04_HM hwp

The Korean Journal of Microbiology (2010) Vol. 46, No. 4, pp. 334-340 Copyright c 2010, The Microbiological Society of Korea 흡연자와비흡연자간의구강내세균분포및항균제감수성 정현자 1 김수정 2 * 1 대구보건대학치위생과, 2 대구보건대학임상병리과 Distribution and Antimicrobial Susceptibility of Bacteria in the Oral Cavity of Smokers or Non-Smokers Hyun-Ja Jeong 1 and Su Jung Kim 2 * 1 Department of Dental Hygiene, Daegu Health College, Daegu 702-722, Republic of Korea 2 Department of Clinical Pathology, Daegu Health College, Daegu 702-722, Republic of Korea (Received October 22, 2010/Accepted December 6, 2010) It is well known that smoking as well as drinking is a factor of stomatopathy, however there are few investigations about comparison of oral flora between smokers and non-smokers. In this study, we isolated the oral flora of 30 smokers and 30 non-smokers and cultured them on blood agar plates. The isolated pathogenic microorganisms were tested for antibiotic susceptibility and resistance using the Kirby-Bauer antibiotic testing method. Each colony was stained using the Gram staining method and was identified by an automatic identifier, known as the VITEK system. We isolated 41 colonies from smokers oral cavity, and they were sorted as 63% of Gram-positive cocci, 29% of Gram-negative cocci, 3% of Gram-positive bacilli, and 5% of Gram-negative bacilli by gram staining, whereas 38 colonies were isolated from non-smoters oral cavity, and their proportions were 55% of Gram-positive cocci, 26% of Gram-negative cocci, 3% of Gram-positive bacilli, and 16% of Gram-negative bacilli. The VITEK system revealed specific distribution of bacteria species that Streptococcus mutans (6/41), Gemella morillorum (6/41), Streptococcus oralis (2/41), Streptococcus pneumoniae (1/41), Staphylococcus aureus (3/41), Streptococcus anginosus (1/41), Streptococcus intermedius (1/41), Streptococcus uberis (1/41), and Streptococcus sanguinis (1/41) in smokers oral cavity whereas Streptococcus sanguinis (8/38), Staphylococcus aureus (1/38), Staphylococcus auricularis (1/38), Streptococcus uberis (1/38), Streptococcus intermedius (1/38), Streptococcus mutans (1/38), and Streptococcus oralis (1/38) in those of non-smokers. Three cases of Staphylococcus aureus from smokers produced Beta-lactamase and were identified methicillin-resistance Staphylococcus aureus (MRSA). However one case of Staphylococcus aureus from non-smoker did not