1.An update on Gardneralla vaginalis associated bacterial vaginosis in Malaysia
Younus Khairi NADA ; Gopinath RENU ; Jegasothy RAVINDRAN ; Nordin Amin SYAFINAZ ; Belkum van ALEX ; Mary NARCISSE ; Neela Kumari VASANTHA
Asian Pacific Journal of Tropical Biomedicine 2017;7(9):831-835
Objeetive:To update the status of Gardnerella vaginalis (G.vaginalis) as a causative agent of bacterial vaginosis (BV) in Malaysia and to define its epidemiology,metronidazole resistance and virulence properties.Methods:It is a single-centre (Gynaecology clinic at the Hospital Kuala Lumpur,Malaysia) prospective study with laboratory-based microbiological follow up and analyses.Vaginal swabs collected from the patients suspected for BV were subjected to clinical BV diagnosis,isolation and identification of G.vaginalis,metronidazole susceptibility testing,vaginolysin and sialidase gene PCR,Piot's biotyping and amplified ribosomal DNA restriction analysis genotyping.Results:Among the 207 patients suspected for BV,G.vaginalis was isolated from 47 subjects.G.vaginalis coexisted with Trichomonas vaginalis and Candida albicans in 26 samples.Three G.vaginalis isolates were resistant to metronidazole.Biotyping revealed 1 and 7 as the common types.Amplified ribosomal DNA restriction analysis genotype Ⅱ was found to be more common (n =22;46%) than Ⅰ (n =12;25.53%) and Ⅲ (n =13;27.6%).All genotype Ⅰ and Ⅲ isolates carried the sialidase gene,while 91.6% and 84.6% contained the vaginolysin gene.Genotype Ⅰ was significantly associated with postgynaecological surgical complications and abortions (P =0.002).Conclusions:The existence of pathogenic G.vaginalis clones in Malaysia including drug resistant strains should not be taken lightly and needs to be monitored as these may bring more complications especially among women of child bearing age and pregnant women.
2. An update on Gardneralla vaginalis associated bacterial vaginosis in Malaysia
Nada Khairi YOUNUS ; Syafinaz Amin NORDIN ; Narcisse MARY ; Vasantha Kumari NEELA ; Renu GOPINATH ; Ravindran JEGASOTHY ; Alex VAN BELKUM
Asian Pacific Journal of Tropical Biomedicine 2017;7(9):831-835
Objective To update the status of Gardnerella vaginalis (G. vaginalis) as a causative agent of bacterial vaginosis (BV) in Malaysia and to define its epidemiology, metronidazole resistance and virulence properties. Methods It is a single-centre (Gynaecology clinic at the Hospital Kuala Lumpur, Malaysia) prospective study with laboratory-based microbiological follow up and analyses. Vaginal swabs collected from the patients suspected for BV were subjected to clinical BV diagnosis, isolation and identification of G. vaginalis, metronidazole susceptibility testing, vaginolysin and sialidase gene PCR, Piot's biotyping and amplified ribosomal DNA restriction analysis (ARDRA) genotyping. Results Among the 207 patients suspected for BV, G. vaginalis was isolated from 47 subjects. G. vaginalis coexisted with Trichomonas vaginalis and Candida albicans in 26 samples. Three G. vaginalis isolates were resistant to metronidazole. Biotyping revealed 1 and 7 as the common types. ARDRA genotype II was found to be more common (n = 22; 46%) than I (n = 12; 25.53%) and III (n = 13; 27.6%). All genotype I and III isolates carried the sialidase gene, while 91.6% and 84.6% contained the vaginolysin gene. Genotype I was significantly associated with post-gynaecological surgical complications and abortions (P = 0.002). Conclusions The existence of pathogenic G. vaginalis clones in Malaysia including drug resistant strains should not be taken lightly and needs to be monitored as these may bring more complications especially among women of child bearing age and pregnant women.
