1.In-vitro determination of minimum inhibitory concentration (MIC) and contact time of povidone-iodine against Staphylococcus aureus and Klebsiella aerogenes using micro suspension test, colorimetric resazurin microplate assay, and Dey Engley neutralizer assay.
Azita Racquel G. LACUNA ; Micaella C. DATO ; Loisse Mikaela M. LOTERIO ; Geraldine B. DAYRIT ; Sharon Yvette Angelina M. VILLANUEVA ; Maria Margarita M. LOTA
Acta Medica Philippina 2025;59(4):113-124
BACKGROUND AND OBJECTIVE
The human nasal passages host major human pathogens. Recent research suggests that the microbial communities inhabiting the epithelial surfaces of the nasal passages play a key factor in maintaining a healthy microenvironment by affecting both resistance to pathogens and immunological responses. Colonization of the nasal cavity by different pathogens such as Staphylococcus aureus and Klebsiella aerogenes, is associated with a higher postoperative infection morbidity. Povidone-iodine (PVP-I) as an antiseptic has been proven to display high antibacterial, antiviral, and antifungal properties even at low concentrations, and was shown to be effective in the control of infections to limit their impact and spread. It can be used as a topical antiseptic for skin decontamination and wound management, as a nasal spray, or as a gargle. There are different methods in testing the efficacy of potential antimicrobial suspensions. This study aimed to determine the concentration of PVP-I that is most effective in nasal decolonization using microsuspension test and colorimetric minimum inhibitory concentration (MIC) determination assays, resazurin microtiter assay (REMA), and Dey-Engley (D/E) neutralizer assay. The findings of this study will contribute to knowledge regarding the intended use of PVP-I in microbial control, particularly in bacterial infections.
METHODSSeveral dilutions (2.0%, 1.0%, 0.5%, 0.25%, 0.1% and 0.09%) of commercially bought 10% (10 mg per 100 ml) povidone-iodine were prepared and tested against a standardized inoculum (1x105) of Staphylococcus aureus and Klebsiella aerogenes at different contacttimes (5 seconds, 10 seconds, 30 seconds, 1 minute, and 5 minutes). Microdilution suspension test was performed to determine the log reduction per variable, while REMA and D/E neutralizer assay were used to determine the MIC. A value of greater than or equal to 5 log reduction was considered effective for microdilution suspension test. Estimates of agreement statistics were used to interpret the results of the assay in which the overall percent agreement (OPA), positive percent agreement (PPA), negative percent agreement (NPA), and Cohen’s kappa statistics were calculated.
RESULTSPovidone-iodine concentration of 0.25% exhibited ?5 log reduction against K. aerogenes at the minimum contact time of 5 seconds. On the other hand, a slightly higher PVP-I concentration was required to achieve ?5 log reduction for S. aureus at 0.5% concentration and a minimum contact time of 1 minute. There was an observed concordance of the results of REMA and D/E neutralizer as MIC colorimetric indicators, which yielded an overall test percent agreement of 90.30% (95% CI: 84.73–94.36), and a strong level of agreement (? = 0.8, pCONCLUSION
Low povidone-iodine concentrations (i.e., 0.5% against S. aureus and 0.25% against K. aerogenes) were observed to have bactericidal activity of at least 5 log reduction as rapid as the minimum contact time of 5 seconds. Furthermore, D/E and REMA, as colorimetric indicators, had comparable performance (OPA = 90.30%; ? = 0.8, p
Human
;
Bacteria
;
Povidone-iodine
;
Microbial Sensitivity Tests
;
Anti-infective Agents, Local
;
Enterobacter Aerogenes
;
Staphylococcus Aureus
2.Surveillance Culture of Carbapenemase-Producing Enterobacteriaceae in a Tertiary-Care Hospital.
