International Journal of Current Research and Review
ISSN: 2231-2196 (Print)ISSN: 0975-5241 (Online)
Bootstrap Slider

Indexed and Abstracted in: SCOPUS, Crossref, CAS Abstracts, Publons, CiteFactor, Open J-Gate, ROAD, Indian Citation Index (ICI), Indian Journals Index (IJINDEX), Internet Archive, IP Indexing, Google Scholar, Scientific Indexing Services, Index Copernicus, Science Central, Revistas Medicas Portuguesas, EBSCO, BOAI, SOROS, NEWJOUR, ResearchGATE, Ulrich's Periodicals Directory, DocStoc, PdfCast, getCITED, SkyDrive, Citebase, e-Print, WorldCat (World's largest network of library content and services), Electronic Journals Library by University Library of Regensburg, SciPeople.

Search Articles

Track manuscript

Readers around the world

Full Html

IJCRR - vol 06 issue 01, January, 2014

Pages: 01-05

Date of Publication: 30-Nov--0001


Print Article   Download XML  Download PDF

USE OF CHROMOGENIC MEDIUM FOR SPECIATION OF CANDIDA ISOLATED FROM CLINICAL SPECIMENS

Author: Dharmeswari T., Sheela Devi Chandrakesan, Nagaraja Mudhigeti, Anitha Patricia, Reba Kanungo

Category: General Sciences

Abstract:Aim: To explore the usefulness of chromogenic medium in speciating clinical isolates of Candida and to determine their antifungal susceptibility. Methodology: A total of 50 Candida isolated from various clinical specimens were included in the study. Speciation of Candida was done based on the growth on chromogenic medium and other conventional methods. Antifungal susceptibility testing was performed against fluconozole and amphotericin B. Results: Among the 50 clinical Candida isolates, 45 could be speciated with the help of chromogenic medium. Only 30% of the Candida isolates were identified as Candida albicans and the rest were non albicans Candida species. Among the non albicans species Candida tropicalis was the commonest isolate followed by C. glabrata and C. parapsilosis. None of the strains were resistant to amphotericin B or fluconazole. Conclusion: Use of chromogenic medium with the morphology on corn meal agar provides rapid identification of commonly isolated Candida species from clinical specimens. This will be useful to initiate appropriate antifungal therapy thereby reducing morbidity and mortality.

Keywords: Candida albicans, Chromogenic medium, Antifungal susceptibility, speciation, Non albicans Candida

Full Text:

INTRODUCTION

The incidence of various fungal pathogens has increased dramatically over the past few decades. Candida species are the most common of these pathogens and have emerged as important opportunistic pathogens in the last decade. Infections by this group are often severe, rapidly progressive and refractory to therapy. Suppressed host defense and exposure to multiple risk factors are responsible for their emergence. Use of newer and broad spectrum antibacterial agents, increasing patient population living with AIDS, haematological malignancies or solid organ transplantation have been the contributory factors for the emergence of Candida species as important pathogens (1,2) Though Candida albicans is considered to be the commonest species causing human diseases, there are increasing reports of non-albicans Candida species emerging as important pathogens. (1,3) A rise in less common Candida sp. has been associated with significant morbidity and mortality. (4) The emergence of drug resistant Candida sp. which is largely attributed to use of prolonged and irrational empirical therapy has further complicated patient management. Some of the non-albicans Candida sp. are known to have varied susceptibility pattern to the routinely used antifungal agents especially the azoles. Conventional methods available to speciate the non albicans Candida sp. are time consuming and cumbersome. (5) To overcome this problem CHROM agar, a chromogenic differential culture medium was introduced. The manufacturers claimed that this chromogenic agar can facilitate the isolation and presumptive identification of certain clinically important Candida species. With the current scenario where non albicans Candida species are frequently isolated from clinical specimens, an early identification and speciation of positive Candida cultures is important . The present study was undertaken to explore the usefulness of chromogenic medium for speciation of Candida isolated from clinical specimens and to determine their drug susceptibility patterns.

