关键词: Antibiotics Cnidarian Culturable bacteria Gene expression Microbiome Nematostella vectensis Settlement

Mesh : Animals Anti-Bacterial Agents / pharmacology Sea Anemones / genetics drug effects Larva / microbiology drug effects genetics Ampicillin / pharmacology Neomycin / pharmacology Streptomycin / pharmacology Rifampin / pharmacology Gene Expression / drug effects

来  源:   DOI:10.7717/peerj.17349   PDF(Pubmed)

Abstract:
UNASSIGNED: Antibiotics are commonly used for controlling microbial growth in diseased organisms. However, antibiotic treatments during early developmental stages can have negative impacts on development and physiology that could offset the positive effects of reducing or eliminating pathogens. Similarly, antibiotics can shift the microbial community due to differential effectiveness on resistant and susceptible bacteria. Though antibiotic application does not typically result in mortality of marine invertebrates, little is known about the developmental and transcriptional effects. These sublethal effects could reduce the fitness of the host organism and lead to negative changes after removal of the antibiotics. Here, we quantify the impact of antibiotic treatment on development, gene expression, and the culturable bacterial community of a model cnidarian, Nematostella vectensis.
UNASSIGNED: Ampicillin, streptomycin, rifampicin, and neomycin were compared individually at two concentrations, 50 and 200 µg mL-1, and in combination at 50 µg mL-1 each, to assess their impact on N. vectensis. First, we determined the impact antibiotics have on larval development. Next Amplicon 16S rDNA gene sequencing was used to compare the culturable bacteria that persist after antibiotic treatment to determine how these treatments may differentially select against the native microbiome. Lastly, we determined how acute (3-day) and chronic (8-day) antibiotic treatments impact gene expression of adult anemones.
UNASSIGNED: Under most exposures, the time of larval settlement extended as the concentration of antibiotics increased and had the longest delay of 3 days in the combination treatment. Culturable bacteria persisted through a majority of exposures where we identified 359 amplicon sequence variants (ASVs). The largest proportion of bacteria belonged to Gammaproteobacteria, and the most common ASVs were identified as Microbacterium and Vibrio. The acute antibiotic exposure resulted in differential expression of genes related to epigenetic mechanisms and neural processes, while constant application resulted in upregulation of chaperones and downregulation of mitochondrial genes when compared to controls. Gene Ontology analyses identified overall depletion of terms related to development and metabolism in both antibiotic treatments.
UNASSIGNED: Antibiotics resulted in a significant increase to settlement time of N. vectensis larvae. Culturable bacterial species after antibiotic treatments were taxonomically diverse. Additionally, the transcriptional effects of antibiotics, and after their removal result in significant differences in gene expression that may impact the physiology of the anemone, which may include removal of bacterial signaling on anemone gene expression. Our research suggests that impacts of antibiotics beyond the reduction of bacteria may be important to consider when they are applied to aquatic invertebrates including reef building corals.
摘要:
抗生素通常用于控制患病生物体中的微生物生长。然而,早期发育阶段的抗生素治疗可能会对发育和生理产生负面影响,从而抵消减少或消除病原体的积极影响。同样,由于对耐药和易感细菌的效力不同,抗生素可以改变微生物群落。虽然抗生素的应用通常不会导致海洋无脊椎动物的死亡,对发育和转录效应知之甚少。这些亚致死效应可能降低宿主生物体的适应性,并在去除抗生素后导致负面变化。这里,我们量化抗生素治疗对发展的影响,基因表达,和模型刺胞动物的可培养细菌群落,线虫。
氨苄西林,链霉素,利福平,和新霉素分别在两种浓度下进行比较,50和200µgmL-1,每个组合为50µgmL-1,评估它们对N.vectensis的影响。首先,我们确定了抗生素对幼虫发育的影响。接下来,使用扩增子16SrDNA基因测序来比较在抗生素处理后持续存在的可培养细菌,以确定这些处理如何针对天然微生物组进行差异选择。最后,我们确定了急性(3天)和慢性(8天)抗生素治疗对成年海葵基因表达的影响.
在大多数曝光下,随着抗生素浓度的增加,幼虫沉降的时间延长,并且在联合治疗中最长延迟3天。可培养细菌在大多数暴露中持续存在,其中我们鉴定了359个扩增子序列变体(ASV)。最大比例的细菌属于γ变形菌,最常见的ASV被鉴定为微杆菌和弧菌。急性抗生素暴露导致与表观遗传机制和神经过程相关的基因差异表达,而与对照组相比,持续应用导致伴侣的上调和线粒体基因的下调。基因本体论分析确定了两种抗生素治疗中与发育和代谢相关的术语的总体消耗。
抗生素可显著增加叶枯病幼虫的沉降时间。抗生素处理后的可培养细菌种类在分类学上是不同的。此外,抗生素的转录效应,去除后,基因表达会产生显著差异,这可能会影响海葵的生理,这可能包括去除细菌信号对海葵基因表达的影响。我们的研究表明,在将抗生素应用于包括造礁珊瑚在内的水生无脊椎动物时,除减少细菌外,抗生素的影响可能很重要。
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