Annals of Clinical Microbiology 2018;21(1):8-11
BACKGROUND: Carbapenem-resistant Enterobacteriaceae (CRE) are increasingly being reported throughout the world, which is a significant problem for patient treatment and infection control. Carbapenem-resistance in Enterobacteriaceae is mainly due to carbapenem-hydrolyzing β-lactamase, which tends to spread through genetic mobile elements. Therefore, the detection of carbapenemase-producing Enterobacteriaceae (CPE) carriers is particularly important for the prevention and epidemiological monitoring of these infections. In this study, we performed surveillance cultures for CPE in patients admitted to the hospital and evaluated the prevalence of CPE. METHODS: Stool cultures were obtained from a total of 228 patients at our tertiary-care hospital between March and May 2017. Stool specimens were inoculated on ChromID CARBA agar (bioMérieux, France) and incubated for 18-24 hours. Suspicious colonies with pink or bluish-green color were screened for CPE by the modified Hodge test (MHT) and carbapenemase inhibition test (CIT). We performed PCR to detect five carbapenemase genes, bla(KPC), bla(IMP), bla(VIM), bla(NDM), and bla(OXA-48). RESULTS: Among 228 isolates, seven were suspicious for CPE: four Klebsiella pneumoniae, one Escherichia coli, one Enterobacter aerogenes, and one Serratia marcescens. Two K. pneumoniae isolates showed positive reactions in both the modified Hodge test and inhibition test with phenylboronic acid. By PCR, bla(KPC) was identified in these two K. pneumoniae isolates. CONCLUSION: Our results showed a very low prevalence (2/228, 0.9%) of CPE in our tertiary-care hospital based on surveillance culture in a recent three month period.
Agar
;
Enterobacter aerogenes
;
Enterobacteriaceae*
;
Epidemiological Monitoring
;
Escherichia coli
;
Humans
;
Infection Control
;
Klebsiella pneumoniae
;
Pneumonia
;
Polymerase Chain Reaction
;
Prevalence
;
Serratia marcescens
3.Performance of MALDI Biotyper for Species Identification of Carbapenem-Resistant Enterobacteriaceae by Media Types and Incubation Time.
Young Eun CHO ; Byoung Hu CHOI ; Jeonghyun CHANG ; Heungsup SUNG ; Mi Na KIM
Journal of Laboratory Medicine and Quality Assurance 2018;40(3):155-160
BACKGROUND: This study was conducted to evaluate the impact of the media type used for direct identification of colonies on the surveillance culture of carbapenem-resistant Enterobacteriaceae (CRE) by matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). METHODS: CRE surveillance culture isolates were subjected to species identification using the MALDI Biotyper (Bruker Daltonics, Germany) for 2 months starting in March 2017. Four types of media were evaluated: blood agar (BA), Mueller Hinton agar (MH), MacConkey agar (Mac), and MacConkey agar containing imipenem of 1 µg/mL (IMP-Mac). CRE-like colonies on IMP-Mac and their subculture colonies on the other media were tested after overnight incubation and extended incubation for one additional day. The percent identification and score value were analyzed for each media types and incubation time when the identification was correct at the genus level. RESULTS: A total of 117 isolates were identified as 84 Klebsiella pneumoniae, 12 Escherichia coli, 9 Enterobacter cloacae, 5 Klebsiella oxytoca, 4 Enterobacter aerogenes, and 2 Raoultella ornithinolytica. The successful identification rates (SIR) for BA and MH were 98.3% and 97.4% (P=0.9), respectively, while those for Mac and IMP-Mac were 82.1% (P < 0.001) and 70.9% (P < 0.001), respectively. After extended incubation, SIRs were decreased to 96.6%, 96.6% (P=1.0), 61.5% (P < 0.001), and 58.1% (P < 0.001) on BA, MH, Mac, and IMP-Mac, respectively. The average score values were significantly lower for Mac (2.017±0.22) and IMP-Mac (1.978±0.24) than for BA (2.213±0.16) (P < 0.001). CONCLUSIONS: The low performance of the MALDI Biotyper applied directly to the colonies grown on Mac or IMP-Mac indicates that subculture on BA or MH is preferable before identification by MALDI-TOF MS.
Agar
;
Enterobacter aerogenes
;
Enterobacter cloacae
;
Enterobacteriaceae*
;
Escherichia coli
;
Imipenem
;
Klebsiella oxytoca
;
Klebsiella pneumoniae
;
Mass Spectrometry
;
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization*
4.Tracheal Infection Resulting from High Endotracheal Tube Cuff Pressure in an Unconscious Patient with Brain Trauma.