MATERIALS AND METHODS

Isolation and identification of yeast

A total of 50 Candida isolated from various specimens (blood, clean-voided mid stream urine, pus, broncho-alveolar lavage and tracheal aspirates) submitted to the clinical microbiology laboratory from different clinical units of Pondicherry Institute of Medical Sciences (PIMS), a tertiary care hospital, were included in the study. All suspected yeast colonies on sheep blood agar were confirmed by Gram staining. All such yeast isolates that grew on sheep blood agar (HiMedia, Mumbai, India) were sub cultured on chromogenic medium (HiChrom agar Candida; HiMedia, Mumbai, India) and incubated at 37o C. The isolates grew well and developed distinctive colored colonies after overnight incubation. The plates were further incubated for 24 h to observe well distinguished colonies. Presumptive identification was made by n o t i n g t h e color of the colonies as per the manufacturer's Instructions. All isolates were further inoculated on corn meal agar (CMA) by slide culture method to determine microscopic morphological features of various Candida species. (6) Antifungal susceptibility testing by disc diffusion method was performed as per CLSI M44A (7) document against fluconazole discs (Hi Media, Mumbai, India) for all the 50 isolates. Briefly the yeast suspension was prepared in 5ml of sterile normal saline and turbidity was adjusted to 0.5McFarland standard. Modified Muller Hinton agar (Muller-Hinton agar + 2% glucose + 0.5 µg/ml of Methylene blue, pH 7.2) plates were inoculated and fluconazole (25 µg) discs (Hi Media, Mumbai, India) was placed on each plate. Results were obtained after 24 hours of incubated at 37 o C. MIC of Amphotericin B was determined by agar dilution method for 37 isolates. Amphotericin B (Hi Media, India) stock solution and working concentration were prepared as per CLSI guidelines. (8) As the study did not involve human subjects, obtaining informed consent was waived by the Institute Ethics Committee. Throughout the study C.albicans (ATCC90028), C.parapsilosis (ATCC90018), and C.krusei (ATCC6258), ATCC standard strains were used as Quality controls.

RESULTS

Only 15/50 (30%) of the strains were identified as C. albicans. Nearly half of the remaining 35 strains were C. tropicalis (43%), followed by C.glabrata (25%) and C.parapsilosis ( 17%). Candida species identified by the color on the chromogenic medium is shown in fig 1. Out of 50 isolates, 45 isolates showed characteristic morphology for that particular Candida species identified by CHROM agar. Three Candida isolates did not correlate with morphology on corn meal agar and hence did not fit with any Candida species. However on CHROM agar t h e s e were identified as Candida glabrata, Candida albicans and Candida krusei. One Candida albicans which showed green colored colonies on CHROM agar and was positive for Germ tube test (GTT) did not show any chlamydospores on Corn meal agar, but only elongated yeast cells. The other two isolates, one identified as Candida glabrata on CHROM agar resembled Candida parapsilosis in morphology on corn meal agar (short pencil shaped yeast cells) and the one identified as Candida krusei on CHROM agar gave a typical morphology of Candida tropicalis (oval blastoconidia and long pseudohyphae) on corn meal agar. Two other isolates did not produce any color on Chrom agar and showed round to oval yeast cells on corn meal agar. Antifungal susceptibility of all the 50 Candida isolates for fluconazole was performed by disc diffusion method. None of the isolates in the study were resistant to fluconazole. Only 3 Candida isolates were intermediately susceptible showing that they were dose dependent susceptible (two Candida glabrata and one Candida tropicalis). The two Candida glabrata strains were isolated from high vaginal swab and one Candida tropicalis was isolated from blood. All the other Candida isolates were sensitive to fluconazole. Minimum Inhibitory concentration (MIC) was performed by agar dilution method for amphotericin B for thirty seven Candida isolates. It ranged from <0.062μg/ml to 1μg/ml for the 37 Candida isolates tested, which were sensitive.