3. Comparative analysis of current diagnostic PCR assays in detecting pathogenic Leptospira isolates from environmental samples
May-Ling YAP ; Zamberi SEKAWI ; Hui-Yee CHEE ; Vasantha NEELA ; Han Alan ONG
Asian Pacific Journal of Tropical Medicine 2019;12(10):472-478
Objective: To compare the efficiency of routine diagnostic PCR assays in detecting pathogenic Leptospira isolated from water and soils. Methods: Seven routine assays targeting six genes (lipL32, flaB, gyrB, lfb1, secY and ligB) were evaluated and compared on the cultures of two groups of pathogenic Leptospira from different sources. One group included 19 described reference strains recovered from infected human or animals, and another group included 22 environmental isolates from recreational and residential sites in Malaysia. The latter have been confirmed for presence of pathogenic Leptospira DNA. PCR positivity or detection sensitivity of each assay was determined and compared between the two groups. Results: Validation on reference strains showed 100.0% PCR sensitivity for all assays except ligB-PCR (95.0%) that failed to amplify Leptospira interrogans serovar Pomona. In marked contrast, there was a notable decline in sensitivity in the environmental isolates (lipL32-PCR, 95.5%;flaB-PCR, 90.9%; gyrB-PCR, 77.3%; lfb1-PCR, 59.1%; secY-PCRs, 40.9% G1/G2-PCR, 36.4%; ligB-PCR, 13.6%), implying a large genetic distance between the two groups, as well as nucleotide polymorphism among environmental isolates. Conclusions: High proportion of false-negative PCR results suggests a need of prudent selection of primers in detecting environmental pathogenic Leptospira. These findings offer valuable insights on the extensive biodiversity of genus Leptospira and its impact on the efficacy and development of molecular detection tool.
4.Antibacterial and Wound Healing Activity of 2% Formulation of 2-Medpy-3-CN on Infected Burn Wounded Animal Model
Vasantha Kumari Neela ; Nor Maziah Ahmad ; Surajo Adamu Wada ; Arun Kumar Karunanidhi
Malaysian Journal of Medicine and Health Sciences 2023;19(No.3):5-12
Introduction: Humans have learned to recognize and process plants into medicinal forms through centuries. Burns
can spread to other tissues, especially when infected with bacteria such as Methicillin-Resistant Staphylococcus
aureus (MRSA). The study aimed to assess the in vivo antibacterial and wound healing activity of 2% formulation
of 2-Medpy-3-CN on infected burn wounded animal model. Methods: In vitro antibacterial activity of the Alsti was
done by broth dilution and disc diffusion methods. Alsti 2% ointment was prepared for the infected burn wound
treatment. A total of 18 rats are grouped into A, B, C, and D, the first three groups (A-C) were injured thermally, and
Group D was used as healthy controls. The three test Groups were exposed to MRSA ATCC 43300 at 105 CFU/mL.
Group A was treated with 2% Alsti, Group B with Silver sulfadiazine 1% (SSD), and Group C was untreated. Wounds
healing was assessed by the healed area and microscopic identification of hematoxylin and eosin (H&E)-stained skin
tissue. Results: Wound healing progresses with application of Alsti 2% ointment as observed through wound diameter and histopathological changes of the skin. Wound diameter decreases with treatments, while the contrary was
observed in the non-treated group. Microscopic observation of the stained skin showed that epidermal development,
and collagen formation progress with treatment days. Untreated wounds showed marked inflammation, progressive ulceration, and necrosis. Conclusion: Alsti 2% formulation showed antibacterial and wound healing activities,
hence, can be used as alternative in burn wound infections.
5.Detection of Pathogenic Leptospira in Rats and Phylogenetic Analysis Using Outer Membrane Lipoprotein Lipl32 Gene at Two Major Public Markets
Steffi Julan Wan ; Nurshahira Sulaiman ; Nurul Farhana Zulkifli ; Vasantha Kumari Neela ; Hui Yee Chee ; Siti Norbaya Masri ; Nurul Huda Mohamad Ikbal ; Subha Bhassu ; Hasmahzaiti Omar ; Mazen M. Jamil Al-Obaidi ; Mohd Nasir Mohd Desa
Malaysian Journal of Medicine and Health Sciences 2022;18(No.2):1-5
Introduction: Limited studies have been documented on the presence of pathogenic Leptospira in public markets
serving the community in sub-districts of Selangor. The aim of this study was to detect the presence of pathogenic
Leptospira in rats using a gene encoding an outer membrane lipoprotein LipL32. Methods: Polymerase chain reaction (PCR) was performed using LipL32 primers on sixty kidney samples of rats trapped at two locations of study;
Pasar Borong Selangor in Seri Kembangan and Pasar Basah Bandar Baru Bangi in Bangi. Results: Out of 60 samples
analysed, 36.7% were positive for the presence of LipL32. All positive samples highly matched (>94%) nucleotide
sequence for LipL32 of pathogenic Leptospira and related to the pathogens through phylogenetic analysis. Conclusion: The detection of LipL32 indicates the potential presence of pathogenic Leptospira species at public markets.
Although only 60 rats were successfully trapped, the rats are mobile and might further transmit the pathogenic organisms to other areas.