Soil microbial analysis are one of an important study with chemical and physical analysis, to indicate safety and healthy for plants, field animals and human. The numbers of soil microorganisms are greatly differences; it is primarily helpful for the healthy growth and development of field flora, animals, and humans, as it is made up of numerous kinds of microbes such as actinomycetes, bacteria, fungus, and microalgae. The goal of this study is to assess the quality of soils that have been irrigated with secondary-treated municipal wastewater during a 20-year period in Serapuim Forest, which is located in Egypt's Ismailia Governorate. The assessment will be based on a set of biological indications. In this study 28 sand soil samples were collected from 6 sites namely: A, B, C, D, E and P on Serapuim Forest, Ismailia Governorate. Several techniques are involved to isolate and identify these microbes have been described. The total microbial count and how many microorganisms are present in samples is one of the key indicators in the field of health of soils. Because the count of microorganisms and their types should not exceed specific guidance values, it is vital to keep track of the overall number of microbes and their types. In this study, the total count of microbes was estimated, in addition to identify a number of pathogenic microbes. Concluded results that the Serapuim Forest soil is a contamination land and this could affect adversely the fertility of the soil and there are containing a wide range of pathogenic microbial populations which ensure that this region dangerous, unhealthy and unsafely.