Indian Journal of Microbiology Research

295-298

Authors: Lekshmi Balaraman, Saritha N, Geetha Raveendran

Introduction and Objectives: Antibiotics, immunodeficiencies and advances in medical practices have all contributed to increased incidence in fungal infections. In recent times Non albicans Candida species are increasingly being isolated from clinical specimens. The present study was aimed at the characterization and antifungal susceptibility of Non albicans Candida species from clinical specimens received in Microbiology Laboratory for one year from June 2015–May 2016. Materials and Methods: A total of 181 Non albicans Candida isolates were included in the study. Candida was identified from clinical samples by gram stain, culture on Saboraud’s Dextrose Agar (SDA) and germ tube test. The organisms were further speciated by cornmeal agar morphology, sugar assimilation and fermentation tests and growth on CHROM agar (Himedia). Antifungal sensitivity was performed by using disc diffusion method as per CLSI guidelines. Results: The most common isolate was Candida tropicalis (59.11%) followed by C.glabrata (28.18%), C.parapsilosis (8.83%) and C.krusei (3.88%). Highest number of isolates were from urine (74/181). Resistance to Fluconazole in clinical isolates of C.tropicalis has increased. C.krusei was the most resistant species. Conclusion: Use of CHROM agar is rapid, simple and cost effective compared to time consuming technically demanding expensive conventional methods. Antifungal susceptibility testing has to be carried out before starting therapy because of varying susceptibility pattern in different species of Candida.

303-306

Authors: Suresh Ahirwar, Vikramjeet Singh, Manodeep Sen, Vineeta Mittal, Anupam Das

Introduction: Superficial mycoses refer to the disease of skin, hair and nail caused by fungi, are a serious problem due to deficient sanitation and education. This group includes dermatophytoses, non dermatophytic molds and Candida spp. Microscopic detection of fungal elements using wet mount preparation of 10% and 40% potassium hydroxide (KOH) is a rapid and precise tool for laboratory diagnosis. Objectives: This retrospective study was conducted to evaluate the sensitivity of wet preparation direct microscopy using potassium hydroxide for early detection of fungal elements from various clinical specimens and thereby helping the clinicians in initiating the appropriate antifungal treatments. Materials and Methods: All clinical specimen like skin and nail scrapping from patients suspected of superficial mycoses were subjected to KOH examination for screening and detection and positive samples were cultured on SDA (Sabouraud’s Dextrose Agar) medium for identification. Results: A total of 5734 samples of clinically suspected patients of superficial mycoses attended the tertiary care teaching hospital between September 2013 and August 2017 were included. 3165 patients (55%) were found to be positive by KOH examination. In the present study the most common clinical type was tinea cruris (74.3%) followed by tinea corporis (22.6%) in the patients attending tertiary health care centre. Conclusion: Clinical finding and wet mount with KOH can be believed upon as the single most important rapid and sensitive tool for early diagnosis of superficial mycoses. Since there is a high incidence of superficial mycoses in rural people coming to the tertiary heath care facilities therefore preventive and early screening measures are need of hours.

313-317

Authors: Anjana Gopi, Faiza Samreen, Madhulatha C.K

Introduction: Tuberculosis (TB) is still a leading cause of death in many parts of the world. Adept techniques of rapid detection, isolation, identification and susceptibility testing of Mycobacterium tuberculosis (M.tb) are essential for management. Our aim was to compare the sensitivity and rapidity of smear microscopy, concentration method, rapid slide culture, Lowenstein - Jensen (LJ) culture and MGIT semiautomated culture methods in detection of Mycobacterium tuberculosis among smear negative pulmonary TB and extra pulmonary TB cases and to detect the drug resistance pattern to first line drugs among M.tb isolates. Materials and Methods: A prospective study was conducted for one year between January 2016 to January 2017 in the Department of Microbiology at a tertiary care hospital. Sputum smear negative pulmonary and extra pulmonary specimens were subjected to direct microscopy by Zeihl-Neelsen and fluorescent staining. Culture was done by Rapid slide culture, Lowenstein-Jensen media (LJ media) and BACTEC MGIT system. Positive isolates were confirmed by appropriate biochemical reactions and rapid immunochromatographic test. Drug susceptibility testing was done for first line drugs. Results: The sensitivity of direct microscopy was 3% and after concentration it was 4%. Overall culture positivity was 12%. Detection rate by LJ method, rapid slide culture method and MGIT methods was 6%, 4% and 12% respectively. The mean detection time was 22.5 days by MGIT and 31.5 days by conventional LJ method. Conclusion: This study highlights the importance of culturing suspected smear negative pulmonary and extra pulmonary Tuberculosis cases prior to empirical therapy. Newer automated culture methods aid in earlier detection of cases.