Ja Myoung LEE ; In Sung PARK ; Chul Hee LEE ; Kwang Ho LEE ; Dong Hyun CHUN ; Ji yoon KIM ; Young Seok LEE
Korean Journal of Neurotrauma 2018;14(2):155-158
Deep neck infections (DNIs) are mainly caused by dental caries, tonsillitis, and pharyngitis; however, DNIs can also occur after head and neck trauma. A 79-year-old male patient underwent a craniectomy due to an acute subdural hematoma. The patient was unconscious and continued to have a fever, but no clear cause was found. On postoperative day 9, he suddenly showed redness and swelling on the anterior neck. Enhanced computed tomography of the pharynx revealed tracheal necrosis and an abscess in the surrounding area. An incision and drainage were performed and Enterobacter aerogenes and E. faecalis were identified. The infection was controlled after antibiotic treatment. High endotracheal tube cuff pressure was suspected as the cause of the tracheal infection. Although DNIs are difficult to predict in patients who cannot report their symptoms due to unconsciousness, prevention and rapid diagnosis are important, as DNIs have serious side effects.
Abscess
;
Aged
;
Brain Injuries*
;
Brain*
;
Dental Caries
;
Diagnosis
;
Drainage
;
Enterobacter aerogenes
;
Fever
;
Head
;
Hematoma, Subdural, Acute
;
Humans
;
Intubation, Intratracheal
;
Male
;
Neck
;
Necrosis
;
Palatine Tonsil
;
Pharyngitis
;
Pharynx
;
Tonsillitis
;
Unconsciousness
5.Detection of mcr-1 Plasmids in Enterobacteriaceae Isolates From Human Specimens: Comparison With Those in Escherichia coli Isolates From Livestock in Korea.
Eun Jeong YOON ; Jun Sung HONG ; Ji Woo YANG ; Kwang Jun LEE ; Hyukmin LEE ; Seok Hoon JEONG
Annals of Laboratory Medicine 2018;38(6):555-562
BACKGROUND: The emerging mobile colistin resistance gene, mcr-1, is an ongoing worldwide concern and an evaluation of clinical isolates harboring this gene is required in Korea. We investigated mcr-1-possessing Enterobacteriaceae among Enterobacteriaceae strains isolated in Korea, and compared the genetic details of the plasmids with those in Escherichia coli isolates from livestock. METHODS: Among 9,396 Enterobacteriaceae clinical isolates collected between 2010 and 2015, 1,347 (14.3%) strains were resistant to colistin and those were screened for mcr-1 by PCR. Colistin minimum inhibitory concentrations (MICs) were determined by microdilution, and conjugal transfer of the mcr-1-harboring plasmids was assessed by direct mating. Whole genomes of three mcr-1-positive Enterobacteriaceae clinical isolates and 11 livestock-origin mcr-1-positive E. coli isolates were sequenced. RESULTS: Two E. coli and one Enterobacter aerogenes clinical isolates carried carried IncI2 plasmids harboring mcr-1, which conferred colistin resistance (E. coli MIC, 4 mg/L; E. aerogenes MIC, 32 mg/L). The strains possessed the complete conjugal machinery except for E. aerogenes harboring a truncated prepilin peptidase. The E. coli plasmid transferred more efficiently to E. coli than to Klebsiella pneumoniae or Enterobacter cloacae recipients. Among the three bacterial hosts, the colistin MIC was the highest for E. coli owing to the higher mcr-1-plasmid copy number and mcr-1 expression levels. Ten mcr-1-positive chicken-origin E. coli strains also possessed mcr-1-harboring IncI2 plasmids closely related to that in the clinical E. aerogenes isolate, and the remaining one porcine-origin E. coli possessed an mcr-1-harboring IncX4 plasmid. CONCLUSIONS: mcr-1-harboring IncI2 plasmids were identified in clinical Enterobacteriaceae isolates. These plasmids were closely associated with those in chicken-origin E. coli strains in Korea, supporting the concept of mcr-1 dissemination between humans and livestock.
