title: Detection of biofilm formation and nanobacteria under long-term cell culture conditions in serum samples of cattle, goats, cats, and dogs.
abstract: OBJECTIVE
  To determine the prevalence of biofilm formation under long-term cell culture conditions in serum samples of dairy cattle, goats, cats, and dogs, and to determine whether there is an association between nanobacteria and biofilm formation.
  
  
  METHODS
  Serum samples of clinically normal animals (313 dairy cattle, 48 goats, 140 dogs, and 44 cats) and animals with various medical conditions (60 dogs and 116 cats).
  
  
  METHODS
  Serum was incubated under cell culture conditions and observed for biofilm formation by use of light microscopy, electron microscopy, and spectroscopy. A polymerase chain reaction assay was developed to identify 16S rRNA gene sequences of nanobacteria.
  
  
  RESULTS
  Biofilm formation developed in serum samples of 304 of 313 (97%) cattle, 44 of 48 (92%) goats, 44 of 44 (100%) cats, and 126 of 140 (90%) dogs. Prevalence of serum samples with positive results for biofilm formation was not significantly different between cats or dogs with and without medical conditions associated with pathologic extraskeletal calcification processes. Scanning electron microscopy and spectroscopy of biofilm samples revealed small coccoid particles consisting mainly of calcium and phosphate. Polymerase chain reaction assay failed to amplify sequences of nanobacteria.
  
  
  CONCLUSIONS
  Under long-term cell culture conditions, biofilm made up of aggregates of calcium and phosphate crystals does form in serum samples of clinically normal dairy cattle, goats, cats, and dogs. Disease, however, does not predispose to biofilm formation in serum samples of dogs and cats. Our findings did not support the existence of nanobacteria in serum samples of cattle, goats, cats, and dogs.
authors.affiliation: Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Barr
authors.forename: Rebecca A
authors.initials: RA
authors.lastname: Linke
authors.forename: Dorothee
authors.initials: D
authors.lastname: Janssen
authors.forename: Charles L
authors.initials: CL
authors.lastname: Guard
authors.forename: Mary C
authors.initials: MC
authors.lastname: Smith
authors.forename: Carole S
authors.initials: CS
authors.lastname: Daugherty
authors.forename: Janet M
authors.initials: JM
authors.lastname: Scarlett
mesh_terms: D000818:Animals; D001419:Bacteria / Q000235:genetics / Q000254:growth & development* / Q000302:isolation & purification* / Q000648:ultrastructure; D018441:Biofilms / Q000254:growth & development*; D002415:Cats; D002417:Cattle; D018929:Cell Culture Techniques; D004285:Dogs; D005260:Female; D005798:Genes, Bacterial / Q000235:genetics; D006041:Goats / Q000097:blood*; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D013056:Spectrophotometry, Ultraviolet
chemical_list: D012336:RNA, Ribosomal, 16S