produce Beta-lactamase and was sensitive to methicillin. In conclusion, the distribution of oral flora was different between smokers and non-smokers oral cavity, especially Gemella morillorum and MRSA were predominantly found in smoker s oral cavity. These results are useful in the treatment and prevention of patients with stomatopathy caused by smoking. Keywords: antimicrobial susceptibility, non-smokers, oral cavity, smokers 우리나라흡연인구는보건가족부의조사결과, 20% 정도이며흡연시발생하는연기는약 400여가지의화학물질을포함하고있으며주성분으로는니코틴, 타르, 일산화탄소, 황화수소및암모니아등으로이루어져있다 (6). 흡연시다양한화학물질을흡입함으로써구강내치주병, 치아우식, 궤양성치주염, 치아마모증및치아변색, 구취, 수술후창상치유의지연과반흔형성, 그리고백반증을유발할수있으며, 더나아가구 * For correspondence. E-mail: sjkim@mail.dhc.ac.kr; Tel: +82-53-320-1303; Fax: +82-53-320-1450 강암으로도발전한다 (6, 18). 구강내에발생할수있는생물학적기전은치아우식증 (Dental caries), 치주질환 (Periodontal disease), 그리고결손치 (Missing teeth) 로나뉜다 (17). 다른연구진에따르면흡연은심각한치주염과매우밀접하게연관된환경적위험요소라고보고하였다 (14). 이런세균성질환은다량의 Viridans Streptococci와유산균이구강내에남아있는당성분을이용하여산을생성하고생성된산은치아에부착되어무기물질을용해또는탈회하여치아조직을파괴함으로써발생한다 (7). Viridans Streptococci은 4그룹으로나누어지며

흡연자와비흡연자간의세균분포및항균제감수성 335 Anginosus (S. nginosus, S.constellatus, S. intermedius), Mitis (S. oralis, S. mitis, S. gordonii, S. sanguinis, S. parasanguis), Salivarius (S. vestibularis, S. salivarius) 그리고 Mutans (S. mutans, S. sorbinus) 로구성되어있으나아직까지도분류는통일되어있지않다. Viridans Streptococci은구강, 장관, 여성생식기의상재균이나면역이약화된환자에서는균혈증및심내막염을일으키는것으로알려져있다 (4, 17). 주로구강내에서만분리되는것으로알려진 Viridans Streptococci가임상에서환자의혈액배양에서도빈번히분리되고있고점차내성을획득한다고보고하였다 (16). 흡연으로인해숙주와병원체간의반응으로세포매개성면역반응 (Cell-mediated immune response) 및체액성면역반응을저해하는것으로알려져있다 (15). 인체내면역반응의저해는다양한질환을유발할수있으며구강내에서는바이러스성질환으로헤르페스감염 (Herpes infection), 만성육아조직성감염질환인결핵감염 (Mycobacterium tuberculosis), 그리고면역이떨어진사람에게기회감염균으로알려진캔디다가이상증식하여캔디다증 (Candidiasis Moniliasis) 을유발하는것으로알려져있기에직접적으로구강환경을악화시키는흡연을지속할경우구강내상재효모균인캔디다가이상증식하여캔디다증을일으킬수있고, 다양한바이러스성질환과세균성질환을악화하는것으로알려져있다 (8, 11, 12). 흡연이구강내환경을악화하여치아우식및치주질환을유발한다는보고는있었으나실험적조사를통한논문은매우미비한상태이기에본연구진은정상인에서지속적인흡연을통해구강내환경이악화되고더나아가전반적인면역력이약화될수있는흡연한그룹과전혀흡연을하지않은그룹으로나누어구강내세균의분포를조사및비교하고, 조사된세균들중병원성을유발할수있는세균을중심으로항균제감수성검사를하고자한다. 그리고항균제의내성, 중간내성및감수성의정도를조사하여향후흡연자의구강질환을예방하고치료하는데기초자료로제공하고자한다. 대상및방법대상본연구는 2010년대구시소재, D대학의 20대초반의남녀학생을대상으로하였으며, 최근 6개월이내치과진료 ( 스케일링포함 ) 를받지않은학생을대상으로하였다. 흡연자는적어도 5년이상현재흡연을하고있는학생 30명을선택하였고비흡연자는전혀흡연한경험이없는학생 30명을선택하였다. 흡연자와비흡연자의구강내세균의분리배양멸균된 1회용브러쉬를이용하여구강내하악전치부설측부위와상악좌 우측협측부위의협점막에있는타액과치아사이의치태 (Dental plaque) 부위를닦은후 0.45% Saline 10 ml 에희석하고, 희석액 100 μl를혈액한천배지 (Becton, Dickinson and Company, USA) 에도말배양하였다. 