Prevalence and genotypes of Enterocytozoon bieneusi in weaned beef calves on cow-calf operations in the USA.

To determine the prevalence and genotype distribution of Enterocytozoon bieneusi in weaned beef calves in the USA, fecal samples were collected from 819 calves (6-18 months of age) from 49 operations. Feces were sieved and subjected to density gradient centrifugation to remove fecal debris and to concentrate spores. DNA extracted from each sample was subjected to the polymerase chain reaction (PCR) to amplify the complete internal transcriber spacer (ITS). All PCR-positive specimens were sequenced to determine the genotype(s) present. Overall, E. bieneusi was detected in 34.8% of the 819 fecal samples. The highest prevalence was found in the Midwest region (42.7%) followed by the South (35.8%) and the West (23.2%). The prevalence of E. bieneusi varied considerably from operation to operation (0-100%). A prevalence of 100% was observed in three operations, one in the Midwest and two in the South; E. bieneusi was not found in six operations, three in the South and three in the West. Sequence analysis revealed the presence of six genotypes, four previously reported (I, J, BEB4, and type IV) and two novel genotypes (BEB8 and BEB9). Mixed infections were identified in five specimens, three contained I and BEB4 and two contained J and BEB4. Most of the positive calves (238 of 285) harbored genotypes with zoonotic potential including I (59), J (108), BEB4 (65), type IV (1), mixed I/BEB4 (3), and mixed J/BEB4 (2).