Pallares-Vega Rebeca, Macedo Gonçalo, Brouwer Michael S M, Hernandez Leal Lucia, van der Maas Peter, van Loosdrecht Mark C M, Weissbrodt David G, Heederik Dick, Mevius Dik, Schmitt Heike
Wetsus, European Centre of Excellence for Sustainable Water Technology, Leeuwarden, Netherlands.
Department Biotechnology, Delft University of Technology, Delft, Netherlands.
Front Microbiol. 2021 Jul 19;12:656250. doi: 10.3389/fmicb.2021.656250. eCollection 2021.
Plasmid-mediated dissemination of antibiotic resistance among fecal in natural ecosystems may contribute to the persistence of antibiotic resistance genes in anthropogenically impacted environments. Plasmid transfer frequencies measured under laboratory conditions might lead to overestimation of plasmid transfer potential in natural ecosystems. This study assessed differences in the conjugative transfer of an IncP-1 (pKJK5) plasmid to three natural strains carrying extended-spectrum beta-lactamases, by filter mating. Matings were performed under optimal laboratory conditions (rich LB medium and 37°C) and environmentally relevant temperatures (25, 15 and 9°C) or nutrient regimes mimicking environmental conditions and limitations (synthetic wastewater and soil extract). Under optimal nutrient conditions and temperature, two recipients yielded high transfer frequencies (5 × 10) while the conjugation frequency of the third strain was 1000-fold lower. Decreasing mating temperatures to psychrophilic ranges led to lower transfer frequencies, albeit all three strains conjugated under all the tested temperatures. Low nutritive media caused significant decreases in transconjugants (-3 logs for synthetic wastewater; -6 logs for soil extract), where only one of the strains was able to produce detectable transconjugants. Collectively, this study highlights that despite less-than-optimal conditions, fecal organisms may transfer plasmids in the environment, but the transfer of pKJK5 between microorganisms is limited mainly by low nutrient conditions.
质粒介导的抗生素耐药性在自然生态系统中的粪便菌群间传播,可能导致抗生素耐药基因在受人为影响的环境中持续存在。在实验室条件下测得的质粒转移频率,可能会高估自然生态系统中质粒的转移潜力。本研究通过滤膜杂交法,评估了IncP-1(pKJK5)质粒向三种携带超广谱β-内酰胺酶的天然菌株的接合转移差异。杂交在最佳实验室条件(丰富的LB培养基和37°C)、与环境相关的温度(25、15和9°C)或模拟环境条件及限制的营养条件(合成废水和土壤提取物)下进行。在最佳营养条件和温度下,两个受体菌株产生了高转移频率(5×10),而第三个菌株的接合频率低1000倍。将杂交温度降至嗜冷范围会导致转移频率降低,尽管所有三种菌株在所有测试温度下均能发生接合。低营养培养基导致转接合子显著减少(合成废水条件下减少3个对数;土壤提取物条件下减少6个对数),其中只有一种菌株能够产生可检测到的转接合子。总体而言,本研究强调,尽管条件并非最优,但粪便微生物仍可能在环境中转移质粒,不过pKJK5在微生物间的转移主要受低营养条件限制。