Prevalence of toxigenic Escherichia coli (E.Coli) and rotavirus among neonatal calves in south Gurarat region (India) using classical and PCR based methods

Title: Prevalence of toxigenic Escherichia coli (E.Coli) and rotavirus among neonatal calves in south Gurarat region (India) using classical and PCR based methods

Authors: Sanjay G Malaviya, IH Kalyani, KK Sharma, PS Sakhare and DR Patel

Source: Ruminant Science (2017)-6(1):139-144.

Cite this reference as: Malaviya Sanjay G, Kalyani IH, Sharma KK, Sakhare PS and Patel DR (2017). Prevalence of toxigenic Escherichia coli (E.Coli) and rotavirus among neonatal calves in south Gurarat region (India) using classical and PCR based methods. Ruminant Science 6(1):139-144.


A total of 62 faecal samples were collected from diarrhoeic bovine calves aging upto 8 weeks of age and processed for presence of rotavirus and E.coli. The enteropathogens detected were E. coli in 45/62(72.58%) samples via bacteriological and molecular methods and rotavirus in 7/62 (11.29%) samples via Latex agglutination test (LAT), respectively and their co- presence were found in 5/62 (8.06%) cases. All the LAT positive rotavirus samples were though found negative by RT-PCR. Among the 25 samples sent for serotyping, ten isolates were serotyped as O25, O136 (04 isolates each), O1 and O34 (01 isolate each), while 15 isolates could not be serotyped with available antisera. PCR based detection of bacteria confirmed the presence of E. coli. PCR of toxigenic genes revealed absence of heat labile (LT) and heat stable (ST) toxins but 4 isolates (8.89%) were found positive for verotoxin (VT) genes. where one isolate (2.22%) possessed VT1 gene and 3 isolates (6.67%) VT2 gene. When E.coli isolates were subjected to antibiotic sensitivity test, Enrofloxacin, Gatifloxacin and Trimethoprim were found most effective antibiotics and respectively, 4.44 %, 11.11 % and 13.33 % isolates showed resistant against them. On the other hand maximum resistance was shown against Cephalexin (93.33%) and Kanamycin (88.89%). Conclusively, verotoxigenic E.coli was detected as a major enteropathogen of neonatal calf diarrhoea, along with rotavirus and fluroquinoles group of antibiotics were found as effective antibacterial drugs.


Anand SK, Fatima S, Joshi CG and Purohit JH (2006). Molecular characterization of E. coli using PCR-SSCP analysis of 16S rRNA gene. Indian Journal of Poultry Science 41(1):1-5.

Arya G (2006). Isolation and identification of Escherichia coli from diarrhoeic calf faeces by biochemical tests, antibiogram pattern and PCR based detection of toxigenic genes. MVSc Thesis submitted to AAU, Anand, Gujrat, India.

Basera SS, Singh R, Vaid N, Sharma K, Chakravarti S and Malik YPS (2010) Detection of Rotavirus Infection in Bovine Calves by RNA-PAGE and RT-PCR. Indian Journal of Virology 21(2):144-147.

Bauer AW, Kirby WMM, Sherris JC and Turck M (1966). Antibiotic susceptibility testing by a standardized single disk method. American Journal of Clinical Pathology 45:493-496.

Bradford PA, Petersen PJ, Fingerman IM and White DG (1999). Characterization of expanded-spectrum cephalosporin resistance in E. coli isolates associated with bovine calf diarrhoeal disease. Journal of Antimicrobial Chemotherapy 44:607-610.

Chattopadhyay UK, Gupta S and Dutta S (2003). Search for Shiga toxin producing Escherichia coli (STEC) including O157:H7 strains in and around Kolkata. Indian Journal of Medical Microbiology 21:17-20.

Cho Y and Yoon KJ (2014). An overview of calf diarrhea -infectious etiology, diagnosis, and intervention. Journal of Veterinary Science 15(1):1-17.

Constable PD (2004).  Antimicrobial use in the treatment of calf diarrhea. Journal of Veterinary Internal Medicine 18:8-17.

Dadawala AI, Chandel BS, Chauhan HC and Bhagat AG (2013). Molecular detection of bovine rotavirus from north Gujarat, India. Ruminant Science 2(2):131-137.

Edwards R and Ewing WN (1972). Identification of Enterobacteriaceae. 3rd Edn, Burgess Publishing Co, Minnesota.

Hussain I, Saikia GK and Rahman H (2003). Virulence factors of Esherichia coli isolated from diarrhoeic calves. Indian   Journal of Comparative Microbiology Immunology and Infectious Diseases 24:63-66.

Hariharan H, Mada C, Poole D and Page R (2004). Antibiotic resistance among enterotoxigenic Escherichia coli from piglets and calves with diarrhea. Canadian Veterinary Journal 45:605-606.

Kapadiya DK, Joshi DV, Chaudhary SS, Shah NM, Panchasara HH and Patel BJ (2012a). Diagnosis of rotavirus infection in neonatal calves by ribose nucleic acid polyacrylamide electrophoresis (RNA-PAGE) and double antibody sandwich enzyme linked immunosorbent assay (DAS-ELISA). Ruminant Science 1(1):15-18.

Kapadiya DK, Joshi DV, Shah NM and Panchasara HH (2012b). Serotyping and in vitro drug sensitivity of Escherichia coli isolated from neonatal buffalo calves with diarrhoea. Ruminant Science 1(2):141-143.

Minakshi, Pandey R, Prasad G and Malik Y (2001). VP4 gene specific RT-PCR for detection of buffalo group A rotaviruses. Indian Journal of Animal Science 71:611-613.

Nishikawa Y, Zhou Z, Hase A, Ogasawara J, Kitase T, Abe N, Nakamura H, Wada T, Ishii E, Haruki K and the Surveillance Team (2002). Diarrheagenic Escherichia coli isolated from stools of sporadic cases of diarrhoeal illness in Osaka city, Japan between 1997 and 2000 : Prevalence of Enteroaggregative E. coli heat stable enterotoxin 1 gene- possessing E. coli. Japanese Journal of Infectious Diseases 55:182-190.

Pachaury R and Kataria AK (2012). Serogrouping and colicin-related properties of Escherichia coli isolated from neonate diarrhoeic ruminants. Ruminant Science 1(1):9-13.

Patel BB (2009). Isolation and identification of Escherichia coli with special reference to colicinogeny, toxin detection and molecular characterization. MVSc Thesis submitted to SDAU, Sardarkrushinagar, Gujrat.

Pollard DR, Jhonson WM, Lior H, Tyler SD and Rozee KR (1990). Rapid and specific detection of verotoxin genes in Escherichia coli by polymerase chain reaction. Journal of Clinical Microbiology 28:540-545.

Shah NM and Jhala VM (1992). Epizootiological investigations on rotavirus infection in cases of neonatal calf diarrhoea. Indian Veterinary Journal 69:5-7.  

Sharma DK, Soni SS, Kashyap SK and Shringi BN (2004). Seroprevalence, antibiotic sensitivity pattern and transfer of plasmid coded characters of E. coli. Indian Veterinary Journal 81:6-8.

Sharma KK, Kalyani IH and Patel DR (2015). Detection of Rotavirus and associated bacteria from outbreak of neonatal calf diarrhoea in colostrums deprived calves. Veterinary Practitioner 16(1):88-89.