Muziasari Windi I, Pitkänen Leena K, Sørum Henning, Stedtfeld Robert D, Tiedje James M, Virta Marko
Department of Food and Environmental Sciences, University of Helsinki Helsinki, Finland.
Department of Food Safety and Infection Biology, Norwegian University of Life Sciences Oslo, Norway.
Front Microbiol. 2017 Jan 6;7:2137. doi: 10.3389/fmicb.2016.02137. eCollection 2016.
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 (), trimethoprim (), tetracycline [], aminoglycoside (), chloramphenicol (), and efflux-pumps resistance genes (). The detected genes also included class 1 integron-associated genes (Δ) and transposases (). 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 and genes were significantly correlated ( = 0.001, = 0.71). In addition, we analyzed the mucosal skin and gill filament resistomes of the farmed fish but only one multidrug-efflux resistance gene () 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.
我们之前的研究表明,在芬兰波罗的海北部养鱼场下方的沉积物中,特定的抗生素抗性基因(ARGs)在当地富集,即便来自抗生素的选择压力可忽略不计。我们推测养殖鱼类粪便的持续流入可能是养殖场沉积物中ARGs富集的合理来源。在本研究中,我们分析了来自波罗的海养殖场的20条鱼肠道内容物的抗生素抗性组组成。我们采用了一种高通量方法,即具有364个引物组的WaferGen qPCR阵列,来检测和定量ARGs、移动遗传元件(MGE)以及16S rRNA基因。尽管qPCR引物组的选择范围相当广泛,但在肠道内容物中仅检测到了(28)个基因。检测到的基因包括编码对磺胺类药物()、甲氧苄啶()、四环素[]、氨基糖苷类()、氯霉素()耐药的ARGs以及外排泵耐药基因()检测到的基因还包括1类整合子相关基因(Δ)和转座酶()。重要的是,大多数检测到的基因与养殖场沉积物中富集的基因相同。这项初步研究表明,尽管养殖场没有同时进行抗生素处理,但养殖鱼类的粪便导致了养殖场沉积物中ARGs的富集。我们观察到,单个养殖鱼类的肠道内容物有其自身的抗性组组成。我们的结果还表明,转座酶和基因的总相对丰度显著相关( = 0.001, = 0.71)。此外,我们分析了养殖鱼类的粘膜皮肤和鳃丝抗性组,但仅检测到一个多药外排耐药基因()。据我们所知,这是第一项使用非培养方法报告养殖鱼类抗性组的研究。确定ARGs尤其是可移动ARGs的可能来源,对于控制养鱼场设施中ARGs的出现和传播以及降低ARGs从养殖场扩散到周围环境的风险至关重要。