배양조건은혐기적상태를유지하기위해 Gaspak Anaerobe Container System (Becton, Dickinson and Company) 를이용하여 35 C에서 48 시간배양하였다. 집락의성상이녹색을띄는 α 용혈을일으키는그룹과그밖의집락을순수분리배양하고자 Brain heart infusion agar (Becton, Dickinson and Company) 에재배양하였다. 재배양된집락을그람염색을실시하여그람양성균과그람음성균으로분리하였다. 흡연자와비흡연자의구강내세균의동정순수분리된집락들을이용하여그람염색을실시한후, 그람양성균으로확인된집락들은 Catalase test를실시하고, 순수분리된집락을 0.45% Saline에 OD값이 600 nm가되도록한후미생물동정을위해고안된 Gram Positive Identification Card 내로균액을주입후 35 C, 24시간배양한다. Gram Positive Identification Card는다양한생화학적반응을일으키며세균의동정을컴퓨터내테이터베이스를통해확인한다. 이는미생물동정을위한자동화기기인 VITEK System (Bio merieux, Marcyl etoile, France) 을이용하였다 (3, 9, 10). 표준균주로는 S. aureus ATCC 29213, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853을사용하였다. 그람음성균으로확인된집락은 Oxidase test를실시한후 Gram Negative Identification Card를이용하여그람양성균과동일한방법으로세균을동정하였다. 흡연자와비흡연자의구강내세균의항균제감수성검사분리동정된세균중그람양성균과그람음성균으로나누어항균제감수성검사를실시하였다. 그람양성균중 Viridans Streptococci는치과시술시예방적항생제투여의권장용법에근거를두고 Ampicillin, Clindamycin, Cefazolin, Penicillin, Tetracycline에대한항균제감수성검사를실시하였다 (13). 검사방법으로는 Clindamycin, Cefazolin, Tetracycline은 disk diffusion법을실시하였고, Ampicillin, Penicillin은액체희석법과항균제에대한최소억제농도를알수있는 E-test (PDM Episilometer; AB Biodisk, Solna, Sweden) 을실시하였다 (16). 그밖의그람양성균인 S. aureus는 Gram Positive Susceptibility Card를이용하여항균제감수성검사를실시하였고항균제종류는 Cephalothin, Ciprofloxacin, Clindamycin, Erythromycin, Gentamicin, Habekacin, Linezolid, Nitrofurantoin, Oxacillin, Penicillin-G, Rifampin, Teicoplanin, Tetracycline, Trimeth-sulfa, Vancomycin, Beta-lactamase이다. 그람음성균들은 Gram Negative Susceptibility Card를이용하였고항균제종류로는 Amikacin, Ampicillin, Ampicillin/Sulbactam, Aztreonam, Cefepim, Ceftazidime, Ceftriaxone, Cephalothin, Ciprofloxacin, Gentamicin, Imipenem, Piperacillin, Piperacillin/ Tazobactam, Ticarcillin/CA, Tobramycin, Trimeth-fula이며항균제감수성검사결과는 2010년 Clinical and Laboratory Standards Institute Guideline으로정의하였다.

336 Hyun-Ja Jeong and Su Jung Kim 70.0% 60.0% Isolated % by gram stain 50.0% 40.0% 30.0% 20.0% 10.0% 0.0% Gram positive cocci Gram negative cocci Gram positive bacilli Gram negative bacilli Fig. 1. Proportion of microorganisms isolated from oral cavity between smoker and non-smoker. Porportion of the Gram-positive (+/-) or Gram-negative (+/-) organisms classified by Gram stain. Samples were collected on June, 2010. 결과흡연자와비흡연자간의구강내세균분포흡연자와비흡연자의구강내하악전치부설측면, 상악좌, 우측협측면의타액과인근부위치아와치아사이의치태부분을채취하여혈액한천배지에접종한후혐기적조건하에서 35 C, 48시간배양하였다. 