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
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Microbial Sensitivity Tests
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Anti-infective Agents, Local
;
Enterobacter Aerogenes
;
Staphylococcus Aureus
2.Artemisia argyi extract subfraction exerts an antifungal effect against dermatophytes by disrupting mitochondrial morphology and function.
Le CHEN ; Yunyun ZHU ; Chaowei GUO ; Yujie GUO ; Lu ZHAO ; Yuhuan MIAO ; Hongzhi DU ; Dahui LIU
Chinese Journal of Natural Medicines (English Ed.) 2024;22(1):47-61
Artemisia argyi (A. argyi), a plant with a longstanding history as a raw material for traditional medicine and functional diets in Asia, has been used traditionally to bathe and soak feet for its disinfectant and itch-relieving properties. Despite its widespread use, scientific evidence validating the antifungal efficacy of A. argyi water extract (AAWE) against dermatophytes, particularly Trichophyton rubrum, Trichophyton mentagrophytes, and Microsporum gypseum, remains limited. This study aimed to substantiate the scientific basis of the folkloric use of A. argyi by evaluating the antifungal effects and the underlying molecular mechanisms of its active subfraction against dermatophytes. The results indicated that AAWE exhibited excellent antifungal effects against the three aforementioned dermatophyte species. The subfraction AAWE6, isolated using D101 macroporous resin, emerged as the most potent subfraction. The minimum inhibitory concentrations (MICs) of AAWE6 against T. rubrum, M. gypseum, and T. mentagrophytes were 312.5, 312.5, and 625 μg·mL-1, respectively. Transmission electron microscopy (TEM) results and assays of enzymes linked to cell wall integrity and cell membrane function indicated that AAWE6 could penetrate the external protective barrier of T. rubrum, creating breaches ("small holes"), and disrupt the internal mitochondrial structure ("granary"). Furthermore, transcriptome data, quantitative real-time PCR (RT-qPCR), and biochemical assays corroborated the severe disruption of mitochondrial function, evidenced by inhibited tricarboxylic acid (TCA) cycle and energy metabolism. Additionally, chemical characterization and molecular docking analyses identified flavonoids, primarily eupatilin (131.16 ± 4.52 mg·g-1) and jaceosidin (4.17 ± 0.18 mg·g-1), as the active components of AAWE6. In conclusion, the subfraction AAWE6 from A. argyi exerts antifungal effects against dermatophytes by disrupting mitochondrial morphology and function. This research validates the traditional use of A. argyi and provides scientific support for its anti-dermatophytic applications, as recognized in the Chinese patent (No. ZL202111161301.9).
Antifungal Agents/chemistry*
;
Arthrodermataceae
;
Artemisia/chemistry*
;
Molecular Docking Simulation
;
Mitochondria
;
Microbial Sensitivity Tests
3.Antimicrobial consumption and resistance of restricted antibiotics in a Level III government hospital.
Mary Anne Abeleda ; Imelda Peñ ; a ; Roderick Salenga ; Francis Capule ; Shiela Mae Nacabu-an ; Pamela Nala
Acta Medica Philippina 2024;58(16):68-76
OBJECTIVES
The objectives of the study were to determine the antibiotic consumption of restricted antibiotics and to correlate this with resistance rate.
METHODSA retrospective review of pharmacy dispensing records was conducted in the adult internal medicine wards of a tertiary level teaching hospital in the Philippines between March 2019 to February 2020. Antibiotic consumption was determined using Defined Daily Dose (DDD) per 1000 patient-days (PD). Correlations between antibiotic consumption and antibiotic resistance of restricted antibiotics were done. Outcomes were compared between Ward 1 (with the presence of a unit-dose pharmacist) and Ward 3 (without a unit-dose pharmacist).
RESULTSBoth wards showed decreasing trends of piperacillin-tazobactam consumption and increasing trends of ceftazidime consumption from quarter 1 to quarter 4. It was observed that levofloxacin was the most prescribed fluoroquinolone with the highest consumption recorded from March to May 2019 in Ward 3 of 350.2 DDD/1000 PD as compared with ciprofloxacin which has the highest consumption (23.3 DDD/1000 PD) during the period June to August 2019 in Ward 1. Antibiotic resistance of Acinetobacter baumannii against ciprofloxacin, levofloxacin, and piperacillin-tazobactam were statistically significantly different between the wards. In Ward 1, ciprofloxacin consumption was strongly positively correlated with Escherichia coli resistance (r = 0.90). In Ward 3, a significantly moderately positive association was observed for ceftazidime consumption and A. baumannii resistance (r = 0.61), positive correlation between piperacillin-tazobactam and E. coli resistance (r = 0.65), and a strong positive correlation in Ward 3 between levofloxacin and Pseudomonas aeruginosa resistance (r = 0.71).
