The present investigation deals with spatial distribution of nitrogen content and nitrogen fixing bacteria, such as Azotobacter sp., Azospirillum sp., Clostridium sp. and cyanobacteria in rhizosphere of Triflium alexandrium (Egyptian clover) from Damietta Governorate centers, Egypt. Twenty different samples were collected, five samples for each center: Damietta, Kafr Saad, Faraskour and El-Zarka. Nitrogen content was determined and N-fixing bacteria were isolated and counted in each soil sample. Also all isolates were identified based on morphological and physiological characteristics and the nitrogenase activities were examined assayed for each isolate. The highest value of nitrogen content was 448 mg/kg that found in El-Zarka center (S4). El-Zarka and Kafr Saad centers contained the highest count of Azotobacter sp. and cyanobacteria with ratios of 35% and 32%, respectively. El-Zarka center is considered the dominant center in terms of Azospirillum sp. (41%). While Damietta center is considered the highest count with ratio 36% in case of Clostridium sp. The high N2-ase activity value of Azotobacter spp. (M.A.M-5) was 130nmole C2H2/h/l while the high value of Azospirillum spp. (N.D.M-5) was 51nmole C2H2/h/l. The high N2-ase activity value of Clostridium spp. (W.A.M-4) was 135nmole C2H2/h/l.