배양된집락의성상에따라분리한후그람염색한결과, 흡연자에서세균으로염색된것은총 41 개이며, 그람염색분리율은그람양성구균 63%, 그람음성구균 29%, 그람양성간균 3%, 그리고그람음성간균 5% 로나타냈고, 비흡연자에서는총 38개로나타냈으며분리율은그람양성구균 55%, 그람음성구균 26%, 그람양성간균 3%, 그리고그람음성간균 16% 로나타냈다 (Fig. 1). 두집단모두에서그람양성구균이많이검출되었고, 특이하게비흡연자에서는흡연자보다그람음성간균이많이검출되었다. 흡연자와비흡연자간의구강내세균동정그람염색후그람양성으로염색된균집락은미생물동정을위해미생물자동화동정기기인 VITEK system을이용하였고그중 Gram Positive Identification Card를이용하여그람양성균을동정하였으며, 흡연자에서는총구균 26개와간균 1개를동정하였으며그결과, S. mutans (6), Gemella morillorum (6), S. oralis (2), S. pneumoniae (1), S. aureus (3), S. anginosus (1), S. intermedius (1), S.uberis (1), S. sanguinis (1) 로나타났으나구균 26개중 22개만동정되었고간균 1개는동정되지않았다. 비흡연자에서는총구균 21개와간균 1개를동정하였으며그결과, S. sanguinis (8), S. aureus (1), S. auricularis (1), S. uberis (1), S. intermedius (1), S. mutans (1), S. oralis (1) 로동정되었으나구균 21개중 14개만동정되었고간균은동정되지않았다 (Table 1). 흡연자와비흡연자간의세균종의분포에는별차이가없으나흡연자에서 S. mutans와 Gemella morillorum가우세하게검출되었고, 비흡연자에서는 S. sanguinis (8) 가우세하게검출되었다. Gram Negative Identification Card를이용하여그람음성균을동정한결과, 흡연자에서는그람음성간균 2개를동정한결과 K. pneumoniae, Pasteurella pneumotrpica로나타났고, 비흡연자에서는그람음성간균 6 개를동정한결과 K. pneumoniae (2), Citrobacter koseri (1), Pasteurella multocida (1), Flavimonas oryzihabitans (1), Pantoea spp. (1) 이동정되었다 (Table 1). 그람음성세균에서는흡연자와비흡연자간의큰차이점은발견할수없었다. 흡연자와비흡연자간의구강내세균의항균제감수성검사그람양성균중 Viridans Streptococci은구강, 장관, 여성생식기의상재균이나면역이약화된환자에서는균혈증및심내막염을일으키는것으로알려져있으며주로구강내에서만분리되는것으로알려져있고혈액한천배지상에서알파용혈을일으키며녹색환을형성하고현재 4개의그룹으로나눠지나분류는통일되어있지않다 (17). Viridans Streptococci은 Ampicillin, Clindamycin, Cefazolin, Penicillin, Tetracycline에대한항균제감수성검사를실시한결과, 흡연자에서는 Ampicillin 에 22% 가내성, 중간내성이 18% 이며감수성이 60% 이다. Clindamycin에는내성이 30% 이고중간내성이 15% 이며감수성이 55% 이었다. Cefazolin에는내성이 25%, 중간내성이 10% 이며감수성이 65% 이었고 Penicillin에서는내성이 35% 이며감수성이 65% 이다. Tetracycline에는내성 15%, 중간내성 30% 이며감수성이 55% 이었다. 비흡연자에서는 Ampicillin에 14.8% 가내성, 중간내성이 7.4% 이며감수성이 77.8% 이다. Clindamycin에는내성이 33.3% 이고감수성이 66.7% 이었다. Cefazolin에는내성이 3.7%, 중간내성이 3.7% 이며감수성이 92.6% 이었고 Penicillin에는내성이 14.8%, 중간내성이 7.4% 이며감수성이 77.8% 이다. Tetracycline에는내성 7.4%, 중간내성 3.7% 이며감수성이 88.9% 이었다 (Table 2). 그람양성균

흡연자와비흡연자간의세균분포및항균제감수성 337 Table 1. Identification of microorganisms isolated from oral cavity between smoker and non-smoker using the VITEK system Source Identification Disease Streptococcus mutans Dental caries, Gingivitis Gemella morillorum Endocarditis Streptococcus oralis Streptococcus pneumonia Pneumonia, Otitis Staphylococcus aureus Septic arthritis, endocarditis, Pneumonia Smoker Streptococcus anginosus Endocarditis, Gingivitis