CHEMOTHERAPY MAR Table 1. Number of isolates of Providencia sitrailii by sources ( ) sisomicin (SISO), gentamicin (GM), chloramphenicol
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1 Providencia stuartii mphenicol, tetracycline sulfamethoxazole fosfomycin, ampicillin, cefazolin streptomycin, kanamycin, gentamicin, dibekacin, sisomicin tobramycin aminoglycoside cefmenoxime, cefotiam, amikacin, fortimicin, norfloxacin, trimethoprim Key words: Providencia stuartii, R plasmid, Minitek system (Becton Dickinson and fortimicin, amikacin (AMK), HAPA-B, tobramycin (TOB), dibekacin (DKB),
2 CHEMOTHERAPY MAR Table 1. Number of isolates of Providencia sitrailii by sources ( ) sisomicin (SISO), gentamicin (GM), chloramphenicol (CP), tetracycline (TC), minocycline (MINO), streptomycin (SM), kanamycin (KM), ampicillin (ABPC), cefazolin (CEZ), cefotiam (CTM), cefmenoxime (CMX), fosfomycin (FOM), norfloxacin (NFLX), sulfamethoxazole-trimethoprim (ST), sulfamethoxazole (SMZ), trimethoprim (TMP) 4. Plasmid profile Plasmid DNA VI KADO L LIL iris-borate buffer (89 mm tris-aminomethan, 89 plasmid DNA P. stuartii Proteus mirabilis (rif), Escherihia coli WA (rzf nal lac) recipient
3 VOL.37 NO. 3 Table 2. Characteristics of 20 patients with P. sluarlii isolated from otorrhea, sputum and pus * M: male F : female Table 3. Minimum inhibitory concentration of Providencia stuartii to amikacin, tobramycin, gentamicin, kanamycin, chloramphenicol and tetracycline (20 isolates, 106CFU/m1)
4 CHEMOTHERAPY 258 MAR Table 1. Minimum inhibitory concentration of Providencia sloarlii to ampicillin, celazolin, cefotiam, cefmenoxime, fosfornycin, norfloxacin and sulfamethoxazole trimethoprim (20 isolates. 108CFU/ml) The location and sizes (in megadaltons) of the molecular size markers are indicated at the sides of the gels. The solid arrows show the position of the 30-megadalton plasmid discussed in the text. Fig. 1. Plasmid profiles of 20 isolates of Providencia stuartii obtained by 0.7% agarose gel electrophoresis fortimicin,hapa-bに はAMKと ほ ぼ 同 様 のMIC値 で 大 部 分 の 株 が 感 受 性 で あ っ た が,SISO,DKBに て はTOBな ど と 同 様 多 く の株 が 耐 性 で あ っ た ま た CP,TC,MINO(dataは 略)に で あ つ た つ ぎ にTable4で 系 薬 剤 で は,CTM,CMXに ABPC,CEZに 値 を 示 し耐 性 で あ っ た strain1,2に 対 し は ほ とん ど の株 が 耐 性 明 らか な よ うに,β-lactam は 全 株 感 受 性 で あ っ た が, は 半 数 以 上 の 株 が25μg/ml以 上 のMIC の 産 生(Beta-Lactamase Ltd.使 用)が Detection は β-lactamase papers,oxoid 認 め ら れ た そ の 他 の 抗 菌 剤 で は,FOM に は 半 数 以 上 の 株 が 耐 性 で あ っ た が,NFLXに が 感 受 性,ST合 剤 に は2/3以 た 表 に は 示 さ な か っ た が,TMPに 受 性 で あ り,SMZに は 全株 上 の株 が 感 受性 であっ は大 部 合 の株 が感 は ほ と ん ど の 株 が 耐 性 で あ っ た
