Abstract
Cryptosporidiosis is a disease that affects calves, causing acute or chronic gastrointestinal disorders, and death. It is also considered one of the main causes of diarrhea in humans. The diagnosis of this disease is through the identification of oocysts by conventional methods, however, there is a lack of information about the performance of this methods in the field, so the objective of this study was to determine the most sensitive method, as well as to determine the frequency of the disease in Holstein cattle. Three diagnostic techniques were compared: Ziehl-Neelsen Cold Staining, Safranin-Methylene Blue Stain and Sheather Sugar Flotation. The results indicate that the best diagnostic method for the identification of Cryptosporidium spp. oocysts in feces was Ziehl-Neelsen. It is easy to perform and has the best sensitivity. A total frequency of cases was 43.4%, indicating a high prevalence of the disease, in calves it was 63.4%. A significant association (p<0.05) was observed between the presence of diarrhea and the presence of oocysts, so it is important to establish control and biosecurity measures in the production units to reduce the presence of this disease.
References
Aboelsoued, D., & Abdel, K.N. (2022). Diagnosis and control of Cryptosporidiosis in farm animals. Journal of Parasitic Diseases, 46(4), 1133–1146. https://doi.org/10.1007/s12639-022-01513-2
Amer, S., Zidan, S., Adamu, H., Ye, J., Roellig, D., Xiao, L., & Feng, Y. (2013). Prevalence and characterization of Cryptosporidium spp., in dairy cattle in Nile River delta provinces, Egypt. Experimental Parasitology, 135(3), 518-23. https://doi.org/10.1016/j.exppara.2013.09.002
Ares-Mazás, M.E., Fernández-da Ponte, B., Vergara-Castiblanco, C.A., Freire-Santos, F., Quílez-Cinca, J., Causapé-Valenzuela, A.C., & Sánchez-Acedo, C. (1999). Oocysts, IgG levels and immunoblot patterns determined for Cryptosporidium parvum in bovine examined during a visit to a farm (northeastern Spain). Veterinary Parasitology, 81(3), 185-93. https://doi.org/10.1016/s0304-4017(98)00245-3
Baxby, D., Blundell, N., & Hart, C,A. (1984). The development and performance of a simple, sensitive method for the detection of Cryptosporidium oocysts in faeces. The Journal of Hygiene, 93(2), 317-23. https://doi.org/10.1017/s0022172400064858
Brook, E., Hart, C.A., French, N., & Christley, R. (2008). Prevalence and risk factors for Cryptosporidium spp., infection in young calves. Veterinary Parasitolgy, 152(1-2), 46-52. https://doi.org/10.1016/j.vetpar.2007.12.003
Castelán-Hernández, O.O., Romero-Salas, D., García-Vázquez, Z., Cruz-Vázquez, C., Aguilar-Domínguez, M., Ibarra-Priego, N.D.J., & Muñoz-Melgarejo, S. (2011). Prevalencia de Criptosporidiosis bovina en tres regiones ecológicas de la zona centro de Veracruz, México. Tropical and Subtropical Agroecosystems, 13(3), 261-267. https://www.redalyc.org/articulo.oa?id=93920942024
Chalmers, R.M., & Davies, A.P. (2010). Minireview: Clinical Cryptosporidiosis. Experimental Parasitology, 124(1), 138-46. https://doi.org/10.1016/j.exppara.2009.02.003
Current, W.L., & Garcia, L.S. (1991). Cryptosporidiosis. Clinical Microbiology Reviews, 4(3), 325–358. https://doi.org/10.1128/CMR.4.3.325
Díaz de Ramírez, A., Ramírez, I., Lilido, N., Morillo, L., José, G., Barreto, B., & Alejandro, J. (2007). Infección con Cryptosporidium spp., y su asociación con diarrea en becerros de ganadería de doble propósito. Zootecnia Tropical, 25(1), 29-36. http://ve.scielo.org/scielo.php?script=sci_arttext&pid=S0798-72692007000100005&lng=es&tlng=es.
