scholarly journals Study on Microbiome of Chronic Non Healing Diabetic Ulcers with Special Reference to Biofilm and Multidrug Resistant Strains

Author(s):  
P Ramaprabha ◽  
CP Ramani ◽  
R Kesavan

Introduction: Diabetic Foot Ulcers (DFU) represent a silent epidemic and are the leading cause of 80% of non traumatic lower-limb amputations. Anaemia in diabetes may have adverse effects on systemic diseases and predict the progression of diabetes complications. Biofilms act as a mechanical barrier to antimicrobials and immune system cells and contribute to Multidrug Resistance (MDR). Aim: To determine the bacteriome and mycobiome of diabetic ulcers and the associated biofilm formation and anti-microbial resistance profile of the pathogens. Also, to determine the molecular characterisation of biofilm-forming resistant isolates by Polymerase Chain Reaction (PCR). Materials and Methods: This cross-sectional study was done on 150 diabetic patients with non healing ulcers and was chosen and studied from January-December 2019. Pus and tissue bit samples were processed as per standard microbiological procedures. Antimicrobial susceptibility test was performed as per Clinical and Laboratory Standards Institute (CLSI) guidelines. Biofilm formation was detected by the tissue culture plate method. Molecular characterisation of resistant pathogens was done by PCR. Variables were expressed as proportions or percentages. Results: Out of 150 diabetic patients, 17.3% of patients underwent amputation. A 90% of patients were associated with anaemia. Most ulcers were polymicrobial in nature. Predominantly isolated pathogens were Pseudomonas aeruginosa 37 (17.1%) and Staphylococcus aureus 33 (15.2%) among aerobic bacteria, Peptostreptococcus 10 (4.6%) among the anaerobes and Candida albicans 20 (9.2%) in fungus. Gram negative bacteria showed high sensitivity to piperacillin-tazobactam, meropenem, and gram positive cocci to vancomycin and linezolid. A 82% of bacterial isolates and 50% of fungal isolates were biofilm producers. Staphylococcus aureus was a strong biofilm producer. On molecular characterisation, blaCTX-M, blaTEM, blaNDM-1, blaOXA-23, mecA genes were present in resistant biofilm-forming isolates. Conclusion: Polymicrobial wound infection and biofilm formation in DFU confers antibiotic resistance and contributes to Multidrug Resistant Organisms (MDRO’s). However, proper antibiotic surveillance and antibiotic policy, and preventive strategies can curtail the spread of resistant strains.

2021 ◽  
Vol 6 (1) ◽  
pp. 1-8
Author(s):  
Kazmi A

Background: Nosocomial infections are great threat for hospitalized patients and Pseudomonas aeruginosa has emerged as one of the most potent nosocomial pathogens along with its diverse mechanisms to counter the various antimicrobial agents such as aminoglycosides, fluoroquinolones, monobactems, third generation cephalosporins, carbapenams and broad- spectrum penicillins. P. aeruginosa is one of the well-known pyogenic bacteria and is 3rd leading cause of pyogenic infections with the variable frequency depending on geographical region and clinical setting. P. aeruginosa is intimately associated with pyogenic nosocomial infections. Objectives: Since multidrug resistant strains of P. aeruginosa have posed serious threats and are frequently implicated in nosocomial infections. Methods: Pus swab were sampled under aseptic conditions and cultured on blood and Muller Hinton agar. Gram reaction, pigment production, Oxidase, indole reaction and citrate test were used to confirm isolate. Antibiotic susceptibility was performed b Kirby Bauer technique. Results compiled by us in this cross sectional study, showed 58 cases of P. aeruginosa out of 289 cases. This included 43% males and 57% females. Majority of the patients were of young age, with mean age 38 years. Antibiotic sensitivity revealed resistance to gentamicin was 50%, amikacin was 64%, ciprofloxacin and Aztronem 66%, Cefaparazone 69%, Tzaocin 71% and meropenem and sulzone was 79%. While Colistin and Ceftazidime were the most effective in 85% and 89% of cases respectively. The multidrug resistant strains of P. aeruginosa infections accounted for 32.76% of total P. aeruginosa infections. This study reveals high prevalence of multidrug resistant organisms at the set of our study. Based on this study, we suggest adopting the strategies to minimize the risk of nosocomial infections to slow down the rapidly growing multidrug resistance. These strategies may include, stricter antiseptic measures, fastening the recovery process and reducing the hospital stay and considering other alternates. Besides this, we would like to suggest the precise use of antibiotic susceptibility facility to reduce the nosocomial infection associated complications.