DISCUSSION

The incidence of fungal infections particularly those caused by Candida species, has increased markedly over the past decade, especially in immuno-compromised patients. Although the recent use of combination anti-retro viral therapy in AIDS patients may help to reverse the trend, Candida infection remain by far the commonest opportunistic infection in AIDS patients. There is a striking increase in the frequency with which non albicans species are isolated primarily Candida tropicalis, Candida glabrata, Candida krusei, and Candida parapsilosis. (9) . In the present study 70% were non albicans compared to 30% of the Candida isolates were Candida albicans. Among the non albicans, Candida tropicalis was the most commonly isolated (30%). This indicates that it is no longer sufficient to confirm or exclude only Candida albicans from clinical specimens. The conventional test used for speciation are time consuming and cumbersome. Moreover the germ tube test, which is the commonest test employed and gives rapid results, is not very accurate as more than 5% Candida albicans can be negative. Some non albicans species like Candida tropicalis and Candida parapsilosis are occasionally germ tube positive.(10) . It is necessary to evaluate simple, cost effective and rapid method like chromogenic medium to identify Candida to species level. Chromogenic agar is a newer and more rapid method to speciate Candida, contains enzymatic substrates that are linked to chromogenic compounds. When specific enzyme cleaves the substrate, the chromogenic substances produce colour. The action of different enzymes produced by yeast species results in color variation which is useful for the presumptive identification of some yeast (11) In our study 45 isolates could be speciated based on the color on the chromogenic medium and the morphology on Corn meal agar blocks. In the present study CHROM agar could identify, Candida albicans, Candida tropicalis, Candida glabrata and Candida paraopsilosis. Similar findings have been reported by different workers in the literature. (5, 12, 13) . Some of these studies have shown that Candida krusei could be identified by CHROM agar, but in our study 2 isolates identified as Candida krusei on CHROM agar did not show the typical morphology on corn meal agar. The Chromogenic medium facilitates presumptive identification of yeast isolates upto the species level within 24hrs of incubation. Primary inoculation of the clinical specimen on chromogenic medium which are positive for yeast cells on direct gram stain can hasten the presumptive species identification of yeast in clinical specimens. This will allow early initiation of appropriate therapy. The antifungal susceptibility of the Candida isolates to Amphotericin B revealed that all the isolates were sensitive to it. But some studies have stated that amphotericin B resistance is on the rise. A study from South India has shown that 100% of Candida albicans isolates and 88.9% of Candida parapsilosis were sensitive to Amphotericin B (14) The fluconazole susceptibility was tested by disc diffusion method and showed that 47(94%) Candida isolates were sensitive. The three isolates (2 identified as Candida glabrata and 1 Candida tropicalis) were dose dependant susceptible. Studies from India have shown that fluconazole resistance varies from zero to 30.6%. (14-16) The limitation in this study was that no other method was used to confirm the identity of the Candida isolates like the conventional carbohydrate fermentation and assimilation tests or the Vitek system.

CONCLUSION

Use of chromogenic medium with the morphology on corn meal agar provides rapid identification of commonly isolated Candida species from clinical specimens. This will be useful to initiate appropriate antifungal therapy thereby reducing morbidity and mortality. The findings of this study have helped our clinical laboratory to use chromogenic agar for presumptive identification of Candida species from clinical specimens.

ACKNOWLEDGEMENT

This study was supported by the Indian Council of Medical Research (ICMR) as a short term research project awarded to Ms.Dharmeswari T, under the guidance of Dr Sheela Devi C (ICMT STS project No.2011-01728).

References:

REFERENCES

1. David Trofa, Attila Gácser and Joshua D. Nosanchuk. Candida parapsilosis, an Emerging Fungal Pathogen. Clin. Microbiol. Rev. 2008; 21(4):606.

2. Jacqueline M. Achkar and Bettina C. Fries. Candida Infections of the Genitourinary Tract. Clin. Microbiol. Rev. 2010; 23(2):253.

3. Wingard JR. Importance of Candida species other than Candida albicans as pathogens in oncology patients. Clin. Infect. Dis.1995;20:115-125.

4. Chakrabarti. A, Singh. S and Das. S. Changing pace of nosocomial candidemia. Indian. J.Med. Microbiol.1999;17:160-166.

5. Li-Ung Huang,Chi-Hsiang Chen,Chu –Fang Chou, Jang- Jih Lu,Wei-Mingchi, Wei-Hwa Lee. A comparison of methods for yeast identification including CHROM agar Candida , Vitek system YBC and a traditional biochemical tests. Chinese Medical Journal (Taipei.).2001;64:568-574.

6. Agarwal S, Manchanda V, Verma N, Bhalla P. Yeast identification in routine clinical microbiology laboratory and its clinical relevance. Ind J of Med Microbiol 2011; 29 : 2:172-177.

7. Wayne Pa. National committee for antifungal disk diffusion susceptibility testing yeast: approved guidelines; CLSI;M-44A;2004.

8. Wayne Pa. Clinical and laboratory Standards Institute Reference method for broth dilution antifungal susceptibility testing of yeasts; approved standard-third edition; CLSI document M27-A3;2008(a)

9. Malini R Capoor, Deepthi Nair, Manorama Deb, Pradeep Kumar Verma, Lakshmi Srivastva and Pushpa Aggarwal. Emergence of Non albicans Candida species and antifungal resistance in a tertiary care hospital. Jpn J Infect Dis.2005; 58:344- 348.

10. J.E. Hoppe, P.Frey: Evaluation of six commercial tests and the germ tube test for the presumptive identification of Candida albicans. Eur. J. Clin. Microbiol. Infect. Dis.1999; 18:188-191.

11. Odds, F. C., and R. Bernaerts. 1994. CHROM agar Candida, a new differential isolation medium for presumptive identification of clinically important Candida species. J. Clin. Microbiol. 32:1923-1929.

