WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES Al-Jebouri et al. World Journal of Pharmacy and Pharmaceutical Sciences SJIF Impact Factor 2.786 Volume 3, Issue 6, 1720-1733. Research Article ISSN 2278 – 4357 ANTIBIOTICS RESISTANCE AMONG ANAEROBIC PATHOGENS CAUSING HUMAN PERIODONTITIS *Mohemid M Al-Jebouri1 and Hadeel M Al-Hadeethy2 1 Department of Microbiology,College of Medicine,University of Tikrit,Tikrit,Iraq 2 Department of Microbiology,College of Dentistry,University of Tikrit Article Received on 11 April 2014, Revised on 06 May 2014, Accepted on 24 May 2014 ABSTRACT Background:Periodontal disease could be defined as a disorder of supporting structures of teeth, including the gingiva, periodontal ligament and alveolar bone. Periodontal disease develops from a preexisting gingivitis. However, not every case of gingivitis develops into *Correspondence for Author Dr. Mohemid M Al-Jebouri Department of Microbiology,College of a periodontal disease. The inflammation of gingiva alone is termed gingivitis, and the severe inflammation of the periodontal ligament with destruction of alveolar bone is called periodontal disease. Medicine,University of Methods: One hundred eighteen samples were examined in the present Tikrit,Tikrit,Iraq study. Samples were obtained from periodontal pockets A plaque sample was inoculated onto different agar plates selected for anaerobic isolations. Periodontal pathogens were identified utilizing conventional methods, API RapID ANA system II , API 20A ,and Vitek2 ANC systems. Fifteen antibiotics were tested for their effectiveness on the various anaerobic bacteria isolated utilizing brucella blood agar plates disc diffusion methods. Results: peptostreptococcus prevotii , prevotella intermedia, prevotella melani , prevotella disiens, Bifidobacterium sp., Fusibacterium mortiferum peptostreptococcus tetradius ,and Wolinella sp. fusibacterium varium ,vellionella sp., campylobacter gracilis, capnocytophaga sp. ,peptostreptococcus magnus ,peptostreptococcus micros ,peptostreptococcus niger,peptostreptococcus anaerobius , staphylococcus saccharolyticus , streptococcus consellatus , and gemella morbillorum were the most common anaerobic periodontal pathogens isolated from patients in the present study.The bacteria isolated were generally highly sensitive to Imipeneme ,erythromcin , doxycycline, amoxycillin ,pipracillin, rodgyle ,spiromycin ,clindamycin , cephalexin ,cephalothin ,tetracycline and augmentin. Conclusion:The present study showed that 19 different species were responsible for the periodontal infections and the most common pathogens were 1720 Vol 3, Issue 6, 2014. www.wjpps.com Al-Jebouri et al. World Journal of Pharmacy and Pharmaceutical Sciences Peptostreptococcus prevotii, Vellionella sp. and Fusibacterium varium.The antibiotic sensitivity testing showed that these pathogens were generally sensitive to a wide range of chemotherapeutic antibiotics including Imipeneme ,erythromcin , doxycycline, amoxycillin ,pipracillin, rodgyle ,spiromycin ,clindamycin , cephalexin ,cephalothin ,tetracycline and augmentin.Periodontal anaerobic pathogens were very difficult to be isolated and timeconsuming work but it worths doing for the seek of oral health and economy. Key words: Periodontitis,anaerobic pathogens,antibiotics susceptibility. 