Synthesis of benzoylthiourea derivatives and analysis of their antibacterial performance against planktonic Staphylococcus aureus and its biofilms.

Following the appearance of several antimicrobial agents to control the spread of infections, two major challenges have emerged: (i) the occurrence and blowout of multiresistant bacteria and the increase of chronic diseases and (ii) difficult-to-eradicate infections. In this study, we tested five benzoylthiourea derivatives for their ability to inhibit and stop bacterial growth and evaluated the possible influence of 1,2,4-triazolyl-benzoylthiourea derivative 4 on the formation and eradication of Staphylococcus aureus biofilms. Benzoylthiourea derivatives 4, 6, 10, 11 and 13 were obtained in one or two steps with low cost and subjected to tests to identify their minimum inhibitory concentration (MIC) and minimum bactericidal concentration. In vitro tests were also performed to assess their effects on biofilm formation and in preformed biofilms and scanning electron microscopy was used to visualize the effects on biofilm formation. The 1,2,4-triazolyl-benzoylthiourea derivative 4 showed bacteriostatic activity against the S. aureus HU25 clinical strain with an MIC of 16 µg ml-1 , which is below the toxic concentration (at 2500 µg ml-1 , 62·25% of the cells remained viable). Compound 4 also effectively prevented biofilm formation at the three subinhibitory concentrations tested (1/2 MIC, 1/4 MIC and 1/8 MIC) as confirmed by scanning electron microscopy. For breakdown of formed biofilms, the main influence was at a subinhibitory concentration (1/2 MIC). These findings make compound 4 a strong candidate for studies on the development of new antimicrobial and antibiofilm agents.

The use of an Allonais inaequalis reproduction test as an ecotoxicological bioassay.

Ecotoxicological bioassays have been widely utilized to evaluate the toxicity of substances to organisms. However, the main challenge for researchers is finding native species to assess the effects of pollutants on aquatic biota. The tropical Oligochaeta, Allonais inaequalis, can be used as a test organism in bioassays to understand the effects of toxicants on aquatic ecosystems and their impact on native aquatic biota. In this study, we tested four methodological designs to validate the use of our "Allonais inaequalis reproduction test" as an ecotoxicological bioassay. For each sample, the assay consisted of a bottle containing 10 mg of sterilized fine sand, 60 mL of dechlorinated tap water and 6 organisms, fed at the beginning of the test and again after 5 days. The assay was first established in a controlled environment and then used to evaluate a stressed environment containing one of the following three toxicants suggested by the OECD (2008) and Corbi et al. (2015): zinc chloride, copper sulfate, or potassium chloride. Our results showed that the best experimental design for reproduction analysis was a static, long-term bioassay, which lasted 10 days without aeration and allowed for the reproduction of multiple generations (10 ± 5 new organisms). The observed inhibition reproduction by toxicants (EC50 ranging between 0.2 mg L-1 and 1.36 g L-1) validated the methods used in this paper. The use of a reproduction endpoint is a new contribution to the ecotoxicological toolbox, examining responses from a native organism to predict the effects of pollutants in an aquatic environment.