2018 ◽  
Vol 84 (15) ◽  
Author(s):  
Yufeng Zhang ◽  
Mengjun Cheng ◽  
Hao Zhang ◽  
Jiaxin Dai ◽  
Zhimin Guo ◽  
...  

ABSTRACT Treatment of infections caused by staphylococci has become more difficult because of the emergence of multidrug-resistant strains as well as biofilm formation. In this study, we observed the ability of the phage lysin LysGH15 to eliminate staphylococcal planktonic cells and biofilms formed by Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, and Staphylococcus hominis. All these strains were sensitive to LysGH15, showing reductions in bacterial counts of approximately 4 log units within 30 min after treatment with 20 μg/ml of LysGH15, and the MICs ranged from 8 μg/ml to 32 μg/ml. LysGH15 efficiently prevented biofilm formation by the four staphylococcal species at a dose of 50 μg/ml. At a higher dose (100 μg/ml), LysGH15 also showed notable disrupting activity against 24-h and 72-h biofilms formed by S. aureus and coagulase-negative species. In the in vivo experiments, a single intraperitoneal injection of LysGH15 (20 μg/mouse) administered 1 h after the injection of S. epidermidis at double the minimum lethal dose was sufficient to protect the mice. The S. epidermidis cell counts were 4 log units lower in the blood and 3 log units lower in the organs of mice 24 h after treatment with LysGH15 than in the untreated control mice. LysGH15 reduced cytokine levels in the blood and improved pathological changes in the organs. The broad antistaphylococcal activity exerted by LysGH15 on planktonic cells and biofilms makes LysGH15 a valuable treatment option for biofilm-related or non-biofilm-related staphylococcal infections. IMPORTANCE Most staphylococcal species are major causes of health care- and community-associated infections. In particular, Staphylococcus aureus is a common and dangerous pathogen, and Staphylococcus epidermidis is a ubiquitous skin commensal and opportunistic pathogen. Treatment of infections caused by staphylococci has become more difficult because of the emergence of multidrug-resistant strains as well as biofilm formation. In this study, we found that all tested S. aureus, S. epidermidis, Staphylococcus haemolyticus, and Staphylococcus hominis strains were sensitive to the phage lysin LysGH15 (MICs ranging from 8 to 32 μg/ml). More importantly, LysGH15 not only prevented biofilm formation by these staphylococci but also disrupted 24-h and 72-h biofilms. Furthermore, the in vivo efficacy of LysGH15 was demonstrated in a mouse model of S. epidermidis bacteremia. Thus, LysGH15 exhibits therapeutic potential for treating biofilm-related or non-biofilm-related infections caused by diverse staphylococci.


Pathogens ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 136
Author(s):  
Nicholas T. K. D. Dayie ◽  
Mary-Magdalene Osei ◽  
Japheth A. Opintan ◽  
Patience B. Tetteh-Quarcoo ◽  
Fleischer C. N. Kotey ◽  
...  

