A Comparison of Tissue versus Swab Culturing of Infected Diabetic Foot Wounds

Objective. To compare the efficacy of swabbing versus tissue biopsy for microbiological diagnosis of diabetic foot infection.Methods. This was a prospective trial. Fifty-six patients with diabetic foot infection were divided into the following 3 groups according to the PEDIS grading system: grade 2 (n=10), grade 3 (n=29), and grade 4 (n=17). Two specimens were collected from each wound for microbial culturing after debridement, including a superficial swab and a deep tissue punch biopsy specimen.Results. Swab culturing identified all of the microorganisms isolated from the corresponding deep tissue specimens in 9/10 of grade 2 wounds (90.0%), and this proportion decreased to 12/29 (41.4%) and 7/17 (41.2%) for grades 3 and 4 wounds, respectively (p=0.02). Moreover, the sensitivity for identifying Gram-negative bacteria, such asE. coliandCitrobacter, by swabbing was low (33.3%). In addition, some Gram-negative bacteria, such asSerratiaandRalstonia pickettii, were isolated from deep tissues but not from swabs.Conclusions. Swab culturing may be reliable for identification of pathogens in diabetic foot wounds classified as grade 2. However, it is advisable to culture deep tissue specimens for wounds of grade ≥3 because swab culturing is associated with a high risk of missing pathogens, especially Gram-negative bacteria.

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Identification and Drug Sensitivity of Aerobic Bacterial Isolates from Diabetic Foot Ulcers of Sudanese Patients: a Cross sectional study

10.21203/rs.2.22115/v1 ◽

2020 ◽

Author(s):

saada Mohamed nour ◽

Maowia Mukhtar ◽

Mohamed Elmakk ◽

Elshibli Mohamed Elshibli ◽

Walyeldin Elnour Elfakey ◽

...

Keyword(s):

Diabetic Foot ◽

Diffusion Method ◽

Foot Ulcers ◽

Aerobic Bacteria ◽

Diabetic Patients ◽

Gram Negative Bacteria ◽

Female Ratio ◽

Gram Negative ◽

The Mean ◽

Grade 3

Abstract Background Diabetic foot ulcer infection cause great morbidity and mortality among diabetic patients and is a major cause of lower extremity amputation worldwide. This study aimed to determine the profile of aerobic bacteria and their antibiotic sensitivity patterns in diabetic foot infections (DFI) among different Wagner's grades. Methods This study was conducted during December 2017 - March 2018 in a Diabetic Center, Sudan. A total of 152 diabetic patients with different grades of foot ulcers were randomly enrolled in the study. The patients were grouped using Wagner's classification. Tissue biopsies and deep swabs were collected from the ulcers for aerobic cultures. The cultured isolates were identified using phenotypic and biochemical properties and their sensitivity to commonly used antibiotics, Colistin, Aikacin, Ciprofloxacin, Augmentin, Ceftazideme, Gentamicin, Clindamycin, Ceftriaxone Meropenum. Cotrimoxazole, Erythomycin, Oxacillin and Vancomycin. Fusidic acid, Imepenem, and Piperacillin was tested using the Kirby Bauer disk diffusion method. Results The mean age of the patients was 54.31 (SD ± 12.1) years, male to female ratio of 8: 1. The mean duration of diabetes was 14 (SD ± 8) years. The ulcers varied in duration from 1 day to 10 years. of 152 samples 181 aerobic bacteria were isolated. Cultures yielded 1-3 isolate per culture. The maximum number was isolated from grade 3 group followed by long standing ulcer LSU group 50.8% and 28% respectively. Polymicrobial infection was higher in LSU (30.4%). The isolates were mostly Gram-negative bacteria. The most frequent were proteus spp. (35.3%), S. aureus MRSA 14.4% and Coliform 12.2% respectively. The most common isolates in grade 3 were P. Mirablis, Staphylococcus and Coliform and in long standing ulcers were P. Mirablis, S. aureus MRSA and Coagulase negative staphylococcus respectively. Conclusion Gram-negative bacteria were more prevalent and the most frequent pathogens were Proteus spp. The most common polymicrobial infections were due to P. mirablis with; P. aerginosa, S. MRSA and Coliform respectively . Gram negative rods were sensitive to Amikacin, (80.6) %) while the highest sensitivity of Gram positive was to Imepnem (85%). Most of the isolates were sensitive to Meropenem. No significant relation between Wagner grades and neuropathy was detected.

