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.Prevalence of rectal carbapenem-resistant organism colonization among neonates admitted in the neonatal intensive care unit of the Philippine General Hospital
Krizia Joy A. Co ; Anna Lisa T. Ong-lim
Pediatric Infectious Disease Society of the Philippines Journal 2025;26(1):12-21
OBJECTIVE
To determine the prevalence of rectal colonization with carbapenem-resistant organisms (CRO) among PGH neonatal intensive care unit (NICU) patients.
METHODOLOGYA prospective single-center observational study conducted over a 1-month period included all NICU 3 and cohort area patients admitted on April 24, 2024. Rectal swabs were collected for multidrug-resistant organism (MDRO) screening and repeated weekly for 1 month while admitted. Swabs were inoculated on chromogenic media, and isolates were identified and tested for antimicrobial sensitivity by disk diffusion. Clinical characteristics and outcomes were collected for 30 days from initial MDRO screening. Descriptive statistics were used to summarize the data.
RESULTSThe point prevalence of CRO colonization was 37% (14 of 38) at initial screening. There were 14 incident colonizations, hence the 4-week period prevalence of CRO colonization was 72.5% (29 of 40). The patients were mostly very preterm, very low birth weight neonates, majority were tested within the first 2 weeks of life, and half were exposed to meropenem at initial screening. Nosocomial infection developed in 29% and 64%, and 30-day mortality rate was 8% and 21% among initially non-CRO-colonized and CRO-colonized patients respectively. Despite high CRO colonization, no culture-proven CRO infection was observed. Surveillance screening documented persistent CRO colonization in 37%, but no decolonization. Escherichia coli, Klebsiella spp. and Serratia spp. were the most common colonizers.
CONCLUSIONThe high prevalence of rectal CRO colonization in the NICU emphasizes the burden of antimicrobial resistance, but despite the high CRO colonization, no CRO infection was documented from the limited sample and study period.
Human ; Infant, Newborn ; Carbapenem-resistant Enterobacteriaceae ; Multidrug Resistance ; Drug Resistance, Multiple
3.Antimicrobial resistance profile of Escherichia coli isolated from raw chicken meat in a selected wet market in Manila City, Philippines
Lyder Kyle A. Dimaapi ; Angela Lorraine G. Dela cruz ; Roger Andrei D. Francisco ; Rei Gilian D. Noble ; Hayley Emerald G. Sabangan ; Azita Racquel Gavino-lacuna ; Maria Margarita M. Lota
Acta Medica Philippina 2025;59(9):107-122
BACKGROUND AND OBJECTIVE
Antimicrobial resistance (AMR) is a leading global public health concern as it resulted in more difficult-to-treat infections and fatalities. In the Philippines, drug-resistant E. coli, including multidrug-resistant (MDR), extended-spectrum beta-lactamase (ESBL)-producing, carbapenemase-producing carbapenem-resistant (CP-CR) E. coli, have been isolated from common food animals, increasing the risk of cross-contamination between humans, animals, and the environment. However, there is a lack of data on the distribution of E. coli in chicken meat in public wet markets. This study aims to describe the AMR profile of E. coli in raw chicken meat from retail stalls in a selected wet market in Manila City.
METHODSThis quantitative descriptive study characterized the AMR profile of E. coli isolated from 25 raw chicken meat samples from a wet market in Manila City. Antimicrobial susceptibility was determined through disk diffusion method against 23 antimicrobial agents in 16 antimicrobial classes. MDR E. coli were identified based on the resistance patterns. ESBL- and carbapenemase-producing capacities of the bacteria were tested through double disk synergy test and modified carbapenem inactivation method, respectively.
RESULTSTwenty-four out of 25 (96%) chicken samples contained E. coli isolates. Of these, 23 (96%) were classified as MDR. High resistance rates were observed against ampicillin (92%), tetracycline (88%), trimethoprim-sulfamethoxazole (83%), chloramphenicol (79%), ampicillin-sulbactam (75%), amoxicillin-clavulanic acid (67%), fosfomycin (67%), and streptomycin (54%). The majority of the E. coli isolates were still susceptible to a wide range of selected antimicrobial agents, including carbapenems (100%), ceftriaxone (100%), cefepime (100%), cefuroxime (96%), cefotaxime (96%), ceftazidime (96%), piperacillin-tazobactam (96%), aztreonam (96%), cefoxitin (92%), and nitrofurantoin (83%), among others. Meanwhile, none of the 24 isolated E. coli samples were classified as ESBL- and CP-CR E. coli.
