**3.1 Re-growth and identification of** *Bacillus megaterium*

Besides using selective media, Sperber Medium, **Table 4** shows the Test of Morphological, and **Table 5** represents the biochemical tests.

## **3.2** *Bacillus megaterium* **hydrolyzes and bacteria growth**

## *3.2.1 Growth of* B. megaterium

In **Figure 1**, the results show that the highest *B. megaterium* growth was in (two months) for both (5, 25) ppm (0.164, 0.167) sequentially, while the 15 ppm show in 60 days, the highest growth is (0.215) in comparison with others when used glyphosate as carbon sources.

## *3.2.2 Degradation rate%*

The highest degradation rate% for Glyphosate by *B. megaterium* in comparison among concentration was for both the 5-25 ppm in 60 days reached (70.01-70.9) %, **Figure 2**.


#### **Table 4.**

*The tests of morphological.*


#### **Table 5.**

*The tests of biochemical.*

**Figure 1.** *Culture of* B. megaterium *on Mineral Salt Media with Glyphosate.*

### *3.2.3 Glyphosate residues by HPLC test*

The *B. megaterium* growth on different concentration in MSM at 30 °C, in comparison with control in **Figures 3** and **4**. Decreasing in glyphosate in 30 days for concentration (5,10) ppm (7, 8)% is the best peak area, while the 25 ppm showed 28%, while the results glyphosate peak area two months incubation, led to (20, 15) ppm in comparative with control. When comparing among HPLC, **Figures 5** and **6**

Bacillus megaterium *Biodegradation Glyphosate DOI: http://dx.doi.org/10.5772/intechopen.96919*

**Figure 2.**

*The degradation rate of Glyphosate in MSM in Comparative with control.*

#### **Figure 3.**

*Peak height, peak area and retention time of Glyphosate after incubation B.M. 30 days in MSM.*

improved that the *Bacteria* degradation ratio% positive results with all glyphosate concentrations via incubating two months incubation. However, the best was for (5, 25) ppm for each degradation ration% and the HPLC.

Organophosphorus pesticides microbial degradation and the bioremediation progress development for contaminated soils depend on the introduction of microbe's biodegrading [16]. *B. megaterium* shows the degradation rate% and highest growth in two months cultivation time for both five and twenty five concentration. The degradation pesticide ability like Chlorpyrifos (600 mg-L1) via *B. megaterium* in ten days incubation was 81% [17] and 73% for 20 ppm during three weeks. On the other hand, *Bacillus megaterium* 99% in seven days, degradation ratio towards atrazine (50 ppm) and Chlorpyrifos in (1-2) weeks [18, 19]. Monocrotophos (MCP), 83% degradation ratio reached led sub-products to CO2, NH4, and (HPO3) [20]. Each bacterium produces enzymes to analyze

#### **Figure 4.**

*Peak height, peak area and retention of Glyphosate after incubation B.M. 60 days in MSM.*

#### **Figure 5.**

*Comparative among the degradation ratio% and Glyphosate concentrations via HPLC in 30 days incubation on MSM.*

compounds and supplying its self with elements in need for re-build enzyme and cell body. One of *Bacillus megaterium* enzyme is β-Amylase [21] that consist from carbon and nitrogen mainly, so the bacteria most often organic-phosphors compounds, Glyphosate used to supply a single element (carbon, phosphorus or sulfur) and [22]. Due to that the concentration of glyphosate generally reduces along with bacteria growth. Phosphate ester and phosphonate are the analysis hydrolysis results of phosphorus. The esters groups have they have many vulnerable to hydrolysis sites. Beside hydrolysis, the oxidation also one of the major reactions, and the alkylation,dealkylation [23]. Detoxification is one of microbial degradation principles via hydrolysis phosphor bonds with oxygen (P-O-alkyl, P-O-aryl [24].

Bacillus megaterium *Biodegradation Glyphosate DOI: http://dx.doi.org/10.5772/intechopen.96919*

#### **Figure 6.**

*Comparative among the degradation ratio% and Glyphosate concentration via HPLC in 60 days incubation on MSM***.**
