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Based on this thesis work two papers were published in major scientific microbiology journals and one paper has been presented in National conference held in November 2000 and later has been also submitted for publication in Diagnostic Microbiology and Iifectious Disease. The title of these papers and other details are given below.

1. Prashanth K, Madhava Ranga MP, Vasudevanand Rao, Reba Kanungo.

Corneal perforation due to Acinetobncter junii: a case report. Dingtt Microbial

Infect Dis 2000; 37: 215-217.

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Acit~ctobnclcr spp. and their antimicrobial susceptibility status. ] Med

Microbtol. 2000; 49: 773-8.

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Indian Association of Medical Microbiologists National confcrcnce, 2000.

Reprints of the above works are enclosed.

D I A G ~ C ~ ~ I O L J X Y AND m m

Dlapnuatr M~crohielugy and Inleci~~,ur Dnesse DLSWSE

17 f2(XWI) 215-217 www elwv~cr .u~ml l~roreld~u~~n~~mh~~~

Corneal perforation due to Acinetobacter junii: a case report

K. Prashanth"." M. P. M a d h a v a Rangab, V a s u d e v A n a n d Raoh, R e b a K a n u n g o "

'"l)e~~~!r!m~~nl rr/ M~~.nrbr,irai J,~whurlul lorlrlul~~ J Piuri.,uduulr M~,d!l.ul Eduriir!nn and Hrrrdn 11 (JIPMERI, Puodrhrm. hO.5 fillfi, hdlo 'Drlsr.!r~war 01 Oj~hihiisoli~pi. ~ a ~ ~ u l t o r l ~ ~ l 1,iisiurr ro/ Riilg,itd~ruir M<,dr<al Eduioiraa and Xn?lm.h iJIPMER1. Paulrr b r n MIS INXI. Qdu

Recencd 6 Dececnkr 19'4% rccejlled b Mrrch ?!YN

A t in~,rohocrrr \pp. 15 cmcrglng a\ a conltnmm cau*e ol n,rvromral infccuon+. Cnmmuntty acquired ocular infections due h Acmr'roh ~r r r x c . Only onc case ofpcrtnrrl~on of corner har hecn ~rcported previou4y. wherc old notnmclaturc war u\ed lo descnhr the causal agent. 111 rcpun 11 caw i r l elmedl pcrl~lratlon duc lu Ac~~rrt~lruocrjunli lor which a lherapcut~c penetrallng Ltdoplaat) wils conducted and the froma cvctitually reco\crcd O ?[I00 El\evler Science Inc. All nghis re\ervcd.

I. Intruductinn

Artnrrobnclrr as a cause of ocular infection thal I\ hctng 'crrasingly recogni~ed I t has been ;tssociaIed with an ~ l e a ~ o n after trauma (Melki el al.. 1992). Expoaurc of ncileu, use of convact Icns. penetrating leratoplaity, and nmunusuppre\sion hakc been implicated as predispr~r~ng litor\ 10 ~nfectinn by Acinrroi?uctrr (Zabcl et al.. 1989; hca n al.. 1990). Rarc rcpons of Leratit~s have also heen

' \ladied (Kcnl et a]., 1W.k Marcovich el al. 1994). Th~s ~o$anism is a Gram-negative shon hacilli. rometimes teen .i~coccuhacillnry k~rms and is hund ubiquitously in nature. ' laentyu~~c DNA-DNA homology groups wilhin this genus II~I\C now hecn r e c o ~ n i ~ e d based on the DNA-DNA hybrid- /,Illon mcthods (Schreckenbcrger et al.. 1999). Seven of hne groups have k e n given for~nal species names (Bouvet hCrimont, 1986). To the best of our knowledge, only one :r!e of pertbration of cornea by this organism has been r:pinted (Wand er al., 1975). We report a case of perforation ''1 curnca secondary to Ar innol~ur~lrr keratitis.

