Indian Journal of Microbiology Research

348-387

Authors: Shweta Sharma, Biswaroop Chatterjee, Anurag Bhargava, Aarti Kotwal, Girish Sindhwani.

Aims: Rapid Slide Culture (RSC) was compared with the standard diagnostic strategy of simultaneous culture in Löwenstein-Jensen medium (L-J) and Mycobacterial Growth Indicator Tubes (MGIT™) with regard to diagnostic accuracy, turnaround time and cost, for isolating M. tuberculosis (MTB) from bronchoalveolar lavage (BAL) specimens. Materials and Method: Centrifuged deposits of 75 consecutive smear-positive and 75 randomly selected smear-negative BAL specimens were subjected to RSC as well as culture in both L-J and MGIT. Isolates were identified as MTB by standard methods. Results: One hundred and ten (73.3%) specimens were culture-positive by L-J and / or MGIT; 99 (90%) of these were RSC-positive too. No RSC-positive specimen was culture-negative on both L-J and MGIT. Thus, RSC demonstrated an overall sensitivity, specificity, PPV and NPV of 90%, 100%, 100% and 78% respectively. Significantly, 25 (33%) smear-negative specimens were RSC-positive, indicating the ability of RSC to isolate MTB from specimens with low bacillary loads. The mean time to positivity was seven, 35 and 24 days for RSC, L-J, and MGIT respectively. The cost per test was the equivalent of USD 1.82, 1.34, and 10.42 for RSC, L-J and MGIT respectively. Conclusion: Rapid Slide Culture proved to be a rapid, effective and low-cost method for culturing MTB from bronchoalveolar lavage fluid, when compared with conventional culture in L-J medium and MGIT.

358-362

Authors: Arthi E, Anitha Raj M, Abarna V, Rajkiran R.

Aim: Diarrheal disease is a frequent disease among paediatric age group especially in developing countries. Dysentery, a sign of invasive enteric infections, is associated increased morbidity and mortality. This study was undertaken to study the bacterial and parasitic causes of childhood diarrhoea and dysentery. Materials and Method: The study was conducted in tertiary care centre in children aged between 1-11 years of age, attending the out-patient clinic with complaints of diarrhoea or dysentery. Fresh stool samples was collected and presence of bacterial infections were identified by culture in MacConkey medium, Sorbitol MacConkey medium (SMAC) and Deoxycholate citrate agar (DCA). Parasites were identified by saline and iodine mount preparations. Results: Out of the 60 stool specimens, 8(13%) showed presence of bacterial pathogens with diarrheogenic E.coli 4 (6.6%), Salmonella typhi 2(3.3%) and Shigella flexneri 2(3.3%). Among the 9(15%) parasites isolated, Ascaris lumbricoides, 3(5%), Giardia lamblia, 2(3.3%), Enterobius vermicularis, 2(3.3%), Ancylostoma duodenale 1(1.7%), and cysts of E.histolytica, 1(1.7%) were obtained. The bacterial isolates showed increased resisistance to Ampicillin(60%), Ciprofloxacin (50%) and Cotrimoxazole(50%). Both E.coli & Shigella flexneri, yielded one ESBL producer each. Conclusion: This study has highlighted the common enteropathogens in pediatric age in our region and hence the necessity to undertake control measures by personal hygiene and environmental sanitation.

367-372

Authors: Nawaz Umar, Bilal Ahmad, S.B. Siddish.

Introduction: Gonorrhea infection is caused by a Sexually Transmitted Infection (STI) that infects both men and women. Based on the clinical manifestations it is difficult to diagnosis Neisseria gonorrhoeae. Conventional diagnosis techniques like gram stain and culture for the diagnosis of gonorrhea are inexpensive but have poor sensitivity especially in asymptomatic patients. Rapid diagnosis is important for the control of the disease, PCR method is one such rapid method. Materials and Method: A total of 611 specimens were collected from different anatomical sites in men and women of reproductive age group who attend the STD and non-STD clinic at Khaja Bandanawaz Institute of Medical Sciences, Gulbarga, Karnataka, during the period of June 2013 to June 2016. Direct smears for gram staining, Culture, Biochemical tests and molecular method (PCR) were performed. Conclusion: in the present study, the comparison between conventional and Molecular methods was done; gram staining was less sensitive than culture and PCR. Culture on chocolate agar [CA] was not showing 100% results compare to the Modified Thayer-Martin Media [MTM] on which all 55 specimens were grown, but PCR showed 94.5% result even though there was less specimen in the swabs. PCR technique for the detection of orf1 gene of Neisseria gonorrhoeae directly from clinical samples is a promising investigative method. However, culture on selective media (MTM) is the method of choice for the identification of Neisseria gonorrhoeae and susceptibility testing, particularly in developing countries like India.

377-379

Authors: Sagarika Dhal, Gitanjali Sarangi, Dhanitri Mohapatra, Bimoch Projna Paty, Nirupama Chayani

Coagulase negative staphylococci (CONS) species identification is difficult for most clinical laboratories. The schemes proposed by Kloos and Schleifer is the reference method used for the identification of staphylococcal species and subspecies. It requires the utilization of a large number of biochemical tests. Antibiotic susceptibility testing was done by Kirby-Bauer disk diffusion method. Out of the total 51 CONS isolates subjected for species identification the most common species identified was S. epidedermidis (58.82%). S. cohinii and S. haemolyticus being the second and third most frequently encountered species. Common source of most species was pus followed by blood and urine. S. epidermidis and S. cohnii showed resistant to multiple antibiotics. The most effective antibiotic for all the species of CONS were linezolid, teicoplanin and vancomycin. So CONS isolated from clinical samples should always be considered as potentially significant pathogen. The species identification with antibiogram will be of benefit for both epidemiological and patient care purpose.

388-393

Authors: Disha A Patel, Dipa M. Kinariwala, Ami U Mistry, Bhavikakumari C. Patel, Mahendra M Vegad.

Purpose: Influenza is a widespread viral infection and a major cause of morbidity and mortality worldwide. The present study was performed to investigate the Influenza A H1N1 2009 pandemic (influenza A(H1N1)pdm09) virus outbreaks, its pattern and its clino-epidemiological correlation in and around the region of Gujarat, during the period of August 2009 to March 2016 with special reference to outbreak of 2015. Materials and Method: This is a retrospective study. A total of 17206 patients attending various hospitals in Gujarat were included from August 2009 to March 2016. Nasal swabs, nasopharyngeal swabs, or throat swabs were collected and tested by real time Reverse Transcriptase PCR (rRTPCR) for presence of influenza A(H1N1)pdm09 virus. Results: Out of 17206 samples, 6078 clinical samples were positive for swine flu and 2605 were positive for seasonal Influenza A flu. Over all people in the age group between 30 years to 60 years were highest affected and females were affected more than males. Case fatality rate of the patients admitted in our hospital in 2015 outbreak was 23.07% with highest mortality in pregnant women. Conclusions: Influenza A(H1N1)pdm09 is still active six years after 2009 pandemic and unable to replace seasonal influenza A virus unlike previous pandemic strains. Middle age patients and pregnant women were highly affected with high mortality in pregnant women.

399-402

Authors: MP Saraswathy, Aruna B. Patil, Raj Prakash.

Aims and Objectives: Otomycosis is a fungal infection of middle ear with infrequent complications involving middle ear. In an era of modern medicine generous use of antibiotics and advanced management give way to many opportunistic infections. Though otomycosis is a localised infection, it might leads to prolonged morbidity and disability like hearing loss. Hence, this study was aimed to find out aetiology, clinical presentation and various predisposing factors of otomycosis in a tertiary care hospital. Materials and Method: A descriptive study was carried out in the department of Microbiology and ENT in a tertiary care hospital. About 129 samples were taken from clinically suspected otomycosis patients and cultured for fungal pathogens. Results and Discussion: About Fifty four percentage (54%) of cases were mycologically confirmed as otomycosis and Aspergillus niger (57%) was the predominant isolate followed by Candida albicans (16%). Ear block and Ear pain were common symptoms in otomycosis. Usage of Ear buds and Ear drops were the common predisposing factors to otomycosis.

408-411

Authors: Divakara SV, Irfan, Meera Meundi.

Introduction: This study has been under taken to find out the drug resistance patterns exhibited by Escherichia coli (E. coli) originating Urinary Tract Infection (UTI) in comparison to intestinal commensal E.coli. Materials and Method: One hundred UTI patients who had E. coli isolates were selected for this study. A whole of 100 isolates of E. coli from stool samples of apparently healthy individuals were included as controls. Antibiotic susceptibility testing (AST) was carried out by Kirby Bauer disk diffusion method on Mueller Hinton agar (Hi media). Result: In the present study E. coli showed more resistance to commonly used antibiotics such as ciprofloxacin, norfloxacin, cefotaxime, ceftriaxone. There is also increasing resistance to gentamicin. In comparison, the antibiotic resistance between cases and controls showed a small difference. Conclusion: E. coli is the most common cause of UTI, and microscopic examination of the urine is an important screening method for UTI cases. High resistance to widely used antibiotics for treatment may be attributed to inadvertent and indiscriminate use.

