Poster session 4: Microbiology P4.01 MRSA and presence of inflammation are associated with increased overall mortality in patients with diabetic foot ostemyelitis Carlo Tascini, Pisa University Hospital, Pisa, Italy Elisabetta Iacopi, Pisa University Hospital, Pisa, Italy Alberto Coppelli, Pisa University Hospital, Pisa, Italy Chiara Goretti, Pisa University Hospital, Pisa, Italy Alberto Piaggesi, University of Pisa, Pisa, Italy Aim: Diabetic foot osteomyelitis (DFO) may be caused by multi-drug resistant microorganisms (MDRO) and is associated with an high incidence of mortality. To check if MDRO are predictors of mortality in DFO we analyzed retrospectively our microbiological database, from 2001 to 2013. Methods: These characteristics were studied: 1) Methicillin-Resistant Staphylococcus Aureus (MRSA) vs Methicillin-Susceptible Staphylococcus Aureus (MSSA); 2) Ciprofloxacin resistant Pseudomonas aeruginosa (CiproRPA) vs Cipro- susceptible Pseudomonas aeruginosa (CiproSPA); 3) Carbapenem resistant Pseudomonas aeruginosa (CRPA) vs Carbapenem suspceptible Pseudomonas aeruginosa (CSPA); 4) Ciprofloxacin resistant enterobacteria (CiproRE) vs Ciprofloxacin susceptible enterobacteria (CiproSE); 5) Extended spectrum Beta Lactamase (ESBL) producers enterobacteria (ESBL+) vs non ESBL producers enterobacteria (ESBL-); 6) Candida parapsilosis (CP) vs Candida non parapsilosis (CNP). Presence of polymorphonuclear leukocytes (PMN) at Gram stain of DFO was recorded. Clinical characteristics studied were: overall mortality, and time to healing (TH) of DFO. Results: 401 patients with DFO were found. From bone specimens analyzed in microbiology laboratory were recovered 625 isolates. Redundant isolates were eliminated. Staphylococcus aureus was found in 140 DFO (78 MRSA – 55.7%); Enterobacteria in 117 DFO (Cipro RE 46 - 39.5%, ESBL+ 34 – 34.3%); Pseudomonas aeruginosa in 95 DFO (53 CiproRPA – 55.2%, 31 CRPA – 32.1%); Yeasts in 32. DFO caused by MRSA was associated with increased overall mortality (p= 0.005) and increased TH (p= 0.024). Presence of PMN was associated with increased overall mortality (p= 0.008) and TH (p= 0.026). Conclusion: MRSA and presence of PMN at gram stain of bone obtained from DFO was associated with increased overall mortality and increased TH in diabetic patients with DFO. www.diabeticfoot.nl Page 1 of 10 P4.02 Differences between etiopathogeny of diabetic foot infections in two different clinics, diabetes and surgery Eduard Lucian Catrina, UMF Carol Davila, Bucharest, Romania Andra Andreescu, I.Cantacuzino Hospital, Bucharest, Romania Raluca Maria Popescu, UMF Gr.T.Popa, Iasi, Romania Development of an infection in diabetic foot lesions is the leading cause of admissions of patients with diabetic foot. Aim: The paper compare the etiopathogenic aspects of diabetic foot infections (DFI) in patients admitted in two different clinics: Diabetes and General Surgery ones. Methods: The study compares in a observational manner and analizes retrospectively two groups of patients, both consisting of 25 patients, consecutively admitted in each of two clinics. Every patient has DFI confirmed in terms of bacteriology. The microbiological sample was collected by swab in Diabetes Clinic (DC), while all patients in the Surgery Clinic (SC) were operated and bacteriological sample was harvested, during the operation, by excision of infected tissue (soft tissue or bone). Results: In SC prevailed extended soft tissue infections (phlegmons, necrotizing infections, extensive gangrene) classified as infection gr. 3 and 4, in PEDIS clasification. Regarding etiopathogeny it prevailed infections with a single germ comparing with polymicrobial infections (17 cases vs 8 cases). The most common germ was Staph aureus, which was found to be involved in 15 from all cases (4 cases MRSA) and only in 9 monomicrobial infections (2 MRSA). From all 25 cases, it was necessary minor amputation in 7 and major amputation in 4 cases. In DC were hospitalized patients with limited lesions, as infected neuropathic ulcers and limited toe gangrene, classified as mild infections (PEDIS 2) and just fewer mild/severe infections (PEDIS 3). There were more frequently polymicrobial than monobacterial infections (14 vs 11 cases) and Staph aureus in 19 from all cases, of which was 7 monobacterial infections. In only 2 cases it was identified MRSA. In 16 cases the evolution was favorable under antibiotics. 