!, Ci~se report

A 35-year-old woman presented with a 20-day hiatory of 'Nunla to the left eye with a thorn. Afler the trauma she had

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'I omehpondlng uuthol. Tcl.. 10198430-97955 (Mobile)

i - lull addrust. Prwh12LPyahw.i.om (K. Prdshnnthl.

complatnts of pain, watering, redness, photophobia, and dtmtnished v i w n in the affected eye. She had received natlvc medictne in the fo rn~ o l plant juice heing inrtllled in the affccted eye. The patient was otherwise healthy. On examination there war lid edema, hlepharospasm, with cir- cum corneal congestion The cornea showed a h to 7 mm ulcer with irregular margins and slough with deep stromal infiltrate\ and dercemetococle. There was a 5-mm hy- popynn. Her visual acuity was limited to perception of hand mo\emmts, close to the lace. with projection of light rays heisg \enred accurately. Ulcer scrapings were taken at pre- sentation Ibr microbiologtc examination. No organisms were detected either by the Gram strain or KOH examina- tions. Heavy growth of nonlactore fermenting coloniea on MacConkey agar was obtained after over night incubation at 37". Gram staining, activity in oxidationlfermentation (OF) medium, absence of motility. a negative oxidase, and positive catalase reilctions, a i well a i hydrolysis of casein and Tween 20 (Barrow et al., 1993) identified the organism as Aonurobucler species. Further characterization of the bacterium led to its idrntification as A, junii; based on the extended phenotypic identification scheme (Gerner-Smidt el al.. 1991). Briefly, test for gruwth at 37'C, 41°C and 44" were performed in brain heart infusion broth with a water bath. Sheep blood agar (56) plates were uved to detect hemolysia. Oxidation of glucose was tested in Hugh and Leifson's medium containing 1 % glucose. Gelatin stab method was performed to test for gelatin liquefaction. As- similation testa were performed in a liquid medium contain-

'll!.l B93/W$ - = lion1 mauu O 2lXi!l Elsevler Sclencc Inc. All rlghti mewed S 1732.8891!00)00142-5

I. Msd. Minob~ol. - Vol. 49 (2000). 773-778 2MO The Ruhologtwl Saeiery of Great BnUm and Inland

tSSN W22.2615

DIAGNOSTIC MICROBIOLOGY

Simplified phenotypic tests for identification of Acinetobacter spp. and their antimicrobial susceptibility status

K. PRASHANTH and S. BADRINATH

Department of Mrcrobrology. Jawaharlal insfrfufe of Posfgraduate Medrcal Educarron and Research lJlPMER) Pondrcherrv, India

Acinefobacler spp. have been found to be responsible for an increasing number of nosocomial infections. During a 16-month period, 22 patients hospitalised mainly in the respiratory intensive care unit (RICU), paediatric and other medical w r d s were investigated either for infection or colonisation by Acinelobacler spp. O f the 45 isolates of Acinetobacter detected among the total of 425 non-fermenters encountered, 24 representative Isolates were selected for extended phenotypic identifleation. Four environmental isolates were also included in the study. These 28 isolates were typed by biotyping and antibiotyping, which helped in delineating the Acinelobacfer spp. into 12 phenotypes and hvo distinct antibiotypes respectively. A sudden increase of cases of acinetobacter infection suggested that three outbreaks during the study period were due to phenotypes I and 2 of A. calcoaceticus-A. baumannii complex (Acb). Strains of Acb- complex showed multiple drug resistance and were sensitive only to netilmicin. A comparatively high proportion of resistance to amihcin (48%) was also detected among these strains by the agar dilution method. The RICU environment was recognised as an important reservoir for the resistant outbreak strain (Acb-1) which was probably leading to persistent colonisation and recurrent infections.