CONCLUSIONThe restriction and pre-authorization strategy of the AMS program has greatly contributed to the decrease in the consumption of almost all restricted antibiotics. This strategy has been helpful in minimizing unnecessary antibiotic use associated with inappropriate drug therapy. The success of the AMS program has been based on the collective efforts of the AMS team with the implementation of hospital policies, such as the AMS program, across the different sites in the hospital in order to achieve optimum patient health outcomes. It was noted that the resistance rates of A. baumannii against ciprofloxacin, levofloxacin, and piperacillin-tazobactam were higher in Ward 3 compared to Ward 1 which makes infections very difficult to treat which may result to prolonged hospital stay, increased health-care costs and increased mortality rate. This study has supported the involvement of pharmacists in the AMS team by conducting auditing activities that promote safe compliance of restricted antibiotic use among patients. Pharmacists can greatly participate on either prospective or retrospective review of antibiotic utilization and analyze trends of antibiotic consumption data to provide feedback to prescribing physicians on prescribing patterns and possible correlation with occurrence of antibiotic resistance.
Antibiotic Resistance ; Drug Resistance, Microbial
4.Evaluation of Microsphere-based xMAP Test for gyrA Mutation Identification in Mycobacterium Tuberculosis.
Xi Chao OU ; Bing ZHAO ; Ze Xuan SONG ; Shao Jun PEI ; Sheng Fen WANG ; Wen Cong HE ; Chun Fa LIU ; Dong Xin LIU ; Rui Da XING ; Hui XIA ; Yan Lin ZHAO
Biomedical and Environmental Sciences 2023;36(4):384-387
5.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
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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
6.Advances in genomics of multi-drug resistant Stenotrophomonas.
Yuhang TANG ; Shiqi FANG ; Linlin XIE ; Chao SUN ; Shanshan LI ; Aiping ZHOU ; Guangxiang CAO ; Jun LI
Chinese Journal of Biotechnology 2023;39(4):1314-1331
Stenotrophomonas species are non-fermentative Gram-negative bacteria that are widely distributed in environment and are highly resistant to numerous antibiotics. Thus, Stenotrophomonas serves as a reservoir of genes encoding antimicrobial resistance (AMR). The detection rate of Stenotrophomonas is rapidly increasing alongside their strengthening intrinsic ability to tolerate a variety of clinical antibiotics. This review illustrated the current genomics advances of antibiotic resistant Stenotrophomonas, highlighting the importance of precise identification and sequence editing. In addition, AMR diversity and transferability have been assessed by the developed bioinformatics tools. However, the working models of AMR in Stenotrophomonas are cryptic and urgently required to be determined. Comparative genomics is envisioned to facilitate the prevention and control of AMR, as well as to gain insights into bacterial adaptability and drug development.
Stenotrophomonas/genetics*
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Drug Resistance, Bacterial/genetics*
;
Anti-Bacterial Agents/pharmacology*
;
Gram-Negative Bacteria
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Genomics
;
Microbial Sensitivity Tests
7.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
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Tigecycline/pharmacology*
;
Escherichia coli/metabolism*
;
Reactive Oxygen Species/therapeutic use*
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Plasmids
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Anti-Bacterial Agents/metabolism*
;
Escherichia coli Infections/microbiology*
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Bacteria/genetics*
;
Microbial Sensitivity Tests
8.Polyethylene biodegradation: current status and perspectives.
Liting ZHANG ; Bo ZHANG ; Weidong XU ; Zhongli CUI ; Hui CAO
Chinese Journal of Biotechnology 2023;39(5):1949-1962
Polyethylene (PE) is the most abundantly used synthetic resin and one of the most resistant to degradation, and its massive accumulation in the environment has caused serious pollution. Traditional landfill, composting and incineration technologies can hardly meet the requirements of environmental protection. Biodegradation is an eco-friendly, low-cost and promising method to solve the plastic pollution problem. This review summarizes the chemical structure of PE, the species of PE degrading microorganisms, degrading enzymes and metabolic pathways. Future research is suggested to focus on the screening of high-efficiency PE degrading strains, the construction of synthetic microbial consortia, the screening and modification of degrading enzymes, so as to provide selectable pathways and theoretical references for PE biodegradation research.
Polyethylene/metabolism*
;
Bacteria/metabolism*
;
Plastics/metabolism*
;
Biodegradation, Environmental
;
Microbial Consortia
9.Engineering microbial consortia through synthetic biology approach.
