Indian Journal of Microbiology Research

1-6

Authors: Aaftab G.P, Asha B Patil, Shobha Medegar.

Introduction: Extended spectrum β-lactamase (ESBL) and AmpC producing organisms pose unique challenges to clinical microbiologists, clinicians, infection control professionals. The objective of the present study was to analyze the risk factors that will help in identifying those at risk of acquiring infection by ESBL and/or AmpC producing organisms thereby making appropriate changes in the choice of antimicrobial therapy and minimizing treatment failures. Materials and Methods: Risk factors were evaluated using a retrospective Case control design. 100 patients with infections due to ESBL and/ or AmpC producing E.coli or K. pneumoniae were employed as cases. These were matched with 100 patients with non ESBL- non AmpC producing organisms (Controls). Univariate and Multivariate logistic regression analysis was performed to identify variables associated with becoming a case. Results: Out of the total 100 cases, E.coli 61 (61%) was the predominant ESBL and/or AmpC producer, followed by Klebsiella pneumonia 39 (39%). In the multivariate analysis, we found that duration of hospital stay for more than 5 days (OR=0.16, P=0), stay in ICU (OR=2.98, P=0.02) and prior treatment with antibiotics (OR=2.81, P=0.003) were statistically significant risk factors associated with acquisition of ESBL/AmpC producers. We also found invasive procedures like usage of urinary catheter (OR=3.34, P<0.005), central venous catheter (OR=3.35, P=0.018), intravenous catheter (OR=7.38, P<0.005) and other surgical procedures (OR=3.14, P=0.0002) contributed significantly to the spread of ESBL/AmpC producers. Conclusion: ESBL and AmpC producing strains were found to show higher rates of resistance to various class of antibiotics when compared to non ESBL and AmpC producers. Knowledge on risk factors of infection helps physicians in framing appropriate empirical antibiotic policies.

14-19

Authors: Jaya Lalwani, Riddhi Hathiwala, Pavan Pandey.

Introduction: Irrational use including non-compliance with prescribed antibiotics is most crucial determinants for the emergence and spread of antimicrobial resistance. The aim of the present study was to assess the prevalence of ‘unused’ antibiotics among households. Materials and Methods: A cross-sectional study was conducted among 2400 households of District Bhopal, Madhya Pradesh using a structured questionnaire. Descriptive statistics were used to describe the study results. Results: The prevalence of unused antibiotics among surveyed households was 17.9%. Most common age group for which unused antibiotics were initially prescribed was 0-5 year of age. The major source of unused antibiotics was a public hospital (59.9%), followed by self-medication (29.6%). A total of 1,470 tablets/capsules of antibiotics were found lying unused in the surveyed homes. Conclusion: Non-compliance with prescribed medicines and self-medication of antibiotics is a troublesome problem. More sophisticated research is needed to evaluate factors involved in the irrational use of antibiotics.

24-30

Authors: Charu Kaushik, Namratha Kurunji Gangadhar, Subrahmanya Bhat K., Subbannayya Kotigadde.

Introduction: Urinary tract infection (UTI) is one of the important causes of morbidity in the community.Development of antibiotic resistance among uro-pathogensposeproblem in the treatment of UTI.Hence it is essential to examine the prevalence of such uro-pathogens and study theiranti-biogram. Materials and Methods: Bacteria isolated from midstream urine samples collected aseptically from 266 patients over a period of 6 months from November 2016 to April 2017 were identifiedbystandard microbiological procedures. Anti-biogram was done by Kirby–Bauer disc diffusion method and interpreted as per the CLSI guidelines. Result: Out of 266 urine samples, 136 (51.12%) were found to be positive for microbial isolates, of which 96 (71.3%) samples were from females and 40 (29.4%) samples were from males. Escherichia coli was the predominant organism (47.05%) followed by Pseudomonas species (13.2%), Enterococcus species (11.7%), Klebsiella species (7.3%), Citrobacter koseri (4.4%) and Proteus species (2.9%). Enterobacteriaceae shows highest sensitivity towards nitrofurantoin (86%), amikacin (68%) and gentamicin (59%) and resistance towards norfloxacin (74%). Sensitivity of Pseudomonas spp was 72.2% for piperacillin/tazobactam, 44.4% for amikacin and 66% resistance to ceftazidime. Sensitivity pattern of Enterococci spp to nitrofurantoin 96.6%, aminoglycosides 78%, and fluroquinolones 50%. An attempt was made to study ESBL production among Enterobacteriaceae members. The isolates were 54.4% multidrug resistant (MDR) and 38.8% were ESBL producers. Conclusion: In our study E.coli was the most common uropathogen isolated. Patterns of antibiotic resistance in a wide variety of pathogenic organism were noted. To prevent the development of resistance, periodic evaluation of antibiotic susceptibility patterns is necessary.

