title: Microbiota Analysis of an Environmental Slurry and Its Potential Role as a Reservoir of Bovine Digital Dermatitis Pathogens.
abstract: At present, very little information exists regarding what role the environmental slurry may play as an infection reservoir and/or route of transmission for bovine digital dermatitis (DD), a disease which is a global problem in dairy herds. To investigate whether DD-related bacteria belong to the indigenous microbiota of the dairy herd environment, we used deep amplicon sequencing of the 16S rRNA gene in 135 slurry samples collected from different sites in 22 dairy farms, with and without DD-infected cows. Both the general bacterial populations and digital dermatitis-associatedTreponemawere targeted in this study. The results revealed significant differences in the bacterial communities between the herds, with only 12 bacterial taxa shared across at least 80% of all the individual samples. These differences in the herd microbiota appeared to reflect mainly between-herd variation. Not surprisingly, the slurry was dominated by ubiquitous gastrointestinal bacteria, such asRuminococcaceaeandLachnospiraceaeDespite the low relative abundance of spirochetes, which ranged from 0 to 0.6%, we were able to detect small amounts of bacterial DNA from DD-associated treponemes in the slurry. However, the DD-associatedTreponemaspp. were detected only in samples from herds with reported DD problems. These data indicate that treponemes involved in the pathogenesis of DD are not part of the normal environmental microflora in dairy herds without clinical DD and, consequently, that slurry is not a primary reservoir of infection.IMPORTANCEBovine digital dermatitis (DD), a dermal disease which causes lameness in dairy cattle, is a serious problem worldwide. To control this disease, the infection reservoirs and transmission routes of DD pathogens need to be clarified. The dairy herd slurry may be a pathogen reservoir of DD-associated bacteria. The rationale for the present study was, therefore, to examine whether DD-associated bacteria are always present in slurry or if they are found only in DD-afflicted herds. The results strongly indicated that DDTreponemaspp. are not part of the indigenous slurry and, therefore, do not comprise an infection reservoir in healthy herds. This study applied next-generation sequencing technology to decipher the microbial compositions of environmental slurry of dairy herds with and without digital dermatitis.
authors.affiliation: National Veterinary Institute, Technical University of Denmark, Frederiksberg, Denmark kksc@vet.dtu.dk.
authors.forename: Kirstine
authors.initials: K
authors.lastname: Klitgaard
authors.affiliation: National Veterinary Institute, Technical University of Denmark, Frederiksberg, Denmark.
authors.forename: Mikael L
authors.initials: ML
authors.lastname: Strube
authors.affiliation: National Veterinary Institute, Technical University of Denmark, Frederiksberg, Denmark.
authors.forename: Anastasia
authors.initials: A
authors.lastname: Isbrand
authors.affiliation: National Veterinary Institute, Technical University of Denmark, Frederiksberg, Denmark.
authors.forename: Tim K
authors.identifier: http://orcid.org/0000-0002-8048-4695
authors.initials: TK
authors.lastname: Jensen
authors.affiliation: National Veterinary Institute, Technical University of Denmark, Frederiksberg, Denmark.
authors.forename: Martin W
authors.initials: MW
authors.lastname: Nielsen
mesh_terms: D000818:Animals; D001419:Bacteria / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D002417:Cattle; D002418:Cattle Diseases / Q000382:microbiology*; D058066:Digital Dermatitis / Q000382:microbiology*; D004197:Disease Reservoirs / Q000382:microbiology*; D064307:Microbiota; D010802:Phylogeny; D012988:Soil Microbiology
keywords: Treponema; deep amplicon sequencing; digital dermatitis; slurry