12. Ann P Koehler, Kai Cheong Chu, Elizabeth T S Houang and Augustine F B Cheng. Simple, reliable and cost effective yeast identification scheme for the clinical laboratory. J. Clin. Microbiol.1999;37(2):422- 426.

13. Pfaller M A, Houston A and Coffmann S. Application of CHROM agar Candida for rapid screening of clinical specimens for Candida albicans, Candida tropicalis, Candida krusei and Candida glabrata. J Clin Microbiol.1996;34:58-61.

14. L Srinivasan and J Kenneth. Antibiotic susceptibility of Candida isolates in a tertiary care hospital in Southern India .Indian J Med Microbiol.2006;24(1):80-81

15. Adikary.R. Joshi.S. Species distribution and anti-fungal susceptibility of candidemia at a multi- super specialty center in southern India. Indian J Med Microbiol.2011; 29(3):309- 311.

16. Rizvi MW, Malik A, Shahid M, Singhal S. Candida albicans infections in a North Indian tertiary care hospital: Antifungal resistance pattern and role of SDS-PAGE for characterization. Biology and Medicine 2011; 3(2):176-181.

Research Incentive Schemes

Awards, Research and Publication incentive Schemes by IJCRR

Best Article Award: 

One article from every issue is selected for the ‘Best Article Award’. Authors of selected ‘Best Article’ are rewarded with a certificate. IJCRR Editorial Board members select one ‘Best Article’ from the published issue based on originality, novelty, social usefulness of the work. The corresponding author of selected ‘Best Article Award’ is communicated and information of award is displayed on IJCRR’s website. Drop a mail to editor@ijcrr.com for more details.

Women Researcher Award:

This award is instituted to encourage women researchers to publish her work in IJCRR. Women researcher, who intends to publish her research work in IJCRR as the first author is eligible to apply for this award. Editorial Board members decide on the selection of women researchers based on the originality, novelty, and social contribution of the research work. The corresponding author of the selected manuscript is communicated and information is displayed on IJCRR’s website. Under this award selected women, the author is eligible for publication incentives. Drop a mail to editor@ijcrr.com for more details.

Emerging Researcher Award:

‘Emerging Researcher Award’ is instituted to encourage student researchers to publish their work in IJCRR. Student researchers, who intend to publish their research or review work in IJCRR as the first author are eligible to apply for this award. Editorial Board members decide on the selection of student researchers for the said award based on originality, novelty, and social applicability of the research work. Under this award selected student researcher is eligible for publication incentives. Drop a mail to editor@ijcrr.com for more details.


