Skip to main content

Periurban outbreaks of bovine calf scours in Northern India caused by Cryptosporidium in association with other enteropathogens

  • A. P. S. BRAR (a1), N. K. SOOD (a1), P. KAUR (a2), L. D. SINGLA (a2), B. S. SANDHU (a1), K. GUPTA (a1), D. NARANG (a3), C. K. SINGH (a1) and M. CHANDRA (a3)...

Bovine calf scours reported to be caused by multiple aetiologies resulting in heavy mortality in unweaned calves and huge economic loss to the dairy farmers. Among these, cryptosporidiosis is an emerging waterborne zoonoses and one of the important causes of neonatal calf diarrhoea. Poor immune response coupled with primary cryptosporidial infections predispose neonatal calves to multiple secondary infections resulting in their deaths. In the present study, faecal samples from 100 diarrhoeic calves randomly picked up out of 17 outbreaks of bovine calf diarrhoea in periurban Ludhiana, Punjab in Northern India were subjected to conventional (microscopy, modified Zeihl–Neelsen (mZN) staining) and immunological and molecular techniques (faecal antigen capture ELISA and PCR) for detection of primary Cryptosporidium parvum infection as well as other frequently reported concurrent pathogens, viz. rotavirus and coronavirus, Salmonella spp., Escherichia coli, Clostridium perfringens and Eimeria spp. The faecal antigen capture ELISA and PCR revealed 35% prevalence of C. parvum in contrast to 25% by mZN staining with a relatively higher prevalence (66·7%) in younger (8–14-day-old) calves. The detection rate of the other enteropathogens associated with C. parvum was 45·71% for C. perfringens followed by Salmonella spp (40·0%), rotavirus (36·0%), coronavirus (16·0%), E. coli (12·0%) and Eimeria spp (4·0%) The sensitivity for detection of C. parvum by ELISA and mZN staining in comparison to PCR was 97·14% and 72·72%, respectively. An important finding of the study was that C. parvum alone was found in only 10% of the diarrhoeic faecal samples, whereas, majority of the samples (90%) showed mixed infections ranging from a combination of two to five agents. This is the first documentary proof of C. parvum and associated pathogens responsible for severe periurban outbreaks of bovine calf diarrhoea culminating in heavy mortality from Northern India.

