Antibiotic usage S633

hospital. It included for each type of procedure, the antibiotic agent andthe dosage.Results: Before the intervention 153 procedures, from differentspecialities were observed (in 2 series). Eight different antibiotics indifferent dosages were used. In 22% of the procedures, PAP was givenafter the incision. Two months after the intervention 147 procedures wereobserved (in 3 series). Three different antibiotics, 135 times cefazoline(92%), 11 times metronidazol (7.5%) and one time cefuroxime, weregiven. All antibiotics were used in the correct dosage. In 12% of theprocedures, PAP was given after the incision. This was a significantimprovement (p = 0.026).Conclusion: This project shows that a uniform, simple and clear protocolimproves not only the choice and dosage of antibiotics but also thetiming. Furthermore, the switch to cefazoline resulted in net savings ofat least €40.000 a year.

R2186 Trends in antibiotic consumption in 5 Hellenic hospitalsover a three-year period: results of the Hellenic Networkfor Nosocomial Antibiotic Consumption for the years2003–2004–2005

M. Lelekis, C. Loupa, K. Katsimpris, I. Tzannou, G. Xylomenos,C. Anitsakis, A. Provis, C. Efthymiou, P. Gargalianos (Athens, Rhodes,Lesvos, Zakynthos, GR)

Objectives: Hellenic Network for Nosocomial Antibiotic Consumption(HENNAC) is a network of 5 hospitals located in different parts ofGreece, established for monitoring antibiotic consumption in Hellenichospitals: “G. Gennimatas” General Hospital (GGH) of Athens (tertiarycare, 700 beds), “A. Fleming” General Hospital (AFH) of Athens(300 beds), “A. Papandreou” General Hospital of Rhodes Island (RH,Southeastern Greece, 335 beds), “Vostaneio” General Hospital of LesvosIsland (LH, Northeastern Greece, 225 beds) and Zakynthos IslandGeneral Hospital (ZH, Western Greece, 125 beds).Methods: The antibiotic consumption for the years 2003, 2004 and 2005was calculated in DDDs per 1000 patient days (ABC calc 3.0).Results: Compared to 2003, in 2005: (1) Total consumption increasedin AFH (from 710 to 892), LH (from 1148 to 1299) and GGH (from 906to 1117), decreased in ZH (from 1268 to 1060) and remained practicallystable in RH (from 1371 to 1346). (2) Concerning consumption of majorclasses of antibiotics: A. Penicillins: increase in AFH and LH, stable inRH and decrease in ZH, GGH. B. Cephalosporins + aztreonam: increasein AFH, RH, decrease in GGH and stable in ZH, LH. C. Carbapenems:increase in all but LH where it was stable. D. Penicillins+inhibitors:increase in RH, LH, GGH, decrease in AFH, ZH. E. Aminoglycosides:increase in LH, GGH, stable in AFH, decrease in ZH and RH. F.Macrolides: increase in all but ZH. G. Glycopeptides: increase in allbut GGH. H. Quinolones: increase in AFH, GGH, decrease in ZH, RH,LH. (3) Cephalosporins + aztreonam were the most popular antibioticsfor the year 2005 (being first in 3 hospitals and second in other 2)followed by penicillins + inhibitors, while for the year 2003 exactly theopposite was the case.Conclusions: A. While there is no uniform trend for antibioticconsumption in the hospitals of the network, total consumption is highin all of them, compared to hospitals of other European countries. B.Cephalosporins + aztreonam are now the most popular antibiotic class. C.Further studies are under way in order to explain local peculiarities inantibiotic prescribing in the hospitals of the network.

