title: Prevalence, species identification and genotyping Cryptosporidium from livestock and deer in a catchment in the Cairngorms with a history of a contaminated public water supply.
abstract: BACKGROUND
  The apicomplexan parasite Cryptosporidium represents a threat to water quality and public health. An important zoonotic species involved in human cryptosporidiosis from contaminated water is Cryptosporidium parvum (C. parvum), the main reservoirs of which are known to be farm livestock particularly neonatal calves, although adult cattle, sheep, lambs and wildlife are also known to contribute to catchment loading of C. parvum. This study aimed to establish Cryptosporidium prevalence, species and genotype in livestock, deer and water in a catchment with a history of Cryptosporidium contamination in the public water supply.
  
  
  METHODS
  A novel method of processing adult ruminant faecal sample was used to concentrate oocysts, followed by a nested species specific multiplex (nssm) PCR, targeting the 18S rRNA gene, to speciate Cryptosporidium. A multilocus fragment typing (MLFT) tool was used, in addition to GP60 sequencing, to genotype C. parvum positive samples.
  
  
  RESULTS
  A very high prevalence of Cryptosporidium was detected, with speciation identifying a predominance of C. parvum in livestock, deer and water samples. Four GP60 subtypes were detected within C. parvum with the majority IIaA15G2R1 which was detected in all host species and on all farms. Multilocus fragment typing further differentiated these into 6 highly related multilocus genotypes.
  
  
  CONCLUSIONS
  The high prevalence of Cryptosporidium detected was possibly due to a combination of the newly developed sample processing technique used and a reflection of the high rates of the parasite present in this catchment. The predominance of C. parvum in livestock and deer sampled in this study suggested that they represented a significant risk to water quality and public health. Genotyping results suggested that the parasite is being transmitted locally within the study area, possibly via free-roaming sheep and deer. Further studies are needed to verify particular host associations with subtypes/MLGs. Land and livestock management solutions to reduce Cryptosporidium on farm and in the catchment are planned with the aim to improve animal health and production as well as water quality and public health.
authors: Moredun Research Institute, Pentlands Science Park, Penicuik, Midlothian, EH26 0PZ, UK. beth.wells@moredun.ac.uk.
authors: Beth
authors: B
authors: Wells
authors: Moredun Research Institute, Pentlands Science Park, Penicuik, Midlothian, EH26 0PZ, UK. Hannah.shaw@moredun.ac.uk.
authors: Hannah
authors: H
authors: Shaw
authors: Moredun Research Institute, Pentlands Science Park, Penicuik, Midlothian, EH26 0PZ, UK. Emily.hotchkiss@moredun.ac.uk.
authors: Emily
authors: E
authors: Hotchkiss
authors: Moredun Research Institute, Pentlands Science Park, Penicuik, Midlothian, EH26 0PZ, UK. Janice.gilray@moredun.ac.uk.
authors: Janice
authors: J
authors: Gilray
authors: Moredun Research Institute, Pentlands Science Park, Penicuik, Midlothian, EH26 0PZ, UK. Remedios.ayton@moredun.ac.uk.
authors: Remedios
authors: R
authors: Ayton
authors: Scottish Water, Juniper House, Heriot Watt Research Centre, Edinburgh, EH14 4AP, UK. J.Green@scottishwater.co.uk.
authors: James
authors: J
authors: Green
authors: Moredun Research Institute, Pentlands Science Park, Penicuik, Midlothian, EH26 0PZ, UK. Frank.katzer@moredun.ac.uk.
authors: Frank
authors: F
authors: Katzer
authors: The Crown Estate, 6 Bells Brae, Edinburgh, EH4 3BJ, UK. andrew.wells@thecrownestate.co.uk.
authors: Andrew
authors: A
authors: Wells
authors: Moredun Research Institute, Pentlands Science Park, Penicuik, Midlothian, EH26 0PZ, UK. lee.innes@moredun.ac.uk.
authors: Elisabeth
authors: E
authors: Innes
mesh_terms: D000818:Animals; D002417:Cattle; D003457:Cryptosporidiosis / Q000453:epidemiology / Q000469:parasitology*; D003458:Cryptosporidium / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D003670:Deer / Q000469:parasitology*; D005243:Feces / Q000469:parasitology; D005618:Fresh Water / Q000469:parasitology*; D005838:Genotype; D058751:Livestock / Q000469:parasitology*; D015995:Prevalence; D012756:Sheep; D006113:United Kingdom / Q000453:epidemiology; D014876:Water Pollution / Q000032:analysis; D014881:Water Supply
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