1.Optimized Tube Dilution Technique and Sole Carbon Utilization Assay for Anti-leptospiral In Vitro Screening of Plant Extracts
Ailyn G. Manglicmot-Yabes ; Sharon Yvette Angelina M. Villanueva ; Nina G. Gloriani
Pediatric Infectious Disease Society of the Philippines Journal 2020;21(2):3-12
Introduction:
Leptospirosis is one of the neglected reemerging zoonoses that is of public health concern globally. The need to discover novel therapeutic alternatives for leptospirosis through screening for and elucidating the mechanism/s of the anti-leptospiral activity of plant extracts is therefore necessary. This study analyzes the optimized tube dilution technique and the BiologTM sole carbon utilization phenotype microarray as screening tool for anti-leptospiral activity of plant extracts.
Methods:
The suitability of the optimized tube dilution technique was evaluated by determining the minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and motility inhibition property of a plant extract and an antimicrobial control (pen G) against 4 dominantly circulating Leptospira serovars/serogroup in the Philippines. Likewise, the suitability of the BiologTM sole carbon utilization assay was evaluated using a plant extract and selected antimicrobials against L. interrogans serovar Manilae strain K64 and L. interrogans serovar Losbanos strain K37.
Results:
The MIC, MBC, and motility inhibition property of a plant extract and the antibiotic controls as well as its effect on the carbon utilization phenome of the Leptospira serovars gave consistent results, within and between several runs. With standard deviation = 0 for all serovars. The MIC and MBC of the antimicrobial control (pen G), the positive control, was 10 ug/ml. The growth control (leptospires without treatment), the negative control, showed presence of motile leptospires. The MIC and the MBC of the test plant extract was 250 ug/ml – 500 ug/ml. Results of the carbon utilization phenome or pattern of carbon utilization were consistent within the 3 replicates and between two runs.
Conclusion
The optimized tube dilution technique and the BiologTM sole carbon utilization assay is a potential in vitro screening tool for determining anti-leptospiral activity of plant extracts.
Leptospira
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Serogroup
2.Identification of nontuberculous mycobacteria in patients with multidrug-resistant tuberculosis in Quezon City, Philippines, using multiplex PCR.
Michelle M. CABANATAN ; Alice Alma C. BUNGAY ; Sharon Yvette Angelina M. VILLANUEVA ; Marohren C. TOBIAS-ALTURA ; Dario D. DEFENSOR ; Maria Margarita M. LOTA
Acta Medica Philippina 2025;59(4):103-112
BACKGROUND AND OBJECTIVE
Nontuberculous mycobacteria (NTM) lung disease appears like tuberculosis infection but is resistant to primary anti-tuberculosis drugs. Hence, patients whose sputum sample tests positive for acid-fast bacilli (AFB) and bacterial culture for several times should be assessed for colonization or infection with NTM in a damaged lung secondary to TB. In such cases, though drug-resistant TB may be adequately treated, treatment may need to be directed towards the NTM as well. In NTM therapy, the duration and choice of treatment agent is based upon the specific organism and disease extent. This study used one-step multiplex PCR (mPCR) assay for rapid differentiation of solid cultures in Ogawa medium as Mycobacterium tuberculosis (MTB) and/or NTM.
METHODSA total of 80 stocked isolates obtained from the Lung Center of the Philippines from January to December 2018 were screened for NTM in terms of growth in Ogawa medium, acid fastness, and MPT64 TB antigen test result. These were from sputum specimens of multidrug-resistant tuberculosis (MDR-TB) patients. DNA was extracted from cultures (n=55) grown in Ogawa medium and one-step mPCR was performed to identify NTM to the species level.
RESULTSOut of 80 samples screened, a total of 55 isolates were identified as NTM. One-step mPCR identified 12.73% (7/55) as M. abscessus, 34.55% (19/55) as M. massiliense, 1.82% (1/55) as M. kansasii, and 50.91% (28/55) were identified only up to genus Mycobacteria spp. Neither M. avium complex nor M. intracellulare was identified among the samples tested.
CONCLUSIONOne-step mPCR was able to identify isolates as MTB or NTM coinciding with the initial screening using MPT64 TB antigen test. Multiplex PCR has given a more specific identificati on to the species level. The use of mPCR in identifying MTB and clinically significant NTM’s is suitable for the adequate treatment of mycobacterial infection.