CONCLUSIONAmong the 25 chicken samples, 24 E. coli colonies were isolated that exhibited 0% to 92% resistance rates against selected antimicrobial agents. Most isolates were classified as MDR, but none were considered ESBLand CP-CR E. coli. This study suggests that chickens in wet markets can potentially serve as reservoir hosts for drugresistance genes, which could transfer to other bacteria and contaminate humans, animals, and the environment within the food production and supply chain. These findings emphasize the need for AMR surveillance and strategies to combat AMR in the Philippines through the One Health approach.
Human ; Drug Resistance ; Drug Resistance, Multiple ; Carbapenemase ; Escherichia Coli
4.Antimicrobial resistance profile of Escherichia coli isolated from raw chicken meat in a selected wet market in Manila City, Philippines
Lyder Kyle A. Dimaapi ; Angela Lorraine G. Dela Cruz ; Roger Andrei D. Francisco ; Rei Gilian D. Noble ; Hayley Emerald G. Sabangan ; Azita Racquel Gavino-Lacuna ; Maria Margarita M. Lota
Acta Medica Philippina 2024;58(Early Access 2024):1-16
Background and Objective:
Antimicrobial resistance (AMR) is a leading global public health concern as it resulted in more difficult-to-treat infections and fatalities. In the Philippines, drug-resistant E. coli, including multidrug-resistant (MDR), extended-spectrum beta-lactamase (ESBL)-producing, carbapenemase-producing carbapenem-resistant (CP-CR) E. coli, have been isolated from common food animals, increasing the risk of cross-contamination between humans, animals, and the environment. However, there is a lack of data on the distribution of E. coli in chicken meat in public wet markets. This study aims to describe the AMR profile of E. coli in raw chicken meat from retail stalls in a selected wet market in Manila City.
Methods:
This quantitative descriptive study characterized the AMR profile of E. coli isolated from 25 raw chicken meat samples from a wet market in Manila City. Antimicrobial susceptibility was determined through disk diffusion method against 23 antimicrobial agents in 16 antimicrobial classes. MDR E. coli were identified based on the resistance patterns. ESBL- and carbapenemase-producing capacities of the bacteria were tested through double disk synergy test and modified carbapenem inactivation method, respectively.
Results:
Twenty-four out of 25 (96%) chicken samples contained E. coli isolates. Of these, 23 (96%) were classified as MDR. High resistance rates were observed against ampicillin (92%), tetracycline (88%), trimethoprim-sulfamethoxazole (83%), chloramphenicol (79%), ampicillin-sulbactam (75%), amoxicillin-clavulanic acid (67%), fosfomycin (67%), and streptomycin (54%). The majority of the E. coli isolates were still susceptible to a wide range of selected antimicrobial agents, including carbapenems (100%), ceftriaxone (100%), cefepime (100%), cefuroxime (96%), cefotaxime (96%), ceftazidime (96%), piperacillin-tazobactam (96%), aztreonam (96%), cefoxitin (92%), and nitrofurantoin (83%), among others. Meanwhile, none of the 24 isolated E. coli samples were classified as ESBL- and CP-CR E. coli.
Conclusion
Among the 25 chicken samples, 24 E. coli colonies were isolated that exhibited 0% to 92% resistance rates against selected antimicrobial agents. Most isolates were classified as MDR, but none were considered ESBLand CP-CR E. coli. This study suggests that chickens in wet markets can potentially serve as reservoir hosts for drugresistance genes, which could transfer to other bacteria and contaminate humans, animals, and the environment within the food production and supply chain. These findings emphasize the need for AMR surveillance and strategies to combat AMR in the Philippines through the One Health approach.