1.INTRODUCTION Periodontal disease could be defined as a disorder of supporting structures of teeth, including the gingiva, periodontal ligament and alveolar bone. Periodontal disease develops from a preexisting gingivitis. However, not every case of gingivitis develops into a periodontal disease. The inflammation of gingiva alone is termed gingivitis, and the severe inflammation of the periodontal ligament with destruction of alveolar bone is called periodontal disease[1]. Healthy gingival sulcus has a flora dominated by equal proportions of gram positive cocci, especially Streptococcus spp, and Actinomuces sp. Later, plaque "matures" resulting in a flora consisting from facultative anaerobic microorganisms, spirochetes and motile rods. The proportions of strict anaerobic, Gram negative organisms increase significantly in accordance with increasing severity of disease. Disease activity in periodontal disease may range from slow, chronic, progressive destruction to brief and acute episodic bursts with varying intensity and duration[2]. The composition of the subgingival microbial flora and the level of pathogenic species differ from subject to subject as well as from site to site. The search for the pathogens of periodontal diseases has been underway for more than 100 years, and continues up today[2].More than 300 species of bacteria colonize subgingival area and their cell wall components can trigger immune activation[3]. The currently recognized key Gram negative periodontopathogens include: Porphyromonas gingivalis, Prevotella intermedia, Bacteroides forsythus ,Aggregatibacter actinomycetemcomitans , Fusobacterium spesies ,Capnocytophaga species , Campylobacter sp.,wollenilla sp.,and Bifidobacterium[3,4,5,6,7]. Also, the following bacteria could be isolated: Eubacterium spp, Peptostreptococcus species Streptococcus consellatus, and Staphylococcus saccharolyticus[2]. The clinical signs of periodontitis are changes in the morphology of gingival tissues, bleeding upon probing as well as periodontal pocket formation. This pocket provides an ideal environment for the growth and proliferation of anaerobic pathogenic bacteria[8]. The choice of the antimicrobial 1721 Vol 3, Issue 6, 2014. www.wjpps.com Al-Jebouri et al. World Journal of Pharmacy and Pharmaceutical Sciences agents in periodontal diseases must be based on the bacterial etiology of the infection . Several antibiotics have been tested for their clinical and microbiological efficacy in periodontal diseases[9]. Multiple factors contribute to the development of resistance to antibiotics. Their widespread and unnecessary use is the greatest contributing factors. 2.MATERIALS AND METHODS 2.1. Patients One hundred eighteen samples were examined in the present study. They were referred to the Laboratory of Microbiology, Department of Microbiology, College of Medicine ,Tikrit University. Patients were of both sexes(140 males and 40 females).Their ages ranged from 16-69 years old. The selection was done randomly among patients in Tikrit University/college of dentistry/dental teaching hospital . The diagnosis of periodontitis was made by the clinical examination including taking dental, medical and family history from the subjects involved in this study, so patients fulfilling the criteria to be diagnosed as periodontitis. Clinical measurements of periodontal parameters used included dental plaque index, gingival index, bleeding on probing, probing pocket depth and clinical attachment loss using (graduated William’s periodontal probe). Clinical diagnosis in each case was according to the dentist . The interviews were performed for each patient. The questionnaire included general information about the patient e.g.: name, age, sex, smoking, education, residence and treatment if the patients treated or not treated by antibiotics befo 2.2.Sampling Samples were obtained from periodontal pockets after supragingival plaque was removed from the teeth to be collected[10]. The supragingival dental plaque was removed with sterile cotton, and the tooth surface was dried with compressed air to prevent contamination with saliva. The exclusion of moisture in the mouth with sterile cotton rolls and subgingival plaque was collected from the most inflamed sites by inserting a sterile paperpoint into the periodontal pocket for 10 seconds , when the pocket depth was from3-7mm.The sample was mixed with contamination 1 ml thioglycollate broth (transport medium) , sealed tightly to avoid and kept at 4 C0. Samples were processed within 2 days of collection[11,12,13,14]. 