Peer Review #2 of "Virgin and UV-weathered polyamide microplastics posed no effect on the survival and reproduction of Daphnia magna (v0.1)"

Although evidence suggests that microplastic (MP) particles pose a risk to organisms, the effects of virgin and weathered MP should be evaluated separately as their effects may be different.In this work, we provide new information on the toxic potential of virgin and UV-weathered polyamide, one of the commonly used plastics worldwide.Polyamide MP particles were subjected to UV-weathering in wet conditions during 26 days in a customized irradiation chamber equipped with UV-C light tubes (15 W each, max.wavelength 254 nm).The toxicity of virgin and UV-weathered polyamide MP (<180 μm in one dimension, 100 and 300 mg L -1) was evaluated by studying Daphnia magna reproduction in natural lake water spiked with MP, following the 21-day OECD 211 test guideline.In parallel, a nonionic surfactant Tween 20 (7 mg L -1) was added to the test medium to improve the suspendability of the MP.The results of the tests showed no adverse effects of either virgin or UV-weathered polyamide MP on the reproduction of D. magna.In addition, presence of Tween 20 in the test medium had no effects on the test results.These results bring a new perspective on the potential long-term impact of polyamide particles on aquatic organisms, especially considering that the polyamide has received marginal attention in the ecotoxicological research.However, standard test endpoints (survival and reproduction) may still miss long-term adverse effects of insoluble e.g., plastic particles and additional studies may be necessary.