9 patients were transferred for surgery. No anaerobic germ was identified. Discusion: Not to draw a conclusion, there is a difference between severity of cases admitted in each of two clinics. Differences between etiopathogeny, such as higher number of polymicrobial infections and Staph aureus infections in patients from DC may be the result of different methods of bacteriological sample collection or suggests that is a direct relationship between Staph infection and mild/moderate infections. www.diabeticfoot.nl Page 2 of 10 P4.03 Comparison of diabetic foot ulcer culture techniques Yuehong Shen, Zhejiang University School of Medicine, Hangzhou, China Wu Ding, Zhejiang University School of Medicine, Hangzhou, China Aim: Wound cultures is part of the diabetic foot clinical assessment as the basis of medicine application. Swabbing is the ordinary specimen-taking technique in most hospital in China and there is no worldwide standard. This study compares two wound culture techniques in diabetic foot ulcers swab and curetted tissue for microbial recovery, Methods:For each wound, two sampling approaches were applied during the same visit: swab culture and curetted tissue culture. Results:A total of 10 diabetic foot, ulcers were assessed among 10 patients.8 ulcers' culture result is same between two methods. Results: Swab and curetted tissue cultures yielded similar recovery rates for common wound bacteria. Conclusions: The swabbased and curetted tissue culture method for diabetic foot ulcer is similar. www.diabeticfoot.nl Page 3 of 10 P4.04 The bacterial profile of infected and hidden neuropathic diabetic foot ulcer in type II diabetes mellitus in Saudi Arabia Osman Habeeb Basheir, Alrajhi medical college, Albukaryiah, Saudi Arabia Abdallah Algoblan, King Fahad diabetic center, Buraidah, Saudi Arabia Ahmed Gamal Frag, Alrajhi medical collge, Albukariyah, Saudi Arabia Mohamad Saddik, Alrajhi medical college, Albukariyah, Saudi Arabia Mohammad Suliman Alasiri, Alrajhi medical college, Albukariyah, Saudi Arabia Diabetes mellitus and it is foot complications is one of the major health problems in Saudi Arabia with an overall prevalence of 23.7%. Most of the diabetic foot ulcer in Saudi Arabia had a neuropathic character and many of them carries a hidden bacterial load. Aim: To study the bacterial pattern and antimicrobial susceptibility in neuropathic diabetic foot ulcers Methods: A cross-sectional study was conducted over a period of 10 month from January to October 2014. Samplings from neuropathic diabetic foot ulcer (infected and non infected) were assessed for bacteriology and antimicrobial sensitivity using stander microbiological technique. Result: Out of 360 diabetic feet patients; sample were taken from 145 patients with neuropathic ulcers. 63.4% (n=92) of them were male and 36.6% (n=53) were female. The mean age is 56 Stander Deviation (STD 9.9), mean body mass index is 31.2 (STD 3), and the mean HbAc1 is 9.5 (STD 2.1). The duration of diabetes is varies between 6 -15 years in most patients (60.3%). Of the 145 patents with neuropathic diabetic foot ulcer a total of 220 organism was isolated, 63% of these patients showed polymicrobial in the culture and 29.7% with monomicrobilal growth. Gram negative organism (63%) reports prevalence more than the gram negative pathogen (37%)., Staph aureus is the commonest organism isolated (19.5%). Wears klebsiella pneumonia, Escherichia coli and Proteus mirabillis reports as common gram negative isolates 17.7%, 14.5 %, & 11.8% respectively (Table 1). All isolated organism showed a variable degree of resistance to the commonly used