1722 Vol 3, Issue 6, 2014. www.wjpps.com Al-Jebouri et al. World Journal of Pharmacy and Pharmaceutical Sciences 2.3.Cultural Technique A plaque sample(loopful from thioglycollate broth containing subgingival paperpoint which inserted into periodontal pocket was inoculated onto Brucella blood agar kanamycin 100 mg/l,vancomycin 7.5 mg/l Brucella blood agar, Enriched blood agar ,brain heart infusion agar,and trypticas soy agar with haemin and vitamin K1.The inoculated media were immediately incubated in an anaerobic environment generated with the anaerogens gas pack CO2 system Chemical, for 3 -7 days[10]. 2.4. Isolation and Identification of Bacteria Isolates were identified using cultural characteristics , gram -stain and biochemichal tests (convenentional methods) which includes : indole spot test, catalase test , oxidase test , arginine hydrolysis, lipase test, nitrate reduction , motility test ,growth in bile , urease test and growth on kanamycin(1mg),vancomycin(5µg), colistin(10µg) Brucella blood agar [15,16] . 2.5. Rapid identification methods 2.5.1.API RapID ANA II system [remel,USA]. This system was used as a qualitative micromethod employing conventional and chromogenic substrates for identification of anaerobic bacteria.The tests used in RapID ANA II system were based on microbial degradation of specific substrates detected by various indicator system[12]. 2.5.2.API 20A [Biomereiux,France]. The API 20 A system was utilized for the biochemical identification of anaerobes according to manufacturer instructions[10]. 2.5.3..VITEK 2 ANC System [Biomereiux,France] The VITEK 2 as an automated microbiological system was utilized for the identification of anaerobes according to manufacturer instructions[17]. 5.4.Antibiotic Sensitivity Testing A loopfull from colonies of anaerobic periodontal pathogens was inoculated into BHI broth and incubated in an anaerobic environment generated with the anaerogas pack (CO2)system chemical, for 2-3 days at 37Co . The bacterial suspension poured on the surface of the brucella blood agar plates, and left for 10 minutes to settle the bacteria. The excess of the 1723 Vol 3, Issue 6, 2014. www.wjpps.com Al-Jebouri et al. World Journal of Pharmacy and Pharmaceutical Sciences bacterial suspension was discarded using Pasteur pipette. The plates were left for one hour at room temperature to dry [16]. The antibiotic discs were placed by sterile forceps sterilized with alcohol. The antibiotics used in sensitivity test were shown in Table 6.The diameter of inhibition zone were measured. Interpretation of the zone of inhibition was done according to standard measurements[18,19,20] shown in Table 7. Table 6 : Antibiotics used in sensitivity testing for anaerobic periodontal pathogens isolates. Antibiotic Erythromycin Ampicillin Cephalothin Amoxycillin- clavulanate potassium[augmentin] Clindamycin Vancomycin Tetracycline Amoxycillin Metronidazole Doxycyclline Kanamycin Imipenem Pipracillin Cephalexin Spiromycin Spiromycin/metronidazol(Ro dogyl) Code E AM CEP Disc potency (mg/disc) 15 10 30 Manufacturer Oxoid(England) Himedia(India) Oxoid(England) AmC DA VAN TE AX MET DO K IPM PI CL SP 30 30 30 30 25 30 30 30 10 100 30 100 Himedia(India) Oxoid(England) Oxoid(England) Oxoid(England) Himedia(India) Oxoid (England) Himedia(India) Himedia(India) Himedia(India) Oxoid (England) Oxoid (England) Oxoid (England) ROD 30 Oxoid (England) 3.RESULTS 3.1. Identification of anaerobic periodontal pathogens Periodontal pathogens were identified by conventional methods, API RapID ANA system II , API 20A ,and Vitek2 ANC systems for identification of anaerobic bacteria. Table 11 showed the types of anaerobic periodontal pathogens isolated. In the present study the most common anaerobic periodontal bacteria isolated were peptostreptococcus prevotii which represented 15(8.3%) of all isolates, while prevotella intermedia, prevotella melani , prevotella disiens, Bifidobacterium sp., Fusibacterium mortiferum peptostreptococcus tetradius ,and Wolinella sp. represented only 1 (0.5%) of all isolates. 