R2187 Antibiotic usage and resistance development in an intensivecare unit

M.S. Lapatschek, A. Hartinger (Munich, DE)

Objectives: SARI (Surveillance of antibiotic usage and bacterialresistance in intensive care units) is a German nationwide projectfor monitoring the usage of antibiotics in intensive care units (ICUs)and to compare it with the development of bacterial resistance tothese antibiotics. The long-term goal is to avoid unnecessary usage of

antibiotics and reduce the burden of highly resistant pathogens in theseICUs.Methods: The antibiotic usage of the 44 participating ICUs is registeredby the providing pharmacies and is send monthly to the study referencecentre at the University of Freiburg. There, the density of antibioticusage is calculated as defined daily doses (DDD) per 1000 patientdays. As well, the microbiology labs are sending the resistance data forselected bacteria (selection is conform to §23 Infektionsschutzgesetz) tothe reference centre.The cumulated collected data is send back twice a year to the ICUs, sothat trends over months and years can be visualised.Results: In our ICU, we were able to detect a correlation betweencephalosporin usage and emergence of ESBL-carrying Klebsiella andbetween carbapenem usage and carbapenem resistant Pseudomonasaeruginosa, which led to a change in antibiotic prescription policy.Conclusions: The monitoring of antibiotic usage together with bacterialresistance data can help to identify reasons for the spread of highlyresistant bacteria in the clinical setting and support awareness for thesensible use of antibiotics in ICUs.

R2188 Main trends of outpatient antimicrobial consumption inRussia, 2000–2005

A.A. Fokin, S.A. Ratchina, S.N. Kozlov, E.P. Shal, A.A. Ishmukhametov,M.N. Denisova, L.P. Abramenko (Smolensk, Moscow, RU)

Objectives: To assess main trends of outpatient (OP) antimicrobials(AM) consumption (Cons) in Russia in 2000–2005.Methods: Data containing products names, ATC/DDD codes and values(WHO, version 2006), drug forms, dosages and number of packageswere collected during pharmacy audit. The use of AM was expressed asa number of defined daily doses/1000 inhabitants/day (DID).Results: From 2000 to 2005 OP AM Cons in Russia was 9.28, 8.28,9.51, 9.57, 9.14 and 8.90DID, respectively. The leading groups weresulfonamides and trimethoprim (J01E) in 2000–2002 and penicillins(J01C) in 2003–2005. The proportions of J01C, quinolones (J01M),macrolides, lincosamides (J01F) and other b-lactam antibacterials(J01D) Cons increased during the study period by 7.2, 5.9, 1.3 and1.3%, respectively, while J01E use decreased (10.3%). The latter waspredominantly due to twofold decrease in co-trimoxazole use (from 1.38in 2000 to 0.68DID in 2005). The J01C Cons was characterised byincreasing in amoxicillin and amoxicillin/clavulanate and decreasing inampicillin use (from 0.26, 0.02 and 0.91 DID in 2000 to 1.28, 0.28and 0.52 DID in 2005, respectively). The J01M Cons increase wasmainly due to cipro- and norfloxacin (from 0.46 and 0.15 in 2000 to0.77 and 0.34 DID in 2005, respectively), similar trends were typical fornewer fluoroquinolones (levo- and moxifloxacin) (from 0.00003 in 2000to 0.01 DID in 2005, respectively). As for J01F Cons, erythromycinuse decreased (from 0.27 in 2000 to 0.17 DID in 2005), whereasazithro-, clarithro- and josamycin Cons was characterised by the oppositetrend (from 0.09, 0.02 and 0.012 in 2000 to 0.25, 0.14 and 0.03 DIDin 2005, respectively). Among cephalosporins the proportion of first-,second-, third- and fourth-generation was 75.1%/3.67%/20.9%/0.33% in2000, 78.2%/3.56%/18.1%/0.14% in 2001, 81.1%/2.64%/16.1%/0.16%in 2002, 81.4%/2.4%/16.0%/0.2% in 2003, 76.8%/2.38%/20.6%/0.22%in 2004 and 71.0%/3.67%/25.1%/0.%23% in 2005, respectively.Conclusion: Total OP AM Cons in Russia in 2000–2005 was ratherstable with the prevalence of the cheapest, old poor safety profile drugs.But in the dynamics some positive tendencies like the increasing of theproportion of the newer AM usage were observed.

