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DOI: 10.14260/jemds/2015/1290 ORIGINAL ARTICLE MYCOTIC KERATITIS IN AND AROUND HYDERABAD Prathiba1, Taruni2, V. Sudha Rani3, Aruna Sunder4 HOW TO CITE THIS ARTICLE: Prathiba, Taruni, V. Sudha Rani, Aruna Sunder. “Mycotic Keratitis in and Around Hyderabad”. Journal of Evolution of Medical and Dental Sciences 2015; Vol. 4, Issue 51, June 25; Page: 8913-8917, DOI: 10.14260/jemds/2015/1290 ABSTRACT: CONTEXT: Patients attending with corneal ulcers. AIM OF THE STUDY: To evaluate the specific pathogenic agents and epidemiology of mycotic keratitis at a tertiary care center in Hyderabad. MATERIAL & METHODS: A total of 863 patients of all age groups suffering from corneal ulcers, who attended Sarojinidevi eye hospital from ‘Jan. 2014 to Dec. 2014’ were included in the study. All patients underwent through slit lamp bio-microscopic examination by an ophthalmologist. The material was obtained from active margins and base of ulcer after the debridement of superficial mucus using a sterile No. 15. Bard – parker blade under slit lamp bio-microscope. Samples were inoculated on blood agar, mac-conkey agar sabauraud’s dextrose agar & brain heart infusion broth. The material obtained in next scraping was thinly spread onto labeled slides for direct examination. The specific identification of pathogen was based onmicroscopic morphology, staining characteristics and biochemical properties using standard laboratory protocol. RESULTS: Out of total 863 corneal scrapings collected from confirmed corneal ulcer patients, 563 were from males and 300 were from females. Highest number of males were in the age group of 49-60, whereas more number of females were above 60 years. Highest number of cases were in the month of August. Out of 863 scrapings 363 samples (40.25%) were positive for fungal elements by 10% KOH mount examination and 294 samples (34.25%) were culture positive for fungi. Out of 294 fungal isolates, commonest isolate was Aspergillus species followed by Fusarium species. CONCLUSIONS: The epidemiological pattern of corneal ulceration varies significantly from country to country and even from region to region. The findings of our study show that there is a region wise variation in the predominance of corneal pathogens. This has an important public health implication for the initiation of therapy. KEYWORDS: Keratitis, KOH mount, Aspergillus, Fusarium. INTRODUCTION: Mycotic keratitis (International Nomenclature of Diseases disease number 2100) is a general term for a mycosis of thecornea, and can be caused by a wide variety of fungi.1Thiscondition is usually manifested by severe inflammation, theformation of a corneal ulcer, and hypopyon, with the presence of fungal hyphae within the corneal stroma. Synonyms include ‘keratomycosis’ and ‘oculomycosis’ (In part), but ‘mycotic keratitis’ is recommended in preference to ‘keratomycosis’ so as to have similar names for the diseases caused by fungi, bacteria and viruses.1 If the fungal species causing the infection is identified, a term such as ‘Fusarium keratitis’ (Or, more specifically, ‘keratitis due to Fusariumsolani’) is recommended.1Corneal infection is a leading cause of ocular morbidity and blindness worldwide.2,3,4,5,6,7 Corneal ulceration is a major cause of monocular blindness in developing countries. Surveys in Africa and Asia have confirmed these findings.2,3,4,5,6,7 and a recent report on the causes of blindness worldwide consistently lists corneal scarring second only to cataract as the major aetiology of blindness and visual disability in many of the developing nations in Asia, Africa and the MiddleEast.8 Any microorganism can invade the corneal stroma if the normal corneal defense mechanisms such as lids, tear film and corneal epithelium iscompromised.9 More than 70 species of filamentous fungi are also identified as etiological agents of J of Evolution of Med and Dent Sci/ eISSN- 2278-4802, pISSN- 2278-4748/ Vol. 4/ Issue 51/ June 25, 2015 Page 8913 DOI: 10.14260/jemds/2015/1290 ORIGINAL ARTICLE fungal keratitis.10 The early diagnosis and its treatment is utmost important in preventing complications and loss of vision. The purpose of this study was to evaluate the specific pathogenic agents and epidemiology of mycotic keratitis at a tertiary care center in Hyderabad. MATERIAL AND METHODS: A total of 863 patients of all age groups suffering from corneal ulcers, who attended Sarojinidevi eye hospital from Jan ‘2014 to Dec’ 2014 were included in the study. All patients underwent through slit lamp bio-microscopic examination by an ophthalmologist. A break in continuity of epithelium associated with underlying stormily infiltrate was considered infectious unless proved otherwise. After a detailed ocular examination, corneal scrapings were taken under aseptic condition after installation of 4% preservative–free lignocaine (Lidocaine) drops. The material was obtained from active margins and base of ulcer after the debridement of superficial mucus using a sterile No. 15. Bard–parker blade under slit lamp bio-microscope. Samples were inoculated on blood agar, mac-conkey agar sabauraud’s dextrose agar & brain heart infusion broth. Sabauraud’s dextrose agar (SDA) media were incubated at room temperature and the remaining were incubated at 370C and evaluated after 24hrs. The material obtained in next scraping was thinly spread onto labeled slides for direct examination. Direct examination: 1. 