1724 Vol 3, Issue 6, 2014. www.wjpps.com Al-Jebouri et al. World Journal of Pharmacy and Pharmaceutical Sciences Table 11: Types of anaerobic bacteria isolated from periodontal patients(No. of patients =180) Types of isolates prevotella intermedia prevotella melani prevotella disiens Bifidobacterium sp Fusibacterium nucleatum Fusibacterium varium Vellionella sp. Campylobacter gracilis Wolinella sp. Capnocytophaga sp Peptostreptococcus prevotii Peptostreptococcus magnus Peptostreptococcus tetradius Peptostreptococcus micros Peptostreptococcus niger Peptostreptococcus anaerobius Staphylococcus saccharolyticus Streptococcus conselatus Gemella morbillorum Total 3.2. Antibiotic Sensitivity Testing No. (%) of isolates 1 (0.5%) 1 (0.5%) 1 (0.5%) 1 (0.5%) 1 (0.5%) 10 (5.5%) 14 (7.7%) 3 (1.6%) 1 (0.5%) 2 (1.1%) 15 (8.3%) 2 (1.1%) 1 (0.5%) 3 (1.6%) 4 (2.2%) 2 (1.6%) 5 (2.7%) 4 (2.2%) 2 (1.6%) 73 (40.7%) Table 18 shows the results of antibiotic sensitivity of periodontal pathogens iso lates. It was found that all isolates( 100%) were resistant to kanamycin and metronidazole. The imipeneme ,erythromcin , doxycycline, amoxycillin ,pipracillin, rodgyle ,spiromycin ,clindamycin , cephalexin ,cephalothin ,tetracycline and augmentin were the most effective drugs used in the present study . In the present study ,Prevotella intermedia was highly sensitive (100%) to doxycycline,imipeneme,amoxycillin,and augmentine .While prevotella melani was sensitive (100%) to all antimicrobial agents used . Prevotella disiense and bifidobacterium sp. were 100% sensitive to doxycycline , imipeneme, amoxycillin, augmentine, cephalthin , and pipracillin. wheras fusibacterium nucleatum only sensitive (100%) to doxycycline, imipenem, and pipracillin . Fusibacterium varium were 100% sensitive to amoxycillin, doxycycline, imipenem,ampicillin,cephalothin,and pipracillin. Vellionella sp.,were 100% sensitive to amoxycillin doxycycline, imipenem, augmentin, cephalothin,and pipracillin while 50% were sensitive to rodogyle and spyromycin. Campylobacter gracilis was sensitive to erythromycin (100% ) , but 50% of the isolates were sensitive to clindamycin, cephalothin and pipracillin. Wollinella sp. was 100% sensitive to 1725 Vol 3, Issue 6, 2014. www.wjpps.com Al-Jebouri et al. World Journal of Pharmacy and Pharmaceutical Sciences doxycycline ,clindamycin , cephalothin,and pipracillin. 50% of Capnocytophaga sp. isolates were sensitive to cephalexin , rodogyle ,spiromycin, erythromycin,clindamycin ,and cephalothin, but 100% of them were sensitive to doxycycline , tetracycline and pipracillin. Peptostreptococcs spp. were more sensitive to doxycycline, amoxycillin, cephalothin and pipracillin .Staphylococcus saccharolyticus was 100% sensitive to doxycycline , imipeneme, cephalothin and pipracillin but 50% were sensitive to amoxycillin, augmentin and clindamycin . Also, streptococcus conselatus and gemella morbillorum were 100% sensitive to doxycycline, imipenem, cephalothin and pipracillin . Table 18: Antibiotics sensitivity testing for isolated periodontal pathogens using disc diffusion method Types of periodontal pathogens isolated(No.) prevotella intermedia(1) Prevotella melani(1) prevotella disiens(1) Bifidobacterium sp(1) Fusibacterium nucleatum(1) Fusibacterium varium(10) Vellionella sp.