Corresponding author
*Author for correspondence: Dr A. P. S. Brar, Department of Veterinary Pathology, College of Veterinary Sciences, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana-141004, Punjab, India. (Email:
Hide All
1. Otter A, Cranwell M. Differential diagnosis of diarrhoea in adult cattle. In Practice 2007; 29: 919.
2. Bangoura B, Daugschies A. Influence of experimental Eimeria zuernii infection in calves on electrolyte concentrations, acidbase balance and blood gases. Parasitology Research 2007; 101: 16371645.
3. Gay CC. Intestinal diseases in ruminants. In: Kahn CM, ed. Merck Veterinary Manual. U.S.A, Merck and Co. inc, N.J.: Whitehouse Station, 2005, pp. 220233.
4. Haschek B, et al. Detection of bovine torovirus in neonatal calf diarrhoea in Lower Austria en Styria (Austria). Journal of Veterinary Medicine. B, Infectious Diseases and Veterinary Public Health 2006; 53: 160165.
5. Singh BB, et al. Prevalence of Cryptosporidium parvum infection in Punjab (India) and its association with diarrhoea in neonatal dairy calves. Veterinary Parasitology 2006; 140: 162165.
6. Cho Y, Yoon KJ. An overview of calf diarrhoea – infectious etiology, diagnosis, and intervention. Journal of Veterinary Science 2014; 15(1): 117.
7. Chattopadhyay UKD, et al. Prevalence of cryptosporidiosis in man and animals in and around Calcutta. Journal of Veterinary Parasitology 2000; 14: 167168.
8. Kumar D, Sreekrishnan R, Das SS. Cryptosporidiosis in man and animals in Pondicherry. Indian Journal of Animal Sciences 2004; 74: 261263.
9. Bhat SA, Juyal PD, Singla LD. Bovine cryptosporidiosis: brief review of its distribution in India. Trends in Parasitology Research 2013; 2(2): 513.
10. Xiao L, Fayer R, Ryan U, Upton SJ. Cryptosporidium taxonomy: recent advances and implications for public health. Clinical Microbiology Reviews 2004; 17: 7297.
11. Randhawa SS, Randhawa SS, Zahid UN, Singla LD, Juyal PD. Drug combination therapy in control of cryptosporidiosis in Ludhiana district of Punjab. Journal of Parasitic Diseases 2012; 36: 269272.
12. MacKenzie WR, et al. A massive outbreak in Milwaukee of Cryptosporidium infection transmitted through the public water supply. The New England Journal of Medicine 1994; 331: 161167.
13. Athanassious R, et al. Detection of bovine coronavirus and type A rotavirus in neonatal calf diarrhoea and winter dysentery of cattle in Quebec: evaluation of three diagnostic methods. Canadian Veterinary Journal 1994; 35: 163169.
14. Ruest N, Faubert GM, Couture Y. Prevalence and geographical distribution of Giardia spp. and Cryptosporidium spp. in dairy farms in Quebec. Canadian Veterinary Journal 1998; 39: 697700.
15. Nasir A, et al. Prevalence of Cryptosporidium parvum infection in Lahore (Pakistan) and its association with diarrhoea in dairy calves. International Journal of Agriculture & Biology 2009; 11(2): 221224.
16. Nooruddin M, Sarma DK. Role of Cryptosporidium in calf diarrhoea: in annual report. Livestock Advisor 1987; 12: 49.
17. Dubey JP, Fayer R, Rao JR. Cryptosporidial oocysts in faeces of water buffalo and zebu calves in India. Journal of Veterinary Parasitology 1992; 6: 5556.
18. Das G, et al. Prevalence of Cryptosporidium infection in cattle. Journal of Veterinary Public Health 2005; 2: 1517.
19. Jeyabal L, Ray DD. 2005. Cryptosporidial infection in cattle and buffaloes. Journal of Veterinary Parasitology 2005; 19: 165166.
20. Randhawa SS, et al. Therapeutic management of cryptosporidiosis in cross bred dairy calves. Indian Veterinary Journal 2012; 89: 1719.
21. Roy SS, et al. Observations on the epidemiology of bovine cryptosporidiosis in India. Veterinary Parasitology 2006; 141: 330333.
22. Paul S, et al. Comparative evaluation and economic assessment of coprological diagnostic methods and PCR for detection of Cryptosporidium spp. in bovines. Veterinary Parasitology 2009; 164: 291295.
23. Venu R, et al. Factors influencing on prevalence of Cryptosporidium infection in south Indian dairy calves. Journal of Parasitic Diseases 2013; 37(2): 168172.