10% KOH mount. 2. Grams stain. 3. Acid fast stain was done only when clinician asks for it. OBSERVING FOR FUNGAL ETIOLOGY: KOH (Postassiurn hydroxide) mount-material of the scraping was placed on a glass slide within 10% KOH and a cover slip placed, then observed under the microscope using low power and high power by direct microscopy. SDA culture media were examined daily for 21days if no growth seen then media was discarded, standard operating laboratory protocol was followed for all laboratory methods. Microbial cultures were considered positive only if at least one of the following criteria were met: 1. The growth of the same organism was demonstrated on twoor more solid media on C streak or there was semi confluentgrowth at the site of inoculation on the solid medium. 2. The same organism was isolated on repeated scrapping. 3. If it was consistent with clinical sign. 4. Smear results were consistent. The specific identification of pathogen was based onmicroscopic morphology, staining characteristics andbiochemical properties using standard laboratory protocol. RESULTS: Out of total 863 corneal scrapings collected from confirmed corneal ulcer patients, 563 were from males and 300 were from females. Highest number of males were in the age group of 4960, whereas more number of females were above 60 years. Highest number of cases were in the month of August. (Table1). Out of 863 scrapings 363 samples (40.25%) were positive for fungal elements by 10% KOH mount examination and 294 samples (34.25%) were culture positive for fungi. Out of 294 fungal isolates, commonest isolate was Aspergillus species followed by Fusarium species, 3 isolates were Candida, 20 were unidentified. (Figure 1). J of Evolution of Med and Dent Sci/ eISSN- 2278-4802, pISSN- 2278-4748/ Vol. 4/ Issue 51/ June 25, 2015 Page 8914 DOI: 10.14260/jemds/2015/1290 ORIGINAL ARTICLE Month JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC Total 0-19 20-40 41-59 Above 60 Male Female Male Female Male Female Male Female Total 5 3 26 8 17 5 14 7 85 4 3 9 5 16 14 16 3 70 1 4 9 10 15 11 11 8 69 3 1 13 4 20 2 9 4 56 9 3 13 9 16 6 11 12 79 2 17 13 15 10 16 13 86 6 1 23 8 14 4 18 11 85 5 3 18 6 19 8 20 11 90 4 1 20 7 12 4 10 8 66 5 2 8 8 13 8 7 9 60 2 1 12 5 15 4 10 6 55 2 3 12 8 12 5 9 11 62 48 25 180 91 184 81 151 103 863 Table 1: Showing age and monthwise distribution of total corneal ulcers Fig. 1: Showing different fungal isolates DISCUSSION: Our study revealed that fungal keratitis accounted for 34.2% of the total microbial keratitis patients who presented to our center. This high prevalence of fungal pathogens in South India was not so different from that found in similar studies done by Sharma Amisha, Agrawal Parul et al (35.66%).11 The age distribution showed the incidence of fungal keratitis predominantly between the 4th to 6th decades, reflecting the active working period of life and hence the increased vulnerability to injury during outdoor activities. The incidence of fungal keratitis was significantly higher in males, in individuals from rural area and following corneal injury. The male predominance of fungal keratitis noted in the present study correlates with the studies of Bharathiet al, Srinivasan et al and Chowdhary et al.12,13 Aspergillusspp was the predominant isolate as compared to Fusariumspp as in the various studies by Samar K, Basak et al(59.8%), Jagdish. J of Evolution of Med and Dent Sci/ eISSN- 2278-4802, pISSN- 2278-4748/ Vol. 4/ Issue 51/ June 25, 2015 Page 8915 DOI: 10.14260/jemds/2015/1290 ORIGINAL ARTICLE Chander et al (41.18%), JagdishChander et al (35%), Javadi et al (34.8%), Shokohi et al (33.3%).14,15,16,17,18 In our case highest number of corneal ulcers occurred in the month of August. In a study done by Lin, Charles C. MD*,†; Lalitha, Prajna MD‡; Srinivasan, Muthiah MD†; Prajna, N. et al there was uneven distribution of fungal keratitis throughout the year with peaks in July and January. No significant seasonal trend was observed for the combined bacterial keratitis group.19 CONCLUSIONS: The epidemiological pattern of corneal ulceration varies significantly from country to country and even from region to region. . The findings of our study show that there is a region wise variation in the predominance of corneal pathogens. This has an important public health implication for the initiation of therapy. REFERENCES: 1. Council for International Organizations of Medical Sciences (CIOMS). International Nomenclature of Diseases. Volume II: Infectious Diseases. Part 2 Mycoses, Geneva: CIOMS, 1982. 