(14) Sensitivity of isolates to antibiotics( %) A M IP A D E DO K M ET M M A C AX C L R O D SP 0 0 0 0 0 100 0 0 100 100 100 10 0 10 0 10 0 10 0 10 0 10 0 10 0 100 0 0 100 100 100 0 0 0 0 100 0 0 100 100 100 0 0 0 0 100 0 0 100 100 100 0 0 0 0 0 100 0 0 100 0 0 0 0 0 100 0 0 100 0 50 50 0 100 0 0 33 0 0 10 0 10 0 TE PI 0 0 0 100 10 0 100 100 0 100 0 100 0 100 0 0 0 0 100 100 100 0 100 0 100 100 0 100 0 100 50 100 0 0 0 33 50 50 0 50 0 0 0 0 0 10 0 100 0 100 0 10 0 10 0 Campylobacter gracilis(3) 0 0 33 33 Wolinella sp.(1) 0 0 0 0 0 50 50 50 50 100 0 0 100 0 0 50 50 50 0 50 50 0 100 0 0 50 0 33. 3 33. 3 0 0 0 0 0 100 0 0 100 100 50 50 100 0 100 0 0 0 0 100 0 0 100 100 100 10 0 100 0 100 0 25 25 0 100 0 0 100 100 100 25 100 0 100 Capnocytophaga sp.(2) peptostreptococcus prevotii(15) Peptostreptococcus magnus(2) Peptostreptococcus Tetradius(1) Peptostreptococcus micros(3) 1726 10 0 10 0 10 0 10 0 10 0 CE P Vol 3, Issue 6, 2014. 10 0 33. 3 www.wjpps.com 100 33.3 Al-Jebouri et al. Peptostreptococcus niger(4) Peptostreptococcus anaerobius(2) Staphylococcus saccharolyticus(5) Streptococcus conselatus(4) 10 0 10 0 World Journal of Pharmacy and Pharmaceutical Sciences 0 0 0 0 100 0 0 100 100 100 50 50 0 100 0 10 0 10 0 10 0 0 0 0 0 100 100 10 0 100 10 0 100 50 0 25 25 0 100 0 0 100 50 50 0 100 25 100 50 0 0 0 0 100 0 0 100 50 50 0 100 25 100 Gemella 50 0 0 0 0 100 0 0 100 50 100 0 100 0 100 morbillorum(2) AX: amoxycillin ; CL: cephalexin; ROD: rodogyle ; SP: spiromycin ; E:erythromycin ; DO : doxycycline ; K: kanamycin ;MET: metronidazole ;IPM: imipenem; AM: ampicillin , MC: augmentin ; DA: clindamycin; CEP: cephalothin; TE: tetracycline;PI: pipracillin. 4.DISCUSSION 4.1.Distribution of periodontal pathogens The oral cavity was characterized by harbouring indigenous microbiota. The ability of microorganisms to colonize the different oral surfaces depends mainly on their binding potential[21,22].Various environmental and host factors are involved in the harbouring of microorganisms and microbial composition[23]. Many indigenous microbiota are anaerobes and these microorganisms can be associated with oral infections and be the origin of distant infection[24].The most frequent oral anaerobic infections include gingivoperiodontal diseases, pulpal and periapical infections, peri-implantitis and pericoronarities [25].Gingivoperiodontal diseases, including gingivitis and periodontitis were caused by dental plaque, which was a biofilm[26]. It had been observed that 1g of dental plaque contains more than 1011 microorganism [23].The biofilm present in the gingival crevice, and later in the periodontal pocket, is extremely diverse, with up to 180 culturable species from a single pocket [4].Since such a diverse flora is present, trying to identify the particular species responsible for disease initiation and progression is a very complex, and difficult undertaking. Therefore, it is to be expected that a progressively more diverse and anaerobic flora will be isolated during disease progression. Moore and Moore presented large numbers of anaerobes increase in their overall proportions during disease progression and, conversely, aerobe and facultative species, decrease (changes in the microflora of the biofilm as a function of periodontitis severity) [27,28].The present study demonstrated a different microbiota in periodontal pockets. The finding presented here that Vellionella sp (7.7) Fusibcterium varium (5.5%) , Campylobacter gracilis (1.6) ,Capnocytophaga sp.(1.1) ,Prevotella intermedia(0.5%), Prevotella melani (0.5%) , Prevotella disiens (0.5%) ,Bifidobacterium sp. 1727 Vol 3, Issue 6, 2014. www.wjpps.com Al-Jebouri et al. World Journal of Pharmacy and Pharmaceutical Sciences (0.5%) ,Fusibacterium nucleatum(0.5%),and Wolinella sp.