Streptococcus intermedius Endocarditis, abscesses Streptococcus uberis Bovine mastitis Streptococcus sanguinis Endocarditis Klebsiella pneumonia Pneumonia Pasteurella pneumotrpica Bacteremia Streptococcus sanguinis Dental caries, Endocarditis Staphylococcus aureus Septic arthritis, endocarditis, Pneumonia Staphylococcus auricularis Streptococcus uberis Streptococcus intermedius Non-smoker Streptococcus mutans Dental caries, Gingivitis Streptococcus oralis Klebsiella pneumonia, Pneumonia Citrobacter koseri Pasteurella multocida Pantoea spp. Flavimonas oryzihabitans Peritoneal infection 중 S. aureus는 Gram Positive Susceptibility Card를이용하여항균제감수성검사를실시하였고항균제감수성검사결과는 2010년 Clinical and Laboratory Standards Institute Guideline 에따라정의하였다. 흡연자에서분리된 3균주의 S. aureus는 3균주중 2균주에서 Oxacillin 내성을나타냄으로 MRSA로판독되었고 Cephalothin에 3균주중 2균주에서내성을나타냈고 1균주에서감수성을나타냈으며 Ciprofloxacin에는 3균주중 2균주에서중간내성을나타냈고 1균주에서감수성을나타냈으며 Penicillin-G에모두내성을나타냈으며 3균주모두 Betalactamase 양성으로나타났다. 비흡연자에서분리된 1균주의 S. aureus는 Ciprofloxacin에만중간내성을가지며다른항균제에서는모두감수성을나타냈다 (Table 3). 그람음성균들은 Gram Negative Susceptibility Card를이용하였고흡연자에서분리된그람음성균은 K. pneumoniae와 P. pneumotrpica이며 K. pneumoniae는사용한항균제중 Ampicillin만내성을나타냈고이를제외한모든항균제에감수성을나타냈으며, P. pneumotrpica는 Aztreonam과 Cefepim에내성을나타냈고다른항균제에는감수성을나타냈다. 비흡연자에서분리된그람음성균중 K. pneumoniae는 2균주에서분리되었으며항균제감수성검사에서모든항균제에감수성을나타냈고 C. koseri 는 Ampicillin 에내성을나타냈으며 Ampicillin/Sulbactam과 Cephalothin에중간내성을나타냈고 P. multocida와 F. oryzihabitans에서는 Aztreonam과 Cefepim에내성을나타냈다. 그리고 Pantoea spp. 는사용한항균제모두에감수성을나타냈다 (Table 4). Table 2. Antimicrobial susceptibilities of Viridans Streptococci by disk diffusion method or E-test (PDM Episilometer; AB Biodisk, Solna) Non-smoker Smoker Antibiotics (μg) Antimicrobial susceptibility Antibiotics (μg) Antimicrobial susceptibility R (%) I (%) S (%) R (%) I (%) S (%) Ampicillin (5 μg) 14.8 7.4 77.8 Ampicillin (5 μg) 22 18 60 Cefazolin (15 μg) 3.7 3.7 92.6 Cefazolin (15 μg) 25 10 65 Penicillin (5 μg) 14.8 7.4 77.8 Penicillin (5 μg) 35 0 65 Tetracycline (15 μg) 7.4 3.7 88.9 Tetracycline (15 μg) 15 30 55 Clindamycin (2 μg) 33.3 0 66.7 Clindamycin (2 μg) 30 15 55 R, resistant; I, intermediate; S, susceptible