5 VOL.37 P.stuartiiの NO.3 分 雌 状 況 と薬 剤感 受 性 259 Table 5. Antibiotic susceptibility of the transconjugants resulting from the mating of Providencia st math i with Escherichia cnli or Protens ruirahilis (106 CFU/ml) Abbreviations of antibiotics are defined in the text. lane1, 2, 3, 4, 18をdollorと 5, 6, 7, 10, し,E.coli bilisをrecipientと WA し た 通 類 で,ABPC,SM,SMZ,KM,GM, よ びSISO耐 SMZ, CP, KM, びSISO耐 性 のE.coli2で cipientど し た 場 合 は,上 性 のE.coli GM, AMK, Fig.1に DNAが 記E.coli1お 示 示 し た 様 に20株 は3.2megadaltons以 で 様 々 で あ っ た が,こ 10,11お よ び.12)に 中14株 中 のplasmid 下 か ら166 か らplasmid DNAの 分 子量 megadaltons以 上ま し たABPC単 は 分 子 量 の ほ ぼ 等 し いplasmids (約30megadaltons,Fig,1に 矢 印 で 示 した)が 検出 4.接 合伝 達 plasmidの 検 出 され た14株 plasmid な わ ちFig.1の のtransconjugant した 時 に 得 ら れ た promeをfig.2に artiiのplasmid 示 す donorで profile(lane E.coli(lane た が,接 A)に F)に Md,同 あ るP.stu- は166,62,30, のplasmidbandが 検 出 あ るP.mirabilis(laneE), はplasmid bandは 検 出 さ れ な か っ 合 伝 達 で 得 ら れ たtransconjugantのP.mira- bilisのplasmid profile(lane じ くlane B)に profileで Dに は70Md,30 あ るlaneCに は166Mdのplasmid は140 bandが 検 出 さ れ た Plasmid 全 て,す 2 得 ら れ た 他 に, 剤 耐 性 strain1株(lane1)をdonorと 5.形 され た よ が 得 ら れ た Md,E.coliのplasmid の 内8株(lane:1,3,4,5,7, ABPC, DKBお よ びE.coli と 同 じ 耐 性 パ タ ー ン のtransconlugantが Table5に さ れ た 一 方,recipientで 検 出 さ れ た 14株 1と, TOB, あ る P.mirabilisをre- 1.1megadaltons(Md)の4本 profile 示 す E し た 場 合 に 得 ら れ たtransconju- TOB,DKBお SM, ど ち ら 株(Tables3,4の 耐 性 バ タ ー ン をTable5に coliをrecipientと gantは2種 17, した 時 の み で あ っ た 得 ら れ た trallsconjugantの Plasmid 14, り で あ る transcon- し た 場 合 もlane1の strain1株)をdonorと 3. 13, よ びP.mira- 得 ら れ た の は,E.coli,P.mirabilisの をrecipientと A: donor strain of Providencia stuartii, B: transconjugant of Proteus C, D: transconjugants of Escherichia coli, E: recipient of Proteus F: recipient of Escherichia coli. Fig 2. Plasmid DNA profiles of the transconjugants, obtained by 0.7% agarose gel electrophoresis 12, し て 接 合 伝 達 を 試 み た 選 択 薬 剤 お よび 選 択 濃 度 は 方 法 の 項 に 示 jugantが 11, 921-1お 質 転 換 DNAを 保 有 す る14株 行 な っ た が,transformantは に つ い て 形 質 転 換 を 全 く得 ら れ な か っ た
6 5) KOKKA F, ESRINIVASAN S, MOWJOOD M, KANTOR H S: Nosocomial multiply resistant Providencia stuartii: a long-term outbreak 1) MCHALE P J, WALKER F, SCULLY B, ENGLISH L, KEANE C T: Providencia stuartii infections: a review of 117 cases over an eight year period. J. Hosp. Infect. 2: 155 `165, ) MCHALE P, ENGLISH L, SPEEKENBRINK A, KEANE C: Kanamycin resistance in Providencia stuartii. J. Antimicrob. Chemother. 4: 273 `278, ) OVERTURF G D, WILKINS J, RESSLER R: Emergence of resistance of Providencia stuartii to multiple antibiotics: specification and biochemical characterization of Providencia. J. Infect. Dis. 129: 353 `357, ) FIERER J, EKSTROM M: An outbreak of Providencia stuartii urinary tract infections. JAMA 245: 1553 `1555, 1981 with multiple biotypes and serotypes at one