R2189 Antibiotic usage and resistance patterns in a Saudi tertiaryhospital

M. Al-Jamal, A. Al-Barrak, N. Elkhizzi (Riyadh, SA)

Background: The increasing difficulty in fighting off microbes leads toan increased risk of acquiring infections in hospitals or other settings.One of the most important factors that could affect the increasing patterns

S634 17th ECCMID / 25th ICC, Abstracts accepted for publication only

of resistance in Saudi Arabia as well as the rest of the world is theinappropriate use of antibiotics.Objective: To evaluate the resistance patterns of Staphylococcus aureus,Streptococcus pneumoniae, Escherichia coli, Pseudomonas aeruginosain 2005 at RMH comparing with 1997–1998 antibiogram.Methodology: All isolates that have been reported in 2005 have beenreviewed and compared to 1997–1998. The isolates were classified tooutpatient, inpatient and intensive care unit (ICU). The isolates have beenidentified at Riyadh Military Hospital (RMH) laboratory according to thestandard of the National Committee for Clinical Laboratory Standards(NCCLS).Results: A total of 11,492 isolates have been isolated from all sitesfrom both inpatient and out patient setting in 2005. About 42%of these isolates were Gram-positive and 58% were Gram-negative.Staphylococcus aureus isolates were 43% of isolated Gram-positive.Of these, the Methicillin Sensitive Staphylococcus aureus (MSSA) was73% and the Methicillin Resistant Staphylococcus aureus (MRSA) was27%. Streptococcus pneumoniae isolates was 295 (16% were penicillinresistant and 34% were intermediate). Escherichia coli was isolatedin 34% of Gram-negative isolates where Pseudomonas aeruginosaisolates were 26%. Inpatient isolates have resistant patterns higher thanoutpatients and the highest was in the ICU. These data were comparedwith 1997–1998 antibiogram where we found that the resistant patterns issignificantly increasing (p< 0.05). The antibiotics usage was significantlyhigher in 2005 compared to 1997–1998 period (p< 0.05).Conclusions: The antibiotic resistance has been a problem as long as weare overusing antibiotics. The resistant pattern is higher at hospitals withthe highest reported in ICU. These data was supported by that reportedin the literature. This might be due to poor compliance to infectioncontrol measures were applied in ICU and the relative small, crowdedand busy space. Several measures have been applied in 2006 to controlinfections and to prevent resistance. These include antibiotic restrictionprotocol, in-service education programme, surgical prophylaxis policyand continuing of implementing a good infection control strategy.

Molecular bacteriology

R2190 Comparison of real-time PCR and BDProbeTec ET systemfor rapid diagnosis of tuberculosis patients

S. Bernal, C. Freyre, M.J. Torres, I.J. de la Calle, M. Rodrı́guez-Iglesias,J.C. Palomares, E. Martı́n-Mazuelos (Sevilla, ES)

Objective: To evaluate the efficacy of an “in house” real-time PCRtechnique using the LightCycler system (Roche Biochemicals) comparedto the semiautomated BDProbeTec ET system (BD) (Becton–Dickinson)and to acid-fast bacilli stain and culture to detect and identifyM. tuberculosis in DNA extracts from samples obtained from patientswith high suspicious of having TB. Final diagnosis of TB determinedon microbiological and clinical findings was used as “gold standard”.Methods: The real-time assays amplifies a region from the IS61110sequence and the BD system amplifies a region of the mycobacterial16S rDNA. Conventional mycobacterial culture was performed withBactec MGIT 960 and Lowenstein–Jensen medium. A total of 93 clinicalsamples (77 pulmonary and 16 extrapulmonary specimens) from 81patients with high clinical and/or radiological suspicious of havingtuberculosis were processed by the three methods.Results: TB diagnosis was confirmed in 33 patients. Both methodsshowed a 92.6% concordance. This concordance was of 84.4% for TBcases and of 97.9% for TB negative cases. Twenty-two TB cases weredetected by both methods. There were five discrepancies, all of them insmear negative/culture positive patients and 6 TB cases that were missedby both methods. The global sensitivity, specificity, positive predictivevalue and negative predictive value were 72.7%, 100% 100%, and 84.2%respectively for real time PCR and 75.7%, 97.8%, 96.1% and 85.2%respectively for BD system. For smear negative specimens the resultswere 59%, 100%, 100%, and 84.2% for real time PCR and 63.6%,97.9%, 93.3% and 85.4% for BD system.