Colistin
;
Enterobacter aerogenes
;
Enterobacter cloacae
;
Enterobacteriaceae*
;
Escherichia coli*
;
Escherichia*
;
Genome
;
Humans*
;
Klebsiella pneumoniae
;
Korea*
;
Livestock*
;
Microbial Sensitivity Tests
;
Plasmids*
;
Polymerase Chain Reaction
6.Analysis of Microorganisms and Antibiotics Susceptibility in Dacryocystitis.
Chang Hoon LEE ; Tae Eun LEE ; In Cheon YOU ; Nam Chun CHO ; Min AHN
Journal of the Korean Ophthalmological Society 2017;58(9):1017-1022
PURPOSE: This article analyzes the microorganisms and antibiotics susceptibility in dacryocystitis. METHODS: In this study, patients who were diagnosed with acute and chronic dacryocystitis with nasolacrimal duct obstruction were selected and underwent endoscopic endonasal dacryocystorhinostomy. Cultures were obtained from the lacrimal sac during operation from January 2008 to January 2016, and were used to analyze the microorganisms and antibiotics susceptibility. RESULTS: The 67 patients, 9 were diagnosed with acute dacryocystitis and 58 were diagnosed with chronic dacryocystitis. Among them, 64 cases showed bacterial growth (95.5%). The most frequently detected bacteria was Staphylococcus epidermidis (S. epidermidis) (33.8%), followed by Staphylococcus aureus (S. aureus) (25.4%) and Enterobacter aerogenes (18.3%). S. epidermidis had the most powerful resistance to ciprofloxacin compared to the other bacteria (58.3%, p = 0.02). Except for S. epidermidis and S. aureus, the other bacteria responded to ciprofloxacin and gentamycin. CONCLUSIONS: As a causative microorganism of dacryocystitis, S. epidermidis is becoming more prominent, and it is thought that S. epidermidis may be resistant to quinolones (i.e., broad-spectrum antibiotics). This resistance might be increasing the percentage of present S. epidermidis when viewed as a causal pathogen in dacryocystitis.
Anti-Bacterial Agents*
;
Bacteria
;
Ciprofloxacin
;
Dacryocystitis*
;
Dacryocystorhinostomy
;
Enterobacter aerogenes
;
Gentamicins
;
Humans
;
Nasolacrimal Duct
;
Quinolones
;
Staphylococcus aureus
;
Staphylococcus epidermidis
7.Anti-inflammatory and Anti-bacterial Effects of Aloe vera MAP against Multidrug-resistant Bacteria.
Natural Product Sciences 2017;23(4):286-290
Multidrug-resistant Acinetobacter baumannii and Pseudomonas aeruginosa are highly dangerous nosocomial pathogens, cause the symptoms of skin infections, pressure sores, sepsis, blood stream and wound infections. Unfortunately, these pathogens are immune to the most common antibiotics, such as, carbapenem, aminoglycoside and fluoroquinolone. Therefore, it is imperative that new and effective antibiotics be developed. In the present study, the antimicrobial effects of Aloe vera MAP (modified Aloe polysaccharide) on Staphylococcus aureus and Bacillus subtilis, Escherichia coli and Enterobacter aerogenes, and clinical Pseudomonas aeruginosa and clinical Acinetobacter baumannii were comprehensibly investigated. Prior to the growth inhibition effect measurement and antibiotic disc diffusion assay on gram-positive and gram-negative bacteria and selected multidrug-resistant Pseudomonas aeruginosa and Acinetobacter baumannii, antimicrobial resistance screening was performed for the multidrug-resistant bacteria obtained from clinical isolates. The results for showed the Aloe vera MAP had a concentration-dependent effect on all of examined bacteria, particularly on Pseudomonas aeruginosa. Anti-inflammatory and anti-oxidant experiments were also performed dose dependently effects to confirm the beneficial physiological effects of Aloe vera MAP.
Acinetobacter baumannii
;
Aloe*
;
Anti-Bacterial Agents
;
Bacillus subtilis
;
Bacteria*
;
Diffusion
;
Enterobacter aerogenes
;
Escherichia coli
;
Gram-Negative Bacteria
;
Mass Screening
;
Pressure Ulcer
;
Pseudomonas aeruginosa
;
Rivers
;
Sepsis
;
Skin
;
Staphylococcus aureus
;
Wound Infection
8.Diagnostic Value of Nasopharyngeal Aspirates in Children with Lower Respiratory Tract Infections.