7 hospital. J. Clin. Microbiol. 11: 167 `469, ) KADO C I, LIU S T: Rapid procedure for detection and isolation of large and small plasmids. J. Bacteriol. 145: 1365 `1375, ) MEYERS J A, SANCHEZ D, ELWELL, L P, FALKOW S: Simple agarose gel electrophoretic method for the identification and characterization of plasmid deoxyribonucleic acid. J. Bacteriol. 127: 1529 `1537, ) YOSHIDA T, TAKAHASHI I, TUBAHARA H, SASAKAWA C; YOSHIKAWA M: Significance of filter mating in integrative incompatibility test for plasmid classification. Microbiol. Immunol. 28: 76 `49, ) HANAHAN D: Studies on transformation of Escherichia coli with plasmid. J. Mol. Biol. 166: 557 `580, ) HAWKEY P M, PENNER J L, POTTEN; M R, STEPHENS M, BARTON L J, SPELLER D C E: Prospective survey of fecal, urinary tract, and environmental colonization by Providencia stuartii in two geriatric wards. J. Clin. Microbiol. 16: 422 `426, ) STICKLER D J, FAWCETT C, CHAWLA J C: Providencia stuartii: a search for its natural habitat. J. Hosp. Infect. 6: 221 `223, ) MCHALE P J, KEANE C T, DOUGAN G: Antibiotic resistance in Providencia stuartii isolated in hospital. J. Clin. Microbiol. 13: 1099 ` 1104, ) Yu P K W, WASHINGTON J A II: Comparative in vitro activity of three aminoglycosidic antibiotics: BB-K 8 (amikacin), kanamycin and gentamicin. Antimicrob. Ag. Chemother. 4: 133, ) PRICE K E, PURSIANO T A, DEFURIA M D, WRIGHT G E: Activity of BB-K 8 (amikacin) against clinical isolates resistant to one or more aminoglycoside antibiotics. Antimicrob. Ag. chemother. 5: 143, ) HOLLICK G E, NOLTE F S, CALNAN B J, PENNER J L, BARTON L J, SPELLACY A: Characterization of endemic Providencia stuartii isolates from patients with urinary devices. Eur. J Clin. Microbiol. 3: 521 `525, ) HAWKEY P M, BENNETT P M, HAWKEY C A: Evolution of R plasmid from a cryptic plasmid by transposition of two copies of Tn 1 in Providencia stuartii. J. Gen. Microbiol. 131: 927 `933, 1985
8 CLINICAL SOURCES OF ISOLATION, ANTIBIOTIC SUSCEPTIBILITY AND RESISTANCE PLASMID OF PROVIDENCIA STUARTH KENICHIRO YAMAZAKI Clinical Laboratory, Omiya Red Cross Hospital, Kamiochiai 903, Yono, Saitama 338, Japan RIEKO KANNO, TAKEO KARIBA and SADAO KIMURA Department of Bacteriology, Teikyo University School of Medicine The frequency of isolation of Providencia stuartii from various clinical sources was examined from Sixty-nine strains were isolated from otorrhea and 4 from sputum or pus. This indicates that P. stuartii is clinically important as a causative microorganism of chronic otitis media. Twenty strains, isolated from , were tested for their susceptibility to various chemotherapeutics. The majority of the strains were resistant to chloramphenicol, tetracyclineand sulfamethoxazole. Many were resistant to fosfomycin, ampicillin, cefazolin and several aminoglycoside antibiotics, namely, streptomycin, kanamycin, gentamicin, dibekacin, sisomicin and tobramycin. In contrast, the majority of the strains were susceptible to cefmenoxime, cefotiam, amikacin, fortimicin, norfloxacin, trimethoprim and sulfamethoxazole-trimethoprim. Profiles of plasmid DNA of the 20 isolates were analyzed, and plasmid DNA was found in 14. Conjugation and transformation were carried out for the detection of R plasmid. One of the 14 donors transferred resistance markers by conjugation.
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