This paper presents research upon the impact of ecological measures on the water content of new and used oil. Water and microorganisms are major factors of contamination for engine oils and fuels, and they lead to a significant reduction in the lifetime and performance of engines. The microorganisms occur naturally in the environment, from which they enter into the oil and fuel. Despite various preventive measures, it is not possible to completely remove water from petrochemical products. That is why the protection against and prevention of the various types of contamination of petroleum products, especially microbiological contamination, is very important. Biocides are one example of the agents used for the prevention of contamination; biocides belong to a group of pesticides that are used to eliminate microbial contamination. Due to the fact that currently available methods are ineffective and often have a detrimental effect on the natural environment, research is underway to discover modern and ecological measures to combat the phenomenon of the microbiological contamination of petroleum products. This paper shows the effects of environmentally friendly additives on the water content in lubricating oil, i.e., whether these additives cause the release of water bound in the oil, and whether their composition increases the water content of the oil. Samples of new and used oil were utilized for the tests. Effective microorganisms (EM), in both liquid form and within ceramic tubes, were added to the new and used oil samples. In addition, silver solution and silver compounds were added in the same amounts as the liquid effective microorganisms. In order to confirm and compare the obtained test results, the dynamic viscosity, flash point, acid number, base number, and water content were measured, and these are presented in this study. These measurements were followed by the analysis of the influence of the additives on the water content. It was found that the adding of effective microorganisms to fresh oil in liquid form caused a significant increase in the water content of the oil; in the case of a smaller amount (2.5 mL), the water content more than doubled, and in the case of a larger amount, the water content exceeded the measuring range of the device. Next, an evaluation of the impact of these environmental measures on the water content was carried out. It was found that the adding of liquid effective microorganisms to new oil caused a significant increase in the water content of the oil; the water content more than doubled for the addition of 2.5 mL, while for a larger addition (5 mL), the water content exceeded the measuring range of the device. The same was true for the addition of silver compounds, regardless of their type and amount. The best results were obtained with ceramic effective microorganisms, but the results for silver cannot be presented due to the over-titration of the device (the amount of water exceeded the measuring range). For used oil, the liquid effective microorganism was found to give the best result.