Effects of recycled manure solids bedding on the spread of gastrointestinal parasites in the environment of dairies and milk.

The primary aim of this work was to isolate common bovine digestive tract parasites in recycled manure bedding (RMS), as well as to determine the ability of current RMS preparation procedures to eliminate these pathogens. Other objectives were to assess whether any of the aforementioned parasites could be retrieved in bulk milk from dairies using RMS and to study whether the prevalence of these parasites differed among manure of cows housed on RMS versus on straw bedding. For the study, 27 RMS farms and 61 control farms were recruited. Samples of manure from the pre-pit and milk from the bulk tank were recovered from straw-bedding farms and RMS-based farms. In addition, samples from the manure solid fraction after liquid extraction, RMS before use, and RMS currently in use were recovered from RMS herds. Parasites were first detected by double centrifugation zinc sulfate flotation to enhance isolation of gastrointestinal protozoa, and by modified Wisconsin sugar flotation for the appraisal of gastrointestinal nematodes. Cryptosporidium parasites were confirmed by nested PCR amplification and sequencing of a portion of the gene encoding the small subunit rRNA. Results revealed a high prevalence of Cryptosporidium spp. (C. parvum, C. andersoni, and C. meleagridis, identified by PCR) and Eimeria spp. (mainly E. bovis and E. zuernii) parasites in both types of farms, with a larger proportion of manure samples from RMS-bedded farms testing positive for Cryptosporidium parasites compared with manure from straw-bedded farms. Both Cryptosporidium spp. and Eimeria spp. oocysts were found at every step of RMS preparation and transformation, showing that current RMS preparation strategies do not guarantee the destruction of protozoan parasites. Cryptosporidium parvum, a potential zoonotic risk for professionals in close contact with livestock, was found to be present in 32 out of 61 straw-bedded and 24 of 27 RMS farms. No protozoan parasites were found in any sample derived from bulk milk, neither by microscopy analysis nor by molecular methods.