Abstract
Fusobacterium are compulsory anaerobic gram-negative bacteria, long thin with pointed ends, it causes several illnesses to humans like pocket lesion gingivitis and periodontal disease; therefore our study is constructed on molecular identification and detection of the fadA gene which is responsible for bacterial biofilm formation. In this study, 10.2% Fusobacterium spp. were isolated from pocket lesion gingivitis. The isolates underwent identification depending on several tests under anaerobic conditions and biochemical reactions. All isolates were sensitive to Imipenem (IPM10) 42.7mm/disk, Ciprofloxacin (CIP10) 27.2mm/disk and Erythromycin (E15) 25mm/disk, respectively. 100% of Fusobacterium spp. isolates had 16S rDNA gene (360bp.), whereas two isolates had fadA gene (232bp.)
Highlights
Fusobacterium spp. are the most abundant species found in buccal cavity, in ill and healthy humans and cause severe damage in different forms of periodontal lesions,between mild reversible gingivitis form irreversible periodontitis form, they are viewed like islands in the mouth because of a special habitat in the human body characterized by liquid water near-constant presence, briefly the extreme temperature fluctuation, by an externally exposed hard surface and by wide variation in carbon and nitrogen input, including a basal component (saliva is a complex mixture with limited sources energy for bacteria, the fadA attachment gene was identified to be involved in oral Fusobacterium spp. and highly conservative for them [7]
Isolation and Identification of Bacterial Strains: Swabs from pocket lesion gingivitis and the teeth surface were inoculated on Fusobacterium selective medium Fusobacterium Selective Medium (FSM) (15g agar, 10g peptone, 5g Na2HPO4, 5g glucose, 3g beef extract, 2g soluble starch, 1g NaNO3, 1g yeast extract, 0.5g Lcysteine·HCl.H2O, 0.004g/1ml ethyl violet solution) according to Atlas and Snyder[8] with modification using vancomycin solution (0.1mg/1 ml) instead of bacitracin incubated at 37 °C for 24 hrs.; except Ethyl Violet and vancomycin solutions, all components dissolved in DDW to bring the volume 980.0ml after autoclaving aseptically, sterile Ethyl Violet solution and vancomycin solution 20 ml. were added to make one liter
From this study, 4 (10.2%) of Fusobacterium isolates were detected. this result somewhat was similar to the results of Hamad and Jasim, they obtained 12 % of clinical bacterial isolates from dental diseases belonging to Fusobacterium spp. and our results are in agreement with some researchers and their study on microorganisms in dental plaque and the important role of dental caries and damage of tooth structure. [11]
Summary
The genus Fusobacterium includes several species of obligatory anaerobic, opportunistic pathogens from bacteroidaceae family, that bare a strong resemblance to certain Bacteroides species, under the microscope, they are long, thin, and tapered rods (normally spindle-shaped cells) with sharp ends, non-spore-forming, motile or nonmotile, slender, spindle shape, fusiform or pleomorphic parallel rods, and show negative behavior to gram stain [1].They are found in mucous membranes of humans and animals and infected epithelial cells of mouth and intestine and play an important role in early buccal cavity biofilm development and in plaque biofilm colonization by co-aggregating with other pathogenic bacteria in buccal cavity, the best isolation of Fusobacterium spp. is from saliva or salivary fluid after concentration or from pocket lesion found between gingiva and surface tooth, the members of this microbe are resistant to vancomycin and susceptible to kanamycin and colistin. [2, 3].They play an important role in gingivitis, osteomyelitis, periodontitis, appendicitis, and other invasive infections of many human organs like head, neck, lung, liver, heart and brain and can causes pregnancy complications (preterm birth, stillbirth and neonatal sepsis) because they can pass through umbilical cord [4, 5, 6].Fusobacterium spp. are the most abundant species found in buccal cavity, in ill and healthy humans and cause severe damage in different forms of periodontal lesions ,between mild reversible gingivitis form irreversible periodontitis form, they are viewed like islands in the mouth because of a special habitat in the human body characterized by liquid water near-constant presence, briefly the extreme temperature fluctuation, by an externally exposed hard surface (teeth) and by wide variation in carbon and nitrogen input, including a basal component The genus Fusobacterium includes several species of obligatory anaerobic, opportunistic pathogens from bacteroidaceae family, that bare a strong resemblance to certain Bacteroides species, under the microscope, they are long, thin, and tapered rods (normally spindle-shaped cells) with sharp ends, non-spore-forming, motile or nonmotile, slender, spindle shape, fusiform or pleomorphic parallel rods, and show negative behavior to gram stain [1] They are found in mucous membranes of humans and animals and infected epithelial cells of mouth and intestine and play an important role in early buccal cavity biofilm development and in plaque biofilm colonization by co-aggregating with other pathogenic bacteria in buccal cavity, the best isolation of Fusobacterium spp. is from saliva or salivary fluid after concentration or from pocket lesion found between gingiva and surface tooth, the members of this microbe are resistant to vancomycin and susceptible to kanamycin and colistin.
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