416-418

Authors: A.Rajendra Babu, T. Jaya Chandra, R. Lakshmi Kumari.

Introduction: Gastrointestinal infections are very common in HIV patients, and diarrhea is a common clinical condition seen in them. With this observation, a study was conducted to find the parasites that cause opportunistic infections (OIs) and its association with immune status in the HIV-infected patients. Materials and Method: This study was conducted in the Department of Microbiology, Rangaraya Medical College, Kakinada, Andhra Pradesh, India, for 3 months, that is, from January to March 2013. Stool samples were collected and observed under microscope to detect diarrhea-causing parasites. Direct saline mount, iodine mount, and modified acid fast, modified trichrome stain were carried out. CD4 counts were measured by using FACS counting system. Results: Among the 178 study participants who had diarrhea and who were on antiretroviral treatment, maximum parasites were observed in patients with CD4 counts <200 cells/µL (65%), followed by 200–499 cells/µL (35%). Conclusion: OIs were identified in 54.5% of the HIV patients and the infections were more in patients with CD4 count <200 cells/µL. This would help the clinician to plan a proper treatment schedule.

424-427

Authors: Mahesh Kumar S, Pramod N. Sumbrani, Asha B. Patil, Sheethal S, Akshat Vij.

Introduction: Dengue fever is a seasonal acute febrile arbo-viral illness ranging from asymptomatic infection to dengue fever and the severe dengue haemorrhagic fever dengue shock syndrome. India is one of the seven identified countries in the South-East Asia region regularly reporting dengue fever dengue hemorrhagic fever outbreaks. Objectives: To identify dengue seropositive patients by NS1 antigen testing and anti-dengue IgM antibody detection by ELISA and correlate the changes in epidemiology. Materials and Method: Retrospective study done from January 2013 to December 2015 with blood samples tested from clinically suspected cases of dengue virus infection. Results: A total of 3839 samples were tested. NS1 ELISA detected 196 (5.11%) cases and MAC ELISA registered 488 (12.71%) positives. Maximum number of cases was reported from June to September. Incidence was high in the paediatric age group with an overall male predominance. Mortality rate reported among dengue positive cases during 2013 was 9 (1.35%), 5 (3.12%) during 2014 and 1 (0.65%) during 2015. Conclusion: Prevalence of dengue seropositive cases was 34.20% in 2013, 19.07% and 14.89% in 2014 and 2015 respectively indicating a relative decline in dengue infection which may be attributable to the increase in awareness and preventive measures taken among the people and health services.

431-433

Authors: Raval Payal N, Purav G. Patel.

Introduction: HIV is one of the prevalent infectious disease and major threat to community. As the infection progresses, CD4 count hence, immunity decreases with time. CD4 count plays vital role in prognosis of HIV infection. Materials and Method: CD4 count reports of total of 109 HIV patients who registered and completed 6 months ART treatment during year 2016 were analyzed in this retrospective study. CD4 cell count was done using Flow Cytometry, FACS count (BD Diagnostic). Results: Baseline CD4 counts at diagnosis of infection for study group was 190.11 cell/cmm. Among them Male were having baseline value of 182.09 cells/cmm and female were having baseline value of 206.36 cells/cmm. After the initiation of ART there is consistent rise in CD4 count among study group patients. Conclusion: As the baseline CD4 count is low, patients in high risk group should be encouraged to visit ICTC centre for early detection of HIV. CD4 is mainstay among prognostic marker of HIV infection in India. Also steady intake of ART by HIV patients results in rise in CD4 count, increase in immunity and lowering of viral load.

437-441

Authors: Subi Das, Kalpana George, Girija Kalarikkal Rajagopal.

Introduction: One of the major risk factors for the nosocomial infection among the hospital population was identified as the nasal carriage of Staphylococcus aureus and Methicillin-Resistant Staphylococcus aureus (MRSA). This study was aimed to find the prevalence and factors associated with nasal carriage of S. aureus and MRSA among patients in a tertiary care hospital. Materials and Method: A cross sectional study was designed for S. aureus and MRSA nasal carriage. History of factors for S. aureus and MRSA colonisation was collected using a questionnaire. A total of 750 specimens were streaked directly onto mannitol salt agar. S. aureus was identified and antibiotic susceptibility test was done. All MRSA isolates were subjected to polymerase chain reaction (PCR). Results: Among the total specimens collected, S. aureus isolated were 115 (15%). Out of this 115 isolates, only 5 were found to be MRSA (4.3%). The overall MRSA nasal carriage rate was only 0.6% (5/750). PCR done for all the MRSA isolates showed positive results for Type III SCC mec gene and negative results for Panton Valentine leukocidin gene indicating that they belong to Hospital Acquired MRSA (HA-MRSA). Majority of the patients with MRSA nasal colonisation had no history of hospitalisation, or other factors contributing to colonisation from hospital environment. Conclusion: S. aureus nasal carriage rate was 15% and the MRSA nasal carriage rate was only 0.6%. Association was not found between S. aureus or MRSA nasal colonization with factors such as previous hospitalisation, antibiotics, infections on skin and soft tissues, use of alcohol and smoking.

448-452

Authors: Swathi Akula, Guru Prasad Manderwad, Ravi Shankar Reddy Anukolu, HRV Rajkumar.

Introduction: Acinetobacter spp., a gram negative coccobacillus, were considered to be opportunistic pathogen till their role in hospital acquired infections was described. It has been declared as one of the ESKAPE pathogens which effectively escapes the effects of antimicrobial drugs due to several resistance mechanisms. Of utmost concern is the emerging drug resistance to Carbapenems. Material and Methods: The clinical specimens received were processed according to standard microbiology procedures and the organism was identified as Acinetobacter spp. Antimicrobial susceptibility testing was performed and the organisms were classified as Multidrug resistant (MDR), Extremely drug resistant (XDR) and Pan drug resistant (PDR). Results: Acinetobacter spp were isolated predominantly in endotracheal aspirates (58.1%), suction tip (21.2%) and tracheal tip cultures (4.96%). There was decreased antimicrobial susceptibility to aminoglycosides, β lactam and β lactam inhibitor combination, fluoroquinolones and carbapenems. Out of 143 isolates, MDR were 131 (91.6%) and XDR were 126 (88.1 %). However no isolate was PDR. Majority of MDR and XDR strains were isolated from respiratory intensive care unit (RICU) (65.7% and 63.6% respectively). Conclusion: Rise in carbapenem resistance in Acinetobacter spp. is quite alarming, as it leads to unchecked use of colistin, a last resort antibiotic. To avoid antimicrobial resistance, antibiotics should be used cautiously and empirical therapy should be formulated based on local antimicrobial sensitivity pattern. Antibiotic de-escalation therapy should be practiced based on the culture and sensitivity report. As it is a nosocomial pathogen, the health care workers have to be trained properly in infection control practices to prevent the transmission of this notorious pathogen.

459-463

Authors: Vinod Raveendran, Nivedita Ganesan, Manju M.

Introduction: Rubella is a disease caused by Rubella virus which can affect anyone of any age and is generally a mild disease, rare in infants or those over the age of 40. Rubella is notorious for causing congenital rubella syndrome (CRS). It occurs in newborns born to a woman who gets rubella infection in pregnancy. Women gain immunity against Rubella following either a natural Rubella infection or a Rubella vaccination. Some girls or women reach the child bearing age group without any immunity against Rubella. This makes them vulnerable to giving birth to children with CRS. The available survey data reveals 6 to 47% seronegativity in this age group. Hence the age group of 16- 20 years females was targeted for the study. Aims & Objectives: To find out the immunity status (IgG) against Rubella in the health care students (medical and paramedical) of our teaching hospital. Materials and Method: Serum samples from 180 healthcare students in a tertiary care teaching hospital were tested for Rubella specific IgG antibodies by ELISA. Result: Out of the 180 subjects tested, 21(12%) were susceptible to Rubella infection. The remaining had protective levels of antibodies against Rubella. Conclusion: Seronegativity among child bearing age group was and is being documented regularly. Routine immunization with Rubella vaccine is the only end point to CRS. Inclusion and adherence of the same in the National immunization schedule will help eradicate this disease. Constant serosurveillance is required to assess the immunity status among the child bearing age group.

468-470

Authors: Tejashree A, Suchetha C, Visakha K, Krishna Karthik M.

Phaeohyphomycoses are fungal infections, caused by dematiaceous fungi, manifested as Cutaneous and Subcutaneous infections, Keratitis, Osteomyelitis, Meningitis, Sinusitis, and disseminated infections. This is a case report of a 59 year old diabetic male who presented with noduloulcerative lesion on right leg for two and half months. Biopsy of the lesion revealed septate, dematiaceous fungal hyphae on 10% KOH mount. Fungal culture of the biopsy material on Sabouraud’s Dextrose Agar showed velvety, dark brown to black growth with black reverse and identified as dematiaceous fungi- Rhytidhysteron rufulum.