drug resistance
;
multi-drug resistance
;
drug resistance, multiple
;
carbapenemase
;
Escherichia coli
5.Antimicrobial activity of Ardisia serrata (Cavs.) Pers. ethanolic and aqueous leaf extract on the growth and biofilm formation of selected bacterial isolates
Patrick Josemaria DR. Altavas ; Alfonso Rafael G. Abaya ; Remo Vittorio Thaddeus D. Abella ; Danna Lee A. Acosta ; Angelica C. Aguilar ; Camille Anne V. Aguinaldo ; Katrina Loise L. Aguirre ; Catherine Therese C. Amante ; Karen B. Amora ; Glen Aldrix R. Anarna ; Rafael T. Andrada ; Gere Ganixon T. Ang ; Jeram Caezar R. Angobung ; Angelo V. Aquino II ; Dennielle Ann P. Arabis ; Hannah Luisa G. Awitan ; Mary Faith D. Baccay ; Chryz Angelo Jonathan B. Bagsic ; Tomas V. Baldosano Jr. ; Cecilia C. Maramba-Lazarte
Acta Medica Philippina 2024;58(18):91-97
Background:
Ardisia serrata (Aunasin) is an endemic Philippine plant of the family Primulaceae, with several studiesshowing the genus Ardisia as having potential antibacterial, antiangiogenic, cytotoxic, and antipyretic properties.
Objective:
This study aims to determine the antibacterial and antibiofilm-forming activity of Ardisia serrata ethanolic and aqueous extracts on Escherichia coli, Methicillin-Sensitive Staphylococcus aureus (MSSA), and Methicillin-Resistant Staphylococcus aureus (MRSA).
Methods:
This is an experimental study testing the activity against bacterial strains of E. coli, MSSA, and MRSA using ethanolic and aqueous extracts of A. serrata leaves. Microtiter susceptibility and biofilm inhibition assays were done with two-fold dilutions of the extract against the selected strains using spectrophotometry with optical density (OD) at 600 nm and 595 nm, respectively, to quantify bacterial growth and biofilm inhibition. The bacterial susceptibility and biofilm inhibition activity was reported as percent inhibition (PI). Minimum inhibitory concentration (MIC), and minimum biofilm inhibition concentration (MBIC) values were obtained using logarithmic regression of the PI values.
Results:
A. serrata ethanolic extracts showed weak growth inhibitory activity against MSSA and MRSA with minimum inhibitory concentration (MIC) values of 2.6192 and 3.2988 mg/mL, respectively, but no biofilm inhibition activity was noted, while the aqueous extracts exhibited negligible biofilm inhibition activity against MSSA and MRSA with minimum biofilm inhibition concentration (MBIC) values of 13.5972 and 8964.82 mg/mL, respectively, and with no growth inhibition activity. Both ethanolic and aqueous extracts showed no growth inhibition and biofilm inhibition activities against E. coli.
Conclusion
Staphylococcus aureus is susceptible to the bioactivity of the leaf extracts of A. serrata and has potential to be used as an antibacterial in the treatment of infectious diseases.
Methicillin-resistant Staphylococcus aureus
;
Escherichia coli
;
natural product
;
biological products
6.A case of Proteus Syndrome in a 12-year-old Filipino male
Bea Mergie T. Beltejar ; Maria Jasmin J. Jamora
Journal of the Philippine Dermatological Society 2024;33(Suppl 1):22-22
Proteus syndrome (PS) is a mosaic disorder characterized by asymmetric overgrowth of a variety of tissues. Diagnostic criteria established in 1999 emphasized the mosaic distribution of lesions, progressive course, and disproportionate overgrowth. We present a case of proteus syndrome in a 12-year-old Filipino male with 9 year-history of enlargement of the left foot with soft, non-tender mass on the sole with a brain-like surface. Skin punch biopsy of the mass showed cerebriform connective tissue nevi which is pathognomonic of PS.
PS is a very rare disease with prevalence of less than 1 in 1,000,000 live births. Management of PS is extremely challenging, owing to the combination of the individuality of each case, the severity of the disease, and the risks of complications from procedures. A multidisciplinary clinical approach is strongly recommended to obtain the best possible management plans for individual patients.
Human ; Male ; Child: 6-12 Yrs Old ; Proteus Syndrome ; Mosaicism
7.A Core Genome Multilocus Sequence Typing Scheme for Proteus mirabilis.