antimicrobials except for amikacin. However imipenem and vancomicin showed high sensitivity to most of the isolated pathogens. Most of the patients 67% take more than one antibiotics at one time, however 46% of the patients not completed the course duration of at least one of their drugs. Conclusion: The study shows predominance of gram negative organism in neuropathic diabetic ulcers and the pattern of the bacteriological susceptibility is mainly towards higher cephalosporin expensive groups ( Imipenem) or to the harmful drugs to the kidney (gentamycin, vancomicin.) this make a high economic burden for diabetic ulcer management in the future. www.diabeticfoot.nl Page 4 of 10 P4.06 Microbiology of moderate-severe diabetic foot infection - a prospective study Andre Couto de Carvalho, Centro Hospitalar Porto, Porto, Portugal Teresa Pereira, Centro Hospitalar Porto, Porto, Portugal Susana Garrido, Centro Hospitalar Porto, Porto, Portugal Rui Carvalho, Centro Hospitalar Porto, Porto, Portugal Aim: Our objective is to present the microbiology and antimicrobial susceptibility of moderate-severe diabetic foot infections (DFI) evaluated in a tertiary referral center. Methods: A prospective study of diabetic patients with moderate-severe DFI episodes was performed between October 2012 and June 2014 (21 months). At each infection episode microbiological cultures were taken at deepest level after cleansing the ulcer and antimicrobial susceptibility was performed by the disk diffusion method. Those cases with positive deep tissue or aspirate culture samples were included in this study. Results: Data from a total of 134 patients with moderate-severe DFI was obtained. Fortyseven different subjects (51.1% with neuro-ischaemic ulcer) have provided 65 positive deep tissue, aspirate and/or exudate culture samples. These positive samples were polymicrobial in 57% of cases, providing 112 distinct isolates, with a mean 1.7 microorganism per culture specimen. The predominant isolate was the Staphylococcus aureus, present in 35.4% of DFI samples (61% of which MRSA). Enterococcus spp. was identified in 32.3% (100% vancomycin-sensitive). Importantly, Enterobacteriaceae were recognized in 43.1% of all DFI episode samples (32% of which ESBL producers). Pseudomonas aeruginosa were found in 18.5% (25% ciprofloxacin-sensitive) and anaerobes species in 4.6% of cases. Conclusions: Bacteriology in our moderate-severe DFI prospective cohort was highly heterogeneous with increasing prevalence of multiresistant Gram-negative bacteria. This data may help develop and update local DFI clinical guidelines regarding empirical antibiotic therapy. www.diabeticfoot.nl Page 5 of 10 P4.07 Microbiology of deep diabetic foot infection in surgery patients Tatiana Zelenina, Northwestern Medical University n.a. I.I. Mechnikov, St. Petersburg, Russia Alexandr Zemlyanoy, Northwestern Medical University n.a. I.I. Mechnikov, St. Petersburg, Russia Natalia Vorokhobina, Northwestern Medical University n.a. I.I. Mechnikov, St. Petersburg, Russia Aim: We investigated specificity of pathogens in patients with diabetic foot deep infection who was required hospitalization and surgery intervention. Methods: We analyzed cultures of 201 inpatients who were hospitalized at the diabetic foot surgery department of City Hospital № 14 from September 2012 to May 2013. 152 patients were primary and 49 had already required previous surgery interventions on foot. All patients had deep diabetic foot infections, underwent surgery debridement, and 63.2% and 62.5% of them were amputated. Major amputations were performed in 26.6% and 33.3% patients respectively. Mean duration of hospitalization was 25.3±17.2 days (1-90 days). Cultures were obtained after surgery interventions immediately and on 10-14 days of hospitalization. We also analyzed cultures of 102 diabetic foot outpatients with post surgery wounds after discharge from the Hospital. Results: St. aureus was the most frequently isolated in primary patients (59% of cultures), including MRSA in 