Introduction

Acinerobacrer spp. have been implicated in recent years as important nosocomial pathogens, especially In lntenslvc care settings [I-31. Despite their low patho- genic potential they are being reported increasingly as the causal organism of numerous hospital outbreaks in several countries [4-61. in a recent internat~onal multicentre study, Acinetobocter spp. were ranked amongst the 10 organisms most commonly causing septicaemia in 18 of 44 large European hospitals 171. ilowever, commun~ty-acquired pneumonia due to Acinrmbacrer species is rare, as evidenccd by the paucity of reports [8,9).

because of their confused taxonomic status 191. In Indla, very few studies of Acinetobacter spp. have been reported and, in vlew of their increasing importance in nosocomial infections, further study is warranted in this part of the world 110. 111.

In the present study an attempt was made to type the Acinerobacrer isolates obtained from mrious sources by a simplified phenotypic identification scheme with a numerical approach [12-141 and also to determine their ant~m~crobial susceptibility.

Material and methods

Despite the increasing significance and frequency of B°Cteria' 'Irains

multiresistant acinetobacter infections, many clinicians Fony-five isolates of Acrnetohocler spp. were included and microbiologists still lack an appreciation of the in the study. These had been isolated mainly from Importance of these organisms in hospitals, in part blood tracheal aspirate, pus and other body fluids over

a period of 16 months (Oct. 1996-Feb. 1998), mainly from patients admitted to the w t o r y intensive care unit (RICU; six beds) (34 patients), paediatric ward (5)

Kcccivcd 4 Fcb. 1999; reviscd wmion accepted 20 DCC, and other medical wards (6) in the 700-bed JIPMER

1999. hospital. The major reasons for admission to the RlCU Comerpanding author Dr S. Badrinath. are mechanical ventilation for respiratory failure.

774 K. FRASHANTH AND S. BADRINATH

postoperative critical can and organ support following multiple trauma. Such admissions are usually for 2-4 days. Four Acinetobacter isolates were fmm 10 environmental samples (hands of anend~ng clinic~ans, ventilator monitors, humidifier and endotracheal tube surfaces) obtained during the per~od o f high incidence of ~solat~on o f this bacterium from clinical specimens from patients in the RICU. Environmental samples were collected according to the method of Seifen era/ . [IS]. The reference strains were lncludcd in the study. All isolates were preserved in glycerol broth at -70°C until tested.

Phenotypic idenrifrcation

All isolates were subjected to simplified phenotypic tests as described by Gerner-Smidt cr a/ [12, 141 (Table I). Briefly, tests for growth at 37'C, 30°C and 44°C were performed in brain hean infusion broth in a water bath. Sheep blood (5%) agar plates were used to detect haemolys~s. Glucose oxidation was determined in Hugh and Leifson's medium containing glucose I% and the gelatin stab method was used to tcst for gelatin liquefact~on 1161. The assim~lation tests were per- formed tn a I~quid medium contain~ng a mineral base as described by Stanier er a / . [I71 w~ th the addition of an appropriate carbon source in a 0.1% w/v concen- tration. t-Aconitate, L-histidine, malonate, histamine and citrate were used for assimilation tests. Additional carbon source assimilation tests such as DL-4 amino- butyrate and L-phenylalanine were used for haemolytic isolates (DNA groups 4,6 and 14). Susceptibility to penicill~n G (10 pg) and chloramphen~col (30 pg) was tested by the d~sk diffus~on method as they were used as phenotyp~c tests along with the rest [I)]. The susceptib~lity tests were read after ovem~ght incubation, and the tests for haemolysis and growth at 37" and 44°C were read on the first and second day; the assim~lat~on tests were read afler incubation for 1, 2. and 6 days. All were incubated at 30°C. except for the tests for growth at 37°C and 44'C, and for gelatin liqu~fication the ~ncubation tempemre was 2Z°C.

Antirtlioubial susceptibility

Antimicrobial suscept~bility was determined by both the Kirby-Bauer disk diffusion method [I81 and an agar dilution method for minimum inhibitory concentration (MIC) determination [19]. Escher~chra coil ATCC 25922 was included as 'control. The tests were read after overnight incubation and the following antibiotics were used: cefotaximc, cebzidime, amikacin, genta- micin, ampicillin, piperacillin, netilmicin, cipmflox- acin, norfloxacin, cefazolin and cephalexin. Only the first three antibiotics were used for MIC detection. Surceptibility results were analysed with the WHONET4 programme.