326-330

Authors: Ahmed Yahya Al-Qudari, Maged Ahmed Al-Garadi, Fawaz Ali Al-Monifi

Introduction: Brucellosis is one of the common zoonotic diseases that leading to the extensive economic losses throughout the world. Control and eradication of this disease depends mainly on the early detection. Aims: The present work was achieved between March and September 2016 as a cross-sectional study to determine the sero-prevalence of brucellosis in cattle. Materials and Methods: Three hundred eighty four sera samples which including 359 females and 25 males were collected randomly from unvaccinated cattle, in twelve different regions at Dhamar governorate in Yemen. All sera samples screened for cattle brucellosis using RBT and the positive samples reconfirmed using I-ELISA. A structured questionnaire used to collect epidemiological data that analyzed using SAS program. Results and Discussion: The sero-prevalence of cattle brucellosis was 0.26%. No significant association (P?0.05) between prevalence of cattle brucellosis and region (2.8%), age (3.22%), sex (0.27%) and breed (0.26%) was determined. Conclusion: Study findings showed a low prevalence of cattle brucellosis in Dhamar governorate. Although, the current work form a baseline data for more study of cattle brucellosis, and start point for its control in Yemen.

334-338

Authors: Pritam Pardeshi

troduction: Urinary tract infection (UTI) is one of the most prevalent diseases affecting people of all age groups i.e. from neonate to geriatric age group. Widespread use of antibiotics has led to the emergence of resistant microorganisms. As the antibiogram of the microorganisms are frequently changing hence the present study was done to analyze the recent antibiotic sensitivity pattern of uropathogens in urinary tract infection. Materials and Methods: In present study is a retrospective analysis of culture results of urine samples, was conducted at Microbiology department of tertiary health care hospital in Mumbai. The age & sex of patients, the organism isolated and also the antimicrobial susceptibility profiles were collected from the laboratory registers using a standard data collection form. Results: The overall prevalence of UTI was 33.54% of which 66.78% were females and 33.22% were from males. High prevalence was observed in females as compared to males (2:1). Though the overall prevalence was high in old aged (>45 years) patients, in females high prevalence was seen among middle-aged (31 to 45 years) patients and in male high prevalence was seen among old age (>45 years) patients. From total 584 uropathogens, E.coli (53.77%) was the commonest isolate causing UTI followed by Klebsiella pneumoniae (27.40%). The most effective antimicrobial agents in our study were Meropenem, gentamicin, Nitrofurantoin and Cotrimoxazole whereas higher resistance was observed among Fluoroquinolones, Amoxicillin and third generation Cephalosporins, these are the drugs which are commonly given emperically for UTI. Conclusion: As drug resistance among bacterial pathogens is vary with time to time regular surveillance and monitoring is necessary for giving updated information to physician for most effective empirical treatment of UTIs.

342-347

Authors: Satyajeet K Pawar*,S T. Mohite,R V. Shinde,S R. Patil,G S. Karande

Introduction: In last few years, Gram-negative bacilli are isolated capable of producing various classes of carbapenemases with ability of hydrolyzing the ?-lactam antimicrobial. Carbapenem Resistant Enterobacteriaceae (CRE) have been reported worldwide. There is a serious threat to public health due to the emergence and rapid spread of CRE. Aims: To find prevalence and bacterial profile of CRE in clinical isolates in indoor patients from a rural tertiary care centre. Materials and Methods: The study was conducted in a tertiary care teaching hospital in Western India, June 2016 to April 2018. The clinical specimens received in microbiology laboratory were processed by the standard method. Bacteria were identified by VITEK 2 compact (Biomerieux) automation system, and antimicrobial susceptibility testing was done with the same system to detect minimum inhibitory concentrations for carbapenem group of antimicrobials. CLSI 2016 guidelines were used to detect CRE. Results: Total 535 Enterobactericaeae clinical isolates were included in the study. Of these, 31.77% (n=170) were CRE. Specimens like urine, pus/ wound swab and endotracheal tube secretion were the major contributors for CRE isolates.82% of CRE were Klebsiella pneumoniae (63%) and E.coli (19%). Conclusions: A high prevalence of 31.77% carbapenem resistance was observed among Enterobacteriaceae isolates. Early detection, isolation and contact precaution of CRE patients will help to prevent rapid dissemination of CRE infection