234-238

Authors: Muktikesh Dash, P. Hema Prakash Kumari.

Seasonal influenza viruses in tropical regions may occur throughout the year, causing outbreaks and epidemics more regularly in humans. There are four types or large groupings of seasonal influenza viruses; Influenza A, B, C, and D, but only influenza A and B viruses cause clinically important human disease and seasonal epidemics. It can cause mild to severe illnesses and even deaths, particularly in high-risk individuals. Vaccination is the most effective means of preventing influenza and its complications. Among healthy adults, influenza vaccine provides protection, even when circulating viruses may not exactly match the vaccine viruses. In elderly, it reduces severity of disease and incidence of complications and deaths. Vaccination is especially important for people at higher risk of serious influenza complications, and for people who live with, care for, high risk individuals.

244-247

Authors: Arvind Khodare, Anita Mutha, Manish Purohit.

Escherichia coli is the most common cause of UTI in community as well as in hospitalised patients. Detection of ESBLs producing organisms from samples such as urine may be important because this represents an epidemiologic marker of colonization, and therefore there is potential for transfer of such organisms to others. Aims: The purpose of this study was to know prevalence of extended spectrum β-lactamases (ESBLs) in strains of Escherichia coli isolated from urine specimens and comparison of PCDDT and IMTDT of ESBL detection. Settings and Design: Department of Microbiology, MGM Medical College, Indore. Cross sectional study. Materials and Method: A total of 200 Escherichia coli strains isolated from 1500 consecutive non repeating urine specimens selected for the study. They studied for ESBL production phenotypic confirmatory disc diffusion test (PCDDT) and indirect modified three-dimensional enzyme extract test (IMTDT). Statistical analysis used: Kappa statistics. Results: 86% (171) of the isolates found to be resistant to atleast one of the 3rd generation cephalosporines tested (cefpodoxime, cefotaxime, ceftazidime and ceftriaxone). ESBL detected in 99 (58%) of the Escherichia coli by PCDDT and in 76% (130) by IMTDT. Conclusions: Our study shows prevalence of ESBL producing Escherichia coli in urinary isolates. IMTDT found to be better than PCDDT for the detection of ESBL. The overall prevalence of ESBL producing uropathogenic Escherichia coli found to be 49.5% for our institute.

253-258

Authors: Apurva Parate, Rajesh Karyakarte, Nitin Ambhore.

Introduction: Carbapenemases are one of the common β-lactamases seen in Klebsiella pneumoniae that are responsible for multi drug resistance. Detection of resistance in these bacteria is necessary for formulation of infection control policies. The hidden resistance of this bacteria is critical to diagnose. To address this problem, the present study aims to detect Carbapenemase production and antimicrobial susceptibility patterns in clinical isolates of Klebsiella pneumoniae at the present setting. Materials and Method: A total of 438 strains of Klebsiella pneumoniae were isolated and subjected to Carbapenemase detection by Screening method using Imepenem (10 µg) disc. Modified Hodge test and Combined disc diffusion test along with E-test was done to confirm carbapenemase production followed by antimicrobial susceptibility testing. E-test was considered to be gold standard. Results: Total 34/438 (7.76%) carbapenem resistant isolates were obtained. The carbapenemase positive isolates were predominantly isolated from Burns wards (14.61%). E-test considering it as gold standard test confirmed all 34 of these as carbapenemase producers. Out of 34, MHT detected 31and CDT detected 32 isolates as positive for carbapenemase production. They were highly resistant to cefotaxime, and ceftazidime (91.18%). Conclusion: Screening for Carbapenemase production needs to be carried out routinely in every clinical diagnostic facility. There is a need for rational use and strict adherence to the concept of “reserve drugs” to minimize the misuse of available antimicrobials. The findings of this study emphasize the need for a continuous surveillance in the ICUs and different hospital wards to detect the resistant strains.

263-266

Authors: Biswaroop Chatterjee, Naresh Khanduri, RK Agarwal.

Introduction: Most carbapenem-resistance in Gram-negative bacilli (GNB) is mediated by carbapenemase enzymes. It is therefore important to evaluate methods for their detection. Aims: Comparing Carba NP with Blue Carba test in Enterobacteriaceae and Pseudomonas aeruginosa, and CarbAcineto NP with Blue Carba test in Acinetobacter calcoaceticus baumannii complex (ACBC). Also, assessing modifications of test protocols for possible improvement of performance. Materials and Method: Total 184 carbapenem-resistant GNB were subjected to Carba NP, Blue Carba, and CarbAcineto NP tests as applicable. Variations were attempted in i) Purity of imipenem, and ii) carbapenemase induction by substrate exposure, to improve test performance. Isolates were also screened for carbapenemase genes by PCR. Results: Carba NP and Blue Carba tests performed equally well with a sensitivity of 91.8% in Enterobacteriaceae and 61.2% in P. aeruginosa. CarbAcineto NP and Blue Carba tests were equivalent with 84.6% sensitivity in ACBC. Imipenem purity did not affect test performance. Carbapenemase induction by substrate exposure increased sensitivity by 6.25% only in Enterobacteriaceae. All strains carried the blaNDM-1 gene; some were positive for blaVIM or blaOXA-48 genes too. Conclusion: Our study is the first to demonstrate the equivalence of the Carba NP and Blue Carba tests in Enterobacteriaceae and P. aeruginosa. It is also the first to demonstrate the equivalence of the CarbAcineto NP and Blue Carba tests in ACBC. Minor modifications to test protocols did not improve performance, except for slightly improved sensitivity by enzyme induction in Enterobacteriaceae.

270-273

Authors: Rupali S. Shinde, Aisha M. Parande, B.G. Mantur, Mahantesh V. Parande.

Introduction: Hepatitis B virus (HBV) infection is a global public health problem. HBV infected pregnant mothers act as a constant source of infection to the babies. This leads to increased chronic cases of HBV infection despite effective vaccine & antiviral therapy. Thus it becomes very essential to detect such unsuspected otherwise healthy HBV carriers & cases. Patients and Method: A retrospective study was aimed to study seroprevalence rate of HBV in all pregnant mothers attending the obstetric services from May 2015 to May 2016. All HBsAg positive cases & their babies’ case records were reviewed to study the clinical, demographic data, HBV vaccination & HBV status of the baby. Results: Prevalence rate is 1.68% in 3800 screened cases. Most cases were from the age group 20-24 years. Primigravids & multigravids were equally infected. Out of 14 delivered cases in the hospital, 13 had live babies & only 1 had IUD. 11 delivered by normal labour whereas only 3 delivered by caesarean section. 3 babies had low birth weight (<2.5kg) whereas 10 babies had birth weight more than 2.5kg. All the babies were fully vaccinated. Only 2 babies’ cases records were available for HBsAg testing & were found negative at 1year of age. Conclusion: Screening of HBV infection in pregnant females can help clinicians to prevent the infection transmission to the babies by taking appropriate measures. Prevalence rate is low in our study but still screening is highly recommended to reduce the addition of chronic cases annually

279-282

Authors: Lakshmi K., Chitralekha S., Sharanya K., Aishwarya J.R..

Introduction: Hepatitis B virus (HBV) infection is one of the global health problems which can lead to complications like chronic hepatitis and carcinoma of liver. HBV is an emerging threat to the treating clinicians due to the risk of transmission of the virus, increased mortality rate and disease progression to fatal complications. Aim: The aim of the present study is to determine the baseline prevalence of Hepatitis B surface antigen in patients attending a tertiary care hospital. Materials and Method: A total of 8190 patients were included in the present study. Screening of HBsAg was done by rapid card test. Positive cases were further confirmed by both Enzyme linked Immunosorbant Assay (ELISA) and Enzyme linked fluorescent assay (ELFA) methods. Results: Out of 8190 cases, 0.86% of the cases were diagnosed to be positive for HBsAg. Among the positive cases, 0.92% were males and 0.76% were female patients. The seropositivity was higher in the age group of 40-60 years. Conclusion: The Seroprevalence of hepatitis B surface antigen is more in age group of 40 – 60 years with a male preponderance and is comparatively similar to the previous studies. This indicates the importance of need for strengthening the hepatitis B vaccination in the population, with a high coverage.

287-290

Authors: Mita D. Wadekar, Swarooparani N.B..

Introduction: E. coli exhibits resistance to commonly used beta-lactam antibiotics by production of β-lactamases. Hence, creating awareness regarding these enzymes and its detection becomes necessary for better patient care. The present study was undertaken to detect extended-spectrum β-lactamases (ESBL) producers. Materials and Method: A total of 101 E.coli isolated from different clinical samples were studied. ESBL production was detected by using combination disk method. Results: Of 101 E.coli isolates tested, 77.2% were found to be ESBL producers. They showed highest susceptibility to amikacin (91%) followed by gentamicin (69.2%), meropenem (62.8%), cotrimoxazole (37.1%), cefoxitin (30.7%), ciprofloxacin (19.2%), amoxicillin-clavulanic acid (16.6%), cefepime (10.2%) and ampicillin (3.8%). Conclusions: Reporting of ESBL producers along with routine sensitivity pattern will help the clinicians in prescribing the appropriate antibiotics. Amikacin, gentamicin and imipenem which showed highest sensitivity are helpful for the treatment of infections which are caused by ESBL producing E.coli.