'NOTYPIC IDENTIFICATION OF ACINETOMA(.TER SPP 715

I n all, 45 isolates were obtained dumg the study period of 16 months. Of these, 34 isolates were from multiple sites in I I patants in the RlCU (Table 2). In eight cases, apart from tracheal aspirate. Acrnefobacfer spp. were repeatedly isolated from blood In cases where multiple isolates were obtained from the same patient, after performing preliminary biochemical testing and antibiotyping a representative strain(s1 was subjected to extended phenotypic characterisatton. In two cases, two dtfferent strains were isolated from each patient. Hence the study population consisted o f 28 strains, which included both cltnical (ICU, 13 strains; medical ward 6; paediatric ward 5) and environmental isolates (4) (Table 3).

O f the 28 ~solates. 22 were blovar unir!alus and 6 were btovar /u,oJhi. With the simpltfied phenotypic ident~fi- cation scheme, the 28 stralns tested belonged to only 12 phenotypes (Table 4). The first five phenotypes (20 strains) were identified as membw of the A. colcoacctrcus-A baumannii complex (Acb-complex: phenotype 1-5. phenon 1-2-3-13). One strain was attributed to the Ach.complex (phenotype 6) despite fail in^ to acid~fy glucose. One stratn was identified as A, hoenroiyr<ctrs (phenotype 7) and another as A, lwoffir (phenotype 8). The remaining four phenotypes (9. 10. I I and 12) did not fit the phenotypic descrtptton of any

: single phenon. Phenotype 9 was like the typical A. haento&crrs phenotype, except for asstmilation of

i malonate (A. hoemo1i~iicu.s-like) Phenotype 10 showed j the typical A lwoffii phenotype (A, /n,afir-ltke) except j for assimilatton o f L-histidtne. Phenotype I I, whtch !cotnpr\sed two strains (phenon I I-like), resembled +-I l In every respect except in falltng to grow at 37°C. Phenotype 12 resembled the members o f phenon 8-9-15, as i t grew at 37°C and was susceptible to chloramphetiicol.

Tsblr 3. Environmental isolates of Arrnerobarlrr spp.

Samplc Dste of Organam no irullation Stte ~rullatcd

6 . . Forearm None 7 . . . Monctor8ng surface, Nonc X 26-02.98 Forefinger Ach-l 9 Axilla None

10 . Bedsides None

Anlimiciubiol susceptibili[t~ resling

The MIC results for 28 Acinelobocter isolates arc shown in Table 5. I t also shows the range of the MIC observed as well as the MICSO, MIC90 and percentage of resistant strains among Ach (21) and non-Ach (7) isolates. A slightly higher percentage of resistance to amikacin (48%) was noted among strains of the Ach- complex when compared with that of the other two antibiotlcs tested (43%). Many stralns (14) showed high-level resistance to am~kacin (3256 mg/L). The M1C90 of cefotaxime was Z256mg/L for 10 strains, whlle that of ceftazidime was 128 mg/L for elght strains. The disk diffusion method showed cefazoltn, piperacillin, gentamicin and norRoxacin as having very ltnle activity, with the percentage of resistance ranging from 68 to 96%. Almost all stralns were resistant to cefazol~n and cephalextn, Generally, strains of the Ach- complex were more resistant to all the antibiottcs tested than strains belonging other phenons.

The agar dilutton technique showed that the overall peTcentage of stralns susceptible to cefotaxime and ceftazidime was 50% and 58% respectively. The overall susceptibility for amlkacin was similar to that of

Table 2. Respiratory ~ntcnslve care pat~enis posltive for Arinrrr>hur!er spp.