Sheng Lin CHEN ; Yu Tong KANG ; Yi He LIANG ; Xiao Tong QIU ; Zhen Jun LI
Biomedical and Environmental Sciences 2023;36(4):343-352
OBJECTIVE:
A core genome multilocus sequence typing (cgMLST) scheme to genotype and identify potential risk clonal groups (CGs) in Proteus mirabilis.
METHODS:
In this work, we propose a publicly available cgMLST scheme for P. mirabilis using chewBBACA. In total 72 complete P. mirabilis genomes, representing the diversity of this species, were used to set up a cgMLST scheme targeting 1,842 genes, 635 unfinished (contig, chromosome, and scaffold) genomes were used for its validation.
RESULTS:
We identified a total of 205 CGs from 695 P. mirabilis strains with regional distribution characteristics. Of these, 159 unique CGs were distributed in 16 countries. CG20 and CG3 carried large numbers of shared and unique antibiotic resistance genes. Nine virulence genes ( papC, papD, papE, papF, papG, papH, papI, papJ, and papK) related to the P fimbrial operon that cause severe urinary tract infections were only found in CG20. These CGs require attention due to potential risks.
CONCLUSION
This research innovatively performs high-resolution molecular typing of P. mirabilis using whole-genome sequencing technology combined with a bioinformatics pipeline (chewBBACA). We found that the CGs of P. mirabilis showed regional distribution differences. We expect that our research will contribute to the establishment of cgMLST for P. mirabilis.
Genome, Bacterial
;
Proteus mirabilis/genetics*
;
Multilocus Sequence Typing
;
Molecular Epidemiology
;
Genotype
8.Genotyping Characteristics of Human Fecal Escherichia coli and Their Association with Multidrug Resistance in Miyun District, Beijing.
Wei Wei ZHANG ; Xiao Lin ZHU ; Le Le DENG ; Ya Jun HAN ; Zhuo Wei LI ; Jin Long WANG ; Yong Liang CHEN ; Ao Lin WANG ; Er Li TIAN ; Bin CHENG ; Lin Hua XU ; Yi Cong CHEN ; Li Li TIAN ; Guang Xue HE
Biomedical and Environmental Sciences 2023;36(5):406-417
OBJECTIVE:
To explore the genotyping characteristics of human fecal Escherichia coli( E. coli) and the relationships between antibiotic resistance genes (ARGs) and multidrug resistance (MDR) of E. coli in Miyun District, Beijing, an area with high incidence of infectious diarrheal cases but no related data.
METHODS:
Over a period of 3 years, 94 E. coli strains were isolated from fecal samples collected from Miyun District Hospital, a surveillance hospital of the National Pathogen Identification Network. The antibiotic susceptibility of the isolates was determined by the broth microdilution method. ARGs, multilocus sequence typing (MLST), and polymorphism trees were analyzed using whole-genome sequencing data (WGS).
RESULTS:
This study revealed that 68.09% of the isolates had MDR, prevalent and distributed in different clades, with a relatively high rate and low pathogenicity. There was no difference in MDR between the diarrheal (49/70) and healthy groups (15/24).
CONCLUSION
We developed a random forest (RF) prediction model of TEM.1 + baeR + mphA + mphB + QnrS1 + AAC.3-IId to identify MDR status, highlighting its potential for early resistance identification. The causes of MDR are likely mobile units transmitting the ARGs. In the future, we will continue to strengthen the monitoring of ARGs and MDR, and increase the number of strains to further verify the accuracy of the MDR markers.
Humans
;
Escherichia coli/genetics*
;
Escherichia coli Infections/epidemiology*
;
Multilocus Sequence Typing
;
Genotype
;
Beijing
;
Drug Resistance, Multiple, Bacterial/genetics*
;
Anti-Bacterial Agents/pharmacology*
;
Diarrhea
;
Microbial Sensitivity Tests
9.Expression, purification, and characterization of the histidine kinase CarS from Fusobacterium nucleatum.