295-297

Authors: Selvaraj Stephen, Seetesh Ghose, Velmurugan Anitharaj, Jothimani Pradeep

Introduction: So far, there has been no report of genital Chlamydia trachomatis prevalence in the pregnant women of Puducherry. The aim of our study is to determine the seroprevalence of C. trachomatis infection in healthy antenatal women attending Mahatma Gandhi Medical College & Research Institute, a tertiary care Teaching Hospital at Puducherry. Materials & Method: We screened 266 healthy pregnant women in the first and second trimesters for anti-C. trachomatis IgG antibody by ELISA test. Results: Only one out of 266 sera screened was positive for C. trachomatis IgG antibody (0.38%). Conclusion: Seroprevalence of C. trachomatis in this part of southern India is very low. However, additional screening of more number of both asymptomatic and symptomatic pregnant women from different geographical locations in the union territory of Puducherry might give a clear picture.

305-307

Authors: Sumanta A, Sudhindra KS, Nagarathnamma T, Shubha DS.

Introduction: Current methods for drug susceptibility testing of Mycobacterium tuberculosis are either costly or slow. As the prevalence of multidrug-resistant strains increases, the need for fast, reliable, and inexpensive methods that can also be applied in settings with scarce resources is obvious. Aim: To determine the prevalence of multi drug resistant tuberculosis by a simple, rapid and cost effective technique. Materials and Method: 100 sputum samples from sputum smear positive patients at RNTCP centre were taken and subjected to drug susceptibility testing for all the first line anti tubercular drugs by modified slide culture method. Samples were also subjected to conventional culture on LJ media. Results: From the 100 smear positive sputum samples, culture of M. tuberculosis was obtained by both LJ medium and modified slide culture for all the samples. Out of the 100 samples, 94 were sensitive for all the drugs, 04 were found to be multidrug resistant and 02 resistant only to isoniazid. Conclusions: The modified slide culture technique was found to be a safe, rapid and cost effective technique and can used for drug susceptibility testing especially in resource limited settings.

316-319

Authors: Sweta Prajapati, Sanjay Mehta, Kunjan Kikani, Dipak Panjwani.

Multidrug resistant organisms (MDROs) infections have increased in recent years. The antibiotic resistance development in clinical isolates of Enterobacteriacae is rapid and that spread in the hospital. Objectives: 1)To detect multidrug resistant Enterobacteriacae 2)To determine frequency of MDR Enterobacteriacae isolates by site of infection. 3)To determine antibiotic susceptibility pattern of multidrug resistant Enterobacteriacae. Materials and Method: Antibiotic susceptibilities of bacterial isolates from clinical specimens were determined according to standard guidelines. MDRO detection was based on the joint definition given by the European Center for Disease Prevention and Control (ECDC) and the Centers for Disease Prevention and Control (CDC).(6) Results: Out of 1585 clinical samples, 961(61%) samples had enterobacteriacae isolates. Out of total 961 Enterobacteriacae species studied, 648 (41 %) species were MDR. The commonest MDR Enterobacteriacae species were E. coli 364/519 (54%), followed by Klebsiella sp. 280/431 (45%). The commonest MDR Enterobacteriacae infections were urinary tract infections 400/648 (61%), followed by respiratory infections 133/648 (21%). MDR Enterobacteriacae isolates showed decreased sensitivity towards third generation cephalosporins, aminoglycosides and fluroquinolones. Conclusion: The early detection of MDR bacterial species should be started by all microbiology laboratories to give effective treatment to the patients and to reduce the cross infections to other patients in hospital as well as to reduce threat of antimicrobial resistance which is at present a global problem.

325-327

Authors: Priyanka Shukla, Shadma Yaqoob, Mastan Singh, Vaibhav Shukla, Fareya Haider.

Introduction: “Onychomycosis” is fungal infection of the nail which may be caused by dermatophytes, non-dermatophytes and Candida.(1) It affects toe nails more than finger nail. It may involve any part of the nail including nail bed or nail plate. The incidence of the disease varies from 3 to 5% and it may reach upto 20% in certain subgroups, e.g. miners, sportsmen. Materials and Method: A total of 100 patients attending out patient clinic of Department of Dermatology and Venererology at Medical College in North India were enrolled in study. Nail scrapings and nail clippings were collected by proper aseptic techniques. KOH wet mount was prepared from nail specimens. Culture was done on SDA media in duplicate. Lacto phenol cotton Blue mount was prepared from growth on SDA. Species identification was done by Micro slide culture technique. Results: In this study M:F ratio was 1.7:1. Labourers (49)were most commonly involved among males and housewives(21) were among females. Out of all isolates maximum were Dermatophytes(79),Non dermatophytes(38) and Candida(12).

333-337

Authors: Reshmi Gopalakrishnan, B.V. Chandrashekara Murthy.

Introduction: Urinary tract infection (UTI) is one of the commonest complications occurring during the antenatal period. It can result in adverse outcomes like abortion, premature delivery, low birth weight. Screening of antenatal patients help in early diagnosis of UTI and thus to prevent maternal and fetal morbidity and mortality. Aims: The present study was carried out: 1. To determine the prevalence of UTI in pregnant women. 2. To study the bacteriological profile and antimicrobial susceptibility pattern of the uropathogens. Materials and Method: 269 antenatal patients who attended the antenatal clinic during the study period January 2015 – December 2016 were included in this study. Mid stream urine sample was collected from these patients. Samples were processed and isolates were identified. Antibiotic susceptibility was recorded as per CLSI guidelines. Results: The prevalence of UTI among pregnant women was 16.9%. Majority of the culture positive patients belonged to the age group of 25-29 yrs. 91.3% were Gram negative isolates and 8.7% were Gram positive organisms. E.coli was the most common uropathogen. Ampicillin, amoxicillin clavulanic acid, cotrimoxazole were found to be resistant to most of the Gram negative uropathogens. Most of the Gram negative isolates were sensitive to amikacin, nitrofurantoin, ceftriaxone, quinolones. For Gram positive uropathogens, ampicillin can still be safely prescribed as first line antibiotic. Conclusion: This study reveals the importance of screening of pregnant women for UTI. Increasing resistance observed among the uropathogens emphasizes the need to rationalize the use of antibiotics which would eventually prevent the development of resistant strains.

341-345

Authors: Vijetha Sajjanar, D. E. Premlatha, Halesh L. H..

Background: Urinary tract infection (UTI) is one of the most common infections in both outpatients and hospitalized patients. The increasing prevalence of antimicrobial resistance is a major health problem & is associated with high morbidity & mortality. Objectives of the study: 1. To analyse the antimicrobial resistance pattern of uropathogen during a period of 6 month in patients of UTI. 2. To know the prevalence of uropathogens. Materials and Method: A prospective study was conducted on urine specimens suspected to be having urinary tract infection between July 2016 and December 2016 received at Department of Microbiology, SIMS Shimoga. Fresh midstream urine samples were aseptically collected in sterile containers and plated on Blood agar & MacConkey agar plates using a standard loop technique & the growth was processed by standard bacteriological technique. Antimicrobial sensitivity testing was done using Kirby-Bauer methods on Mueller-Hinton agar. Results were interpreted as per the CLSI guidelines. Result: Out of 510 samples 250 samples showed significant bacteriuria. Most common uropathogen isolated was Escherichia coli 61. 6%(154/250) followed by Staphylococcus aureus 19. 2%(48/250), Enterococcus species 10%(25/250), Klebsiella species 3. 2%(8/250), Pseudomonas species 1. 2%(3/250), CONS 0. 8%(2/250). Gram negative organisms were highly sensitive to Fosfomycin, Nitrofurantoin, Imipenam and highly resistant to Norfloxacin, Ciprofloxacin, Levofloxacin. Gram positive organisms were highly sensitive to Linezolid, Vancomycin, Fosfomycin, Nitrofurantoin and least sensitive to Norfloxacin, Ciprofloxacin, Amoxyclavulanic acid. Conclusion: This study showed that high resistance was observed for commonly used antibiotics. Fosfomycin and Nitrofurantoin were the most sensitive drug for gram negative organisms mainly E. coli which is the most common uropathogen isolated.

349-352

Authors: Meena Dias, Anusha Dias, Sagar Chandrakar.

Mycotic keratitis has emerged as a major ophthalmic problem. Fungal keratitis is usually caused by Aspergillus, Fusarium, Curvularia and Candida species. But one should be aware of other rare environmental pathogens causing keratitis. We report here two cases of corneal ulcer caused by plant pathogens Colletotrichum truncatum and Colletotrichum gloeosporioides following injury. Both the cases were treated successfully with voriconazole and natamycin.