24. Bhat SA, et al. Comparison of nested PCR and microscopy for the detection of cryptosporidiosis in bovine calves. Journal of Parasitic Diseases 2014; 38: 101105.
25. Cho YI, et al. Development of a panel of multiplex real-time polymerase chain reaction assays for simultaneous detection of major agents causing calf diarrhoea in feces. Journal of Veterinary Diagnostic Investigation 2010; 22(4): 509517.
26. Xiao L, Herd RP, Rings DM. Concurrent infections of Giardia and Cryptosporidium on two Ohio farms with calf diarrhoea. Veterinary Parasitology 1993; 51(1–2): 4148.
27. de la Fuente R, et al. Cryptosporidium and concurrent infections with other major enteropathogens in 1–30-day-old diarrhoeic dairy calves in central Spain. Veterinary Parasitology 1999; 80: 179185.
28. García A, et al. Rotavirus and concurrent infections with other enteropathogens in neonatal diarrhoeic dairy calves in Spain. Comparative Immunology, Microbiology and Infectious Diseases 2000; 23(3): 175183.
29. Becher KA, et al. Molecular epidemiology of Giardia and Cryptosporidium infections in dairy calves originating from three sources in Western Australia. Veterinary Parasitology 2004; 123: 19.
30. Fayer R, Santin M, Dumitru M. Detection of concurrent infection of dairy cattle with Blastocystis, Cryptosporidium, Giardia, and Enterocytozoon by molecular and microscopic methods. Parasitology Research 2012; 111: 13491355.
31. O.I.E. Cryptosporidiosis. In: Linnane S, Pearson JE, eds. Manual of Standards for Diagnostic Tests and Vaccines for Terrestrial Animals, 6th edn. OIE Biological Standards Commission; adopted by the International Committee of the OIE. Paris, France: OFFICE INTERNATIONAL DES EPIZOOTIES, 2008, pp. 11971200.
32. Perrin P, Sureau P. A collaborative study of an experimental kit for rapid rabies enzyme immunodiagnosis. Bulletin World Health Organization 1987; 65: 489493.
33. Brar APS, et al. Validation of Romanowsky staining as a novel screening test for the detection of faecal cryptosporidial oocysts. Journal of Parasitic Diseases 2017; 41(1): 260262.
34. Singla LD, et al. Antigen based diagnosis of Cryptosporidium parvum infection in cattle and buffalo faeces. Indian Journal of Animal Science 2013; 83(1): 3739.
35. Bhat SA, Juyal PD, Singla LD. Prevalence of cryptosporidiosis in neonatal buffalo calves in Ludhiana district of Punjab, India. Asian Journal of Animal and Veterinary Advances 2012; 7: 512520.
36. Castro-Hermida JA, González-Losada YA, Ares-Mazás E. Prevalence of and risk factors involved in the spread of neonatal bovine cryptosporidiosis in Galicia (NW Spain). Veterinary Parasitology 2002; 106(1): 110.
37. Trotz-Williams LA, et al. Prevalence of Cryptosporidium parvum infection in southwestern Ontario and its association with diarrhoea in neonatal dairy calves. Canadian Veterinary Journal 2005; 46: 349351.
38. Kvac M, Kouba M, Vitovec J. Age-related and housing dependence of Cryptosporidium infection of calves from dairy and beef herds in South Bohemia, Czech Republic. Veterinary Parasitology 2006; 137: 202209.
39. Fayer R, Santin M, Dargatz D. Species of Cryptosporidium detected in weaned cattle on cow-calf operations in the United States. Veterinary Parasitology 2010; 170: 187192.
40. Van den BD, et al. Comparison of four rapid diagnostic tests, ELISA, microscopy and PCR for the detection of Giardia lamblia, Cryptosporidium spp. and Entamoeba histolytica in feces. Journal of Microbiological Methods 2015; 110: 7884.
41. Settawy MA EL, Fathy GM. Evaluation and comparison of PCR, coproantigen ELISA and microscopy for diagnosis of Cryptosporidium . Journal of American Science 2012; 8(12): 13781385.
42. Xiao L, Ryan UM. Molecular epidemiology. In: Fayer R, Xiao L, eds. Cryptosporidium and Cryptosporidiosis. Boca Raton: CRC Press IWA Publishing, 2008.