This cross-sectional study investigated the Staphylococcus aureus (S. aureus) and methicillin-resistant S. aureus (MRSA) nasopharyngeal carriage epidemiology in Accra approximately five years post-pneumococcal conjugate vaccines introduction in the country. Archived nasopharyngeal swabs collected from 410 children aged under five years old were bacteriologically cultured. The resultant S. aureus isolates were subjected to antimicrobial susceptibility testing and screening for carriage of the mecA and LukF-PV (pvl) genes, following standard procedures. The data obtained were analyzed with Statistical Products and Services Solutions (SPSS) using descriptive statistics and Chi square tests of associations. The isolated bacteria decreased across coagulase-negative Staphylococci (47.3%, n = 194), S. aureus (23.2%, n = 95), Diphtheroids (5.4%, n = 22), Micrococcus species (3.7%, n = 15), Klebsiella pneumoniae (3.2%, n = 13), Moraxella species and Citrobacter species (1.5% each, n = 6), Escherichia coli, Enterobacter species, and Pseudomonas species (0.9% each, n = 2). The MRSA carriage prevalence was 0.49% (n = 2). Individuals aged 37–48 months recorded the highest proportion of S. aureus carriage (32.6%, 31/95). Resistance of S. aureus to the antibiotics tested were penicillin G (97.9%, n = 93), amoxiclav (20%, n = 19), tetracycline (18.9%, n = 18), erythromycin (5.3%, n = 5), ciprofloxacin (2.1%, n = 2), gentamicin (1.1%, n = 1), cotrimoxazole, clindamycin, linezolid, and teicoplanin (0% each). No inducible clindamycin resistance was observed for the erythromycin-resistant isolates. Three (3.2%) of the isolates were multidrug resistant, of which 66.7% (2/3) were MRSA. The pvl gene was associated with 59.14% (55/93) of the methicillin-sensitive S. aureus (MSSA) isolates, but was not detected among any of the MRSA isolates.


2016 ◽  
Vol 82 (13) ◽  
pp. 3892-3899 ◽  
Author(s):  
Xiaohua Ye ◽  
Xiaolin Wang ◽  
Yanping Fan ◽  
Yang Peng ◽  
Ling Li ◽  
...  

ABSTRACTUse of antimicrobials in industrial food animal production is associated with the presence of multidrug-resistantStaphylococcus aureusamong animals and humans. The livestock-associated (LA) methicillin-resistantS. aureus(MRSA) clonal complex 9 (CC9) is associated with animals and related workers in Asia. This study aimed to explore the genotypic and phenotypic markers of LA-MRSA CC9 in humans. We conducted a cross-sectional study of livestock workers and controls in Guangdong, China. The study participants responded to a questionnaire and provided a nasal swab forS. aureusanalysis. The resulting isolates were assessed for antibiotic susceptibility, multilocus sequence type, and immune evasion cluster (IEC) genes. Livestock workers had significantly higher rates ofS. aureusCC9 (odds ratio [OR] = 30.98; 95% confidence interval [CI], 4.06 to 236.39) and tetracycline-resistantS. aureus(OR = 3.26; 95% CI, 2.12 to 5.00) carriage than controls. All 19S. aureusCC9 isolates from livestock workers were MRSA isolates and also exhibited the characteristics of resistance to several classes of antibiotics and absence of the IEC genes. Notably, the interaction analyses indicated phenotype-phenotype (OR = 525.7; 95% CI, 60.0 to 4,602.1) and gene-environment (OR = 232.3; 95% CI, 28.7 to 1,876.7) interactions associated with increased risk for livestock-associatedS. aureusCC9 carriage. These findings suggest that livestock-associatedS. aureusand MRSA (CC9, IEC negative, and tetracycline resistant) in humans are associated with occupational livestock contact, raising questions about the potential for occupational exposure to opportunisticS. aureus.IMPORTANCEThis study adds to existing knowledge by giving insight into the genotypic and phenotypic markers of LA-MRSA. Our findings suggest that livestock-associatedS. aureusand MRSA (CC9, IEC negative, and tetracycline resistant) in humans are associated with occupational livestock contact. Future studies should direct more attention to exploring the exact transmission routes and establishing measures to prevent the spread of LA-MRSA.


