Ritu Goyal

Epidemiology and Virulence Determinants including Biofilm Profile of Candida Infections in an ICU in a Tertiary Hospital in India

The purpose of this prospective study was to isolate, speciate, and determine antifungal susceptibility and virulence patterns of Candida species recovered from the intensive care units (ICUs) in an Indian hospital. Study included 125 medical/postoperative patients admitted to ICU. Identification and speciation of yeast isolates were done by the biochemical methods. Antifungal susceptibility was done by broth microdilution method. Virulence testing of Candida species was done by phospholipase, proteinase, and adherence assay. A total of 103 Candida isolates were isolated; C. tropicalis was the predominant species (40.7%), followed by C. albicans (38.83 %), C. glabrata (11.65%), C. parapsilosis (3.88%), and 1.94% each of C. krusei, C. kefyr, and C. sphaerica. 60 Candida isolates (58.25%) showed resistance to fluconazole, while 7 (6.7%) isolates showed resistance to amphotericin B. Phospholipase and proteinase activities were seen in 73.8% and 55.3% Candida isolates with different species showing a wide range of activities, while 68.9% Candida isolates showed {4

Identification and Antifungal susceptibility testing of Candida species: A Comparison of Vitek-2 system with conventional and molecular methods

Background: Candida infection is a major cause of morbidity and mortality in immunocompromised patients; an accurate and early identification is a prerequisite need to be taken as an effective measure for the management of patients. The purpose of this study was to compare the conventional identification of Candida species with identification by Vitek-2 system and the antifungal susceptibility testing (AST) by broth microdilution method with Vitek-2 AST system. Materials and Methods: A total of 172 Candida isolates were subjected for identification by the conventional methods, Vitek-2 system, restriction fragment length polymorphism, and random amplified polymorphic DNA analysis. AST was carried out as per the Clinical and Laboratory Standards Institute M27-A3 document and by Vitek-2 system. Results: Candida albicans (82.51%) was the most common Candida species followed by Candida tropicalis (6.29%), Candida krusei (4.89%), Candida parapsilosis (3.49%), and Candida glabrata (2.79%). With Vitek-2 system, of the 172 isolates, 155 Candida isolates were correctly identified, 13 were misidentified, and four were with low discrimination. Whereas with conventional methods, 171 Candida isolates were correctly identified and only a single isolate of C. albicans was misidentified as C. tropicalis. The average measurement of agreement between the Vitek-2 system and conventional methods was >94%. Most of the isolates were susceptible to fluconazole (88.95%) and amphotericin B (97.67%). The measurement of agreement between the methods of AST was >94% for fluconazole and >99% for amphotericin B, which was statistically significant (P < 0.01). Conclusion: The study confirmed the importance and reliability of conventional and molecular methods, and the acceptable agreements suggest Vitek-2 system an alternative method for speciation and sensitivity testing of Candida species infections.

Oral Thrush to Candidemia: A Morbid Outcome

A 35-year-old female living with AIDS presented with oral thrush 3 months after the initiation of antiretroviral therapy (ART). On investigation, it was found to be due to Candida albicans. She was started on fluconazole. After 4 months on ART, she presented with fever. Blood culture was performed. After 4 days, she expired due to septic shock. Two days after her death, C albicans strain was isolated from the blood culture which was similar to the C albicans strain (by DNA fingerprinting) isolated from oral thrush. Both strains of C albicans were found to be resistant to fluconazole by broth microdilution method. A high index of suspicion in high-risk patients along with early and aggressive management of the patient with antigen detection would go a long way in the management of these patients. Guidelines for treatment of candidiasis need to be re-evaluated, keeping in mind the increasing emergence of resistance to azoles and its effect on morbidity outcome.

Spectrum of Opportunistic Fungal Infections in HIV/AIDS Patients in Tertiary Care Hospital in India

HIV related opportunistic fungal infections (OFIs) continue to cause morbidity and mortality in HIV infected patients. The objective for this prospective study is to elucidate the prevalence and spectrum of common OFIs in HIV/AIDS patients in north India. Relevant clinical samples were collected from symptomatic HIV positive patients (n = 280) of all age groups and both sexes and subjected to direct microscopy and fungal culture. Identification as well as speciation of the fungal isolates was done as per the standard recommended methods. CD4+T cell counts were determined by flow cytometry using Fluorescent Activated Cell Sorter Count system. 215 fungal isolates were isolated with the isolation rate of 41.1%. Candida species (86.5%) were the commonest followed by Aspergillus (6.5%), Cryptococcus (3.3%), Penicillium (1.9%), and Alternaria and Rhodotorula spp. (0.9% each). Among Candida species, Candida albicans (75.8%) was the most prevalent species followed by C. tropicalis (9.7%), C. krusei (6.4%), C. glabrata (4.3%), C. parapsilosis (2.7%), and C. kefyr (1.1%). Study demonstrates that the oropharyngeal candidiasis is the commonest among different OFIs and would help to increase the awareness of clinicians in diagnosis and early treatment of these infections helping in the proper management of the patients especially in resource limited countries like ours.

