title: The Resistome of Farmed Fish Feces Contributes to the Enrichment of Antibiotic Resistance Genes in Sediments below Baltic Sea Fish Farms.
abstract: Our previous studies showed that particular antibiotic resistance genes (ARGs) were enriched locally in sediments below fish farms in the Northern Baltic Sea, Finland, even when the selection pressure from antibiotics was negligible. We assumed that a constant influx of farmed fish feces could be the plausible source of the ARGs enriched in the farm sediments. In the present study, we analyzed the composition of the antibiotic resistome from the intestinal contents of 20 fish from the Baltic Sea farms. We used a high-throughput method, WaferGen qPCR array with 364 primer sets to detect and quantify ARGs, mobile genetic elements (MGE), and the 16S rRNA gene. Despite a considerably wide selection of qPCR primer sets, only 28 genes were detected in the intestinal contents. The detected genes were ARGs encoding resistance to sulfonamide (sul1), trimethoprim (dfrA1), tetracycline [tet(32), tetM, tetO, tetW], aminoglycoside (aadA1, aadA2), chloramphenicol (catA1), and efflux-pumps resistance genes (emrB, matA, mefA, msrA). The detected genes also included class 1 integron-associated genes (intI1, qacEΔ1) and transposases (tnpA). Importantly, most of the detected genes were the same genes enriched in the farm sediments. This preliminary study suggests that feces from farmed fish contribute to the ARG enrichment in farm sediments despite the lack of contemporaneous antibiotic treatments at the farms. We observed that the intestinal contents of individual farmed fish had their own resistome compositions. Our result also showed that the total relative abundances of transposases andtetgenes were significantly correlated (p= 0.001,R2= 0.71). In addition, we analyzed the mucosal skin and gill filament resistomes of the farmed fish but only one multidrug-efflux resistance gene (emrB) was detected. To our knowledge, this is the first study reporting the resistome of farmed fish using a culture-independent method. Determining the possible sources of ARGs, especially mobilized ARGs, is essential for controlling the occurrence and spread of ARGs at fish farming facilities and for lowering the risk of ARG spread from the farms to surrounding environments.
authors.affiliation: Department of Food and Environmental Sciences, University of Helsinki Helsinki, Finland.
authors.forename: Windi I
authors.initials: WI
authors.lastname: Muziasari
authors.affiliation: Department of Food and Environmental Sciences, University of Helsinki Helsinki, Finland.
authors.forename: Leena K
authors.initials: LK
authors.lastname: Pitkänen
authors.affiliation: Department of Food Safety and Infection Biology, Norwegian University of Life Sciences Oslo, Norway.
authors.forename: Henning
authors.initials: H
authors.lastname: Sørum
authors.affiliation: Department of Civil and Environmental Engineering, Center for Microbial Ecology, Michigan State University Michigan, MI, USA.
authors.forename: Robert D
authors.initials: RD
authors.lastname: Stedtfeld
authors.affiliation: Department of Civil and Environmental Engineering, Center for Microbial Ecology, Michigan State University Michigan, MI, USA.
authors.forename: James M
authors.initials: JM
authors.lastname: Tiedje
authors.affiliation: Department of Food and Environmental Sciences, University of Helsinki Helsinki, Finland.
authors.forename: Marko
authors.initials: M
authors.lastname: Virta
keywords: class 1 integrons; culture-independent method; fish intestinal content; mobile genetic elements; qPCR array; rainbow trout; transposons; whitefish