355-357

Authors: R. K. Agarwal, B. Chatterjee, G. Mittal, T. Bhatla, Ankit Agarwal

Introduction: Despite limitations, the Widal test is widely used in resource-constrained settings because it is fast, inexpensive and user-friendly. Therefore, efforts continue to improve its diagnostic utility. Aims: Using 2-mercaptoethanol (2-ME) to detect the IgM fraction of total agglutinins detected by the Widal test in the serum of febrile patients. Materials and Methods: Single serum specimens were collected from 333 patients presenting to our hospital with fever. One aliquot of serum was tested by standard Widal test; a positive result was defined as simultaneous elevation of ‘O’ and 'H' titres to 1:80 or above. A second aliquot was simultaneously tested by a modified Widal test in which serum was diluted in isotonic saline with added 2-ME (0.12M) instead of in plain isotonic saline. Differences in titre between the two versions of Widal test were noted. A four-fold or greater reduction in titre upon testing with 2-ME, was considered evidence of specific IgM. Results: Standard Widal test was positive in 55 (16.5%) out of 333 patients. A ?4-fold reduction of ‘O’ titre in modified Widal test was found in 38 (69.1%) out of 55 cases, indicating the presence of specific IgM. A similar reduction of 'H' titre was seen in only 6 (11%) out of 55 cases. Conclusion: Simultaneous use of standard Widal test and modified Widal test with 2-ME allows the detection of specific IgM at a low cost. This can increase the diagnostic utility of the Widal test.

364-367

Authors: Hitesh R Ahir,Bhavesh P. Gohil

Introduction: With the increasing number of immuno-compromised patients suffering from different types of infections, invasive fungal infections have emerged as a most common cause of morbidity and mortality in these patients. Candida spp. and Aspergillus spp. are the most common causes of fungal infection in immuno-compromised patients. Different types of Candida non-albicansspecies have emerged as a major cause of candidemia in most of the countries. Aim: The aim of this study is to detect the prevalence of fungal infection in patients and to study their sensitivity pattern to different antifungal drugs. Materials and Methods: The study was carried out from January’17 to December’17, to know the fungal infection prevalence, Candida albicans, Aspergillus, opportunistic fungal infections in patients of our institute. During this period total number of 2867 different clinical samples received in the microbiology laboratory for fungal culture and sensitivity testing. All other specimen received in microbiology laboratory was processed using standard microbiological guidelines for yeast and mold isolation. Susceptibility of different fungal species to antifungal agents was performed by using standard guidelines for broth dilution and antifungal disk diffusion susceptibility test. Results: During the study period total 193 fungal species were isolated from 2867 different clinical specimens received in microbiology laboratory. Candida infection was more common in patient age group between 19-60 yrs., also in male patients and also predominates in admitted patients. Invasive fungal infection was most commonly caused by Candidanon-albicans. C.tropicalis was highest isolated followed by C.albican. Conclusion: All Candida isolated were susceptible to Caspofungin. Candida albicans shows 26% and non-albicans shows 4% resistance to Voriconazole.