2. Chirambo M. C, Tielsch J. M, West KP, Katz J. Blindness and visual impairment in Southern Malawi. Bull WHO 1986; 64: 567-572. 3. Chirambo M. C. Causes of blindness among students in blind school institutions in a developing country. Br J Ophthalmol 1976; 60: 665-668. 4. Rapoza PA, West SK, Katala SJ, Taylor HR. Prevalence and causes of vision loss in Central Tanzania. IntOphthalmol 1991; 15: 123-129. 5. Brilliant LB, Pokhrel RP, Grasset NC, Lepkowski JM, Kolstad A, Hawks W, et al. Epidemiology of blindness in Nepal. Bull WHO 1985; 63: 375-386. 6. Khan MU, Hague MR, Khan MR. Prevalence and causes of blindness in rural Bangladesh. Ind J Med Res 1985; 82: 257-262. 7. Gilbert CE, Wood M, Waddel K, Foster A. Causes of chilhood blindness in East Africa: results in 491 pupils attending 17 school for the blind in Malawi, Kenya and Uganda. Ophthamic Epidemiol 1995; 2: 77-84. [PUBMED] 8. Thylefors B, Negrel AD, Segaram PR, Dadzie KY. Available data on blindness (update 1994). Ophthalmic Epidemiology1995; 2: 5-39. 9. Garg P, Rao GN. Corneal ulcer: Diagnosis and management. Community eye health 1999; 22: 2124. 10. Agarwal PK, Roy P, Das A, Banerjee A, Maity PK, Banerjee AR. Efficacy of topical and systemic itraconazole as a broad-spectrum antifungal agents in mycotic corneal ulcer: A preliminary study. Indian J Ophthalmol 2001; 49: 173-76.[PUBMED] 11. Sharma Amisha , Agrawal Parul , Dhiman D., Maurya A. K. , Baranwal A. K. et al 1, Etiological diagnosis of microbial keratitis in patients of West U.P.study of 272 cases over a period of 2 years; Journal of Advance Researches in Biological Sciences, 2013, Vol. 5 (2) 164-166. 12. M Jayahar Bharathi, R Ramakrishnan, S Vasu, R Meenakshi, R Palaniappan Department of Microbiology, Aravind Eye Care System, Tirunelveli, Tamil Nadu, India Epidemiological characteristics and laboratory diagnosis of fungal keratitis. A three-year study, 2003: 51: 4: 315-321. 13. Thomas PA –Fungal infections of the cornea. Eye 2003; 17: 852-62. J of Evolution of Med and Dent Sci/ eISSN- 2278-4802, pISSN- 2278-4748/ Vol. 4/ Issue 51/ June 25, 2015 Page 8916 DOI: 10.14260/jemds/2015/1290 ORIGINAL ARTICLE 14. Samar K Basak, Sukumar Basak, Ayan Mohanta, Arun Bhowmick–Epidemiological and microbiological diagnosis of Suppurative Keratitis in Gangetic West Bengal, Eastern India. Indian J Ophthalmol 2005; 53: 17-22. 15. 15. Tahereh Shokohi, Kiumars Nowroozpoor, Dailami – Fungal Keratitis in patients with corneal ulcer in San, Northern Iran. Archives of Iranian Medicine 2006; 9: 222 – 227. 16. Jagdish Chander, NidhiSingla, NaliniAgnihotri, Sudesh Kumar Arya,Antariksh Deep Keratomycosis in and around Chandigarh ; A five year study from a north Indian tertiary care hospital. Indian J Patho-Microbiol 2008: 51: 2: 304–306. 17. Chander J, Chakrabarti A, Shama A, Saini JS, Panigarhi D - Evaluation of calcofluor white staining in the diagnosis of fungal corneal ulcer. Mycosis 1993; 36: 243 – 245. 18. Vajpayee R.B et.al – Laboratory diagnosis of keratomycosis: comparative evaluation of direct microscopy and culture results. Ann Ophthalmol, 25(2): 68-71.Copyright 2010 Bio Med Sci Direct Publications IJBMR. 19. Lin, Charles C. MD*,†; Lalitha, Prajna MD‡; Srinivasan, Muthiah MD†; Prajna, N. et al Seasonal trends of microbial keratitis in South India.Cornea. 2012; 31(10): 1123-7. AUTHORS: 1. Prathiba 2. Taruni 3. V. Sudha Rani 4. Aruna Sunder PARTICULARS OF CONTRIBUTORS: 1. Assistant Professor, Department of Microbiology, Sarojini Devi Eye Hospital, Hyderabad, Osmania Medical College. 2. Assistant Professor, Department of Microbiology, SRRIT&CD, Osmania Medical College. 3. Associate Professor, Department of Microbiology, KMC Warangal. FINANCIAL OR OTHER COMPETING INTERESTS: None 4. Professor, Department of Microbiology, Sarojini Devi Eye Hospital, Osmania Medical College, Hyderabad. NAME ADDRESS EMAIL ID OF THE CORRESPONDING AUTHOR: Dr. V. Sudha Rani, Associate Professor, Department of Microbiology, Kakatiya Medical College, Warangal, Telangana. E-mail: [email protected] [email protected] Date of Submission: 05/06/2015. Date of Peer Review: 06/06/2015. Date of Acceptance: 19/06/2015. Date of Publishing: 24/06/2015. J of Evolution of Med and Dent Sci/ eISSN- 2278-4802, pISSN- 2278-4748/ Vol. 4/ Issue 51/ June 25, 2015 Page 8917