Best Article Award

A Study by Juna Byun et al. entitled "Study on Difference in Coronavirus-19 Related Anxiety between Face-to-face and Non-face-to-face Classes among University Students in South Korea" is awarded Best Article for Vol 12 issue 16
A Study by Sudha Ramachandra & Vinay Chavan entitled "Enhanced-Hybrid-Age Layered Population Structure (E-Hybrid-ALPS): A Genetic Algorithm with Adaptive Crossover for Molecular Docking Studies of Drug Discovery Process" is awarded Best article for Vol 12 issue 15
A Study by Varsha M. Shindhe et al. entitled "A Study on Effect of Smokeless Tobacco on Pulmonary Function Tests in Class IV Workers of USM-KLE (Universiti Sains Malaysia-Karnataka Lingayat Education Society) International Medical Programme, Belagavi" is awarded Best article of Vol 12 issue 14, July 2020
A study by Amruta Choudhary et al. entitled "Family Planning Knowledge, Attitude and Practice Among Women of Reproductive Age from Rural Area of Central India" is awarded Best Article for special issue "Modern Therapeutics Applications"
A study by Raunak Das entitled "Study of Cardiovascular Dysfunctions in Interstitial Lung Diseas epatients by Correlating the Levels of Serum NT PRO BNP and Microalbuminuria (Biomarkers of Cardiovascular Dysfunction) with Echocardiographic, Bronchoscopic and HighResolution Computed Tomography Findings of These ILD Patients" is awarded Best Article of Vol 12 issue 13 
A Study by Kannamani Ramasamy et al. entitled "COVID-19 Situation at Chennai City – Forecasting for the Better Pandemic Management" is awarded best article for  Vol 12 issue 12
A Study by Muhammet Lutfi SELCUK and Fatma COLAKOGLU entitled "Distinction of Gray and White Matter for Some Histological Staining Methods in New Zealand Rabbit's Brain" is awarded best article for  Vol 12 issue 11
A Study by Anamul Haq et al. entitled "Etiology of Abnormal Uterine Bleeding in Adolescents – Emphasis Upon Polycystic Ovarian Syndrome" is awarded best article for  Vol 12 issue 10
A Study by Arpita M. et al entitled "Estimation of Reference Interval of Serum Progesterone During Three Trimesters of Normal Pregnancy in a Tertiary Care Hospital of Kolkata" is awarded best article for  Vol 12 issue 09
A Study by Ilona Gracie De Souza & Pavan Kumar G. entitled "Effect of Releasing Myofascial Chain in Patients with Patellofemoral Pain Syndrome - A Randomized Clinical Trial" is awarded best article for  Vol 12 issue 08
A Study by Virendra Atam et. al. entitled "Clinical Profile and Short - Term Mortality Predictors in Acute Stroke with Emphasis on Stress Hyperglycemia and THRIVE Score : An Observational Study" is awarded best article for  Vol 12 issue 07
A Study by K. Krupashree et. al. entitled "Protective Effects of Picrorhizakurroa Against Fumonisin B1 Induced Hepatotoxicity in Mice" is awarded best article for issue Vol 10 issue 20
A study by Mithun K.P. et al "Larvicidal Activity of Crude Solanum Nigrum Leaf and Berries Extract Against Dengue Vector-Aedesaegypti" is awarded Best Article for Vol 10 issue 14 of IJCRR
A study by Asha Menon "Women in Child Care and Early Education: Truly Nontraditional Work" is awarded Best Article for Vol 10 issue 13
A study by Deep J. M. "Prevalence of Molar-Incisor Hypomineralization in 7-13 Years Old Children of Biratnagar, Nepal: A Cross Sectional Study" is awarded Best Article for Vol 10 issue 11 of IJCRR
A review by Chitra et al to analyse relation between Obesity and Type 2 diabetes is awarded 'Best Article' for Vol 10 issue 10 by IJCRR. 
A study by Karanpreet et al "Pregnancy Induced Hypertension: A Study on Its Multisystem Involvement" is given Best Paper Award for Vol 10 issue 09
Late to bed everyday? You may die early, get depression
Egg a day tied to lower risk of heart disease
88 Percent Of Delhi Population Has Vitamin D Deficiency: ASSOCHAM Report

List of Awardees

Awardees of COVID-19 Research

Woman Researcher Award

A Study by Neha Garg et al. entitled "Optimization of the Response to nCOVID-19 Pandemic in Pregnant Women – An Urgent Appeal in Indian Scenario" published in Vol 12 issue 09

A Study by Sana Parveen and Shraddha Jain entitled "Pathophysiologic Enigma of COVID-19 Pandemic with Clinical Correlates" published in Vol 12 issue 13

A Study by Rashmi Jain et al. entitled "Current Consensus Review Article on Drugs and Biologics against nCOVID-19 – A Systematic Review" published in Vol 12 issue 09

Emerging Researcher Award

A Study by Madhan Jeyaraman et al. entitled "Vitamin-D: An Immune Shield Against nCOVID-19" published in Vol 12 issue 09

Study by Dheeraj Kumar Chopra et al. entitled "Lipid-Based Solid Dispersions of Azilsartan Medoxomil with Improved Oral Bioavailability: In Vitro and In Vivo Evaluation" published in Vol 12 issue 19


RSS feed

Indexed and Abstracted in


Antiplagiarism Policy: IJCRR strongly condemn and discourage practice of plagiarism. All received manuscripts have to pass through "Plagiarism Detection Software" test before forwarding for peer review. We consider "Plagiarism is a crime"

IJCRR Code of Conduct: We at IJCRR voluntarily adopt policies on Code of Conduct, and Code of Ethics given by OASPA and COPE. To know about IJCRRs Code of Conduct, Code of Ethics, Artical Retraction policy, Digital Preservation Policy, and Journals Licence policy click here

Disclaimer: International Journal of Current Research and Review (JICRR) provides platform for researchers to publish and discuss their original research and review work. IJCRR can not be held responsible for views, opinions and written statements of researchers published in this journal.



Company name

International Journal of Current Research and Review (JICRR) provides platform for researchers to publish and discuss their original research and review work. IJCRR can not be held responsible for views, opinions and written statements of researchers published in this journal

Contact

148, IMSR Building, Ayurvedic Layout,
        Near NIT Complex, Sakkardara,
        Nagpur-24, Maharashtra State, India

editor@ijcrr.com

editor.ijcrr@gmail.com


Copyright © 2020 IJCRR. Specialized online journals by ubijournal .Website by Ubitech solutions