2020 ◽  
Author(s):  
Yingying Wang ◽  
Jialing Lin ◽  
Junli Zhou ◽  
Zhigang Han ◽  
Zhenjiang Yao

Abstract Background: Staphylococcus aureus ( S. aureus ), particularly methicillin-resistant Staphylococcus aureus (MRSA), remains the predominant cause of infections in drug users. This cross-sectional study aims to elucidate the prevalence, risk factors, phenotypic and molecular characteristics of S. aureus carriage among community-based drug users. Methods: All eligible drug users, with both injection and non-injection route of drug administration , were asked to complete questionnaires and collect nasal swabs by trained personal during the period between May and December 2017 in Guangzhou, China. Swabs were processed for identification of S. aureus . Antimicrobial susceptibility test and polymerase chain reaction assays were used to detect phenotypic and molecular characteristics for identified isolates. Univariate and multivariate logistic regression analyses were used to assess risk factors for S. aureus carriage. Results: Overall, 353 drug users were included in the study and the prevalence of S. aureus carriage was 15.01% (53/353). The prevalence of MRSA carriage was 6.80% (24/353). Cohabitation was a risk factor for S. aureus (adjusted OR=8.80, 95% CI: 1.89-40.99). The proportion of multidrug resistance was 54.72% for S. aureus isolates and most of these isolates were resistant to penicillin, erythromycin and clindamycin. Seventeen MRSA isolates were multidrug resistant. The results of clonal complexes (CCs) and sequence types (STs) for S. aureus were diverse. The three predominant types for CCs were CC5 (64.15%, 34/53), CC59 (11.32%, 6/53), and CC7 (7.55%, 4/53); and for STs were ST188 (20.75%, 11/53), ST5 (11.32%, 6/53), and ST59 (11.32%, 6/53). Conclusion: The prevalence of S. aureus nasal carriage was lower while the prevalence of MRSA carriage was moderate compared to previous studies. Phenotypic and molecular characteristics of S. aureus isolates, particularly MRSA isolates, revealed high proportions of antibiotic resistance, indicating the existence of cross-circulation, and implying high opportunity of virulence-related diseases. Decolonization and antibiotic stewardship might be implemented for drug users with MRSA carriage.


2021 ◽  
Vol 2 (1) ◽  
Author(s):  
Taqi Mohammed Jwad Taher ◽  
Firas Turki Rashed Sarray ◽  
Ammar Shimal Shwekh ◽  
Tariq Kuraidi Mareedh ◽  
Hasanain Faisal Ghazi

  Introduction: Urinary tract infection is a very prevalent disease among humans and it is highly presented among patients with diabetes mellitus. The main aim of the current study was to find out the commonest bacterial organisms causing urinary tract infection among a sample of diabetic and non-diabetic patients in Al-Kut city, Iraq. Methods: A cross-sectional study was conducted among 100 conveniently sampled patients suffering from urinary tract infections and attending Al-Karama Teaching hospital laboratory for urine culture between October and December 2019 were included in the study. The patients were consented to answer a special questionnaire containing data like patients' age, gender, and disease status (diabetic or non-diabetic). Results: From the 100 participated patients there were 29 (29%) with diabetes and the remaining 71(71%) from the sample were non-diabetics. The females represent the majority of the sample (67%) while males represented only (33%). The most common identified bacteria from this sample were Staphylococcus aureus (48%), Escherichia coli (24%), Klebsiella pneumoniae (17%), Enterococcus species (5%), and Pseudomonas aeruginosa (2%). The study result shows a significant association of being diabetic or not >0.001, 0.038with age and gender of the patients (p-value) respectively. While this association was non-significant when considering the type of bacteria between the two . While this association was non-significant when considering the groups (P-value=0.056).type of bacteria between the two groups (P-value=0.056). Conclusion: The urinary tract infection is frequently presented among young non-diabetic females and near half of the urine cultures showed the Staphylococcus aureus bacteria as the commonest cause of infection among them.