126-130

Authors: Norah Saad J. Bin Saad.

Antimicrobial agents are compounds which inhibit the growth, or kill microorganisms. Antimicrobial agents can be used for either treatment or prevention of infectious disease but must be used properly and accurately. The use of antimicrobials in the prevention of disease is called prophylaxis. There are four occasions when prophylactic antimicrobials are indicated and these are: · When the risk of post-operative infection is high. · When wounds are contaminated with soil or dirt (e.g. after road traffic accidents) and there is a risk of infection (e.g. Clostridium tetani). · Where the consequences of infection are serious or life threatening. · When a person’s defences against infection are compromised.

134-137

Authors: Hitesh Assudani, Jigar K. Gusani, Sanjay J. Mehta.

Introduction: Infections remain one of the major problems in pediatric intensive care unit and are leading cause not only of admission but also mortality in developing countries. Mortality from pediatric sepsis ranges from 9% to 35%. Objective: To study the prevalence & bacteriological profile of septicemia in post neonatal (29 days to 1 year) age group and older children (> 1- 12 year). Material and Methods: Venous blood was collected (2 ml in post neonatal and 5 ml in older children) and inoculation in blood culture bottle. Further processing was done by standard microbiological guidelines. Results: In present study out of 84 clinically suspected 52 (61.90%) were males and 32 (38.10%) were females. Among 84 cases, 31 (36.90%) were post neonatal cases and 53 (63.10%) were older children. Out of 31 post neonatal cases, in 6 (19.35%) cases pathogen were isolated and among them 3 were S.aureus & 1(33.33%) was MRSA. Out of 53 older children, in 14 (26.42%) cases pathogen were isolated and among them 5 S.aureus, 3E.coli, 3Klebsiella spp. were isolated out of which 3(60%), 6 (50%) were MRSA & ESBL respectively. Conclusion: Prevalence of septicemia is 9.83% in post neonatal age group and 22.95% in older children. Septicemia is more common in male age group. Most common clinical presentation in post neonatal age group and older children was fever of unknown origin.

144-149

Authors: RV Shinde, RV Mohite, AR Shinde, ST Mohite, GS Karande.

Aims & Objectives: To assess and compare the feasibility, availability, affordability, stability of In-house and commercial third party internal control of HIV antibody ELISA test. Methods: A comparative study was conducted in NABL Accredited Diagnostic Laboratory at Krishna Institute of Medical Sciences Karad during June 2014 to July 2016. Sample size includes total of 457 ELISA tests which were carried out on 11425 patients along with internal, external commercial and in-house control as per NACO guidelines. Descriptive and inferential statistical tests were applied to find out significant difference between in-house and commercial control. Results: There is significant difference in mean values of positive kit control and commercial positive control (p<0.05). There is also significant difference observed in mean values of commercial positive control and in-house positive control (p<0.05). The coefficient variance of positive in -house control is better (3.32) than commercial positive control (4.60) which indicates, in-house control is more precise than commercial control. The kit cut off values remains constant throughout the month indicates best technical competence, less environmental influences and reagent stability. The average cost of screening for HIV with ELISA and in-house control was Rs. 57/ per person. Conclusion: There is no significant difference in mean values of positive kit control and in-house positive control, the diagnostic cost, feasibility and percentage CV% is better observed in in-house control as compared to commercial control indicating in-house control could be considered as third party internal control for HIV antibody ELISA test.

154-158

Authors: Shreevidya Kinila, Mehnaz Zephyr, Vinay Padumane Gopalakrishna.

Background: To study the bacteriological profile of infections in patients admitted to ICU and to determine the antibiotic susceptibility patterns of the bacterial isolates. This study was done as Prospective study of 2 months period with a study population of patients admitted to Intensive care unit of a tertiary care hospital. Materials and Methods: Selection criteria for this study was with an inclusion criteria of patients admitted to ICU for various reasons and developing infection within 48 hours of admission and the exclusion criteria was patients admitted to ICU and not developing infection and patients admitted to ICU with an already existing infection. All the samples were processed as per standard microbiology guidelines. Results: Gram negative bacilli were predominant with 73.35% as compared to Gram positive cocci of 9.97% of the total aerobic bacteria grown from various samples of patients admitted in Intensive care unit. Escherichia coli were more common with 26.67% of the total bacteria isolated. This was followed by Acinetobacter species 16.67%, Pseudomonas aeruginosa 16.67%, Klebsiella pneumoniae 6.67% and Enterobacter species6.67%. Among the Gram positive cocci, Staphylococcus aureus was more commonly isolated with 16.67% followed by Enterococcus species 6.67% and Streptococcus species 3.30%. Antibiotic resistance was observed by most bacteria to Penicillins, third generation Cephalosporins, Fluoroquinolones like Ciprofloxacin, Cotrimoxazole. Conclusion: Multi-drug resistance is a major hurdle in treating patients admitted to ICU setting in a hospital. Regular surveillance of antibiotic susceptibility patterns is very important for setting orders to guide the clinician in choosing empirical or directed therapy of infected patients.

162-166

Authors: Rajesh Bareja, Iqbal Singh, Subhash Chand Jaryal, Savita Mahajan, Hiba Sami.

Background: Importance of non-fermenting gram-negative bacilli (NFGNB) in the aetiopathogenesis of human infections has increased considerably in recent years. Their higher rate of isolation from a wide variety of clinical specimens necessitates their identification and characterization. Material and Method: A total of 5,369 bacterial isolates were obtained from 12,591 various clinical samples. All the samples were processed on Blood agar and MacConkey agar medium and identified according to standard protocol. Result: Out of 5,369 isolates, 478 were identified as non-fermentative bacteria. Out of NFGNB strains isolated, the predominant numbers were that of P. aeruginosa 337 (70.5%) followed by other Pseudomonad’s 63 (13.1%) and other Non-fermenters 78 (16.4%). Out of strains of other non-fermenters (78) predominant number was that of Acinetobacter anitratus (34), followed by Acinetobacter lwoffii (17), Alcaligenes xylosoxidans (10), Alcaligenes denitrificans (10) and Alcaligenes faecalis (7). The most sensitive drugs against non-fermenter isolates were found imipenem followed by amikacin. Conclusion: There is enough reason to believe that they are associated with human disease process. Therefore, careful attempts must be made for their isolation and identification from various clinical specimens.

171-176

Authors: Preethi Venkatesan, Priyadarshini Shanmugam, Shameem Banu Abdul Satter.

Introduction: Bacteremia is a serious infection with high morbidity and mortality. Knowledge of the etiological agents of blood-stream infections and their antibiotic resistance pattern could help in the choice of empirical antibiotics. Aim: To determine the causative agents of blood stream infection along with their antibiotic resistance pattern in our hospital between 2013 and 2016 and also to identify the uncommon blood culture isolates. Materials and Method: A retrospective study that included isolates obtained from blood culture between January 2013 and December 2016. The identification and drug resistance pattern of the organisms were performed using standard microbiological techniques. For those patients whose blood yielded uncommon isolates, the medical records were looked into to identify the risk factors and outcome. Results: Of a total of 9,926 blood samples received for culture, 534 (5.38%) yielded positive results, of which 277 were Gram-positive organisms and 257 were Gram-negative organisms. The most common etiological agents of bacteremia in the present study were Coagulase Negative Staphylococcus species (37.27%) followed by Salmonella typhi (10.11%), Escherichia coli (9.36%) and Staphylococcus aureus (8.05%), of which 53.49% were MRSA. Conclusion: The most effective antibiotics against Gram positive organisms were vancomycin, teicoplanin and linezolid. The Gram negative organisms showed least resistance to antibiotics like amikacin, tobramycin and carbapenems. The uncommon isolates were all non-fermenting Gram negative bacilli and among these Burkholderia species and Stenotrophomonas maltophila were the most frequently isolated.

182-186

Authors: Marcia Jaqueline Mendonca Maciel, Karol de Souza Barbosa, Luciana Araujo Cassa-Barbosa, Spartaco Ast.

New strains of the yeasts Meyerozyma guilliermondi CBA-524 and Pichia kudriavzevii CBA-519, capable of growing on sugarcane bagasse hydrolysate, have a great potential for second-generation bioethanol processes. Meyerozyma guilliermondi CBA-524, achieved a maximum ethanol concentration of 3.01g/L corresponding to a yield of 0.26 g ethanol per g of sugar and a productivity of 0.125 g/l/h, after 24 h of cultivation. Pichia kudriavzevii CBA-519 exhibited a prolonged fermentation time on bagasse hydrolysate resulting in an ethanol yield of 0.26 g/g and a productivity of 0.015 g/l/h after 192 h of fermentation. The ethanol yield and the volumetric productivity using the adapted P. stipitis CBS6054 standard were 0.34 g/g and 0.083 g/l/h, respectively, after 48 h of cultivation. Furthermore, the newly adapted M. guilliermondi CBA-524 demonstrated significantly enhanced tolerance to sugarcane bagasse hydrolysate, exhibiting a maximum in ethanol concentration after 24 h of fermentation. More adapted yeasts and improved fermentation conditions for this yeast can be of great importance for the increased production of ethanol using sugarcane bagasse hydrolysate.