Conclusions: (1) Both methods showed an excellent concordance.(2) The real time PCR diagnosed the 59% of smear negative patientsin the same day of specimen’s reception. (3) A negative result of PCRdiscard the TB diagnosis because the specificity was 100%.

R2191 Analysis of essential gene changes in MRSA and non-MRSAstrains inhibited by methanolic extract correlated tomembrane permeabilising peptide

M. Shamsudin, N. Alipiah, N. Vasantha, F. Yusoff, A. Arshad (Serdang,MY)

Objective: In this study, a methanolic extract derived from a marineorganism was investigated for the effect on the growth of MRSAresistant to b-lactam antibiotics. The nucleotide sequence changes ofessential genes inclusive of mecA, mecR1, 16sRNA, mprF and msR ofthe inhibited MRSA strains were determined. A potential antibacterialpeptide with activity on permeabilising the membrane of MRSA strainswas investigated through a cellular bioassay.Methods: S. aureus isolates (MRSA and non-MRSA) obtained frompatients of hospitals in Malaysia and environmental isolates were studied.Anti-MRSA activity was determined through MIC and disc diffusionassay. Membrane permeabilising property of the methanolic extract wasinvestigated by the penetration of the fluorescence Sytox green dyeinto MRSA cells. The RT-PCR analysis and nucleotide sequencing ofessential genes, followed by gene sequence analysis were conducted fordetermining molecular mechanism of inhibition of S. aureus isolates.Results: The methanolic marine extract showed variation in nucleotidesequence changes of several essential genes namely the mprF membranegene in S. aureus (gene encoding for peptides involved in transportationof lysine to phospholipids bilayer in membrane). The intensities ofSYTOX green fluorescence dye in S. aureus cells treated with theextract at the MIC values increases with time, similar to treatment withpolymyxin, the positive control antibiotic. The negative control fucidicacid accordingly did not affect bacterial cell membrane permeabilisationand thus showed constant fluorescence intensity even after 6 hourof incubation in the antibiotic. The fluorescence intensities increasedrastically first 120 minutes treatments indicating the membranepermeabilising ability is affected and the dye coloured the nucleic acid.Conclusions: This study demonstrates the anti-methicillin-resistantStaphylococcus aureus (MRSA) activity of methanolic marine extractby exhibiting substantial effect at both the molecular and cellular levelselucidated through nucleotide sequence changes and Sytox green dyepenetration. Molecular correlation to cellular activity of extract is aviable strategy to explore alternative anti-MRSA agents.

R2192 Investigation and sequencing of urease gene of H. pyloricocoid forms after exposure to different environmentalconditions

F. Can, Z.C. Karahan, I. Dolapci, M. Demirbilek, A. Tekeli, H. Arslan(Ankara, TR)

Objectives: The aim of this study was to investigate whether convertionto the coccoid form under different environmental conditions resulted insequence differences in the Urease A gene in H. pylori strainsMethods: Helicobacter pylori standart strain NTCC 11637 was usedin this study. Transformation of the bacteria from helical to coccoidform was carried out with different techniques; exposure to amoxicillin,cold starvation, aerobiosis and aging of the culture. Urease activity wasmeasured by rapid urea test. DNA was extracted from all samples byusing Nucleospin DNA extraction kit (Clontech, CA). Urease A genewas amplified by using the primers HPU1 (GCC AAT GGT AAA TTAGTT CC) and HPU2 (GTA AAA ACA ATT AAG GAG). Bi-directionalsequence analysis was performed by using one of the primers in eachrun by ABI Prism 310 DNA sequencer.Results: Urease activity was determined in spiral and coccoid formsof H. pylori. All samples yielded 411 bp amplimer by polymerasechain reaction. For all sequence comparisons, spiral form of H. pylori