338 Hyun-Ja Jeong and Su Jung Kim Table 3. Antimicrobial susceptibilities of S. aureus by disk diffusion method Antibiotics (μg) Smoker Non-smoker 1 strain 2 strain 3 strain 4 strain Instru ment Cephalothin >=32 R <=2 S >=32 R <=2 S Ciprofloxacin 2 I <=0.5 S 2 I 2 I Clindamycin <=0.5 S <=0.5 S <=0.5 S <=0.5 S Erythromycin <=0.5 S <=0.5 S <=0.5 S <=0.5 S Gentamicin <=2 S <=2 S <=2 S <=2 S Habekacin <=4 S <=4 S <=4 S <=4 S Linezolid <=2 S <=2 S 4 S <=2 S Nitrofurantoin <=32 S <=32 S <=32 S <=32 S Oxacillin >=8 R <=0.5 S 4 R 2 S Penicillin-G >=16 R >=16 R >=16 R 0.06 S Rifampin <=0.25 S <=0.25 S <=0.25 S <=0.25 S Teicoplanin <=2 S <=2 S 8 S <=2 S Tetracycline <=1 S <=1 S <=1 S <=1 S Trimeth-sulfa <=10 S <=10 S 40 S <=10 S Vancomycin 2 S <=0.5 S 2 S 2 S Beta-lactamase + + + - R, resistant; I, intermediate; S, susceptible 고찰구강내상재균총은타액, 연령, 숙주의방어능력, 구강위생관리정도, 치아상태, 탄수화물섭취등에의해영향을받을수있다 (6). 구강을직접적으로자극하는것중흡연이있으며, 흡연으로인해타액분비가감소되며, 구강건조증을일으킬수있기에본연구진은흡연자와비흡연자간의구강내세균의분포를통해구강내세균의변화를조사하였다. 연구대상자는적어도 5년이상흡연한자와전혀흡연을경험하지않은비흡연자를대상으로하였다. 구강내치태부위와타액을채취하여세균을분류한결과, 흡연자에서는비흡연자에비해그람양성구균이약 8% 정도많이검출되었고, 그람음성구균은약 11% 정도적게검출되었다. 이는흡연으로인해구강내미생물의분포량에는큰영향을미치지않는것으로조사되었다. Ahn이발표한자료에의하면, 치과치료시이용되는방사선조사는구강내부를자극하여구강균총의변화를가져오며, 특히 S. mutans, Lactobacillus, Staphylococcus spp. 및 Candida 등은증가하는반면, S. sanguinis, Neisseria spp. 및 Fusobacterium spp. 는감소하였으며, 이런변화는방사선조사로인해구강건조증이심화되며, 점차치태의축적이증가됨으로써세균의전체적인수는일정하나세균구성에는변화를초래함을보고하였다 (1, 2). 본연구에서는외부자극중하나인흡연이구강균총의변화에어떠한영향을나타내는지를조사하고자그람염색을실시하여분류하고동정하였다. 흡연자와비흡연자의구강내동정된세균을살펴본결과흡연자에서치아우식을일으키는 S. mutans가많이검출되었고, 비흡연자에서는 S. mutans에길항작용을나타내는 S. sanguinis가많이검출되었다. 치아우식과 S. mutans/s. sanguinis의상관관계를조사한자료에의하면, 구강환경악화로인해치아우식유발균인 S. mutans가우세하게분포하는반면, S. sanguinis는감소하는것으로보고되었다 (5). 이는흡연으로인해구강내환경이악화되면치아우식과직접적관련이있는 S. mutans가 S. sanguinis에비해많이검출됨을확인하였다. 구강에서분리되는세균중 Viridans streptococci는 Ampicillin, Clindamycin, Cefazolin, Penicillin, Tetracycline에대한항균제감수성검사를흡연자와비흡연자로나누어실시하였다. 그결과, 비흡연자에비해흡연자에서항균제내성율이약간높게나타났다. Uh 등이환자혈액에서분리된 Viridans Streptococci의항균제감수성에대해조사한결과를본연구진의구강내분리된 Viridans Streptococci의항균제감수성과비교하였으며그결과, Penicillin은내성 ( 중간내성포함 ) 이 35% 와 35.6%, 감수성이 65%, 64.4% 이었으며, Clindamycin은내성 ( 중간내성포함 ) 이 35%, 24.4% 이고감수성이 65%, 75.6% 이며, Cephem 계열은내성이 35%, 11.1% 이며감수성이 65%, 88.9% 이었다 (16). Penicillin은구강과혈액에서의항균제감수성이비슷하였으나 Clindamycin과 Cephem 계열은내성율에 10% 이상차이가나타났다. 구강에서분리한 Viridans Streptococci는환자혈액으로부터분리한것에비해항균제내성율은다소낮게나타났다. 구강내에서분리된 Viridans Streptococci은항균제를투여받은환자혈액에서분리된 Viridans Streptococci에비해항균제를접할기회가적었기때문에항균제내성획득율이낮은것으로생각된다. 흡연자에서분리한 S. aureus 3균주중에서 2균주에서 MRSA 로확인되었으며, 3균주모두 Beta lactam 계열항균제에내성을나타내는 Beta-lactamase라는효소를생성하는균주로확인되었다. 흡연자에서호흡기질환중폐렴을유발하는 S. pneumoniae가 1균주검출되었다. 그람음성으로동정된결과, 흡연자와비흡연자에서는 K. pneumoniae, P. pneumotrpica, C.