374-377

Authors: Jaya Lalwani*,Camilla Rodrigues,Rajeev Soma

Introduction: In resource-limited countries like India, where resistance testing is often not available, an accurate treatment history can guide doctors in quantifying suboptimal drug exposure and thus anticipate drug resistance Materials and Methods: In the present longitudinal study, a total of 75 patients failing antiretroviral therapy; the effective drug exposure was calculated after taking into account the adherence, appropriateness of prescriptions, and pharmacokinetic interactions. Treatment was modified and patients were followed up. Results: Of the total 75 patients included in the study; 69 (92.0%), 63 (84.0%) and 42 (56.0%) patients had virological, immunological and clinical failure respectively. Suspected reasons for antiretroviral drug failure were non-adherence in 48 (64.0%), improper prescriptions in 12(16.0%) and drug interactions in 12 (16.0%). We observed that following treatment change, CD4 count increased by a mean of 127 cells/?l and 33 (44.0%) patients had fully suppressed viral loads. Conclusion: Our results show that empirical treatment changes based on a comprehensive drug history, followed by good adherence lead to good treatment outcomes in patients showing one or other evidence of drug resistance.

382-386

Authors: Santosh Shankar Patil*,Bipin Munjappa,,Shubhangi A. Gadgil,Amit Gadve

Introduction: Patients on Maintenance Hemodialysis (HD) are at high risk of viral hepatitis due to blood transfusion, prolonged vascular access, exposure to infected patients and contaminated equipments etc. Knowledge of the prevalence of such infections is very important to access magnitude of the problem because these infections play important role in the morbidity and mortality of hemodialysis (HD) patients. Aim and Objectives: This study was done to determine the prevalence of Hepatitis B virus (HBV) and Hepatitis C virus (HCV) infections in patients on hemodialysis at our tertiary care hospital of Western Maharashtra. Materials and Methods: Settings and Design: Hospital based cross sectional study was conducted for six months in HD unit at our tertiary care hospital. All patients (n=135) were screened for Hepatitis B surface antigen and antibodies to HCV (anti-HCV). Prevalence, age and sex wise distribution as well as risk factors for transmission of these infections were observed. Results: Total 135 HD patients were screened for presence of HBV and HCV infections. Of these, 39 (28.9%), 20 (14.8%), 6 (4.4%) were found to be having HCV, HBV and Dual HBV and HCV infection respectively. Conclusions: HCV was the most prevalent among HD patients than HBV. History of multiple blood transfusions, HD outside center or longer duration of HD were all important risk factor noted for high prevalence of sero-positivity for HBV and/or HCV. Strict adherence to universal precautions careful attention to hygiene and strict sterilization of dialysis machines are important to prevent further transmission of these viruses.

393-397

Authors: Sadhna Sodani,Ranjana Hawaldar*

Introduction: Autoimmune diseases (AID) are a manifestation of a self damaging immune response of the body to its own antigen. In any suspected AID, the first test to be done is ANA detection by indirect immunofluoresence using HeP 2 cells due to its sensitivity. A positive ANA test is further evaluated by performing immunoblot test to detect specific antigens causing autoimmunity. The present study was undertaken to study the prevalence of antinuclear antibodies in patient population of Central Madhya Pradesh as well as to study the prevalence of disease specific antigens. Materials and Methods: A total of 650 patients of all ages and both sexes coming to our diagnostic centre between January to June 2018 for ANA testing were included in the study. Serum ANA was determined by indirect immunofluorescence (IIFA) using Hep 2 cell lines (Euroimmun, Germany). A positive result by IIFA was further tested by immunoblot method (Euroimmun, Germany) when requested by the clinician. Result: Out of 650 patients ANA screen was positive in 280 (43.08%) patients and negative in 209(32.15%) patients. ANA blot was requested in 161 patients (57.5%) of the total 280 ANA screen positive patients. Out of the 161 patients, 69 had a positive Immunoblot test for one or more antigens. Total 53.69% patients were positive by both ANA screen and immunoblot test. The total coincidence rate was 44.5% between ANA screen and ANA blot test with a positive coincidence rate of 81.6% and negative coincidence rate of 79.1%. Conclusion: ANA IIF is considered to be the gold standard screening test for detecting autoantibodies and ANA blot is done as a confirmatory test to detect specific autoantibodies against antigens causing autoimmunity. The combination of ANA IIF and ANA blot is an effective method to diagnose autoimmune diseases. If used alone, a possibility of missing a diagnosis is high and so a combinbation of these two tests is of great benefit to both the clinicians as well as laboratorians.