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DIABETIC FOOT INFECTION

The Professional Medical Journal ◽

10.29309/tpmj/2015.22.11.874 ◽

2015 ◽

Vol 22 (11) ◽

pp. 1415-1422

Author(s):

Abdul Ghani Rahimoon ◽

Muhammad Tanveer Alam ◽

Muhammad Saeed Talpur

Keyword(s):

Diabetic Foot ◽

Tertiary Care ◽

Life Forms ◽

Resistance Rate ◽

Care Hospital ◽

Gram Negative ◽

Diabetic Foot Infection ◽

Staph Aureus ◽

Culture Growth ◽

Grade 3

Objectives: To determine the frequency of common bacterial isolates culturedfrom diabetic foot infection in patients with type 2 diabetes mellitus (DM) falling in Wagner’sgrade-2 and grade-3 classification of diabetic foot (DF) infection. Study Design: Descriptivestudy. Period: A six months. Setting: Dow University of health sciences and civil hospital Karachi.Methods: Completed to examine the bacterial identification in cases admitted with the infectiondiabetic foot along with gave Wagner’s evaluation 2 and 3 at tertiary care hospital Karachi.Bacteriological finding and anti-biotic affectability profiles were completed and analyzed withutilizing standard strategies. Results: Out of 115 cases, 82 (71%) were male and 23 (29%) werefemale. The mean age of patients was 51.7 ±9.45 years, mean duration of diabetes was 10.6± 4.73 years, similarly mean length of time of diabetes foot wound was 46.15±23.75 days. 45(39%) patients had Wegner’s evaluation 2 and 70 (61%) patients had Wagner’s evaluation 3. 99cases indicated with culture growth, out of which 65 (65.65%) with gram negative microbes and25 (25.25%) gram-positive microbes. The most successive bacteria’s were Proteus (35.35%),Staph. Aureus (25.25%), Klebsiella (16.16%) and Pseudomonas (15.15%). Both gram positiveand gram negative showed frequent resistance to Cloxacillin, Amoxacillin, Levofloxacin, andLinezolid, gram negative life forms likewise indicated high resistance rate to Clindamycin,Vancomycin, and Cefotaxime. Tienam (Imipenem), Sulzone (salbactam in addition tocefoperazone) and Amikacin were the best effective against gram -ve and gram +vemicrobes.Staph. Aureus and Staph. Epidermidis were profoundly susceptible to Ciprofloxacin, Ceftriaxone,Clindamycin, and Vancomycin. Conclusion: Gram negative microbes were more common thangram positive living beings. Proteus, Staph Aureus, Klebsiella and Pseudomonas aeruginosawere the most widely recognized microorganisms of DF infection. Tienam (Imipenem), Sulzone(salbactam in addition to cefoperazone), and Amikacin were best effective agents.

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Deep tissue culture and hemoculture in dogs with wounds and sepsis

Pesquisa Veterinária Brasileira ◽

10.1590/s0100-736x2017001200020 ◽

2017 ◽

Vol 37 (12) ◽

pp. 1483-1490

Author(s):

Mônica V. Bahr Arias ◽

Flávia N. Padilha ◽

Marcia R.E. Perugini

Keyword(s):