190-193

Authors: Sathya Bhama MC, Saritha N, Jyothi R.

Introduction: Melioidosis is caused by Burkholderia pseudomallei which is endemic in South East Asia. Lack of proper microbiological services in many countries results in under-reporting of cases and the global burden of infection remains underestimated. This is a study to analyse the clinical presentation and risk factors for melioidosis in India. Materials and Method: A retrospective study was carried out on ten culture proven cases of melioidosis in a tertiary care hospital in South India between January 2009 and December 2014. Result: Ten cases were studied in five years of which nine were males. They had varying clinical presentations like loss of weight and loss of appetite(10%), fever(60%), headache(10%), abdominal pain(20%), joint pain(10%), weakness of limbs(10%), swelling of thigh(20%) and faecal and urinary incontinence((10%). The risk factors included diabetes mellitus (100%), alcohol consumption(90%)and tuberculosis(30%). All the isolates were sensitive to Ceftazidime, Piperacillin-tazobactam, Imipenem, Cotrimoxazole, Amoxycillin-clavulanate and Meropenem while only 80% were sensitive to Ciprofloxacin. Of the 10 cases, two expired and the rest responded to treatment. Conclusion: Melioidosis is an emerging infection in India especially in males and diabetes and alcoholism are identified as the commonest risk factors. Control of the disease requires close monitoring, improved clinical laboratory standards and aggressive therapy.

199-202

Authors: Prabha Poonusamy, Radhika Katragadda, Thyagarajan Ravinder.

Introduction: Asymptomatic bacteriuria is more common in pregnant females. Early screening and treatment avoids complications like acute pyelonephritis and chronic kidney failure. Escherichia coli is the most commonly encountered organism among them. Certain virulence factors associated with E.coli have a direct link in pathogenic mechanism of organism like adhesins, toxins and hemolysin. Aim: We aimed at identifying E.coli and demonstrating phenotypic and genotypic presence of mannose resistance by hemagglutination in urinary isolates. Materials and Method: A cross sectional study during 2014 – 2015 among pregnant females was conducted in a tertiary care hospital. About 1000 urine samples were included in study. Urine culture with antimicrobial susceptibility testing was performed as per standard microbiological conditions. Virulence factor were identified using phenotypic method i.e., haemagglutination test and subsequently confirmed by polymerase chain reaction by looking for Pap A gene expression. Results: Out of 1000 urine samples, 118 culture positives were seen. Out 118 culture positivity, 54 (45.76%) Escherichia coli isolates causing asymptomatic bacteriuria was identified. Among the 54 isolates, phenotypic method for mannose resistance haemagglutination showed 9 positivity (16.66%), which was confirmed with PCR for Pap A expression. Conclusion: As asymptomatic bacteriuria is more common in pregnancy, it is a must to screen pregnant females in every trimester even without any specific symptoms. Virulence factors need to be explored in order to understand the pathogenic mechanism and importance of screening the asymptomatic women.

207-209

Authors: Anjana Gopi, Swati Jain, Faiza Samreen.

Introduction: Onychomycosis is a fungal infection of nails caused by dermatophytes, yeasts or non-dermatophyte moulds and represents about 30% of mycotic cutaneous infections. Onychomycosis manifests itself in various forms, notably onychodystrophy, onycholysis, subungual hyperkeratosis, or nail-plate discoloration. The objective of this study was isolation and speciation of various fungi by culture and to determine its prevalence. Materials and Method: The study was conducted from Jan 2014 - June 2016 at a tertiary care hospital, Bangalore. Nail-clipping and subungual debris of clinically suspected patients were subjected to Potassium hydroxide (KOH) preparation. Culture was done on Sabouraud’s dextrose agar (SDA) media. Species identification was done by colony character, pigment production, Lactophenol cotton blue (LPCB) staining, some special tests like germ tube test, inoculation into chrome agar in case of Candida and slide culture wherever necessary. Result: Out of 400 cases, the KOH preparation was positive for fungal elements in 288 (72%) cases and culture grew fungi in 268 (67%) cases. Among those 268 cases, the infective fungal agents were predominantly dermatophytes (59.7%) and the rest were due to moulds (22.38%) and yeasts (17.91%). Among the different species, Trichophyton rubrum (T. rubrum-37.5%) accounted for the majority of dermatophytes, Fusarium (60%) was the commonest mould and Candida albicans (37.5%) the predominant yeast. Conclusion: Direct microscopy and culture are extremely important in the diagnosis of onychomycosis and dermatophytes are the most common etiological agent in our region.

216-219

Authors: Sudhindra KS, Sumanta A, Shubha DS, Narayana Murthy C, Srinivasa Setty TK.

Introduction: Rickettsial diseases though very commonly found in India, are often misdiagnosed due to lack of awareness and non-availability of diagnostic facilities. Aim: The aim of this study was to determine the prevalence of rickettsial infections in patients attending our hospital with complaints of fever. Materials and Method: A total of 150 serum samples were tested for rickettsial infections by Weil Felix test and the results were analysed. 25 serum samples from afebrile patients were also tested as controls. Results: Of the 150 samples, 59 samples showed significant titres, out of which 29 showed significant titres to OX-K, 10 to OX-2, 11 to OX-19 and mixed titres seen in other 09 samples. Conclusions: Clinical suspicion and routine testing for rickettsial diseases will be useful in patients presenting with fever for prompt diagnosis and management of these cases.

224-226

Authors: Ashwini Kacharu Dedwal, Sae Satish Pol, Sujata Sudhir Ranshing, Shobha Dattatray Chitambar, Renu Sat.

Introduction: Acute gastroenteritis is a major cause of illness and death among infants and young children worldwide. According to the World Health Organization (WHO), an estimated 2.5 billion cases of diarrhea occur among children under 5 years of age each year. Nearly one in five pediatric deaths (about 1.5 billion each year) is due to diarrhea. Present study was thus drawn to identify microbial etiology of acute gastroenteritis in pediatric patients in Western India. Materials and Method: This was Prospective laboratory based study approved by Institutional Ethical committee. 100 stool samples were collected from children admitted with acute diarrhea of 72 hours or less duration in 1- 60 months of age over a period of one year. Bacteria were identified by standard Microbiological methods and serotyping of isolated Escherichia coli (E. coli) was done. Rotavirus antigen was detected by Enzyme Linked Immunosorbant Assay (ELISA) followed by genotyping by Reverse Transcriptase Polymerase chain Reaction (RT-PCR) and Multiplex PCR. Parasitic identification was done by microscopy. Results: 74% of children with diarrhea were in the age group of 7 to 12 months. Watery diarrhea (94%) was the commonest clinical presentation, followed by vomiting (78%), fever (78%), and dehydration (74%). Pathogenic bacteria were isolated in 51% of samples. Escherichia coli was most common (48%) followed by Shigella flexneri (2%) and Vibrio cholerae (1%). The most prevalent E. coli type was Enterotoxigenic Escherichia coli (ETEC) (20.8%) followed by Enteropathogenic Escherichia coli (EPEC)(16.7%), Enterohaemorrhagic Escherichia coli (EHEC) (4.1%), and STEC (2.1%). The most prevalent serotypes of ETEC were O27, O23, and O169. Among EPEC most prevalent serotypes were O90, O26. The most prevalent EHEC strain found in this study was O71. Rotavirus was detected in 35% of patients. Most prevalent Rotavirus genotype was G9P[4] (28.6%) followed by G2P[4] (21.4%), G1P[8] (21.4%), G12P[6] (14.3%), G9P[8] (7.1%). Parasitic etiology was detected in 5% of cases. Coinfection of E.coli and Rotavirus was detected in 23% of children. Rotavirus was most commonly associated with EPEC (25.7%) followed by ETEC (17.1%). Conclusion: In the present study, E. coli was the commonest microorganism followed by Rotavirus. Thus, the importance of safe water and food hygiene would be most important intervention to prevent acute gastroenteritis in children along with Rotavirus vaccine.

231-233

Authors: B. Divya.

Sporotrichosis is a chronic pyogranulomatous fungal infection of cutaneous and subcutaneous tissues which remain localised or may occasionally disseminate to other parts of the body. The causative agent is a thermally dimorphic environmental saprotrophic fungus, Sporothrix schenckii which enters the body through scratches or cuts in the skin. Diabetes, alcoholism, AIDS, leukemia, patients on corticosteroids, chemotherapeutic agents and patients with pre-existing lung disease are important predisposing factors. Here, we report a 65 year old female patient with history of diabetes mellitus for the past 12 years, presented with a leg ulcer and pus discharge for the past 2 years, which did not respond to antibiotic therapy. On examination, a single ulcer of 4x3 cm size was present over the plantar aspect of left foot with pus discharge. Pus was sent for bacteriological and fungal culture identification which revealed growth of Staphylococcus aureus and Sporothrix schenckii respectively. Slide culture and histopathological examination confirmed Sporothrix schenckii. Patient was treated with saturated solution of potassium iodide(SSKI) oral drops and itraconazole and followed up for the regression of lesions.