43. Webster KA, et al. Detection of Cryptosporidium parvum oocysts in faeces: comparison of conventional coproscopical methods and the polymerase chain reaction. Veterinary Parasitology 1996; 61: 513.
44. Morgan UM, et al. Differentiation between human and animal isolates of Cryptosporidium parvum using rDNA sequencing and direct PCR analysis. Journal of Parasitololgy 1997; 83: 825830.
45. Morgan UM, Thompson RCA. PCR detection of Cryptosporidium: the way forward? Parasitology Today 1998; 14(6): 241245.
46. Morgan UM, et al. Comparison of PCR and microscopy for detection of Cryptosporidium parvum in human fecal specimens: clinical trial. Journal of Clinical Microbiology 1998; 36(4): 995998.
47. Meyer CL, Palmer CJ. Evaluation of PCR, nested PCR, and fluorescent antibodies for detection of Giardia and Cryptosporidium species in waste water. Applied Environmental Microbiology 1996; 62: 20812085.
48. Balatbat AB, et al. Detection of Cryptosporidium parvum DNA in human faeces by nested PCR. Journal of Clinical Microbiology 1996; 34: 17691772.
49. Persing DH. Diagnostic Molecular Microbiology: Principles and Applications. Washington, DC: Published by American Society for Microbiology, 1993, pp. 5187.
50. Fenner F, MacLachlan NJ, Dubovi EJ, eds. Fenner's Veterinary Virology, 4th edn. Burlington: Academic Press, 2011, pp. 288290.
51. Espy MJ, et al. Real-time PCR in clinical microbiology: applications for routine laboratory testing. Clinical Microbiology Reviews 2006; 19: 165256.
52. Brar APS, Sood NK, Ahuja CS, Sandhu BS, Gupta K and Singh CK. Serum biochemical changes in neonatal diarrhoeic claves of different age groups. Indian Journal of Veterinary Pathology 2014; 38: 1417.
53. Brar APS, Ahuja CS, Sood NK, Sandhu BS and Gupta K. Haematological changes in neonatal diarrhoeic claves of different age groups. Indian Journal of Veterinary Pathology 2015; 39: 7377.
54. Morin M, et al. Diarrhoea of newborn calves. II. Agents responsible for the disease on Quebec dairy farms. Med. VeÂt. Quebec 1980; 10: 6065.
55. Bartels CJM, et al. Prevalence, prediction and risk factors of enteropathogens in normal and non-normal faeces of young Dutch dairy calves. Preventive Veterinary Medicine 2010; 93: 162169.
56. USDA. Dairy 2007 Part II: Changes in the U.S. Dairy Cattle industry, 1991–2007. Fort Collins: USDA-APHIS-VS, CEAH, 2008, pp. 5761.
57. Hur TY, et al. The dairy calf mortality: the causes of calf death during ten years at a large dairy farm in Korea. Korean Journal of Veterinary Research 2013; 53: 103108.
58. Mohammed HO, Wade SE, Schaaf S. Risk factors associated with Cryptosporidium parvum infection in dairy cattle in southeastern New York State. Veterinary Parasitology 1999; 83: 113.
59. José A, et al. Prevalence of and risk factors involved in the spread of neonatal bovine cryptosporidiosis in Galicia (NW Spain). Veterinary Parasitology 2002; 106: 110.
60. Tsunemitsu H, Smith DR, Saif LJ. Experimental inoculation of adult dairy cows with bovine coronavirus and detection of coronavirus in feces by RT-PCR. Archives of Virology 1999; 144(1): 167175.
61. Lin CK, Tsen HY. Use of two 16S DNA targeted oligonucleotides as PCR primers for the specific detection of Salmonella in foods. Journal of Applied Bacteriology 1996; 80: 659666.
62. Yoo HS, et al. Molecular typing and epidemiological survey of prevalence of Clostridium perfringens types by multiplex PCR. Journal of Clinical Microbiology 1997; 35: 228232.
Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

Epidemiology & Infection
  • ISSN: 0950-2688
  • EISSN: 1469-4409
  • URL: /core/journals/epidemiology-and-infection
Please enter your name
Please enter a valid email address
Who would you like to send this to? *



Altmetric attention score

Full text views

Total number of HTML views: 4
Total number of PDF views: 34 *
Loading metrics...

Abstract views

Total abstract views: 176 *
Loading metrics...

* Views captured on Cambridge Core between 8th August 2017 - 15th December 2017. This data will be updated every 24 hours.