Abstract: Background and Objective: Organophosphorus insecticides (OPIs), biocides and bactericides (antimicrobial) residues can accumulate in plant, fish, water, milk, animals and cause adverse health effects. Therefore, new and fast tools for detection and removal OPIs such as chlorpyrifos (CPF) and other xenobiotics are very important. This study was carried out for the first time to evaluate the toxicity of CPF, Malachite Green (MG), leucomalachite green (LMG) and tetracyclines (TC) using Microtox® assay and to remove CPF residues from apple juice by ultrasonic. Methodology: Acute toxicity of standard CPF, CPF residues and removal of residues from apple juice by ultrasonic were studied. Oxytetracycline (OTC), tetracycline (TC), chlortetracycline (CTC), doxycycline (DC), Malachite Green (MG) and leucomalachite green (LMG) were evaluated by Microtox®. Results: CPF and CPF residues are very toxic with EC50, 5 min, 15 min = 30.39 and 54.08 μg L–1 of CPF and 22.35 and 49.855 μg L–1 of CPF residues, respectively. CPF can be effectively and rapidly degraded by ultrasonic and the half-life time account 11.44 min with strongly correlated by sonication times and power. MG is very toxic with EC50, 5 min, 15 min = 365.8 and 192.56 μg L–1. CTC and DC are toxic with EC50, 5 min, 15 min = 9.664 and 4.628 mg L–1 of CTC and 19.888 and 5.208 mg L–1 of DC, respectively, while, OTC and LMG are harmful based on the toxicity categories established in the EU legislation. Conclusion: It is concluded that toxicity of CPF residues and their intermediate products can determined by Microtox® assay. CPF was effectively and rapidly degraded by ultrasonic. CPF, MG and TCTs act as toxic compounds in the environment at low concentration levels since they were detected in the environment at these low concentrations by Microtox® assay.