Jiao ZHANG ; Jiangfeng CHEN ; Yixuan CHEN ; Lei DAI ; Zhuojun DAI
Chinese Journal of Biotechnology 2023;39(5):2517-2545
There are a large number of natural microbial communities in nature. Different populations inside the consortia expand the performance boundary of a single microbial population through communication and division of labor, reducing the overall metabolic burden and increasing the environmental adaptability. Based on engineering principles, synthetic biology designs or modifies basic functional components, gene circuits, and chassis cells to purposefully reprogram the operational processes of the living cells, achieving rich and controllable biological functions. Introducing this engineering design principle to obtain structurally well-defined synthetic microbial communities can provide ideas for theoretical studies and shed light on versatile applications. This review discussed recent progresses on synthetic microbial consortia with regard to design principles, construction methods and applications, and prospected future perspectives.
Microbial Consortia/genetics*
;
Synthetic Biology
;
Microbiota
;
Models, Theoretical
10.Characteristics of Gram-positive cocci infection and the therapeutic effect after liver transplantation.
Xiaoxia WU ; Lingli WU ; Lin SHU ; Chenpeng XIE ; Qiquan WAN
Journal of Central South University(Medical Sciences) 2023;48(5):707-715
OBJECTIVES:
Gram-positive cocci is the main pathogen responsible for early infection after liver transplantation (LT), posing a huge threat to the prognosis of liver transplant recipients. This study aims to analyze the distribution and drug resistance of Gram-positive cocci, the risk factors for infections and efficacy of antibiotics within 2 months after LT, and to guide the prevention and treatment of these infections.
METHODS:
In this study, data of pathogenic bacteria distribution, drug resistance and therapeutic efficacy were collected from 39 Gram-positive cocci infections among 256 patients who received liver transplantation from donation after citizens' death in the Third Xiangya Hospital of Central South University from January 2019 to July 2022, and risk factors for Gram-positive cocci infection were analyzed.
RESULTS:
Enterococcus faecium was the dominant pathogenic bacteria (33/51, 64.7%), followed by Enterococcus faecalis (11/51, 21.6%). The most common sites of infection were abdominal cavity/biliary tract (13/256, 5.1%) and urinary tract (10/256, 3.9%). Fifty (98%) of the 51 Gram-positive cocci infections occurred within 1 month after LT. The most sensitive drugs to Gram-positive cocci were teicoplanin, tigecycline, linezolid and vancomycin. Vancomycin was not used in all patients, considering its nephrotoxicity. Vancomycin was not administered to all patients in view of its nephrotoxicity.There was no significant difference between the efficacy of daptomycin and teicoplanin in the prevention of cocci infection (P>0.05). Univariate analysis indicated that preoperative Model for End-Stage Liver Disease (MELD) score >25 (P=0.005), intraoperative red blood cell infusion ≥12 U (P=0.013) and exposure to more than 2 intravenous antibiotics post-LT (P=0.003) were related to Gram-positive cocci infections. Multivariate logistic regression analysis revealed that preoperative MELD score >25 (OR=2.378, 95% CI 1.124 to 5.032, P=0.024) and intraoperative red blood cell transfusion ≥ 12 U (OR=2.757, 95% CI 1.227 to 6.195, P=0.014) were independent risk factors for Gram-positive cocci infections after LT. Postoperative Gram-positive cocci infections were reduced in LT recipients exposing to more than two intravenous antibiotics post-LT (OR=0.269, 95% CI 0.121 to 0.598, P=0.001).
CONCLUSIONS
Gram-positive cocci infections occurring early after liver transplantation were dominated by Enterococcus faecalis infections at the abdominal/biliary tract and urinary tract. Teicoplanin, tigecycline and linezolid were anti-cocci sensitive drugs. Daptomycin and teicoplanin were equally effective in preventing cocci infections due to Gram-positive cocci. Patients with high preoperative MELD score and massive intraoperative red blood cell transfusion were more likely to suffer Gram-positive cocci infection after surgery. Postoperative Gram-positive cocci infections were reduced in recipients exposing to more than two intravenous antibiotics post-LT.
Humans
;
Daptomycin/therapeutic use*
;
Linezolid/therapeutic use*
;
Teicoplanin/therapeutic use*
;
Gram-Positive Cocci
;
Liver Transplantation/adverse effects*
;
Tigecycline/therapeutic use*
;
End Stage Liver Disease/drug therapy*
;
Gram-Positive Bacterial Infections/microbiology*
;
Severity of Illness Index
;
Anti-Bacterial Agents/pharmacology*
;
Vancomycin/therapeutic use*
;
Microbial Sensitivity Tests


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