Dynamics of excretion and molecular characterization of Cryptosporidium isolates in pre-weaned French beef calves

Studies on excretion and molecular characterization of Cryptosporidium have been mostly conducted in dairy calves, both diarrhoeic and non-diarrhoeic. Little is known about Cryptosporidium in beef calves, especially in non-diarrhoeic ones. This study was conducted in a herd of Parthenais beef cattle (France) with no history of clinical cryptosporidiosis. Twenty-five calves were sampled once a week from birth to one month of age (age range: 5-34 days). At each sampling date, presence of clinical signs of cryptosporidiosis (diarrhoea) was recorded. Oocyst excretion was assessed using the Heine staining method and a direct immunofluorescence method (Merifluor (R) C/G) which allowed quantification (oocysts per gram of faeces, opg). All samples were subjected to a two-step nested PCR protocol to amplify the 18S rRNA gene and amplification products were sequenced. None of the calves presented diarrhoea. Twenty-three of them excreted oocysts at least one sampling date. Prevalence of excretion was maximal when calves were 27-34 days old, with a percentage of excretion of 85% in this age category [95% CI: 70; 1001 Mean excretion was maximal when calves were 20 to 26 days old, with a mean excretion of 7.6 x 10(5) opg(range: 0-8 x 10(6) opg). 32 isolates were successfully identified: 27 as Cryptosporidium bovis, 4 as Cryptosporidium ryanae and 1 as Cryptosporidium parvum. C. bovis was isolated from samples of calves between 11 and 33 days old. C. ryanae was isolated from samples of calves between 17 and 34 days old. C parvum was isolated from one calf aged 13 days. This survey demonstrated the high infection rate of non-diarrhoeic beef calves by Cryptosporidium species other than C. parvum. (C) 2012 Elsevier B.V. All rights reserved.