Page No. 1-6

Authors: Lakshmi K., Santhanam R., Chitralekha S.

Tuberculous Meningitis (TBM) is one of the leading causes of deaths and disabilities in the developing nations like India. Rapid diagnosis of the cases is essential to minimise the mortality and morbidity. Although clinical diagnosis is made based on the presenting symptoms, signs and radiological imaging studies, confirmation of the diagnosis is still difficult and there are significant diagnostic and treatment challenges. Cerebro spinal fluid (CSF) examination shows a lymphocytic-predominant pleiocytosis, elevated protein, and normal or low glucose. Multiple and repeated Culture and CSF examination by acid fast staining and PCR usually may show good results. Empirical treatment with at least four first-line drugs, preferably isoniazid, rifampin, pyrazinamide, and streptomycin or ethambutol is usually started in clinical suspected cases with supportive initial CSF findings. Corticosteroid can be added as adjunctive treatment, but its benefits remains doubtful. Drug interactions, possibility of drug resistant tuberculosis and development of immune reconstitution inflammatory syndrome needs to be kept in mind in initiating the TBM treatment in patients who are HIV positive. This review article throws light into the current practice in the laboratory diagnosis and management of Tuberculous meningitis

Page No. 14-19

Authors: Jyotismita Khasnabis, Paulomi Adhikari, Deblina Chowdhury, Chandan Rai, Arindam Roy

Background: Snacks and sweets from road side eateries are cheap sources of nutrition but occasionally cause numerous public health hazards most importantly gastroenteritis. Various factors including poor infrastructure to prepare foods, post-preparation contamination during storage and handling as well as unhygienic surroundings may contaminate these foods with pathogenic bacteria. Bacillus cereus is one of the common and important food borne pathogens causing spoilage to the foods and producing gastroenteritis to the consumers. This study was carried out to enumerate B. cereus and assess their spoilage potentiality and antibiotic resistance pattern. Methods: Standard plate count was performed using plate count agar while presumptive B. cereus was isolated in Bacillus cereus agar base supplemented with polymyxin B selective supplement and egg yolk emulsion. Confirmation as B. cereus was done by morphological, biochemical and physiological methods. Confirmed isolates were tested for extracellular enzyme production and antibiotic resistance pattern. Result: Standard plate count exceeded 1010 cfu g-1 and B. cereus count was positive in 57% of the samples. All the 43 B. cereus isolates produced amylase and caseinase and were multiple drug resistant. None produced lipase. Conclusion: Presence of B. cereus with spoilage potential and multiple drug resistance require good manufacturing practices (GMPs) and good hygienic practices (GHPs) during food production and storage.

Page No. 26-30

Authors: Gomathi Maniyan, Dillirani Vedachalam, Nithya Chinnusamy

of life threatening nosocomial infections and as multidrug resistant pathogens. Acinetobacter species are the opportunistic pathogens with increasing prevalence in the nosocomial infections. Community acquired infections are also common in Acinetobacter. It accounts for 10% of all community-acquired bacteraemic pneumonias. Aim: To isolate, identify and detect Carbapenem resistance producing Acinetobacter spp., and confirm Oxacillanase (OXA) and Metallo Beta Lactamase (MBL) production by phenotypic& genotypic methods. Materials and Methods: This cross sectional study conducted in a tertiary care hospital for a period of one year and samples collected like pus, urine, endotracheal aspirates, blood, sputum and body fluids were identified using standard protocol, which includes Gram staining, test for motility, catalase test, oxidase test, OF test and various biochemical reactions. The resistant strains of Acinetobacter species as identified by phenotyping were subjected to molecular analysis of OXA-51, VIM and IMP genes. Results: Out of 110 isolates of non-fermenters, Acinetobacter baumannii accounted for 36 (81.8%) and Acinetobacter lwoffi 8 (18.2%). The antimicrobial susceptibility pattern revealed maximum resistance to Gentamycin (50%), Cotrimoxazole (47%), followed by Ciprofloxacin (50%) and Cefotaxime (32.2%). Sensitivity to Polymyxin B (100%) followed by Imipenem and Meropenem (75.5%). MBL production was 20.5%. Molecular characterization of MBL of Acinetobacter species revealed, OXA- 51 (33.3%), 3 (33.3%) for bla IMP. and 2(22.2%) isolates blaVIM positive. Conclusion: Acinetobacter baumannii were the most common isolate in this study. Differences in antimicrobial susceptibility pose a great problem in treating these infections. MBL production by these organisms leads to high morbidity and mortality and left with the only option of treating them by potentially toxic drugs like Colistin and Polymyxin B.

Page No. 36-39

Authors: Rajeswari Jayakumar

Background & Objectives: Decompensated liver disease (DCLD) is defined as irreversible chronic injury of the hepatic parenchyma. Bacterial infections are more common in decompensated liver disease and causes 30%-50% of deaths. Therefore, this study was done to determine the various bacterial agents causing infections in decompensated liver disease patients and to determine the drug susceptibility and resistance pattern. Methods: A prospective study was conducted over a period of one year in a tertiary care hospital. Ascitic fluid, urine, sputum, blood and wound swab were collected. All the samples were processed through Gram’s stain and culture. The organisms were identified by standard protocols and antibiotic susceptibility testing. Results &Conclusion: Out of 150 samples, culture positivity seen in 81(54%). In 81 culture positive isolates, 63(78%) were Gram Negative bacilli (GNB) and 18 (22%) were Gram Positive cocci (GPC). Among Gram negative bacilli, Escherichia coli and in Gram positive cocci, Staphylococcus aureus was the most common isolates. The most common infections were spontaneous bacterial peritonitis (27%) followed by urinary tract infections (26%), Spontaneous bacteraemia (19%), pneumonia (16%), and skin and soft-tissue infections (12%).

Page No. 44-47

Authors: Sathish J.V., Ram Mohan Reddy K., Sujith Pavan

Background: Due to large number of infections caused by biofilm producing organism, a reliable and reproducible method for its detection is necessary. Specific methods like Polymerase Chain Reaction (PCR) are available for detection of biofilm, but these are beyond the scope majority of laboratories in developing nations like India. Objective: To test reliability of Tube Adherence test as a screening tool for detection of biofilm formation among Staphylococcus. Materials and Methods: Various clinical samples were collected with universal precautions from patients with indwelling medical device for more than 48 hours. Screening for biofilm detection was done using Tube adherence method. Results: Tube method was 97.5% sensitive and 100% specific for detection of biofilm formation. Conclusion: Tube method can be used as a general screening method for detection of biofilm formation by Staphylococci

Page No. 52-58

Authors: Soumya Kaup, Sunitha B. U., Jaya Sankarankutty

Introduction: Emergence of multi-drug resistant (MDR) organisms is an important public health concern especially in developing countries. The present study was carried out to assess the current antimicrobial pattern of multi-drug resistant Escherichia coli. Materials and Methods: 346 consecutive, non-duplicate isolates of E. coli isolated during a period of two years were included in the study. The numbers of multi-drug resistant isolates were determined. Their antimicrobial resistance phenotypes and Multiple Antibiotic Resistance (MAR) index were determined. Results: Among the 346 isolates of E. coli, 297 (85.84%) were multi-drug resistant with 61.3% of the isolates being resistant to drugs in more than five antimicrobial categories. The Multiple Antibiotic Resistance index of 82.7% of isolates was found to be greater than 0.2. Carbapenems, aminoglycosides, Chloramphenicol and Nitrofurantoin were found to be the most effective agents against MDR E.coli. Conclusion: Antimicrobial resistance profiles of bacteria are ever-changing and periodic evaluation of resistance phenotypes of isolates is essential for the formulation of appropriate antibiotic policy and initiation of pertinent empirical therapy.

Page No. 64-67

Authors: Vijaya Shivanna, Achut Rao

Background: The extended spectrum β-lactamases(ESBLs), AmpCβ-lactamases and metalloβ-lactamases (MBLs), have emerged worldwide as a cause of antimicrobial resistance in Gram negative bacteria (GNB). So the present study was conducted to detect all three β-lactamases in GNB using disc potentiation tests. Materials and Methods: A total of 200 Gram negative clinical isolates from a tertiary care centre were identified and antibiotic susceptibility testing was done according to Clinical and Laboratory Standards Institute (CLSI) guidelines. The screening and phenotypic confirmatory test recommended by CLSI were carried out for ESBL detection. Detection of AmpCβ-lactamase was done by phenyl boronic acid test. MBL were detected using EDTA disc potentiation test. Result: Of a total of 200 Gram negative isolates, pure ESBL were seen in 50(25%) isolates and pure AmpC were seen in 35(17.5%) isolates. ESBL and AmpC co-existed in 38(19%) isolates. AmpC and MBL co-occurred in a single isolate. Conclusion: we recommend the phenotypic tests using various inhibitors for the detection of β-lactamases.