Patient Survival ◽

Punch Biopsy ◽

Microbial Culture ◽

Gram Negative Bacteria ◽

Deep Tissue ◽

Gram Negative ◽

Gram Positive Bacteria ◽

Enterococcus Spp ◽

Infected Wounds ◽

Staphylococcus Spp

ABSTRACT: Contaminated and infected wounds occur very frequently in veterinary medicine and can cause systemic inflammatory response syndrome, sepsis, and death. This study aimed to test the feasibility of collecting wound material by deep-tissue or punch biopsy for microbial culture, determine the frequency of bacteria in the wound(s) and blood cultures and the susceptibility of these microbes to antimicrobials, and evaluate clinical parameters that could be related to prognosis. Thirty dogs with wounds and signs of SIRS/sepsis were included in this study. Bacteria were isolated from all wounds and 41 bacterial isolates could be identified based on culture of the materials collected by punch biopsy; 53.66% of the isolates were gram-negative, mainly involving Pseudomonas aeruginosa, Klebsiella pneumoniae, and Enterococcus spp., and 46.34% were gram-positive bacteria such as Streptococcus spp., Enterococcus spp., and Staphylococcus spp. The survival rate was 66.67%. Based on blood culture analysis, we identified bacteremia in seven patients, predominantly of gram-negative bacteria, which negatively affected patient survival, as six dogs died. Hypoglycemia (≤60mg/dL) and severe hyperglycemia (≥180mg/dL) also negatively affected survival as 23.33% of the hypo/hyperglycemic dogs died. Factors such as blood lactate level at admission and hematocrit levels, and mean arterial pressure were not significantly correlated with death or survival of the dogs.

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Etiology of Diabetic Foot Infection in Patients with Poorly Controlled Diabetes

Journal of Biomedical and Clinical Research ◽

10.1515/jbcr-2015-0103 ◽

2013 ◽

Vol 6 (1) ◽

pp. 51-56

Author(s):

Sergey D. Iliev ◽

Lyubomir Ts. Beshev ◽

Kiril L. Nedyalkov ◽

Dobromir D. Nguen ◽

Valentina E. Edreva-Besheva ◽

...

Keyword(s):

Diabetic Foot ◽

Bacterial Species ◽

Anaerobic Bacteria ◽

Clinical Signs ◽

Deep Tissue ◽

Tissue Samples ◽

Diabetic Foot Infection ◽

Diabetic Foot Infections ◽

Anaerobic Infections ◽

Tissue Specimens

SummaryThe aim of the study was to define the spectrum and susceptibility of microorganisms, isolated from diabetic foot ulcers in patients with poorly controlled diabetes, treated at the clinic of surgery, and compare microbial findings of specimens collected superficially and from deep tissues. The study included 19 patients with type 1 and 2 diabetes with clinical signs of infection. All patients were with poorly controlled diabetes and staged from 3rd to 5th grade according to the Wagner diabetic foot scale. Swab samples from non-debrided wounds and biopsy samples from deep tissues were collected from each patient. Specimens were inoculated on media for isolation of aerobic and anaerobic bacteria. Identification and susceptibility testing of the isolated oiganisms were performed by conventional methods, and VITEK 2 and mini API Systems (bioMerieux, France). A total of 88 bacterial isolates were cultured, comprising 56 clinical strains. Gram positive bacteria were the most common isolated organisms (53.57%), followed by Gram negative bacteria (26.78%) and anaerobic bacteria (19.64%). Staphylococcus aureus was the most common organism detected (10 strains), followed by Enterococcus spp. (7 strains), Escherichia coli (7 strains), Bacteroides spp. (6 strains) and various other organisms of low incidence. Polymicrobial infection was detected in 17 (89.47%) of the patients. In most of the cases infections were caused by 3 bacterial species. Mixed aerobic/anaerobic infections were detected in 9 (47.3%) patients. In 15 (78.94%) patients, there was a coincidence of bacterial findings from superficial and deep tissue samples. The strains isolated were susceptible to commonly used antimicrobials for treatment of diabetic foot infection. The predominant part of the diabetic foot infections were polymicrobial, caused by association between two or three microbial species. In half of the cases the infection was mixed. There was a good correlation between microbial findings from superficial swabs and deep tissue specimens when they were delivered to the microbiology laboratory immediately after collection.