Díaz, P., Navarro, E., Remesar, S., García-Dios, D., Martínez-Calabuig, N., Prieto, A., López-Lorenzo, G., López, C.M., Panadero, R., Fernández, G., Díez-Baños, P., & Morrondo, P. (2021). The Age-Related Cryptosporidium Species Distribution in Asymptomatic Cattle from North-Western SPAIN. Animals, 11(2), 256. https://doi.org/10.3390/ani11020256
Elliot, A., Morgan, U.M., & Thompson, R.C. (1999). Improved staining method for detecting Cryptosporidium oocysts in stools using malachite green. Journal of General and Applied Microbiology, 45(3), 139-142. https://doi.org/10.2323/jgam.45.139
Fayer, R. (2010). Taxonomy and species delimitation in Cryptosporidium. Experimental Parasitology, 124(1), 90-7. https://doi.org/10.1016/j.exppara.2009.03.005
Fayer, R., Morgan, U., & Upton, S.J. (2000). Epidemiology of Cryptosporidium: transmission, detection, and identification. International Journal for Parasitology, 30(12-13), 1305-22. https://doi.org/10.1016/s0020-7519(00)00135-1
Fayer, R., Santín, 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(2), 105-12. https://doi.org/10.1016/j.vetpar.2005.08.003
Fitz-Sánchez, E., Rosario Cruz, R., Hernández Ortiz, R., Hernández Castro, E., Rodríguez Bataz, E., & García Vázquez, Z. (2013). Cryptosporidium parvum: prevalencia y factores de riesgo en becerros del municipio de Cuajinicuilapa, Guerrero, México. Revista Veterinaria Y Zootecnia, 7(1), 49–61. https://revistasojs.ucaldas.edu.co/index.php/vetzootec/article/view/4404
Fujino, T., Matsuo, T., Okada, M., & Matsui, T. (2006). Detection of a small number of Cryptosporidium parvum oocysts by sugar flotation and sugar centrifugation methods. Journal of Veterinary Medical Science, 68(11), 1191-3. https://doi.org/10.1292/jvms.68.1191
García, A.M., Cruz, C., Quezada, T., Silva, E., Valdivia, A., Vazquez, S., & Ramos, M. (2009). Cryptosporidiosis in Dairy Calves from Aguascalientes, Mexico: Risk Infection in Relation with the Seadon and Months of Sampling. Journal of Advanced Veterinary and Animal Research, 8(8), 1579-1583. https://www.researchgate.net/publication/287032121
García-Romo, D., Cruz-Vázquez, C., Quezada-Tristán, T., Silva-Peña, E., Valdivia-Flores, A., Vázquez-Flores, S., & Ramos-Parra, M. (2014). Prevalence and risk factors associated with infection by Cryptosporidium spp. In suckling calves in Aguascalientes, Mexico. Veterinaria México, 1(1), https://doi.org/10.21753/vmoa.1.1.332
González, M., Gómez, E.S., & Aluja, A.S. (1997). Criptosporidiosis en bovinos lactantes (histopatología, microscopía electrónica y de barrido). Veterinaria México, 14(1), 12-22. https://biblat.unam.mx/es/revista/veterinaria-mexico/articulo/criptosporidiosis-en-bovinos-lactantes-histopatologia-microscopia-electronica-de-transmision-y-de-barrido
Hadfield, S.J., Robinson, G., Elwin, K., & Chalmers, R.M. (2011). Detection and differentiation of Cryptosporidium spp. in human clinical samples by use of real-time PCR. Journal of Clinical Microbiology, 49(3), 918-24. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3067739/
Hawash, Y. (2014) Evaluation of an immunoassay-based algorithm for screening and identification of Giardia and Cryptosporidium antigens in human faecal specimens from saudi arabia. Journal of Parasitology Research, 2014, 213745 https://doi.org/10.1155/2014/213745
Instituto Nacional de Estadística y Geografía [INEGI]. (2010). Compendio de Información geográfica municipal 2010. Pedro Escobedo, Querétaro. https://www.inegi.org.mx/contenidos/app/mexicocifras/datos_geograficos/22/22012.pdf
Instituto Nacional de Estadística y Geografía [INEGI]. (2020). Panorama sociodemográfico de Querétaro: Censo de Población y Vivienda 2020. https://www.inegi.org.mx/contenidos/productos/prod_serv/contenidos/espanol/bvinegi/productos/nueva_estruc/702825197957.pdf
Kehl, K.S., Cicirello, H., & Havens, P.L. (1995). Comparison of four different methods for detection of Cryptosporidium species. Journal of Clinical Microbiology, 33(2), 416–418. https://doi.org/10.1128/jcm.33.2.416-418.1995