Page No. 74-78

Authors: Alice Peace Selvabai, A. S. Shameem Banu, M. Jeya, Priyadarshini Shanmugam

Staphylococcus aureus remains as one of the most potent bacterial human pathogen because of its expression of various virulence factors and also due to its property of multidrug resistance. A total of 468 non –duplicate S.aureus strains obtained from various clinical specimens were included in the study. Methicillin Resistant S.aureus (MRSA) strains were screened for DNase production using DNase agar, hemolytic property of the isolates were detected in 5% sheep blood agar plates, hemagglutination property of the isolates were demonstrated using 1% O group RBC’s,slime production was detected using congo red agar medium and biofilm production was quantitatively assayed by microtitre plate method. Antimicrobial susceptibility profile was studied for the MRSA isolates. Out of 468 S.aureus strains, 114(24%) strains were detected as MRSA. Among the MRSA strains 99(86%) were positive for DNase, 77(67%) were showed beta hemolysis, 49 (42%) strains were positive for hemagglutination, 70 (61%) were slime producers, most of the strains were biofilm producers, 4(3.5%) were non-adherent, 85(74%) were weakly adherent, 23(20%) were moderately adherent and 2(1.7%) were strongly adherent. About 56%, 60% and 63% of the isolates were resistant to cotrimoxazole, erythromycin and ciprofloxacin respectively and about 22% and 33% of the isolates showed resistance towards clindamycin and gentamicin respectively. All the isolates were sensitive to vancomycin, teicoplanin and linezolid. About 10(8.7%) strains showed high level mupirocin resistance (HLMR) and 1(0.8%) strain showed low level mupirocin resistance (LLMR). Both HLMR and LLMR (100%) strains showed susceptibility to fusidic acid. MRSA infections remain a major threat in both community and nosocomial settings. Therefore a thorough understanding of its virulence mechanisms and regular surveillance of antimicrobial susceptibility pattern will help the clinician to choose appropriate treatment options and to control the emergence of multidrug resistant strains.

Page No. 83-86

Authors: Asha Pai KB, Manjunath R Kamath, Rekha Rai, Gopalakrishnan M

Background: Mupirocin has been recognised as the best and the most effective topical antimicrobial agent for decolonisation. With widespread use of mupirocin for decolonisation, increasing number of MRSA have developed resistance to mupirocin. The society of thoracic surgeons recommends a glycopeptide (vancomycin) for cardiac surgical prophylaxis in patients known to be colonised with MRSA. Development of resistance to vancomycin among MRSA is a cause of concern. Currently, there is very limited data regarding prevalence of mupirocin and vancomycin resistance in MRSA colonising the anterior nares of patients undergoing high risk surgeries. Aims: To assess the prevalence of mupirocin and vancomycin resistance in MRSA colonising anterior nares of patients undergoing cardiac surgical procedures in our tertiary care hospital. Material and Methods: Screening for MRSA was done by disc diffusion method using Cefoxitin disc. Screening for MuL and MuH was done by disc diffusion method using mupirocin disc of strength 5 µg and 200 µg respectively. Screening for detection of reduced susceptibility to vancomycin was done by Agar Dilution method. The MIC of mupirocin and vancomycin determined by Agar dilution method. Results: Twenty two percent of MRSA isolates had MuL and eleven percent had MuH. One of the mupirocin sensitive MRSA, had a presumptively reduced susceptibility to vancomycin by BHI agar screen method. Conclusion: Screening of patient populations for MRSA, MuH and VISA/VRSA colonisation, especially those undergoing high risk surgeries should be considered, to prevent self infection and nosocomial transmission of resistant strains.

Page No. 90-95

Authors: Garima, Deepti Chaurasia

Introduction: The multidrug resistant Enterobacteriaceae isolates pose not only therapeutic problems but also serious concerns for infection control management. Early detection of lactamase producers is crucial to establish appropriate antimicrobial therapy and to prevent their interhospital and intrahospital dissemination. Objective: To study the prevalence of ESBL producing bacteria among all the members of Enterobacteriaceae isolated from various clinical specimens collected from all wards across a tertiary care teaching hospital. Methodology: ESBL production was detected among all the Enterobacteriaceae using, Phenotypic Combined Disk Diffusion Test (PCDDT), as recommended by the Clinical Laboratory Standards Institute (CLSI). Results: There is a high prevalence (25.67%) of ESBL producers in our hospital and so, it is essential to report the ESBL production along with the routine sensitivity reports, which will help the clinician in prescribing proper antibiotics. Pediatric ward had the highest number of ESBL positive Blood cultures. The pus isolates culture sensitivity testing from Surgery ward, Orthopedics and Burn and Plastic Surgery ward showed maximum number of ESBL strains. ESBL isolates show coresistance to many other classes of antimicrobials. Carbapenems (Imipenem, Meropenem), BL/BLI combination viz. Cefoperazone-sulbactum and aminoglycoside (amikacin) came out to be effective against these. Conclusion: A high degree of ESBL producers and carbapenem resistant Enterobacteriaceae is concerning; with emerging resistance to colistin, raising the fear of a return to the preantibiotic era. An urgent intervention including creating awareness and establishment of robust infection control and antibiotic stewardship program is the most important need of the hour.

Page No. 99-103

Authors: Mangala Ghatole, Virendra Kashetty, Apeksha Ghule, Avinash Wuike

Background: Control of tuberculosis is hampered by emergence of drug resistance such as MDR, XDR & TDR. Treatment of this disease will be facilitated if early reliable drug susceptibility profile is available. Various commercial systems are available but in resource poor country like India cheaper colorimetric methods are evaluated. One such method is using 2,3,5- Triphenyl tetrazolium chloride (TTC), which is an oxidation–reduction dye indicating cellular viability and growth. Colorimetric methods used in liquid media are shown to give better results than using them in Lowenstein–Jensen medium (LJ). Material and Method: A total of 82 strains isolated from patients suffering pulmonary tuberculosis and identified as M. tuberculosis were subjected to DST for Isoniazid and Rifampicin by Indirect drug susceptibility testing using absolute concentration method. 1% TTC was incorporated in Middlebrook 7H9 broth along with the drugs and compared with drug containing LJ. Results: Among the 82 strains, 9 (10.9%) strains were resistant to INH by both methods. Whereas 06 (7.3%) were resistant only in TTC. (sensitivity-100%, specificity -60%, positive predictive value- 91.8% and negative predictive value of 100%, P≤0.0001). Similarly 11(13.4%) strains showed resistance to RMP by both methods, 08 (9.8%) were resistant by only TTC and 01 (1.2%) only by conventional method. (Sensitivity- 98.4%, specificity- 57.9%, PPV-88.6%, NPV-91.7%, P≤0.0001). Conclusion: Results were available for 60 (73.2%) and 70 (85.4%) of strains by TTC method for INH and RMP respectively in 07 days as against 28 days in conventional method.

Page No. 110-113

Authors: Meenakshi Aggarwal, Sonal Saxena

Urinary tract infections constitute a large share of human systemic infections and are one of the commonest bacterial infections that lead patients to seek medical care. Positive culture results are a prerequisite for timely and effective treatment of UTIs. In resource limited settings, conventional media and biochemicals are used for isolation of pathogens and a positive urine culture sometimes may require more than 48 hours for identification. To overcome this time lag, chromogenic media have been developed which help in early presumptive identification of the uropathogens and timely initiation of therapy. We have evaluated one such media Urichrom–II™ (International Microbio, France) in comparison to in house conventional media for growth of common urinary pathogens.

Page No. 118-125

Authors: Kulal B., Bhat K.S., Meundi M., Kotigadde S.

Aim of the study: To determine the fungi causing otomycosis and the associated bacterial pathogens in a rural set up in clinically suspected cases of otomycosis. Methodology: Ear swabs were collected from 100 clinically suspected cases of otomycosis and from 50 persons who were apparently healthy. All the samples were processed by direct microscopy of KOH mount and Gram staining. Cultures were carried out for both fungal and bacterial isolates, which were identified by standard procedures. The data were statistically analysed. Results: Among 150 samples, 51.3% yielded only fungal growth, 23.3% grew bacteria only and 19.3% showed mixed growth of fungi and bacteria. Major fungal isolates were Aspergillus spp. and Candida spp. One sample grew A. sydowii which is an uncommon agent causing otomycosis. Major bacterial isolates were Staphylococcus aureus and Pseudomonas spp. All pathogenic bacteria were sensitive to routinely used antibiotics. Samples taken from healthy persons did not yield any fungal or bacterial pathogens.