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Numerous pollutants from industrial or agricultural origins are found in the environment, and especially in water. Compounds such as herbicides, pesticides, chlorinated solvents as well as compounds linked to petroleum utilization are frequently found as pollutants of aquifers. These chemicals are generally toxic for humans and animals and their presence in water must be monitored to avoid the contamination of drinking water sources. Classical analytical tools (GC, GC/MS, etc.) provide accurate, reproducible and sensitive determination of contaminant concentrations. Nevertheless, their use requires to take samples on the contaminated sites and to transport the samples to laboratory for analysis. Such handling of samples is time-consuming and expensive. Biosensors are new analytical tools, whose conception has benefited from advances in different scientific areas, and particularly in biology; they now allow the development of highly specific tools. They provide real-time determination and detection of very low concentrations of contaminant and they can be used directly on-site.