Along with traditional ecotoxicological approaches in model organisms, toxicological studies in non-model organisms are being taken into consideration in order to complement them and contribute to more robust approaches. This allows us to figure out the complexity of the exposures involved in natural ecosystems. In this context, in the present research we have used the model species Chironomus riparius (Chironomidae, Diptera) and the non-model species Prodiamesa olivacea (Chironomidae, Diptera) to assess the aquatic toxic effects of acute 4-h and 24-h exposures to 1 μgL⁻¹ of three common environmental pollutants: butyl benzyl phthalate (BBP), bisphenol A (BPA), and benzophenone 3 (BP3). Individuals of both species were collected from a contaminated river (Sar) in Galicia (Spain).

Regarding Chironomus, there are four OECD standardized tests for the evaluation of water and sediment toxicity, in which different species in this genus can be used to assess classical toxicity parameters such as survival, immobilization, reproduction, and development. In contrast, Prodiamesa is rarely used in toxicity studies, even though it is an interesting toxicological species because it shares habitats with Chironomus but requires less extreme conditions (e.g., contamination) and higher oxygen levels. These different requirements are particularly interesting in assessing the different responses of both species to pollutant exposure.

Quantitative real-time PCR was used to evaluate the transcriptional changes caused by xenobiotics in different genes of interest. Since information about P. olivacea in genomic databases is scarce, its transcriptome was obtained using de novo RNAseq. Genes involved in biotransformation pathways and the oxidative stress response (MnSOD, CAT, PHGPx, Cyp4g15, Cyp6a14-like and Cyp6a2-like) were de novo identified in this species. Our results show differential toxic responses depending on the species and the xenobiotic, being P. olivacea the dipteran that showed the most severe effects in most of the studied biomarker genes.

This work represents a multi-species approach that allows us to deepen in the toxicity of BBP, BPA, and BP3 at the molecular level. Besides, it provides an assessment of the tolerance/sensitivity of natural populations of model and non-model insect species chronically exposed to complex mixtures of pollutants in natural scenarios. These findings may have important implications for understanding the adverse biological effects of xenobiotics on P. olivacea, providing new sensitive biomarkers of exposure to BBP, BPA, and BP3. It also highlights the suitability of Prodiamesa for ecotoxicological risk assessment, especially in aquatic ecosystems.

与模型生物中的传统生态毒理方法一起，正在考虑对非模型生物中的毒理学研究，以补充它们并为更可靠的方法做出贡献。这使我们能够弄清楚自然生态系统所涉及的暴露的复杂性。在这种情况下，在本研究中，我们使用模式物种Chironomus riparius（Chironomidae，Diptera）和非模式物种Prodiamesa olivacea（Chironomidae，Diptera）来评估急性4小时和24小时暴露对水生的毒性作用。至1μgL ^-1三种常见的环境污染物：邻苯二甲酸丁苄酯（BBP），双酚A（BPA）和二苯甲酮3（BP3）。两种物种的个体均从加利西亚（西班牙）的受污染河流（萨尔州）收集。

关于Chironomus，有四个OECD标准化测试用于评估水和沉积物的毒性，其中该属的不同物种可用于评估经典的毒性参数，例如存活，固定，繁殖和发育。相比之下，Prodiamesa很少用于毒性研究，尽管它是一种有趣的毒理学物种，因​​为它与Chironomus共享栖息地，但需要的极端条件较少（例如，污染）和较高的氧气水平。这些不同的要求在评估两个物种对污染物暴露的不同响应时特别有趣。

实时定量PCR用于评估异源生物在不同目标基因中引起的转录变化。由于基因数据库中关于寡橄榄假单胞菌的信息稀少，因此使用从头RNAseq获得其转录组。在该物种中从头鉴定了参与生物转化途径和氧化应激反应的基因（MnSOD，CAT，PHGPx，Cyp4g15，Cyp6a14样和Cyp6a2样）。我们的结果表明，取决于物种和异种生物体，不同的毒性反应是P. olivacea，在大多数研究的生物标记基因中显示出最严重影响的二萜。

这项工作代表了一种多物种的方法，使我们能够在分子水平上加深BBP，BPA和BP3的毒性。此外，它还评估了在自然场景下长期暴露于复杂污染物混合物中的模型和非模型昆虫自然种群的耐受性/敏感性。这些发现可能对理解异种生物对橄榄紫假单胞菌的不利生物学影响具有重要意义，为暴露BBP，BPA和BP3提供了新的敏感生物标记。它还强调了Prodiamesa尤其适合在水生生态系统中进行生态毒理学风险评估的适用性。