13.5%., Gram-negative bacilli were found: Acinetobacter baumanii in 20.5% cases, Enterococcus faecalis in 16%, Pseudomonas aeruginosae in 13.6%. St. aureus was the most frequently isolated in patients with previous history of surgery interventions too (75.7% cultures), including MRSA in 39.3%. Acinetobacter baumanii and Enterococcus faecalis were obtained in 35% specimens, Pseudomonas aeruginosae in 27%. After 14 days of hospitalization St. aureus was isolated in 47% and 62% specimens respectively, including MRSA in 33.3% and 61%., Acinetobacter baumanii was obtained in 27 6% and 36%, Enterococcus faecalis in 55% and 59% specimens respectively. Post surgery wounds were clinically infected in 55.9% outpatients. St. aureus was isolated in 78% specimens, including 53.1% MRSA. Acinetobacter baumani was obtained in 24.4% cultures, Enterococcus feacalis in 29.3%, Pseudomonas aeruginosa in 24.4%. Conclusions: The infections of surgery patients are polymicrobial. St. aureus is most frequently isolated pathogen. The prevalence of MRSA and gram-negative bacilli (Acinetobacter baumanii, Enterococcus faecalis) should be taking into account for patients with previous history of surgery interventions and with infected post surgery wounds. www.diabeticfoot.nl Page 6 of 10 P4.08 A hospital based study of multi drug resistance bacteria in diabetic foot infection from North India Shailesh K. Shahi, Banaras Hindu University, Varanasi- 221005, India, Varanasi, India Ashok Kumar, Banaras Hindu University, Varanasi- 221005, India, Varanasi, India Sanjeev K. Gupta, Institute of Medical Sciences, Banaras Hindu University, Varanasi221005, India, Varanasi, India Background: Diabetic foot infection is a common cause of lower extremity amputations. Increasing incidence of antimicrobial resistance is becoming a serious problem in India, there is paucity of data dealing with the incidence of multiple-drug resistance (MDR), bacteria from Diabetic foot ulcers (DFUs). Aim: Isolation and characterization of multiple-drug resistance bacteria from Diabetic foot ulcers. Materials and Methods: The study was conducted at S.S. Hospital, Institute of Medical Sciences and School of Biotechnology, Banaras Hindu University, Varanasi. Antibiotic sensitivity test was performed by disk diffusion method. Identification of bacteria was done by 16S rDNA sequencing. The presence of β-lactamase genes and class-1 integron gene responsible for multiple antibiotic resistance was detected by PCR. Result: Altogether 142 aerobic bacteria were isolated from swab and tissue samples of 42 severe DFUs patients. Antibiotic sensitivity test of was performed against sixteen antibiotics belonging to eight classes. Of the 142 isolates, 38 (26.76%) showed resistance to eight or more than eight antibiotics. 38 isolates with MDR characteristics were identified by 16S rRNA gene sequencing. All the sequences have been deposited to the NCBI GenBank. Majority of isolates were identified as Enterococcus spp., Staphylococcus aureus, Pseudomonas spp., Stenotrophomonas spp., Klebsiella pneumonia, Escherichia coli, and Alcaligenes spp. With a view to understand the mechanism (s) of multi drug resistance (MDR), screening of integron and β-lactamases was made. Multiplex PCR reveled that 89.47 % of MDR isolates carried blaTEM, blaOXA, blaSHV and blaCTX-M like β-lactamase genes. Furthermore, class 1 integrons of size ranging from 0.3 to 4.0 kb were noted in 68.42% of MDR isolates. Conclusion: The high prevalence of MDR bacteria in DFUs from North India is found in the present study. It is felt that proper management of antibiotics must be implemented to lower the incidence of MDR bacterial infection considering the risks of amputation in DFU patients. www.diabeticfoot.nl Page 7 of 10 P4.09 Clinical and microbiological characteristics of infected diabetic ulcers in siriraj hospital, the largest university hospital Thailand Oranich Navanukroh, Siriraj hospital, Bangkok, Thailand Pornpan Koomanachai, Siriraj hospital, Bangkok, Thailand Introduction: Diabetic foot infection (DFI) is one of the most common causes of hospitalization in diabetes mellitus patients. DFI cause both physical and mental morbidity. However, there were no clinical characteristics and causative pathogens of DFI available in Thailand. This study aimed to explore the epidemiological data of DFI in Thai patients who were treated at Siriraj hospital, Thailand. Methods: The retrospective study was conducted at Siriraj hospital, Bangkok, Thailand. All medical records of type 2 diabetic patients with DFI during January 2012 to December 2013 were reviewed. The patients’ demographic data, ulcer characteristics, culture results and treatment outcome were collected and analyzed. Results: One hundred and five patients were enrolled, 63 patients were male. Mean duration of diabetes was 689.4 ± 464.4 weeks. Mean ulcer duration was 36.58 ± 6.02 days. Mean glysated hemoglobin was 8.3%. One hundred and eighty two ulcers were obtained clinical specimens for culture. Poly-microbial infection was 65%. Gram-negative bacteria was common pathogens of DFI, 231 isolates vs. 159 gram-positive bacteria. The most common gram-negative and gram-positive pathogen was Escherichia coli and Staphylococcus aureus, respectively. For E. coli, 74.2% of 31 E. coli isolates were ESBL-producer. Fluoroquinolone-resistance was found 80.6% for E. coli and 60% Klebsiella pneumonia. Previous antibiotic used within 3 months had tendency to associate with drug-resistance organisms. Overall mortality rate was 11 out of 105 patients; 3 were from DFI and 8 were from other organs infection acquired during hospitalization for DFI. Univariate analysis showed severe ulcer grading, poly-microbial infection and inappropriate empiric antibiotics were associated with unfavorable outcome of amputation. Mortality associated with high HDL-cholesterol and high serum creatinine. Conclusion: Multidrug- resistance organisms were demonstrated in DFI. Appropriate antimicrobial therapy, especially in severe ulcer, using pathogens and antibiogram-guided is very important in conjunction with surgical treatment and revascularization to improve outcome of DFI in Siriraj hospital. Main drug-resistant organisms cultured from diabetic foot ulcers www.diabeticfoot.nl Page 8 of 10 P4.10 Management of diabetic foot infections in a tertiary hospital in UK: Microbiological testing strongly influences the antibiotic treatment Danielle Lowry, University Hospital Birmingham NHS Foundation Trust, Birmingham, United Kingdom Robin Chisman, University of Birmingham, Birmingham, United Kingdom Mujahid Saeed, University Hospital Birmingham NHS Foundation Trust, Birmingham, United Kingdom Alok Tiwari, University Hospital Birmingham NHS Foundation Trust, Birmingham, United Kingdom Miruna David, University Hospital Birmingham NHS Foundation Trust, Birmingham, United Kingdom Aim: To characterise the results of microbiological testing and examine its influence on the antibiotic (AB) choice in diabetic foot disease. Methods: Retrospective analysis of all diabetic patients who had deep tissue samples from the foot between January 2012 and December 2013. Using the Infectious Diseases Society of America (IDSA) and International Working Group on the Diabetic Foot (PEDIS) classifications, each patient’s presenting condition was graded 1 (no infection) to 4 (severe infection) with osteomyelitis (OM) as a discrete diagnostic entity. Information was collected on gram stain, culture result and AB therapy. Results: 118 samples from 74 patients (mean age 63 years (range 27-88 years)) were examined. Gram stain showed poor correlation with the final culture, with an overall sensitivity of 76% and specificity 73%. Sensitivity was especially low in detecting the presence of gram negative rods (24%) and specificity was poorest when gram positive cocci were seen (35%). Up to five organisms per sample were isolated in culture. Excluding anaerobic growth, gram positive organisms were isolated in 54% of samples and gram negatives in 41%. There was no