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Bacteriological Profile and Antibiotic Susceptibility Pattern of Diabetic Foot Infection in a Tertiary Care Hospital in Lampung, Indonesia

Malaysian Journal of Medical Sciences ◽

10.21315/mjms2021.28.5.4 ◽

2021 ◽

Vol 28 (5) ◽

pp. 42-53

Author(s):

Iswandi Darwis ◽

◽

Hidayat Hidayat ◽

Gusti Ngurah P Pradnya Wisnu ◽

Sekar Mentari ◽

...

Keyword(s):

Diabetic Foot ◽

Medical Records ◽

Antibiotic Susceptibility ◽

Gram Negative Bacteria ◽

High Susceptibility ◽

Gram Positive ◽

Gram Negative ◽

Diabetic Foot Infection ◽

Gram Positive Bacteria ◽

Wound Culture

Background: Diabetic foot infection (DFI) is a serious complication of diabetes mellitus and identification of the causative bacteria is an essential step in selecting the appropriate antibiotic therapy. This study aimed to evaluate the bacterial pattern and antibiotic susceptibility of the bacteria causing DFI in Lampung Province in Indonesia. Methods: This study is a retrospective study reviewing the medical records of DFI patients admitted to the Dr Hi Abdul Moeloek Regional General Hospital in 2017–2019. DFI patients with complete medical record data were included in this study. Demographic, clinical, laboratory, wound culture and antibiotic susceptibility data were collected from the medical records using a short structural chart. The data obtained then reviewed. Results: In this study, 131 DFI patients met the study criteria and were included. Based on the wound culture results, Gram-negative bacteria were obtained in 112 (85.5%) subjects with Enterobacter spp. as the predominant bacteria. Gram-positive bacteria were found in 19 (14.5%) subjects with Staphylococcus spp. as the predominant bacteria. Gram-negative bacteria found in this study showed high susceptibility to amikacin, meropenem and sulbactam/cefoperazone. Meanwhile, the Gram-positive bacteria showed high susceptibility to meropenem, sulbactam/ cefoperazone and amikacin. Conclusion: The findings of the study revealed Enterobacter spp. as the most predominant bacteria causing DFI in the studied population. The highest antibiotic susceptibility was seen for amikacin, meropenem and sulbactam/cefoperazone.

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Disinfection of Fruits with Activated Water

JOURNAL OF ADVANCES IN AGRICULTURE ◽

10.24297/jaa.v10i0.8462 ◽

2019 ◽

Vol 10 ◽

pp. 1864-1872

Author(s):

Prof. Teodora P. Popova

Keyword(s):

Antimicrobial Activity ◽

Aqueous Solutions ◽

Antimicrobial Properties ◽

The Other ◽

Gram Negative Bacteria ◽

High Antimicrobial Activity ◽

Gram Negative ◽

E Coli ◽

Number Of Microorganisms ◽

Lower Effect

The effect of ionized aqueous solutions (anolytes and catholyte) in the processing of fruits (cherries, morellos, and strawberries) for decontamination has been tested. Freshly prepared analytes and catholyte without the addition of salts were used, as well as stored for 7 months anolytes, prepared with 0.5% NaCl and a combination of 0.5% NaCl and 0.5% Na2CO3. The anolyte prepared with a combination of 0.5% NaCl and 0.5% Na2CO3, as well as the anolyte obtained with 0.5% NaCl, exhibit high antimicrobial activity against the surface microflora of strawberries, cherries, and sour cherries. They inactivate E. coli for 15 minutes. The other species of the fam. Enterobacteriaceae were also affected to the maximum extent, as is the total number of microorganisms, especially in cherries and sour cherries. Even stored for 7 months, they largely retain their antimicrobial properties. Anolyte and catholyte, obtained without the addition of salts, showed a lower effect on the total number of microorganisms, but had a significant effect on Gram-negative bacteria, and especially with regard to the sanitary indicative E. coli.