Khan, S., Khan, A., Khan, I., & Muhammad, I. (2016). Differential techniques used for detection of Cryptosporidium oocysts in stool specimens. Journal of Parasitic Diseases, 1(1), 1-11. https://www.researchgate.net/publication/338823938_DIFFERENTIAL_TECHNIQUES_USED_FOR_DETECTION_OF_CRYPTOSPORIDIUM_OOCYSTS_IN_STOOL_SPECIMENS
Kumar, D., Sreekrishnan, R., & Das, S.S. (2004). Cryptosporidiosis in man and animals in Pondicherry. Indian Journal of Animal Sciences, 74(3), https://epubs.icar.org.in/index.php/IJAnS/article/view/38362
Lombardelli, J.A., Tomazic, M.L., Schnittger, L., & Tiranti, K.I. (2019). Prevalence of Cryptosporidium parvum in dairy calves and GP60 subtyping of diarrheic calves in central Argentina. Parasitology Research, 118(7), 2079-2086. https://doi.org/10.1007%2Fs00436-019-06366-y
López-Torres, L.L, López-Cuevas, O., Vázquez-Vázquez, C., Alvarado-Gómez, O.G., Vázquez-Alvarado, R.E., Rodríguez-Fuentes, H., Chaidez-Quiroz, C., & Vidales-Contreras, J.A. (2020). Prevalence of Cryptosporidium spp. in dairy livestock from the Laguna Region, Mexico. Revista Bio Ciencias 7, e881. https://doi.org/10.15741/revbio.07.e881
Maldonado-Camargo, S., Atwill, E.R., Saltijeral-Oaxaca, J.A., & Herrera-Alonso, L.C. (1998). Prevalence of and risk factors for shedding of Cryptosporidium parvum in Holstein Freisian dairy calves in central México. Preventive Veterinary Medicine, 36(2), 95-107. https://doi.org/10.1016/s0167-5877(98)00084-1
McCluskey, B.J., Greiner, E.C., & Donovan, G.A. (1995). Patterns of Cryptosporidium oocyst shedding in calves and a comparison of two diagnostic methods. Veterinary Parasitology, 60(3–4), 185–190. https://doi.org/10.1016/0304-4017(95)00790-4
Nwosu, A., Berke, O., Pearl, D.L., & Trotz-Williams, L.A. (2019). Exploring the geographical distribution of Cryptosporidiosis in the cattle population of Southern Ontario, Canada, 2011-2014. Geospatial Health, 14(2), https://doi.org/10.4081/gh.2019.769
Omoruyi, B., Nwodo, U., Udem, C., & Okonkwo, F. (2014). Comparative Diagnostic Techniques for Cryptosporidium Infection Molecules, 19(2), 2674–2683. https://doi.org/10.3390/molecules19022674
Rajkhowa, S., Rajkhowa, C., & Hazarika, G.C., (2006). Prevalence of Cryptosporidium parvum in mithuns (Bos frontalis) from India. Veterinary Parasitology, 142(1–2), 146–149. https://doi.org/10.1016/j.vetpar.2006.06.026
Ralston, B., Thompson, R.C., Pethick, D., McAllister, T.A., & Olson, M,E. (2010). Cryptosporidium andersoni in Western Australian feedlot cattle. Australian Veterinary Journal, 88(11), 458-60. https://pubmed.ncbi.nlm.nih.gov/20973292/
Rekha, K.M., Puttalakshmamma, G.C., & D'Souza, P.E. (2016). Comparison of different diagnostic techniques for the detection of Cryptosporidiosis in bovines. Veterinary World, 9(2), 211-5. https://doi.org/10.14202/vetworld.2016.211-215
Rodríguez, A. (2016). Identificación de Cryptosporidium en becerras Holstein de 1 día a 12 meses de edad. [Tesis de Licenciatura, Universidad Autónoma Agraria Antonio Narro]. https://repositorio.uaaan.mx/xmlui/bitstream/handle/123456789/8264/ANA%20JANETH%20RODRIGUEZ%20CABRERA.pdf?sequence=1&isAllowed=y
Rojekittikhun, W., Mahittikorn, A., Prummongkol, S., Puangsa-Art, S., Chaisiri, K., & Kusolsuk, T. (2015). Evaluation of sugar flotation and formalin-ether concentration techniques in the examination of GI parasites of refuge dogs and cats in Kanchanaburi province, Thailand. Tropical Medicine & Parasitology, 38, 17-24. https://ptat.org/uploads/pdf/journalPdf_38-1-2015-e3.pdf
Romero-Salas, D., Alvarado-Esquivel, C., Cruz-Romero, A., Aguilar-Domínguez, M., Ibarra-Priego, N., Merino-Charrez, J.O., Pérez de León, A.A., & Hernández-Tinoco, J. (2016). Prevalence of Cryptosporidium in small ruminants from Veracruz, Mexico. BMC Veterinary Research, 12, 14. https://doi.org/10.1186/s12917-016-0638-3
