HIV/AIDS - Research and Palliative Care Dovepress

Biofilm-Producing Candida Species Causing Oropharyngeal Candidiasis in HIV Patients Attending Sukraraj Tropical and Infectious Diseases Hospital in Kathmandu, Nepal

Keshav Lamichhane,^1,* Nabaraj Adhikari,^1,* Anup Bastola,² Lina Devkota,² Parmananda Bhandari,² Binod Dhungel,¹ Upendra Thapa Shrestha,¹ Bipin Adhikari,³ Megha Raj Banjara,¹ Komal Raj Rijal,¹ Prakash Ghimire¹

¹Central Department of Microbiology, Tribhuvan University, Kathmandu, Nepal; 

²Sukraraj Tropical and Infectious Disease Hospital, Teku, Kathmandu, Nepal; 

³Centre for Tropical Medicine and Global Health, Nuffield Department of Medicine, University of Oxford, Oxford, UK

Introduction: Oropharyngeal candidiasis are the commonest fungal infections among HIV-positive patients. The main objective of this study was to explore biofilm-producing Candida species causing oropharyngeal infections among HIV patients attending Sukraraj Tropical and Infectious Diseases Hospital (STIDH) in Kathmandu, Nepal.

Methods: Oropharyngeal swabs were collected from the HIV-positive patients between July and December 2019. A total of 174 oropharyngeal swabs were cultured on Sabouraud Dextrose Agar (SDA). All samples were inoculated on SDA slants supplemented with chloramphenicol and underwent incubation at 37°C for 24– 48 hours. Any visible growth reported was processed for the identification of the species. Candida species were differentiated based on the growth and colour of the isolates on CHROM agar candida. Biofilm production in Candida species was determined by the microtiter plate method (MPM). Antifungal susceptibility testing was performed using the disc diffusion method.

Results: Among 174 oropharyngeal samples, 23.6% (n=41/174) of them had oropharyngeal infections and 36.6% of the oropharyngeal infections (15/41) had CD4 T-lymphocytes count below 200 cells/mm³ who were also active tobacco users (p< 0.05). Among Candidial growth, 61% (25/41) were Candida albicans and 39% (16/41) were non-albicans. Of 41 Candida spp., 65% (27/41) were biofilm producers. An equal proportion of Candida albicans (4 isolates) and non-albicans (4 isolates) were strong biofilm producers. C. albicans isolates were sensitive towards clotrimazole (96%; 24/25) and fluconazole (92%; 23/25), whereas sensitivity towards ketoconazole was only 48% (12/25). Non-albicans Candida was highly sensitive to amphotericin-B (62.5%; 10/16) followed by clotrimazole (56.2%; 9/16). The biofilm-producing Candida isolates showed the highest resistivity (51.9%; 14/27) to ketoconazole and lowest (22.2%; 6/27) to clotrimazole.

Conclusion: Oropharyngeal candidiasis is a common opportunistic infection among HIV-infected individuals. The majority of cases of oropharyngeal candidiasis are caused by biofilm producers Candida albicans and non-albicans Candida. Biofilm producers Candida were more resistant towards commonly used antifungal drugs.

Keywords: Oral candidiasis, HIV, Candida albicans, Biofilm, Antifungal susceptibility test

HIV/AIDS - Research and Palliative Care 2020:12 211–220

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ACTA SCIENTIFIC MICROBIOLOGY (ISSN: 2581-3226)

Etiology of Ocular Infections and Minimum Inhibitory Concentration of Multidrug- Resistant Staphylococcus aureus Isolates to Vancomycin, Ciprofloxacin and Chloramphenicol

Samina Thapa1, Nabaraj Adhikari2, Binod Dhungel2, Madhu Thapa3, Upendra Thapa Shrestha2, Megha Raj Banjara2, Komal Raj Rijal2* and Prakash Ghimire2

1Kantipur College of Medical Sciences, Sitapaila, Kathmandu, Nepal

2Central Department of Microbiology, Tribhuvan University, Kirtipur, Nepal

3BP Koirala Lions Center for Ophthalmic Studies, Institute of Medicine, Tribhuvan

University, Kirtipur, Nepal

*Corresponding Author: Komal Raj Rijal, Associate Professor, Central Department of Microbiology, Tribhuvan University, Kirtipur, Kathmandu, Nepal.

Abstract

Introduction: Ocular infections constitute a significant portion of curable eye diseases. However, in developing countries like Nepal, bacterial and fungal spectrum is not well established for ocular infections. Therefore, ocular infections still remain as the major cause of blindness. This study was sought determine the bacterial and fungal etiology of various ocular infections and assess the antibiotic susceptibility pattern of the bacterial pathogens.

Methods: This cross-sectional study was conducted at BP Koirala Lions Center for Ophthalmic Studies, Kathmandu, Nepal from June 2012 through April 2013. Various ophthalmic specimens of clinical values such as the corneal scraps, conjunctival swab and eye pus and biopsy materials were collected aseptically and processed for culture. After sufficient incubation, isolates were identified by colony morphology, Gram staining and relevant biochemical tests. Identified bacteria; isolates were then subjected under Antibiotic Susceptibility test (AST) by modified Kirby-Bauer disc diffusion method to determine Multi Drug Resistant (MDR) isolates and Minimum Inhibitory Concentration (MIC).

Results: A total number of 810 ocular specimens, mainly from the case of conjunctivitis (644/810) were processed. Of which, 31.97% (270/810) showed growth on culture medium: 267 (98.85%) were bacterial, 2 (0.77%) were fungal (Penicillium spp and Fusarium spp) and 1 (0.38%) was yeast (Rhodotorula) isolate. Gram positives alone accounted for 96.25% (257/267) among bacterial isolates. Staphylococcus aureus (52.96%) and Escherichia coli (1.48%) were the most common Gram positive and Gram-negative isolates respectively. Gram positive isolates showed resistance to ciprofloxacin (39.29%), cephalexin (38.13%) and amikacin (31.52%) while gram negatives were resistant to cefazolin (80%). All isolates of E. coli, Hemophilus influenzae and Serratia spp together with 48.95% of S. aureus were MDR. Among 5 methicillin resistant S. aureus (MRSA) isolates, 2 isolates were vancomycin intermediate S. aureus (VISA). Further, the minimum inhibitory concentration (MIC) break points for S. aureus to chloramphenicol, ciprofloxacin and vancomycin were also determined.

Conclusion: Conjunctivitis among adults was the most prevalent case found so far, yielding higher culture positivity. Along with MDR bacterial agents, fungal agents were observed to be posing significant threat in the disease management. Early diagnosis, proper management and empirical treatment are advised to prevent infections related blindness.

Keywords: Ocular Infections; Conjunctivitis; AST; MDR; MRSA; VISA; MIC

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Citation: Komal Raj Rijal., et al. “Etiology of Ocular Infections and Minimum Inhibitory Concentration of Multidrug- Resistant Staphylococcus aureus Isolates to Vancomycin, Ciprofloxacin and Chloramphenicol". Acta Scientific Microbiology 3.6 (2020): 134-145.