흡연자와비흡연자간의세균분포및항균제감수성 339 Table 4. Antimicrobial susceptibilities of Gram-negative bacteria by disk diffusion method Smoker Non-Smoker Klebsiella pneumoniae Pasteurella pneumotrpica Citrobacter koseri Pasteurella multocida Flavimonas oryzihabitans Pantoea spp. Klebsiella pneumoniae Klebsiella pneumoniae Antibiotics (μg) Amikacin <=2 S <=2 S <=2 S <=2 S <=2 S <=2 S <=2 S <=2 S Ampicillin >=32 R <=0.25 S >=32 R <=0.25 S <=0.25 S <=0.25 S <=0.25 S <=0.25 S Ampicillin/ Sulbactam <=4 S <=4 S 16 I <=4 S <=4 S <=4 S 8 S <=4 S Aztreonam <=8 S >=32 R <=8 S >=32 R >=32 R <=8 S <=8 S <=8 S Cefepim <=8 S >=32 R <=4 S >=32 R >=32 R <=8 S <=4 S <=8 S Ceftazidime <=4 S <=8 S <=8 S <=8 S <=8 S <=8 S <=8 S <=4 S Ceftriaxone <=2 S <=8 S <=8 S <=8 S <=8 S <=8 S <=8 S <=2 S Cephalothin <=8 S <=8 S 16 I <=8 S <=8 S <=8 S <=2 S <=8 S Ciprofloxacin <=0.5 S <=0.5 S <=0.5 S <=0.5 S <=0.5 S <=0.5 S <=0.5 S <=0.5 S Gentamicin <=0.5 S <=0.5 S <=0.5 S <=0.5 S <=0.5 S <=0.5 S <=0.5 S <=0.5 S Imipenem <=4 S <=4 S <=4 S <=4 S <=4 S <=4 S <=4 S <=4 S Piperacillin <=8 S <=8 S <=8 S <=8 S <=8 S <=8 S <=8 S <=8 S Piperacillin/ Tazobactam <=8 S <=8 S <=8 S <=8 S <=8 S <=8 S <=8 S <=8 S Ticarcillin/CA <=16 S <=16 S <=16 S <=16 S Tobramycin <=0.5 S <=0.5 S <=0.5 S <=0.5 S <=0.5 S <=0.5 S <=0.5 S <=0.5 S Trimeth-fulfa <=10 S <=10 S <=10 S <=10 S <=10 S <=10 S <=10 S <=10 S R, resistant; I, intermediate; S, susceptible

340 Hyun-Ja Jeong and Su Jung Kim koseri, P. multocida, F. oryzihabitans, Pantoea spp. 이동정되었으며, 검출된균종에서는큰차이를발견할수없었고, 비흡연자에서의 K. pneumoniae는상재균총으로분포할수있으나, 정상인에비해호흡기질환발병율이높은흡연자에서의폐렴유발세균은질병으로전환가능성을잠재하고있다. 이는구강내세균의항균제내성획득경로는알수없으나흡연으로인해다양한호흡기감염이유발될경우항균제선택에있어서상당한어려움을초래하게될것이고흡연자에서분리된황색포도구균인 S. aureus는항균제내성획득으로 MRSA 및 Beta-lactamase라는효소를생성하는균주로확인되었다. 흡연자와비흡연자간의세균분포에는큰변화가없지만이런세균중병원성을가진세균과항균제내성을획득한세균들은흡연과더불어구강내환경을악화시킬수있고더나아가인체내상재하는세균들도악화된환경으로인해항균제내성과병원성을획득하여심각한질환으로전이될수있다. 지속적인흡연은구강을직접적으로자극하고비흡연자에비해구강질환을유발할수있는세균이우세하게나타날수있기에흡연이구강질환을악화시키는원인임을시사한다. 적요흡연은음주와더불어치과질환을유발하는원인인자로알려져있으나, 흡연자와비흡연자간의구강내분포하는미생물을조사한연구는현재매우미비한상태이다. 본연구에서는흡연자 30명, 비흡연자 30명으로나누어구강내분포하는세균을분리하고혈액한천배지에서배양하였다. 분리된병원성세균은 Kirby-Bauer 항생제감수성검사방법으로항생제에대한감수성및내성을검사하였다. 각각의콜로니는그람염색을하였고자동동정기인 VITEK을이용하여동정하였다. 그람염색법을이용한결과흡연자의구강에서는총 41개균주가분리되었고, 그중그람양성구균 63%, 그람음성구균 29%, 그람양성간균 3%, 그람음성간균 5% 가나타났고, 비흡연자의구강에서는총 38개균주가분리되었고그중그람양성구균 55%, 그람음성구균 26%, 그람양성간균 3%, 그람음성간균 16% 가나타났다. VITEK system을이용하여세균의특이적분포를조사한결과흡연자의구강에서는 Streptococcus mutans (6/41), Gemella morillorum (6/41), Streptococcus oralis (2/41), Streptococcus pneumoniae (1/41), Staphylococcus aureus (3/41), Streptococcus anginosus (1/41), Streptococcus intermedius (1/41), Streptococcus uberis (1/41), and Streptococcus sanguinis (1/41) 반면비흡연자의구강에서는 Streptococcus sanguinis (8/38), Staphylococcus aureus (1/38), Staphylococcus auricularis (1/38), Streptococcus uberis (1/38), Streptococcus intermedius (1/38), Streptococcus mutans (1/38), and Streptococcus oralis (1/38) 로나타났다. 3 명의흡연자에서분리된 S. aureus는 Beta-lactamase를분비하는 MRSA로동정된반면비흡연자한명에서분리된 S. aureus는 Beta-lactamase를분비하지않고 methicilin에감수성을보였다. 