Rubio-Guarín, D. (2010). Comparación de tres técnicas de tinción para la identificación de Cryptosporidium spp., a partir de muestras coprológicas obtenidas en una población infantil de Calarcá Quindío [Tesis de Licenciatura, Universidad del Quindío, Armenia] https://bdigital.uniquindio.edu.co/server/api/core/bitstreams/18cbece2-2357-49f2-8497-0e41dbc093e4/content
Ruest, N., Faubert, G.M., & Couture, Y. (1998). Prevalence and geographical distribution of Giardia spp. and Cryptosporidium spp. in dairy farms in Québec. The Canadian Veterinary Journal, 39(11), 697-700. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1539484/
Saha, S., Sarkar, S., Batabyal, S., Pramanik, A.K., & Das, P. (2006). Observations on the epidemiology of bovine cryptosporidiosis in India. Veterinary Parasitology, 141(3–4), 330–333. https://doi.org/10.1016/j.vetpar.2006.06.017
Santín, M. (2020). Cryptosporidium and Giardia in Ruminants. Veterinary Clinics of North America: Food Animal Practice, 36(1), 223-238. https://doi.org/10.1016/j.cvfa.2019.11.005
Santín, M., Trout, J.M., & Fayer, R. (2008). A longitudinal study of cryptosporidiosis in dairy cattle from birth to 2 years of age. Veterinary Parasitology, 155(1-2), 15-23. https://doi.org/10.1016/j.vetpar.2008.04.018
Santín, M., Trout, J.M., Xiao, L., Zhou, L., Greiner, E., & Fayer, R. (2004). Prevalence and age-related variation of Cryptosporidium species and genotypes in dairy calves. Veterinary Parasitology, 122(2), 103-17. https://doi.org/10.1016/j.vetpar.2004.03.020
Silverlås, C., Bosaeus-Reineck, H., Näslund, K., & Björkman, C. (2013). Is there a need for improved Cryptosporidium diagnostics in Swedish calves?. International Journal for Parasitology, 43(2), 155–161. https://doi.org/10.1016/j.ijpara.2012.10.009
Sischo, W.M., Atwill, E.R., Lanyon, L.E., & George, J. (2000). Cryptosporidium on dairy farms and the role these farms may have in contaminating surface water supplies in the northeastern United States. Preventive Veterinary Medicine, 43(4), 253–267. https://doi.org/10.1016/S0167-5877(99)00107-5
Smith, R.P., Clifton-Hadley, F.A., Cheney, T., & Giles, M. (2014). Prevalence and molecular typing of Cryptosporidium in dairy cattle in England and Wales and examination of potential on-farm transmission routes. Veterinary Parasitology, 204(3-4), 111-9. https://doi.org/10.1016%2Fj.vetpar.2014.05.022
Starkey, S.R., Kimber, K.R., Wade, S.E., Schaaf, S.L., White, M.E., & Mohammed, H.O. (2006). Risk Factors Associated with Cryptosporidium Infection on Dairy Farms in a New York State Watershed. Journal of Dairy Science, 89(11), 4229–4236. https://doi.org/10.3168/jds.S0022-0302(06)72468-7
Tarekegn, Z.S., Tigabu, Y., & Dejene, H. (2021). Cryptosporidium infection in cattle and humans in Ethiopia: A systematic review and meta-analysis. Parasite Epidemiology and Control, 13,(14), e00219. https://doi.org/10.1016/j.parepi.2021.e00219
Thomson, S., Hamilton, C.A., Hope, J.C., Katzer, F., Mabbott, N.A., Morrison, L.J., & Innes, E.A. (2017). Bovine Cryptosporidiosis: impact, host-parasite interaction and control strategies. Veterinary Research, 48(1), 42. https://doi.org/10.1186%2Fs13567-017-0447-0
Trotz-Williams, L. A., Martin, D. S., Gatei, W., Cama, V., Peregrine, A. S., Martin, S. W., Nydam, D. V., Jamieson, F., & Xiao, L. (2006). Genotype and subtype analyses of Cryptosporidium isolates from dairy calves and humans in Ontario. Parasitology Research, 99(4), 346–352. https://doi.org/10.1007/s00436-006-0157-4
Trotz-Williams, L.A., Jarvie, B.D., Martin, S., Leslie, K.E., & Peregrine, A.S. (2005). Prevalence of Cryptosporidium parvum infection in southwestern Ontario and its association with diarrhea in neonatal dairy calves. Canadian Veterinary Journal, 46(4), 349– 351. https://pubmed.ncbi.nlm.nih.gov/15943123/
Xiao, L., Morgan, U.M., Limor, J., Escalante, A., Arrowood, M., Shulaw, W., Thompson, R.C., Fayer, R., & Lal, A.A. (1999). Genetic diversity within Cryptosporidium parvum and related Cryptosporidium species. Applied and Environmental Microbiology, 65(8), 3386-91. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC91508/
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