significant difference between organisms grown and grade of infection. At the time the sample was taken, only 34% of patients were on an appropriate empirical AB regimen. After the final culture report was received, 70% of patients had a change of AB regimen and 86% of patients were on an appropriate AB regimen. Conclusions: Gram stain is poorly representative of the tissue culture result in this cohort. Considerable adjustment of AB regimens was made in response to the final microbiology culture and sensitivity report. Despite improved guidance both internationally and locally, many diabetic patients are still started on inappropriate empirical ABs for their foot infection by non-specialist clinicians. In many cases, this is represented by the use of a narrowspectrum agent (e.g. flucloxacillin) for grade 4 infections or OM. This may be due to clinicians erroneously considering the treatment of a diabetic foot infection to be the same as that for cellulitis in a non-diabetic patient, misclassification of severity, unfamiliarity with the PEDIS classification and/or non-adherence to local AB guidelines. www.diabeticfoot.nl Page 9 of 10 P4.11 Surgical wound cultures: A subanalysis of a randomized controlled trial Caitlin Garwood, Medstar Georgetown University Hospital, Washington, DC, United States Paul Kim, Medstar Georgetown University Hospital, Washington, DC, United States John Steinberg, Medstar Georgetown University Hospital, Washington, DC, United States Karen Evans, Medstar Georgetown University Hospital, Washington, DC, United States Christopher Attinger, Medstar Georgetown University Hospital, Washington, DC, United States The aim of this study was to perform an in-depth analysis of the pre and post-debridement wound culture data from patients undergoing serial surgical debridements., The data was collected and analyzed as a sub-analysis of a randomized controlled trial. Of the 100 patients enrolled in the study, 35 were excluded for the following: 15 for incomplete culture data sets, 9 with more than 3 operating room (OR) visits, 5 were not covered or closed at final surgical procedure, and 6 had no 30-day follow-up. Specimen counts and semiquantitative culture data, both pre-debridement and post-debridement, were recorded and analyzed for each OR visit. 30-day follow-up was analyzed to compare final OR cultures with outcomes. 65 patients were included in this sub-analysis. Pre-debridement cultures from the 1st OR visit showed 8 patients (12.1%) had no growth, 29 (44.6%) grew 1 species, 19 grew 2 species, and 9 had 3-5 species. Comparing the pre and post-debridement cultures from the 1st OR visit, 56.9% decreased in the number of species., Similar decreases were seen after each debridement. The semiquantitative culture data shows that after 2 debridements all patients either had no growth (35 patients) or scant/rare/ growth in broth (30 patients). 14 patients (21.5%) dehisced by their 30 day follow-up and 51 remained closed. There was no difference in the number of OR visits or the rate of dehiscence. The final post-debridement culture on the date of closure had no growth in 64.6% of patients (6 dehisced, 36 remained closed) and there was no statistically significant difference., 23 patients still had positive cultures with all of them only having scant/rare/broth growth and 8 dehisced (35%) while 15 (65%) remained closed. Surgical wound cultures have long been a staple in the management of wounds., They often serve as a guide for antibiotic selection and determining when a wound is considered clean, which aids surgical decision making. This data demonstrates that surgical debridement significantly decreases number of species and semiquantitative cultures. There was no statistical significance in the rate of dehiscence between those with no growth and those with positive final cultures, but there is a trend suggesting a positive culture may yield a higher rate of dehiscence. www.diabeticfoot.nl Page 10 of 10
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