Ai-Zhen LU ; Peng SHI ; Li-Bo WANG ; Li-Ling QIAN ; Xiao-Bo ZHANG
Chinese Medical Journal 2017;130(6):647-651
BACKGROUNDThe accuracy of nasopharyngeal aspirate (NPA) specimens in detecting lower respiratory pathogens remains controversial. The objective of this study was to evaluate the diagnostic accuracy of aspirates (NPAs) specimen in lower respiratory tract infections (LRTIs) in children.
METHODSThe prospective study was designed to collect the data of paired NPAs and bronchoalveolar lavage fluids from children with acute LRTIs from January 2013 to December 2015. All specimens were subjected to pathogen detection: bacterial detection by culture, Mycoplasma pneumoniae (Mp) detection by polymerase chain reaction assay and virus (influenza A and B viruses, parainfluenza virus [PIV] Types 1 and 3, respiratory syncytial virus, and adenovirus) detection by immunofluorescence assay. The diagnostic accuracy analysis of NPAs was stratified by age ≤3 years (n = 194) and >3 years (n = 294).
RESULTSWe collected paired specimens from 488 children. The positive rate of pathogen was 61.6%. For Streptococcus pneumoniae, NPA culture had the specificity of 89.9% and negative predictive value of 100% in age ≤3 years, the specificity of 97.2% and negative predictive value of 98.9% in age >3 years. For Mp, the positive predictive values of NPA was 77.4% in children ≤3 years, and 89.1% in children >3 years. For PIV III, NPA specimen had the specificity of 99.8% and negative predictive value of 96.5% in children ≤3 years. For adenovirus, NPA had the specificity of 97.8% and negative predictive value of 98.4% in age ≤3 years, the specificity of 98.9% and negative predictive value of 99.3% in age >3 years.
CONCLUSIONSNPAs are less invasive diagnostic respiratory specimens, a negative NPA result is helpful in "rule out" lower airway infection; however, a positive result does not reliably "rule in" the presence of pathogens.
Acinetobacter baumannii ; isolation & purification ; pathogenicity ; Adolescent ; Child ; Child, Preschool ; Clinical Laboratory Techniques ; methods ; Enterobacter aerogenes ; isolation & purification ; pathogenicity ; Escherichia coli ; isolation & purification ; pathogenicity ; Female ; Haemophilus influenzae ; isolation & purification ; pathogenicity ; Humans ; Infant ; Male ; Nasopharynx ; microbiology ; Prospective Studies ; Pseudomonas aeruginosa ; isolation & purification ; pathogenicity ; Respiratory Tract Infections ; diagnosis ; microbiology ; Sensitivity and Specificity ; Staphylococcus aureus ; isolation & purification ; pathogenicity ; Streptococcus pneumoniae ; isolation & purification ; pathogenicity
9.Carbapenem-resistant Enterobacteriaceae: Prevalence and Risk Factors in a Single Community-Based Hospital in Korea.