Zhuting LI ; Xian SHI ; Ruochen FAN ; Lulu WANG ; Tingting BU ; Wei ZHENG ; Xuqiang ZHANG ; Chunshan QUAN
Chinese Journal of Biotechnology 2023;39(4):1596-1608
Fusobacterium nucleatum is an opportunistic pathogenic bacterium that can be enriched in colorectal cancer tissues, affecting multiple stages of colorectal cancer development. The two-component system plays an important role in the regulation and expression of genes related to pathogenic resistance and pathogenicity. In this paper, we focused on the CarRS two-component system of F. nucleatum, and the histidine kinase protein CarS was recombinantly expressed and characterized. Several online software such as SMART, CCTOP and AlphaFold2 were used to predict the secondary and tertiary structure of the CarS protein. The results showed that CarS is a membrane protein with two transmembrane helices and contains 9 α-helices and 12 β-folds. CarS protein is composed of two domains, one is the N-terminal transmembrane domain (amino acids 1-170), the other is the C-terminal intracellular domain. The latter is composed of a signal receiving domain (histidine kinases, adenylyl cyclases, methyl-accepting proteins, prokaryotic signaling proteins, HAMP), a phosphate receptor domain (histidine kinase domain, HisKA), and a histidine kinase catalytic domain (histidine kinase-like ATPase catalytic domain, HATPase_c). Since the full-length CarS protein could not be expressed in host cells, a fusion expression vector pET-28a(+)-MBP-TEV-CarScyto was constructed based on the characteristics of secondary and tertiary structures, and overexpressed in Escherichia coli BL21-Codonplus(DE3)RIL. CarScyto-MBP protein was purified by affinity chromatography, ion-exchange chromatography, and gel filtration chromatography with a final concentration of 20 mg/ml. CarScyto-MBP protein showed both protein kinase and phosphotransferase activities, and the MBP tag had no effect on the function of CarScyto protein. The above results provide a basis for in-depth analysis of the biological function of the CarRS two-component system in F. nucleatum.
Humans
;
Histidine Kinase/metabolism*
;
Fusobacterium nucleatum/metabolism*
;
Automobiles
;
Protein Kinases/genetics*
;
Escherichia coli/metabolism*
;
Colorectal Neoplasms
10.Synergistic effect of β-thujaplicin and tigecycline against tet(X4)-positive Escherichia coli in vitro.
Muchen ZHANG ; Huangwei SONG ; Zhiyu ZOU ; Siyuan YANG ; Hui LI ; Chongshan DAI ; Dejun LIU ; Bing SHAO ; Congming WU ; Jianzhong SHEN ; Yang WANG
Chinese Journal of Biotechnology 2023;39(4):1621-1632
The widespread of tigecycline resistance gene tet(X4) has a serious impact on the clinical efficacy of tigecycline. The development of effective antibiotic adjuvants to combat the looming tigecycline resistance is needed. The synergistic activity between the natural compound β-thujaplicin and tigecycline in vitro was determined by the checkerboard broth microdilution assay and time-dependent killing curve. The mechanism underlining the synergistic effect between β-thujaplicin and tigecycline against tet(X4)-positive Escherichia coli was investigated by determining cell membrane permeability, bacterial intracellular reactive oxygen species (ROS) content, iron content, and tigecycline content. β-thujaplicin exhibited potentiation effect on tigecycline against tet(X4)-positive E. coli in vitro, and presented no significant hemolysis and cytotoxicity within the range of antibacterial concentrations. Mechanistic studies demonstrated that β-thujaplicin significantly increased the permeability of bacterial cell membranes, chelated bacterial intracellular iron, disrupted the iron homeostasis and significantly increased intracellular ROS level. The synergistic effect of β-thujaplicin and tigecycline was identified to be related to interfere with bacterial iron metabolism and facilitate bacterial cell membrane permeability. Our studies provided theoretical and practical data for the application of combined β-thujaplicin with tigecycline in the treatment of tet(X4)-positive E. coli infection.
Humans
;
Tigecycline/pharmacology*
;
Escherichia coli/metabolism*
;
Reactive Oxygen Species/therapeutic use*
;
Plasmids
;
Anti-Bacterial Agents/metabolism*
;
Escherichia coli Infections/microbiology*
;
Bacteria/genetics*
;
Microbial Sensitivity Tests


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