Human ; Bacteria ; Multiplex Pcr ; Multiplex Polymerase Chain Reaction ; Mycobacteria ; Mycobacterium ; Tuberculosis, Multidrug-resistant
3.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
4.Seroprevalence of leptospirosis among water buffaloes, pigs, and dogs in selected areas in the Philippines, 2007 to 2008.
Sharon Yvette Angelina M. VILLANUEVA ; Rubelia A. BATERNA ; Lolita L. CAVINTA ; Yasutake YANAGIHARA ; Nina G. GLORIANI ; Shin-ichi YOSHIDA
Acta Medica Philippina 2018;52(1):109-117
BACKGROUND: Domestic animals are known to be either maintenance or accidental hosts of Leptospira. Determination of seroprevalence of leptospirosis among these animals is of great importance due to their close association with humans, economic loss as well as the public and veterinary health problems caused by the said zoonosis.
OBJECTIVE: This study aimed to determine the seroprevalence of leptospirosis among water buffaloes, pigs, and dogs in selected areas in the Philippines.
METHODS: Microscopic agglutination test (MAT) was used to test for leptospirosis. Testing was done from January 2007 to August 2008.
RESULTS: With the cut-off titer set at 1:80, the MAT-positivity rates were 82%, 67%, and 79% for 190 water buffaloes, 45 pigs, and 106 dogs, respectively. The most common prevailing serovars detected were Hardjo, Tarassovi, and Hebdomadis for water buffaloes; Semaranga, Grippotyphosa, and Patoc for pigs; and, Manilae, Patoc, and Autumnalis for dogs. MAT-positivity rates among these animals in terms of age (except for water buffaloes), sex and sample collection sites were not statistically significant. No Leptospira was isolated from the blood, urine, and kidney samples of these animals.
CONCLUSION: Results indicate a high seroprevalence of leptospirosis among the animals studied and that several pathogenic leptospires are causing infection in these animals.
Animal ; Seroepidemiologic Studies ; Leptospirosis ; Swine ; Philippines
5.Advancing the role of higher education institutions in attaining the health-related sustainable development goals: Proceedings of the 53rd Asia Pacific Academic Consortium for Public Health, 21-23 September 2022, Philippines.
Maria Margarita M. LOTA ; Paul Michael R. HERNANDEZ ; Vivien Fe F. FADRILAN-CAMACHO ; Fresthel Monica M. CLIMACOSA ; Francis Andrew B. CUBE ; Kim Leonard G. DELA LUNA ; Crystal Amiel M. ESTRADA ; Emerito Jose A. FARAON ; Fernando B. GARCIA JR. ; Myra S. MISTICA ; Frederick S. NIETO ; Sharon Yvette Angelina M. VILLANUEVA ; Vicente Y. BELIZARIO JR.
Acta Medica Philippina 2025;59(4):10-13
Higher Education Institutions (HEIs) are acknowledged as key drivers in realizing health-related Sustainable Development Goals (SDGs). The University of the Philippines Manila, College of Public Health (UP CPH) together with the Asia-Pacific Academic Consortium for Public Health (APACPH), hosted the 53rd APACPH International Conference last 21-23 September 2022. The conference discussed current issues relating to the attainment of SDGs and promoted collaboration of leading academic institutions and other stakeholders in addressing various public health challenges. The conference revolved around the challenges and opportunities in attaining health-related SDGs, and the good practices and roles of HEIs in addressing health disparities. The lack of certificati on framework of public health tertiary programs, pedagogy and infrastructure, and ambiguous roles and network of public health professionals were discussed. The conference served as a platform for discussing potential resolutions and ways forward in addressing these challenges. Opportunities for improvement such as updating of policies and curricula, strengthening of internship and community engagement programs, establishment of capacity-building partnerships and programs, and developing multidisciplinary-competent faculty and students were identified. This paper providesthe highlights of the conference focusing on the good practices and roles of HEIs in addressing health disparities, the impact of COVID-19 pandemic, and other issues and challenges in attaining SDGs.
Human ; Sustainable Development ; Sustainable Development Goals ; Public Health