Pailen! l>nr or fiin Phenolypc no. Agrlrex D~ilgllarls Outcomu Sltri poslt~vc nrolst~on ~nolrted

I 40/f POEI-surgery (orrophagarromy) Dlrchagrd Blood'. PF 10-10.96 Alh-2 xpltcaemks spuam. TA, pus

2 JOiM lntncranlal spilcr-occupy~ng lcalon Dlcd Blood' 18.10-96 Ah2 3 4S/F Smnplaed hamla. scptlcaemla Dced Blood". TA 26.10-96 .lc6-6 4 2011: Asptmtlon pncumonln Dled Blood TA 26-10-96 Arb-2 S 47lM Prrilonltls Dwhaged TA. h l d 28-02-97 Acb.2 6 20!F Cardarc *rest Dischutged Blmd TA IR-09.97 Arb4 Acb-2 7 I I/F Dcmyelmstag dlrcase. pneumonta Tmnrfcrrud Blood'. TA. 11-09-97 Ach-2

qputum 8 74/M Mulliple organ dysfunct~on Dled TA, bile. blud' 29-09-91 Arb1 9 63iM /\rp~rs~~on pneumonta Dixharg~d Blwd", TA ; ~ch-S, ~ch-2 10 Z O ~ M Sepilcaemlu Darct!arpd TA, blmd' , A&-I I I 451F Trauma. ARDS Trsnsfcnrd TA. blmd' 2R-02.98 Ach.2

TA, lrachcal arplnle. PF, pleural flutd. ARDS. adull rcrpimlory d~slress ssndromr 'Rma irolst~an fmm b l d . 'Colontmion onk aa there wrrc no related cl~ntcal findlngi

776 K . PRASHANTH AND S. BADRINATH

ceftazldime (58%) and only 3% were of intermediate susceptibility. Strains moderately sensitive (intemedi- ate range) to ctfotaxime (23%) were more common than for other antibiotics tested. The d~sk diffusion method showed netilmicin and amikacin to have the greatest activity against the strains, with overall sensitivity of 64% and 48% respect~vely The A Iwofli. A. Iwojii-like. A. haemolyficus and A. haemo- lyticus-like strains were generally mom susceptible to quinolones, pipemillin, gentamicin and amikacin than the Acb-complex strains.

Discussion

Acinetobacters have been considered members of a single species, A. calcooceticus, with two b i ~ . However, new molecular methods of identification (DNA-DNA hybridisation) have divided the genus Acinelobacrer into 19 DNA-DNA homologous groups [9,20-221. The confusion that the new taxonomy has caused in the clinical laboratory IS compounded by the fact that identification of the various species and DNA groups by phenotypic characteristics, many of which are assimilation of numerous carbon sources, is vety difficult The tests for identification of Acinetobac~er spp. selected by Bouvet and colleagues [20,22] seem to be appropriate, but the large panel of phenotypic tests is tedious and time-consuming to perform. Simple identification schemes such as those of Gerner-Sm~dt el al. [I21 and Tjernberg 1131, along wlth antimicrobial susceptibility testing, may be useful for laboratories with limited resources and can be adapted for typing ~solates, thus avoiding the need for expensive mole- cular methodology.

The ~ncreasing clinical Importance of Acinelobacler spp, is attributed mainly to their capacity to cause nosocomial mfections, particularly outbreaks in ICUs. The most lniponant nosocomial acmetobacters belong to the Ach-complex [9,23]. The present study also identified more strains belonging to the Acb-complex than other phenotypes. Repeated sola at ion of an organism from the blood with related clinical findings is a strong ind~cation of infection 1241. This study reports six patients with infection and two cases of colonisatlon in the ICU. The remaining three cases had a slngle positive blood culture along with positive trachial aspirate. Sporadic infections with Acinetohac- ter spp were punctuated with outbreaks lasting for a month at d~fferent times. Outbreaks encountered during the months of Oct. 1996, Sept. 1997 and Feb 1998 were caused by multidrug-resistant strains of the Acb- complex (phenotypes I and 2). The source of infection was the ICU environment, where the epidemlc Acinerohacrer straip (Arb-I) isolated over a prolonged period. he Arb-l phenotype strain from the environment (humidifier) was phenotypically indis- tingulshable from strains ~solated from patients 8 and 10, and was distinct from all other phenotypes. The

