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Imidacloprid (IMI) is existence in the soil environment with a half-life habitually more than hundred days. This study targets to determine, identify and characterize photo-biodegradation bacteria from neonicotinoids (NEOs) contaminated agricultural field soils. The sub-surface soil had a higher level contamination of NEOs, in specifically greater concentration of IMI (3445.2 ± 0.09 μg/g) and thiacloprid (4084.4 ± 0.09 μg/g) has been found. Three bacteria Ralstonia pickettii (PBMS-2), Bacillus cereus (PBMS-3) and Shinella zoogloeoides (PBMS-4) was identified from soil-free stable enrichment cultures. The biodegradability of IMI (50 mg L−1) by three bacteria under different colors of light-emitting diodes (LEDs) with a constant 12 V power supply was tested and found that the blue-LEDs had greatest efficiency in supporting biodegradation of IMI which is called photo-biodegradation. In specific, the rate of photo-biodegradation of IMI by Ralstonia pickettii (87%), Bacillus cereus (80%) and Shinella zoogloeoides (80%) was measured. Besides this study also tested the effect of aeration (rpm), pH, and temperature on photo-biodegradation of IMI. There were seven intermediate metabolites were measured as biodegradation products of IMI under photo-biodegradation conditions and they are; IMI-urea, IMI-desnitro, 6-chloronicotinic acid, 6-hydroxy nicotinic acid, IMI- aminoguanidine, IMI-nitrosoguanidine and 4,5-hydroxy IMI, these metabolites are may non-toxic to the environment. • Neonicotinoids insecticide such as imidacloprid in the soil environment had greater half-life. • The agricultural field sub-surface soil had a higher concentration of IMI (3445.2 ± 0.09 μg/g). • Photo-biodegradation performance is greater than separate photo degradation and biodegradation. • The bacteria of Ralstonia pickettii with blue-LEDs enhanced IMI degradation rate. [ABSTRACT FROM AUTHOR]