(0.5%) were more frequent as isolated from periodontal pocket as showed in Table 11 .It was ,however, interesting to note that gram positive anaerobic bacteria, especially Peptostreptococcus spp. were isolated in high rates in periodontal pocket which includes Peptostreptococcus prevotii (8.3%), Peptostreptococcus magnus(1.1%), Peptostreptococcus micros (1.6%), Peptostreptococcus tetadius(0.5%), Peptostreptococcus niger (2.2%) , and Peptostreptococcus anaerobius (2.7%) .Moreover , Staphylococcus saccharolyticus , Streptococcus consellatus, and Gemella morbillorum were isolated in the present study in percentages of 2.7%,2.2%,and 1.6% respectively. These results were almost similar to those of Daniluk et al who found that the most common periodontal pathogens were Veillonella species, Fusobacterium, Staphylococcus saccharolyticus ,Streptococcus consellatus, Gemella morbillorum , Peptostreptococcus spp. and Prevotella intermedia[28].Moreover, these results were similar to those of Spartt , Winkelhoff et al , Piovano , Haffajee and Socransky who found that Veillonella species, Fusobacterium, Staphylococcus saccharolyticus consellatus, Gemella morbillorum, Peptostreptococcus spp. ,Streptococcus Prevotella intermedia, Campylobacter gracilis and Bifidobacterium sp.[29]. Furthermore , Mohammad et al found that Actinobacillus actinomycetemcomitans (26.8%), Porphyromonas gingivalis (21.9%), Capnocytophaga sputigena (16.7%), Eikenella corrodens (13.2%), Prevotella intermedia (10.5%), Prevotella disiens (3.1%), Capnocytophaga gingivalis (2.2%), Peptostreptococcus micros (2.9%), Prevotella corporis (1.8%), Peptostreptococcus magnus (1.3%) , and Fusobacterium nucleatum (0.4%)were most common isolates[30].Spratt et al was found that Capnocytophga sp. involvement in some forms of periodontitis[31].According to Gürsoy et al Prevotella intermedia is associated with periodontal disease [32].Mayorga-Fayad et al revealed that the frequency increases Prevotella intermedia in patients with periodontitis[33]. The present results were different from those of Yacoubi et al who found that Aggregatibacterium actinomycetemcomitans , Eikenella corrodens are primary causative agent in periodontitis[14].Moreover , Cisar et al and Ximenez-Fyvie et al found that actinomyces were the most common component of periodontal disease[34].The data from the investigation suggest that there was heterogeneity in the subgingival periodontopathogenic bacteria among subjects. However, as many bacteria in the oral cavity cannot be cultured, it was likely that these still uncharacterized bacteria could play a role in the initiation and progression of periodontal disease. 1728 Vol 3, Issue 6, 2014. www.wjpps.com Al-Jebouri et al. World Journal of Pharmacy and Pharmaceutical Sciences 4.2. Antibiotics Sensitivity Testing of Periodontal Pathogens During several years, an increasing resistance to different antimicrobial agent in anaerobic bacteria has been observed[35].The present study demonstrated that 100% of anaerobic periodontal pathogens isolates were resistant to kanamycin and metronidazole (Table 18) .The imipeneme ,erythromcin , doxycycline, ampicillin ,pipracillin, rodgyle ,spiromycin ,clindamycin , cephalexin ,cephalothin ,tetracycline and augmentin were the most effective . In the present study Prevotella intermedia was highly sensitive (100%) to doxycycline , imipeneme, amoxycillin ,and augmentine .On the other hand , Prevotella melani was also sensitive to all antimicrobial agents used except kanamycin and metronidazole. Prevotella disiense and Bifidobacterium sp. were 100% sensitive to amoxycillin , doxycycline , imipeneme, amoxycillin, augmentine, cephalthin ,and pipracillin. Whereas Fusibacterium nucleatum only sensitive to doxycycline, imipenem, and pipracillin . Fusibacterium varium was mostly sensitive to amoxycillin, doxycycline, imipenem, ampicillin, cephalothin, and pipracillin. Vellionella sp. was sensitive to amoxycillin ,doxycycline, imipeneme ,augmentin,cephalothin,and pipracillin while 50% of its isolates were sensitive to rodogyle and spyromycin.These results were almost similar to those of Brook who found