흡연자와비흡연자의상재균은다른분포를보였고특히 G. morillorum와 MRSA가흡연자의구강에서흔히발견됨을알수있었다. 이러한결과들은흡연자의구강질환을예방및치료에유용할것으로사료된다. 참고문헌 1. Ahn, K.D., G.T. Kim, Y.S. Choi, and E.H. Whang. 2007. Effect of irradiation on the Streptococcus mutans. Korean J. Oral Maxillofac. Radiol. 37, 35-43. 2. Brown, L.R., S. Dreizen, S. Handler, amd D.A. Johnston. 1975. Effect of radiation-induced xerostoma on human oral microflora. J. Dental Res. 54, 740-750. 3. Choi, G.W., W.Y. Jang, J.W. Lee, and S.J. Kim. 2010. Microorganism contamination from wearing one-day disposable contact lenses according to wearing time. Korean J. Microbiol. 46, 152-156. 4. Cho, E.H. and N.Y. Lee. 2003. Clinical significance of viridans Streptococcal bacteremia. Korean J. Lab. Med. 23, 246-250. 5. Ge, Y., P.W. Caufield, G.S. Fisch, and Y. Li. 2008. Streptococcus mutans and Streptococcus sanguinis colonization correlated with caries experience in children. Caries Res. 42, 444-448. 6. Han, G.S. 2007. Periodontal status in smokers and the effects of professional periodontal care. Hanyang university Doctor of health education degree. 7. Jeon, E.H., J.H. Han, and T.Y. Ahn. 2007. Comparison of bacterial composition between human saliva and dental unit water system. J. Microbiol. 45, 1-5. 8. Kim, O.J. 2010. Oral Candidiasis. J. Korean Dental Association. 48, 355-364. 9. Kim, S.J. 2009. Identification and distribution of the pathogenic microorganisms isolated from edible ice in North Area of Daegu, Korea. Korean J. Microbiol. 45, 86-90. 10. Kown, J.L and J.S. Park. 2002. Comparison of automated systems for identification of Vibrio species. Korean J. Microbiol. 38, 62-66. 11. Lee, S.A., S.Y. Yoo, K.S. Kay, and J.K. Kook. 2004. Detection of hepatitis B virus and Mycobacterium tuberculosis in Korean dental patients. J. Microbiol. 42, 239-242. 12. Lee, S.S. and S.K. Lee. 2010. Herpes infection. J. Korean Dental Association 48, 365-370. 13. Nam, J.H. 2010. Considerations in dental management and medication for the medically compromised patients. J. Korean Dental Association 48, 27-37. 14. Page, R.C. and J.D. Beck. 1997. Risk assessment for periodontal diseases. Int. Dent. J. 47, 61-87. 15. Sliepen, I., J. Van Damme, M. Van Essche, G. Loozen, M. Quirynen, and W. Teughels. 2009. Microbial interactions influence inflammatory host cell responses. J. Dent. Res. 88, 1026-1030. 16. Uh, Y., G.Y. Hwang, I.H. Jang, K.J. Yoon, and H.Y. Kim. 2002. Antimicrobial susceptibilities of viridans Streptococci isolated from blood cultures during recent period. J. Lab. Med. Quality Assuarance 24, 225-230. 17. Yoo, S.Y., P.S. Kim, H.K. Hwang, S.H. Lim, K.W. Kim, S.J. Choe, B.M. Min, and J.K. Kook. 2005. Identification of nonmutans Streptococci organisms in dental plaques recovering on mitis-salivarius bacitracin agar medium. J. Microbiol. 43, 204-208. 18. van Winkelhoff, A.J., C.J. Bosch-Tijhof, E.G. Winkel, and W.A. van der Reijden. 2001. Smoking affects the subgingival microflora in periodontitis. J. Periodontol. 72, 666-671.