PHENOTYPIC IDENTIFICATION OF ACINEMBACTER SPP 177

Tmble 5. MIC values of three antibiotics sgatnst 28 Acrnerobocrrr isolates

MIC (mg/Ll

Number o f Geomc~r~r Reslrunce Antibtotrc Phenotype lsolalcs Rnngc MICSO MICOO mean (%I

CrRmdlmc Aebsampln (1-6) 21 2-118 16 12 14.98 41 Other phcnotypcs' 7 1-4 2 4 2 0

Amihcln Acb.complrr (1-6) 21 1-2256 32 216 23 48 Other phsnotps 7 0.5- 128 I 128 1.28 28.6

Ccfour!mc Acbzampln (1-6) 21 8-128 32 M 29.% 43 Mhcr phcnotypcs 7 0.5-8 2 8 2 21 0

'Includes phenotype nor 7. 8. 9. 10. 11 and 12,

inability to trace the source o f the Acb phenotype 2 strain, which was responsible for most of the infection occurring during Oct. 1996 was probably due to inadequate envtronmental sampling. The outbreak strain Acb-I was distinguished from other Acb phenotypes by the inability o f the latter to u t~ l~se malonate. Although both outbreak phenotypes I and 2 of Acb-complex showed resistance to almost all the antibiotics tested by both methods, strains o f Acb-l phenotype were distinct because of their susceptibility to amikacin and netilmicin, unlike strains o f Acb phenotype.2. The simplified identification scheme for the asstmilation tests showed similarity between DNA groups 1, 2, 3 .and 13 aqd failed to speciate exactly w~thin the Acb-complex. This is understandable because they are closely' related genotypically 1121. Despite this inadequacy, phenotypic identification was possible for two species within the genus (201. One strain was A. haernolyficus, isolated fmm an 86-year- old male patient suffering from recurrent bronchiecta- sis, the other was A. Iwolfii, responsible for septicaemla in a 5-year-old female patient. Both A. haernoly/icus and A. IwofJii are known to cause systemic infections [25,26]. Four isolates were closely related to pheno- type A. lwolfii and one resembled the typical A. haemo(vricus phenotype. O f four A. Iwolfii-like organ- tsms, one was isolated from a humidifier and indtcated colonisation in ventilators in the ICU. The other three were responsible for systemic infections in elderly debilitated patients.

Tradttional typing methods like phenotyping and antibiogram typing have an advantage over genotyping as they are readily available in all clinical microbiology laboratories. The biotyping procedure o f Bouvet and Grimont 1s definitive [20]. However, genotyping meth- ods such as ribotyplng, pulse-field gel electrophoresis and PCR [h, 15, 27,281 have a definitive advantage over phenotyping, as they clearly distinguish isolates within the Achcomplex. In the present study, all reference strains of DNA groups I, 2, 3 and 13 showed almost identical phenotypic results. Laboratories with limited resources can still rely on a combination ofdata from two of the above-mentioned traditional ap- proaches to typing. New molecular techniques have shown the genetic diversity within the Acb-complex to be greater than hitherto perceived [29]. Consideration

should be given to placing the whole Acb-complex in a single species, .4 calcoacericus, which has priority over A. baurnannii, and biotyping them further only for ep~demiological purposes. As a large panel of assim- ilatlon tests is cumbersome, the simplified scheme followed in the present study is useful, as the discriminatory power IS almost the same for both the methods (121.