that Vellionella sp. was generally susceptible to betalactam but resistant to tetracycline and erythromycin[36]. Campylobacter gracilis were 100% sensitive to erythromycin , but 50% sensitive to clindamycin,cephalothin and pipracillin. Wollinella sp. isolates were sensitive to doxycycline ,clindamycin,cephalothin,and pipracillin. Furthermore , Capnocytophaga sp revealed a sensitivity to cephalexin , rodogyle ,spiromycin, erythromycin,clindamycin ,and cephalothin, and 100% of its isolates were sensitive to doxycycline , tetracycline and pipracillin .Moreover , Peptostreptococcs spp. were more sensitive to doxycycline, amoxycillin, cephalothin and pipracillin .Staphylococcus saccharolyticus were sensitive to doxycycline , imipeneme,cephalothin and pipracillin 100% but 50% of its isolates were sensitive to amoxycillin,augmentin and clindamycin . Streptococcus conselatus and Gemella morbillorum were sensitive to doxycycline , imipenem,cephalothin,and pipracillin as showed in Table 18. These results were almost similar to those of Thomas who found that Prevotella sp , Fusibacterium sp, Campylobacter sp. and Streptococcus conselatus were sensitive to amoxycillin, ampicillin ,doxycycline, and pipracilin and also anaerobic periodontal pathogens resist to metronidazole [37]. Sally et al found that metronidazole ,pipracillin ,cephalexin, amoxycilin, augmentin remain good empirical choices for anaerobic bacteria[38]. Nyfors et al found that Fusibacterium sp. was susceptible to penicillin ,amoxycillin ,augmentin,cephalexin, and metronidazole[39].In addition , pierre et al found that anaerobic 1729 Vol 3, Issue 6, 2014. www.wjpps.com Al-Jebouri et al. World Journal of Pharmacy and Pharmaceutical Sciences periodontal pathogens were susceptible to spiromycin and rodogyle[40]. Maria et al found that 70% of Prevotella sp.and 30% of Fusibacterium sp. produce b-lactamases are making penicillin inactive [41] . Boyanova et al found that 21% of gram positive anaerobic cocci were resistant to penicillin and 16% of them were resistant to clindamycin [42]. These results were different from those of Schwach et al who found that metronidazole most common selective efficacy against obligate anaerobic periodontal pathogens[8]. Hagashi et al and Sauvetre et al found that clindamycin used for treatment periodontal disease [43] . Moreover , Louise et al found that 50% of the isolates were inhibited by metronidazole when tested anearobically[44] .Selcuk et al found that metronidazole had a good effect on anaerobes[45]. Ahmed found that metronidazole was not inhibiting aerobic facultative, microearophillic organisms such as Actinomyces, Streptococcus , and Capnocytophaga[46]. The diffusability of the antimicrobial agents depends on it’s composition , hydrophilicity and its rate of release from soluble matrix in which bound. Therefore , the size of the inhibition zone may not be related entirely to the differences in the chemical effect of different antimicrobial agents[47]. 5.CONCLUSION The present study revealed that 19 different anaerobic species were isolated from periodontal infection site and the most dominant types were Peptostreptococcus prevotii, Vellionella sp. and Fusibacterium varium and their frequencies of isolation were 8.3,7.7 and 5.5 respectively.Fifteen different antibiotics were tested against the different isolates and Imipeneme ,erythromcin , doxycycline, amoxycillin ,pipracillin, rodgyle ,spiromycin ,clindamycin , cephalexin ,cephalothin ,tetracycline and augmentin were the most effective drugs for eradication of these periodontal pathogens. REFERENCES 1. Jayamathi G , Pamela E and Puvanakrishnan R. Therapeutic effect of proanthocyanidins on the pathogenesis of periodontitis . Indian J Experim Biol.2011;49:83-93. 2. Kesic L, Milasin J, Igic M and Obradovic R. 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