Study of TH1/TH2 Cytokine Profiles in HIV/AIDS Patients in a TertiaryCare Hospital in India

Background and Objectives: Switch of cytokines from a T helper 1 (Th1) to a Th2 is an important factor in the progression of HIV infection to AIDS. Hence, our objectives were to analyze the levels of Th1 (IL-2, IFN-γ) and Th2 (IL-4, IL-10) cytokines and their correlation with clinical and immunological profiles in HIV/AIDS patients. Methods: We studied 234 symptomatic HIV positive patients (case group) attending OPDs, ART clinic and medical wards of the Hospital. CD4+T-cell count was determined by flow cytometry using Fluorescent Activated Cell Sorter Count system. Quantitative determination of cytokines (Th1 subtype: IL-2, IFN-γ and Th2 subtype: IL-4, IL-10) was done by ELISA. Results: Patients (08-68 years) had CD4+T-cell counts ranged from 16-1033 cells/μl. The median CD4+T cell count was 204.50 cells/μl and the mean was 265.48 cells/μl. The concentration of IL-2 and IFN-γ were significantly lower in case group compared to asymptomatic HIV patients (P<0.001) while the IL-4 in symptomatic HIV negative was higher than healthy controls and case group compared to asymptomatic HIV patients. Concentration of IL-10 was also higher in case group compared to asymptomatic HIV patients. A positive correlation was found between IL-4, IFN-γ, IL-2 with CD4+T-cells and a negative correlation between IL-10 and CD4+T-cells among the case group. Conclusions: Hence a decline in type-1 cytokines (IFN-γ, IL-2) and rise in type-2 cytokines (IL-4, IL-10) was observed in symptomatic patients with HIV infection in comparison to asymptomatic HIV patients suggesting a shift from Th1 to Th2 type cytokine response.

Concurrent Cryptococcal and Pneumocystis Pneumonia along with Pulmonary Tuberculosis in an HIV-Positive Patient: Lessons Learned for Early Management

Case: We are presenting a 50-year-old patient of pulmonary tuberculosis, on anti-tuberculosis therapy (ATT) for last 2 months who presented with fever, cough, breathlessness, anorexia, and weight loss. The case was found to be HIV reactive. His sputum sample showed Candida albicans and Pneumocystis jirovecii. Fluconazole and cotrimoxazole + sulphamethoxazole were added. The index case did not respond to the treatment and his clinical condition started to deteriorate and he developed headache, vomiting, and dysphagia. Repeat sputum sample and cerebrospinal fluid (CSF) showed Cryptococcus neoformans which was found to be sensitive to Amphotericin B. Amphotericin B was added to the treatment and patient clinically responded to treatment. In conclusion, emphasis should be given to correct etiological identification, allowing appropriate treatment and decreasing the morbidity and mortality in these patients as concomitant opportunistic infections may cause diagnostic problems.

Is antifungal resistance a cause for treatment failure in dermatophytosis: A study focused on tinea corporis and cruris from a tertiary centre?

Background: Dermatophytoses are one of the most common skin diseases that have been largely simple to treat. However, in recent years, these infections have become recalcitrant to treatment which can possibly be due to antifungal resistance. Aim: To analyze the resistance pattern of patients with recalcitrant dermatophytoses. Materials and Methods: A cross-sectional evaluation was undertaken of 40 consecutive patients with recalcitrant tinea corporis/cruris/both who had taken systemic antifungal treatment and did not respond completely to therapy or had recurrent lesion within 1 month of stopping the therapy. Terbinafine, fluconazole, itraconazole, ketoconazole, amphotericin B, and voriconazole were the antifungals tested using broth microdilution assay for antifungal susceptibility testing of dermatophytes, and MIC50, 90 values were recorded. Results: KOH mount was positive in 18 (45%) patients, culture was positive in 28 (70%) patients. Trichophyton mentagrophytes (35%) and T. rubrum (27.5%) were the predominant isolates. Overall, activity of terbinafine and itraconazole were significantly higher than the other drugs tested. For terbinafine, both T. mentagrophytes and T. rubrum were inhibited at MIC90of 0.125 μg/ml. Itraconazole-inhibited T. mentagrophytes and T. rubrum at MIC90of 0.0625 and 0.25 μg/ml, respectively. All isolates had reduced susceptibility to fluconazole. Conclusion: While MIC seen were higher than western data, in-vitro resistance (>1 μg/ml) to antifungals was not seen and probably may not be a cause of treatment failure. Possibly, treatment failure lies in the intricate host fungal interaction and virulence of species which help it to evade host immune response.