2020 ◽  
Vol 27 (01) ◽  
pp. 1-4
Author(s):  
Sadaf Razzak ◽  
Hina Faisal ◽  
Amir Bux Detho ◽  
Muhammad Asif Durrani ◽  
Fakhuruddin ◽  
...  

Objectives: D-Test as a tool to detect the frequency of clindamycin resistance in community acquired and hospital acquired methicillin resistant Staphylococcus aureus infections. Study Design: A cross-sectional study. Setting: Microbiology department of BMSI, JPMC, Karachi. Period: January 2015 till December 2015. Material & Methods: Pus samples from deep wounds, skin lesions, abscesses, postoperative wounds from surgical, medical wards and OPDs were collected. MRSA testing and susceptibility testing for antibiotics was done according to CLSI2014. The frequency of inducible clindamycin resistance was detected by D-Test of the CA-MRSA and HA-MRSA. Result: In a total of 402 S. aurous isolates, 253 (62.93%) were methicillin-sensitive and 149 (37.06%) were methicillin-resistant. Out of 149 MRSA, 106 (71.14%) were HA-MRSA and 43(28.85%) were CA-MRSA. Among the HA-MRSA, 63(59.8%) were resistant to clindamycin while with D-Test, it increased to 78(73.58%). Out of 43 CA-MRSA, 9 (21.6%) were clindamycin resistant, while with D-Test, the resistance to clindamycin increased to 13 (30.23%). Conclusion: Inducible clindamycin-resistant strains may lead to clindamycin treatment failure in patients with S. aureus infection. Therefore, D-test should be done in priority to detect inducible clindamycin resistance in S. aurous.


2019 ◽  
Author(s):  
Yingying Wang ◽  
Jialing Lin ◽  
Junli Zhou ◽  
Zhigang Han ◽  
Zhenjiang Yao

Abstract Background: Staphylococcus aureus (S. aureus), particularly methicillin-resistant Staphylococcus aureus (MRSA), remains the predominant cause of infections in drug users. The cross-sectional study aims to elucidate the prevalence, risk factors, phenotypic and molecular characteristics of S. aureus carriage among community-based drug users.Methods: All eligible drug users, both intravenous and oral drug users, were asked to complete questionnaires and collect nasal swabs during the period between May and December 2017 in Guangzhou, China. Swabs were processed for identification of S. aureus. Antimicrobial susceptibility test and polymerase chain reaction assays were used to detect phenotypic and molecular characteristics for identified isolates. Univariate and multivariate logistic regression analyses were used to assess risk factors for S. aureus carriage.Results: Overall, 353 drug users were included in the study and the prevalence of S. aureus carriage was 15.01% (53/353). The prevalence of MRSA carriage was 6.80% (24/353). Cohabitation was a risk factor for S. aureus (adjusted OR=8.80, 95% CI: 1.89-40.99). The proportion of multidrug resistance was 54.72% for S. aureus isolates and most of these isolates were resistant to penicillin, erythromycin and clindamycin. Seventeen MRSA isolates were multidrug resistant. The results of clonal complexes (CCs) and sequence types (STs) for S. aureus were diverse. The three predominant types for CCs were CC5 (64.15%, 34/53), CC59 (11.32%, 6/53), and CC7 (7.55%, 4/53); and for STs were ST188 (20.75%, 11/53), ST5 (11.32%, 6/53), and ST59 (11.32%, 6/53). Conclusion: The prevalence of S. aureus nasal carriage was lower when the prevalence of MRSA carriage was moderate. Phenotypic and molecular characteristics of S. aureus isolates, particularly MRSA isolates, revealed serious antibiotic resistance, indicating the existence of cross-circulation, and implying high opportunity of virulence-related diseases. Decolonization might be considered for drug users with MRSA carriage.