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Structure-Activity Relationship and Antimicrobial Evaluation of N-Phenylpyrazole Curcumin Derivatives

Current Bioactive Compounds ◽

10.2174/1573407215666190124115010 ◽

2020 ◽

Vol 16 (4) ◽

pp. 481-488

Author(s):

Heli Sanghvi ◽

Satyendra Mishra

Keyword(s):

Antibacterial Activity ◽

Gram Negative Bacteria ◽

Pyrazole Derivatives ◽

Gram Positive ◽

Gram Negative ◽

E Coli ◽

Curcumin Derivatives ◽

Structure Activity ◽

Electron Donating Groups ◽

Derivatives Of

Background: Curcumin, one of the most important pharmacologically significant natural products, has gained significant consideration among scientists for decades since its multipharmacological activities. 1, 3-Dicarbonyl moiety of curcumin was found to be accountable for the rapid degradation of curcumin molecule. The aim of present work is to replace 1, 3-dicarbonyl moiety of curcumin by pyrazole and phenylpyrazole derivatives with a view to improving its stability and to investigate the role of substitution in N-phenylpyrazole curcumin on its antibacterial activity against both Gram-positive as well as Gram-negative bacteria. Methods: Pyrazole derivatives of curcumin were prepared by heating curcumin with phenyhydrazine/ substituted phenyhydrazine derivatives in AcOH. The residue was purified by silica gel column chromatography. Structures of purified compounds were confirmed by 1H NMR and Mass spectroscopy. The synthesized compounds were evaluated for their antibacterial activity by the microdilution broth susceptibility test method against gram positive (S. aureus) and gram negative (E. coli). Results: Effects of substitution in N-phenylpyrazole curcumin derivatives against S. aureus and E. coli were studied. The most active N-(3-Nitrophenylpyrazole) curcumin (12) exhibits twenty-fold more potency against S. aureus (MIC: 10μg/mL)) and N-(2-Fluoroophenylpyrazole) curcumin (5) fivefold more potency against E. coli (MIC; 50 μg/mL) than N-phenylpyrazole curcumin (4). Whereas, a remarkable decline in anti-bacterial activity against S. aureus and E. coli was observed when electron donating groups were incorporated in N-phenylpyrazole curcumin (4). Comparative studies of synthesized compounds suggest the effects of electron withdrawing and electron donating groups on unsubstituted phenylpyrazole curcumin (4). Conclusion: The structure-activity relationship (SAR) results indicated that the electron withdrawing and electron donating at N-phenylpyrazole curcumin played key roles for their bacterial inhibitory effects. The results of the antibacterial evaluation showed that the synthesized pyrazole derivatives of curcumin displayed moderate to very high activity in S. aureus. In conclusion, the series of novel curcumin derivatives were designed, synthesized and tested for their antibacterial activities against S. aureus and E. coli. Among them, N-(3-Nitrophenylpyrazole curcumin; 12) was most active against S. aureus (Gram-positive) and N-(2-Fluoroophenylpyrazole) curcumin (5) against E. coli (Gram-negative) bacteria.

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Quality Improvement of the DNA extracted by boiling method in Gram negative bacteria

International Journal of Bioassays ◽

10.21746/ijbio.2017.04.004 ◽

2017 ◽

Vol 6 (04) ◽

pp. 5347 ◽

Cited By ~ 3

Author(s):

Omar B. Ahmed* ◽

Anas S. Dablool

Keyword(s):

Dna Extraction ◽

Control Method ◽

Extraction Procedure ◽

Cost Effective ◽

High Yield ◽

Gram Negative Bacteria ◽

Bacterial Dna ◽

Gram Negative ◽

E Coli

Several methods of Deoxyribonucleic acid (DNA) extraction have been applied to extract bacterial DNA. The amount and the quality of the DNA obtained for each one of those methods are variable. The study aimed to evaluate bacterial DNA extraction using conventional boiling method followed by alcohol precipitation. DNA extraction from Gram negative bacilli was extracted and precipitated using boiling method with further precipitation by ethanol. The extraction procedure performed using the boiling method resulted in high DNA yields for both E. coli and K. pneumoniae bacteria in (199.7 and 285.7μg/ml, respectively) which was close to control method (229.3 and 440.3μg/ml). It was concluded that after alcohol precipitation boiling procedure was easy, cost-effective, and applicable for high-yield quality of DNA in Gram-negative bacteria.