Hyo Jin LEE ; Jae Ki CHOI ; Sung Yeon CHO ; Si Hyun KIM ; Sun Hee PARK ; Su Mi CHOI ; Dong Gun LEE ; Jung Hyun CHOI ; Jin Hong YOO
Infection and Chemotherapy 2016;48(3):166-173
BACKGROUND: Carbapenemase-producing Enterobacteriaceae (CPE) are Gram-negative bacteria with increasing prevalence of infection worldwide. In Korea, 25 cases of CPE isolates were reported by the Korea Centers for Disease Control and Prevention in 2011. Most CPE cases were detected mainly at tertiary referral hospitals. We analyzed the prevalence and risk factors for carbapenem-resistant Enterobacteriaceae (CRE) in a mid-sized community-based hospital in Korea. MATERIALS AND METHODS: We retrospectively analyzed all consecutive episodes of Enterobacteriaceae in a mid-sized community-based hospital from January 2013 to February 2014. CRE was defined as organisms of Enterobacteriaceae showing decreased susceptibility to carbapenems. Risk factors for CRE were evaluated by a case–double control design. Carbapenemase was confirmed for CRE using a combined disc test. RESULTS: During 229,710 patient-days, 2,510 Enterobacteriaceae isolates were obtained. A total of 41 (1.6%) CRE isolates were enrolled in the study period. Thirteen species (31.7%) were Enterobacter aerogenes, 8 (19.5%) Klebsiella pneumoniae, 5 (12.2%) Enterobacter cloacae, and 15 other species of Enterobacteriaceae, respectively. Among the 41 isolates, only one (2.4%) E. aerogenes isolate belonged to CPE. For evaluation of risk factors, a total of 111 patients were enrolled and this included 37 patients in the CRE group, 37 in control group I (identical species), and 37 in control group II (different species). Based on multivariate analysis, regularly visiting the outpatient clinic was a risk factor for CRE acquisition in the control group I (P = 0.003), while vascular catheter and Charlson comorbidity index score ≥ 3 were risk factors in control group II (P = 0.010 and 0.011, each). Patients with CRE were more likely to experience a reduced level of consciousness, use a vasopressor, be under intensive care, and suffer from acute kidney injury. However, CRE was not an independent predictor of mortality compared with both control groups. CONCLUSION: In conclusion, the prevalence of CRE was higher than expected in a mid-sized community-based hospital in Korea. CRE should be considered when patients have a vascular catheter, high comorbidity score, and regular visits to the outpatient clinic. This study suggests the need for appropriate prevention efforts and constant attention to CRE infection control in a mid-sized community-based hospital.
Acute Kidney Injury
;
Ambulatory Care Facilities
;
Carbapenems
;
Centers for Disease Control and Prevention (U.S.)
;
Comorbidity
;
Consciousness
;
Critical Care
;
Drug Resistance
;
Enterobacter aerogenes
;
Enterobacter cloacae
;
Enterobacteriaceae*
;
Gram-Negative Bacteria
;
Humans
;
Infection Control
;
Klebsiella pneumoniae
;
Korea*
;
Mortality
;
Multivariate Analysis
;
Prevalence*
;
Retrospective Studies
;
Risk Factors*
;
Tertiary Care Centers
;
Vascular Access Devices
10.Antibacterial activity of some actinomycetes from Tamil Nadu, India.
Pachaiyappan Saravana KUMAR ; John Poonga Preetam RAJ ; Veeramuthu DURAIPANDIYAN ; Savarimuthu IGNACIMUTHU
Asian Pacific Journal of Tropical Biomedicine 2012;2(12):936-943
OBJECTIVETo isolate novel actinomycetes and to evaluate their antibacterial activity.
METHODSThree soil samples were collected from Vengodu (village) in Kanchipuram district, Tamil Nadu, India. Actinomycetes were isolated using serial dilution and plating method on actinomycetes isolation agar.
RESULTSTotally 35 isolates were obtained on the basis of colony characteristics on actinomycetes isolation agar. All the isolates were screened for antibacterial activity by cross streak method. Medium and optimization of day were done for the potent strains using Nathan's agar well diffusion method. Isolation of bioactive compounds from significant active isolates was done by using different media. The most active isolate VAS 10 was identified as Actinobacterium Loyola PBT VAS 10 (accession No. JF501398) using 16s rRNA sequence method. The hexane, ethyl acetate, dichloromethane and butanol extracts of VAS 10 were tested against bacteria. The maximum antibacterial activity was observed in dichloromethane and ethyl acetate; maximum zones of inhibition were observed against Enterococcus durans. The rRNA secondary structure and the restriction sites of Actinobacterium Loyola VAS 10 were predicted using Genebee and NEBCutter online tools respectively.
CONCLUSIONSThe present study showed that among the isolated actinomycetes, Actinobacterium Loyola PBT VAS 10 (accession No. JF501398) showed good antibacterial activity against the tested bacteria.
Actinobacteria ; chemistry ; isolation & purification ; physiology ; Animals ; Anti-Bacterial Agents ; pharmacology ; Antibiosis ; physiology ; Bacillus subtilis ; drug effects ; Enterobacter aerogenes ; drug effects ; Escherichia coli ; drug effects ; India ; Microbial Sensitivity Tests ; Phylogeny ; RNA, Ribosomal, 16S ; genetics ; Soil Microbiology ; Species Specificity ; Vibrio parahaemolyticus ; drug effects


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