The strains of Ach-complex could be clearly distin- guished from non-Ach phenotypes by their resistance to antimicrobial agents [9]. However, within the Acb- complex phenotypes I and 2 were more resistant than other Acb phenotypes, as the other phenotypes were susceptible to three or more antibiotics tested and there is no major increase in the overall percentage of resistance in Acb group. Variable resistance patterns like this have been observed by other workers [S], showing that not ail phenotypes of Acb are multidrug- reststant

The dlsk dtffuslon technique showed chat members of the Ach-complex were resistant to almost all antibiotics except for newer aminoglycosides and cephalosporins. i.e., amikactn, netilmlcin and ceftazidime. Six strains of the Acb-complex (all Acb-2) were resistant to netilmicin, the sole anttbiotic that was effective against all isolates o f other phenotypes. With the agar dilution method 14 strains had an MIC 2256mg/L for amikacin. Thls high-level resistance is probably due to extensive use of this antibiotic in the ICU. Ceftazidime was one of the second4ine antibiotics used in this hospltal to treat infections caused by multi- drug-resistant gram-negative bacteria. Ceftazidtme was also the moa efficacious drug in the present study, as only 27% of strains were resistant, which can be attnbuted to its jud~c~ous use. In concordance with earlier studies 15.61, the present study also encountered strains o f the Ach-complex showing resistance to all drugs tested, whlch la o f concern as infections caused by these organisms pose a therapeutic problem. I t 1s surprising that the level of resislanc to cefotaxime was moderate even though,it was the c h o n e s t antibiotic used in the hospital. One environmental isolate (Acb-I) was from the hands o f an attending clinician (in the ICU) and was phenotypically indistinguishable fmm the outbreak stratn and moderately resistant, suggesting

778 K. PRASHANTH AND S. BADRINATH

colonisation by the Ach-complex strain and a source of infection for the patients.

Simple identification schemes and antimicrobial sus- ceptibility testing provide a cost-effective approach to typing Acinetobacter spp. Although the above systems have certain lim~tat~ons when compared to molecular methodologies, the distinction between resismt and susceptible acinetobacters at least, is useful for the effective clinical management of the infections caused by this group of organisms.

We Lank h P. Gemcr-Smldt (Statens Serum Instttut. Copenhagen, Denmark) for gcncmuly provldlng Acmeroboer~r reference stram and the faculty o l L e Depanmcnt of M~crobtology (JIPMER. Ind~a) for their sslslance and suggestions in the preparation o f the manuscript

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( lnslltuuon : Jawahanl InrUtulc of Postgraduale Medial Educalion and Rsrearch (JIPMER), Pondicherry I 1

- . Acinelobaclersp. is an irnporlanl cause ol nosocomial inleclms. Clinical isolates 01 this genus are ollen

resistant lo many antibiotics. In this hospital il is increasingly assodaled with inleclions in patients admlted to Respiratory inlens'we care Unl (RICU). A total bl66 isolates which includes 49 clinical and 17 environmental ones were characterized. Susceplibili panem using disc diHusion methad mlh 12 antimicrobial agents were checked. MlCs were determined using agar dilulim method lor only second line broad.spectrum anlibiotics such as celolaxime. celtatidime, amikacin, ciprolloxacin and olloxacin, lsolales belonging lo species other lhan A, baumannii were more susceptible to most ol Ihe anlibiolics tested, as Multi Drug Resistance (MDR) was only seen in A. baumanii amincglycosides such as amikaun, netelmicin were most aclive against most 01 the MDR isolates lested (60F susceptibility). Celtazidime was more active than celolaxime. MDR A . baummanii strains showed susceplibility only to amikacin, nelelmicin and cellazidime. Ciprolloxacin had poor acti\lly irrespeclive ol isolates belonging to diHerent DNA grwps tested (58% resistance over all, 7% among A. baurnman~l), lsolales ol biolypes 6 & 19 of

A baummanii exhibited broader multi resislance than those of biolype 10 and other biolypes.

Title : ' IN WRO SUSCEPllBlLllY PAlTERN OF CLINICALLY SIGNIFICANT AClNETOBACrER SPECIES TO COMMONLY USED CEPHALOSPORINS, OWNOLONES AND AMtNOGLYCOSlDES

liuhora : l BADRINATH, S