02.005
Plutzer J, Lassen B, Jokelainen P, Djurkovic´-Djakovic´ O, Kucsera I et al (2018) Review of Cryptosporidium and Giardia in the eastern part of Europe, 2016. Euro Surveill 23(4):16. https://doi.org/10.2807/1560-7917.es.2018.23.4.16-00825
Quintero-Betancourt W, Gennaccaro AL, Scott TM, Rose JP (2003) Assessment of methods for detection of infectious Cryptosporidium oocysts and Giardia cysts in reclaimed effluents. Appl Environ Microbiol 69:5380–5388. https://doi.org/10.1128/aem.69.9.5380-5388.2003
Santin M (2020) Cryptosporidium and Giardia in Ruminants. Vet Clin North Am Food Anim Pract 36(1):223–238. https://doi.org/10.1016/j.cvfa.2019.11.005
Schreiber GH and Schreiber RD (2003) The interferon- c. In: Thomson AW, Lotze MT (eds) Chapter 24, The cytokine handbook, 4th Ed. London. ISBN 0-12-689663-1
Takeda K, Omata Y, Koyama T, Ohtani M, Kobayashi Y et al (2003) Increase of Th1 type cytokine mRNA expression in peripheral blood lymphocytes of calves experimentally infected with Cryptosporidium parvum. Vet Parasitol 113(3–4):327–331. https://doi.org/10.1016/s0304-4017(03)00080-3
Tessema TS, Schwamb B, Lochner M, Forster I, Jakobi V, Petry F (2009) Dynamics of gut mucosal and systemic Th1/Th2 cytokine responses in interferon-gamma and interleukin-12p40 knockout mice during primary and challenge Cryptosporidium parvum infection. Immunobiology 214(6):454–466. https://doi.org/10.1016/j.imbio.2008.11.015
Thomson S (2015) Cryptosporidiosis in farmed livestock. Ph.D. thesis. University of Glasgow, Glasgow, United Kingdom Thomson S, Hamilton CA, Hope JC, Katzer F, Mabbott NA et al (2017) Bovine cryptosporidiosis: impact, host-parasite interaction and control strategies. Vet Res 48(1):42. https://doi.org/10.1186/s13567-017-0447-0
Whitmire WM, Harp JA (1991) Characterization of bovine cellular and serum antibody responses during infection by Cryptosporidium parvum. Infect Immun 65:185–190 Zambriski JA, Nydam DV, Bowman DD, Bellosa ML, Burton AJ et al (2013) Description of fecal shedding of Cryptosporidium parvum oocysts in experimentally challenged dairy calves. Parasitol Res 12(3):1247–1254. https://doi.org/10.1007/s00436-012-3258-2
Copyright © 2025 Schobot.net All rights reserved.