404-407

Authors: Neelesh Naik,Anusuya Devi D*,Veena Krishnamurthy

Introduction: Staphylococcus have become common cause of skin and soft tissue infections. Resistance to a number of drugs have increased and methicillin resistant Staphylococcus aureus (MRSA) and inducible clindamycin resistance (iMLSB) have become a major problem for the treatment of Staphylococcal infections. This study was undertaken to detect MRSA and iMLSB and to determine the antibiotic susceptibility pattern of the isolates. Materials and Methods: 150 isolates of Staphylococcus were studied for detecting the antibiotic resistance pattern and also to detect MRSA using cefoxitin disc and oxacillin E test. iMLSB resistance among MRSA strains was detected using D test. Results: Out of 150 isolates of Staphylococcus, 117(78%) isolates were of Staphylococcus aureus and 33(22%) isolates were of Coagulase negative Staphylococci. Staphylococcus was most sensitive to vancomycin followed by linezolid and clindamycin. Penicillin was the least sensitive antibiotic. 29 (24.7%) strains of Staphylococcus aureus were MRSA. Among them, 16(44.8%) were erythromycin resistant and 4(13.7%) of erythromycin resistant strains were found to be inducible clindamycin resistant. Conclusion: Testing of all the isolates of Staphylococcus for antibiotic resistance and to test Staphylococcus aureus for MRSA and for iMLSB resistance is important in determining the antibiotic sensitivity which will prevent treatment failure.

415-420

Authors: Aruna Solanki,Arun More,Jyoti Tomar*

Introduction: Microbiota of respiratory tract may be responsible for acute exacerbation of bronchial asthma and chronic bronchitis. So the present study was carried out to find out specific bacterial etiological agents’ involvement in acute exacerbation of these diseases. Materials and Methods: Sputum samples from patients of bronchial asthma, chronic bronchitis during acute exacerbation and endotracheal aspirates from control cases were processed to isolate the microbiota of respiratory tract using quantitative methods after liquefying and diluting the samples. Significant and insignificant bacterial counts from the samples were determined. Grading of pus cells and bacterial morphology was seen by Gram's staining. Results: Pus cell grading of 1+ to 3+ were seen in the both cases of bronchial asthma and chronic bronchitis whereas pus cells were not found in control cases. Percentage of microorganisms by Gram's staining in asthmatics, chronic bronchitis, and controls cases were 93.33%, 86.67%, 76.67% respectively. Among the pathogenic bacteria, percentage of significant count of Staphylococcus aureus and Klebsiella pneumonia were found to be high in chronic bronchitis cases when compared these pathogens in asthmatics. In bronchial asthma cases, the percentage of significant count of Streptococcus pyogenes, Proteus mirablis and Pseudomonas aeruginosa were high as compared to chronic bronchitis cases. In chronic bronchitis patients and asthmatics, significant count of non-pathogens were 16.67% and 13.33% respectively. Conclusion: The quantitative cultures showing significant numbers as 104/ml and 107 /ml of pathogenic and nonpathogenic bacteria respectively are found to be associated with acute exacerbation of bronchial asthma and chronic bronchitis.

425-429

Authors: Muzafar Amin,Vidya Pai,,Sumaira Qayoom*,Syed Arshi,Syed Khurshid Volume : 5

Purpose: To study the biofilm formation and to examine the correlation between antibiotic resistance and biofilm formation among the clinical isolates of Acinetobacter. Materials and Methods: A total of 43 isolates of Acinetobacter collected from samples like peripheral venous catheter tips, urine from Foley’s catheter, central venous catheter tips and endotracheal tube aspirates were preserved and processed. The tube method was performed to qualitatively detect Biofilm production. Antimicrobial susceptibility was done as per CLSI guidelines. Place of Study and Study Period: Yenepoya medical college, Mangaluru and June 2015 to December 2015. Study Design: Prospective Results: Out of 43 isolates of Acinetobacter, 26(60%) showed biofilm production, among which 19 isolates (73%) were Multidrug resistant (MDR) and only 7 isolates (27%) were Non-MDR. Highest sensitivity was seen to colistin (100%) followed by imipenem (73.7%) and least sensitivity to ampicillin (0%) followed by ciprofloxacin (3.8%). Conclusion: Antibiotic resistance was found to be significantly higher among biofilm producing Acinetobacter isolates than nonbiofilm isolates. Routine and advanced studies of the biofilm production will help in making better usage of the invasive devices with less critical complications.