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Clonal Clusters, Molecular Resistance Mechanisms and Virulence Factors of Gram-Negative Bacteria Isolated from Chronic Wounds in Ghana

Antibiotics ◽

10.3390/antibiotics10030339 ◽

2021 ◽

Vol 10 (3) ◽

pp. 339

Author(s):

Denise Dekker ◽

Frederik Pankok ◽

Thorsten Thye ◽

Stefan Taudien ◽

Kwabena Oppong ◽

...

Keyword(s):

Molecular Epidemiology ◽

Virulence Factors ◽

Iron Uptake ◽

Chronic Wounds ◽

Sub Saharan Africa ◽

Gram Negative Bacteria ◽

Beta Lactamase ◽

Secretion Systems ◽

Gram Negative ◽

E Coli

Wound infections are common medical problems in sub-Saharan Africa but data on the molecular epidemiology are rare. Within this study we assessed the clonal lineages, resistance genes and virulence factors of Gram-negative bacteria isolated from Ghanaian patients with chronic wounds. From a previous study, 49 Pseudomonas aeruginosa, 21 Klebsiellapneumoniae complex members and 12 Escherichia coli were subjected to whole genome sequencing. Sequence analysis indicated high clonal diversity with only nine P. aeruginosa clusters comprising two strains each and one E. coli cluster comprising three strains with high phylogenetic relationship suggesting nosocomial transmission. Acquired beta-lactamase genes were observed in some isolates next to a broad spectrum of additional genetic resistance determinants. Phenotypical expression of extended-spectrum beta-lactamase activity in the Enterobacterales was associated with blaCTX-M-15 genes, which are frequent in Ghana. Frequently recorded virulence genes comprised genes related to invasion and iron-uptake in E. coli, genes related to adherence, iron-uptake, secretion systems and antiphagocytosis in P. aeruginosa and genes related to adherence, biofilm formation, immune evasion, iron-uptake and secretion systems in K. pneumonia complex. In summary, the study provides a piece in the puzzle of the molecular epidemiology of Gram-negative bacteria in chronic wounds in rural Ghana.

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Bacterial Diversity, Antibiotic Resistance, and the Risk of Lower Limb Amputation in Younger and Older Individuals With Diabetic Foot Infection

The International Journal of Lower Extremity Wounds ◽

10.1177/1534734621992290 ◽

2021 ◽

pp. 153473462199229

Author(s):

Stefan Dörr ◽

Ann-Kathrin Holland-Letz ◽

Gregor Weisser ◽

Apostolos Chatzitomaris ◽

Ralf Lobmann

Keyword(s):

Bacterial Diversity ◽

Diabetic Foot ◽

Care Center ◽

Bacterial Load ◽

Limb Amputation ◽

Older Individuals ◽

Foot Care ◽

Gram Positive ◽

Gram Negative ◽

Diabetic Foot Infection

Introduction A diabetic foot infection (DFI) contributes to high mortality and morbidity in diabetics due to its often rapid progressive and intricately treatable infection. DFIs are usually a polymicrobial infection and characterizing the entire bacterial load is still challenging. Prompt and effective treatment of DFI is nevertheless mandatory to safe limbs and lives. It is therefore crucial to know the local pathogen spectrum and its antibiotic susceptibility. Methods and Material For a 12-month period, we investigated 353 individuals with infected diabetic foot ulcer, their bacterial diversity, and antimicrobial susceptibility at fist-time visit in a Diabetic Foot Care Center in southern Germany. Results Cultures yielded 888 species, most of them gram-positive cocci (primary Staphylococcus aureus). The gram-negative sector was mainly formed by Pseudomonas aeruginosa and Enterobacteriacae. Because the prevalence of multiresistant species was surprisingly low (0.9% of isolated strains), we suggest penicillins with β-lactamase inhibitor in case of gram-positive–dominated infection or piperacillin/tazobactam or rather carbapenems with equal efficacy when gram-negative species are involved.

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