2019 ◽  
Vol 2019 ◽  
pp. 1-7 ◽  
Author(s):  
Laila Chaoui ◽  
RajaaAit Mhand ◽  
Fouad Mellouki ◽  
Naima Rhallabi

Nosocomial infections (NIs) are known worldwide and remain a major problem despite scientific and technical advances in the field of health. The severity of the infection depends on the characteristics of the microorganisms involved and the high frequency of resistant pathogens in the hospital environment. The aim of this study is to determine the distribution of pathogenic bacteria (and their resistance to antibiotics) that spread on hospital surfaces, more specifically, on those of various departments in the Provincial Hospital Center (PHC) of Mohammedia, Morocco. A cross-sectional study was conducted from March 2017 to April 2018. Samples were collected by swabbing the hospital surfaces, and the isolated bacteria were checked for their susceptibility to antibiotics by the Kirby–Bauer disk diffusion method following the standards of the Clinical and Laboratory Standards Institute (CLSI). Among 200 swab samples, 176 (88%) showed bacterial growth. Gram-negative isolates were predominant at 51.5% (101/196), while the Gram-positives were at 48.5% (95/196). The main isolates are Enterobacteria weighted at 31.6% (62/196), Staphylococcus aureus reaching 24% (47/196), Pseudomonas aeruginosa at 9.2% (18/196), and Acinetobacter spp. with 3.3% (6/196). Moreover, the antimicrobial susceptibility profile of the isolates showed that about 31.7% (32/101) of the Gram-negative isolates were found to be MDR. This resistance is also high among isolates of S. aureus of which 44.7% (20/47) were methicillin-resistant Staphylococcus aureus (MRSA). Contamination of hospital surfaces by MDR bacteria is a real danger to public health. The concept of environmental bacterial reservoir is a reality that requires strict compliance with current guidelines and recommendations for hand hygiene, cleaning, and disinfection of surfaces in hospitals.


Author(s):  
Dayane Otero Rodrigues ◽  
Laís da Paixao Peixoto ◽  
Erica Tatiane Mourao Barros ◽  
Julianne Rodrigues Guimaraes ◽  
Bruna Clemente Gontijo ◽  
...  

Aims: The hospital environment is an important reservoir of microorganisms, including multidrug-resistant pathogens, which can cause in-patient contamination and healthcare-related infections.  The objective of this study was to describe the epidemiology of bacterial contamination (contaminated sites, pathogen species and their antimicrobial susceptibility, and identifying of multidrug-resistant microorganisms - MDR) of inert hospital surfaces and medical equipment in two public hospitals in Northern Brazil. Methods: This was a cross-sectional study with 243 samples (n = 208, from Hospital A; and n = 35, from Hospital B) collected by friction with humidified swabs from inert surfaces and equipment. Sequentially the samples were cultivated and bacterial species were identified by culture-based methods and tested for their susceptibility through agar diffusion assay according to the Clinical and Laboratory Standards Institute (CLSI). Results: Most inert surfaces and equipment analyzed presented bacterial contamination (95.5%). Staphylococcus aureus was the main pathogen of clinical significance detected both in Hospital A (61.8%) and B (68.6%). Hospital A showed higher rates of isolated MDR bacteria than Hospital B, especially in the Adult Intensive Care Unit, which included methicillin-resistant Staphylococcus aureus (MRSA) (52.7%), Enterobacteria resistant to 4th generation cephalosporins (19.4%), and multidrug-resistant Pseudomonas aeruginosa (2.8%). Conclusion: The failures in the control of bacterial contamination of inert surfaces and equipment in the two hospitals analyzed reinforce the need for a revised protocol for cleaning and disinfection of the inert surfaces and equipment, and for regulation of antibiotic dispensing, mainly in the AICU of Hospital A, which was found to be a reservoir of MDR pathogens.


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