38-43

Authors: Arunagiri Ramesh, Anand B Janagond, Sundaramurthy Raja, Sorakka Pandurangan Gobinathan, Jhansi Charl.

Introduction: Catheter-associated urinary tract infections (CAUTI) being the commonest hospital-acquired infection (HAI) [40%], its burden, aetiology along with antibiotic susceptibility is essential to implement and monitor an effective infection control program. Aims: To assess the incidence and rate of CAUTI in a tertiary care hospital and to identify the common organisms and their antibiotic sensitivity pattern. Settings and Design: Prospective hospital based observational study was carried out in a tertiary care hospital in South Tamilnadu for a period of 2 months (June 2016 to July 2016) after obtaining the Institutional Ethics committee approval. Materials and Methods: Total of 100 catheterized adult ICU patients were observed for signs of UTI on a daily basis. On clinical suspicion of UTI, urine sample was analysed microbiologically. CAUTI among various gender and age groups; pathogenic isolates, antibiotic sensitivity pattern and associated co-morbid conditions were expressed as percentages Results: Incidence of CAUTI was 16/100 catheterization. Device utilization ratio was 0.72. CAUTI rate was 36.2/1000 catheter days. CAUTI was commonly seen among patients with >80 years, females with altered sensorium. Diabetes mellitus (35.7%) was the most common comorbid condition followed by TB (25%). Risk of CAUTI after 3-5 days of catheterization was 11.7% and proportionately increased with prolonged catheterization (100% if >20 days). Escherichia coli (37.5%) followed by Candida Spp. (25%) were common pathogens among 3-5days of catheterization. Pseudomonas Spp. was the commonest isolate after one week of catheterization. Imipenem, colistin resistance was highest among pseudomonas. Conclusions: CAUTI rate was higher than the benchmark set by NHSN (0-4). Appropriate health education, strict infection control practices, appropriate antibiotic policy, proper guided bundle care of catheter are need of the hour to prevent the CAUTI.

47-51

Authors: T. S. Shailaja, A. Mohan Kumar, Rajan J. Payyappilly.

Introduction: Antimicrobial resistance (AMR) is a global menace of recent times which does not spare the uropathogenic bacteria also. We aim to study the changes in bacteriological profile and antibiotic resistance pattern among the bacterial isolates of Urinary tract infections (UTI) in a multi-centric study from two different tertiary care hospitals located in rural parts of Kerala in two different time periods. Materials and Methods: First study was conducted from Jul 2008 – Jun 2009 and the second from May 2016 - Apr 2017. Results: Total numbers of urine samples processed in the first and second study are 3262 and 3772, of which 1096 and 1265 respectively showed significant bacteriuria. The predominant isolate is E.coli in both, with a percentage of 50.8 and 48.9 respectively followed by Klebsiella sp. and Enterococcus sp. There is a marked increase in the resistance rate of E.coli to cefotaxime, piperacillin tazobactam and imipenem. In case of first line drugs, there is a minimal fall in resistance rate except for ciprofloxacin. Similarly for aminoglycosides there is no significant increase in resistance. Conclusion: The high prevalence of resistance to empirical drugs has long been appreciated in this area which precludes their use. Resistance to higher antibiotics also is on the rise. Very high rate of AMR is exhibited by the bacterial agents of UTI even in rural areas which necessitates routine culture and sensitivity mandatory in all cases. In order to stop this growing menace, judicious use of antibiotics has to be implemented.

57-60

Authors: Monika Sekar, Swapna M., Joshy M Easow.

Introduction: Health care workers (HCWs) are constantly associated with generation, segregation and disposal of biomedical wastes (BMW). Knowledge, attitude and practice (KAP) of BMW management among HCWs are the three determinants used to evaluate the effective functioning of BMW management system of the Institution. Aims: This study aims to detect the degree of knowledge, attitude and practice of BMW management among HCWs in a tertiary care hospital. Materials and Methods: This study included 150 HCWs from five groups (postgraduates, interns, nurses, technicians and sanitary staffs), each group comprising of 30 individuals. They were instructed to tick their response in the questionnaire containing 30 questions (10 questions about each parameter) and were graded as good, average and poor based on individual score. Results: Order of decreasing knowledge and practice seen among the five groups was postgraduates followed by laboratory technicians, interns, nurses and sanitary staffs. Decreasing order of attitude was postgraduates, interns and laboratory technicians with same score, followed by nurses and sanitary staffs. In our study, 81.33%, 86.7% and 69.33% of participants had above average KAP values respectively. Conclusion: Postgraduates and interns had better understanding of BMW management than other groups. Laboratory technicians were better than nurses and sanitary staffs. Sanitary workers were highly ignorant regarding BMW management. So a continuing medical education program on BMW management should be conducted on yearly basis to train and update newly appointed as well as existing HCWs.

66-70

Authors: Anna Cherian, Jyothi R..

Introduction: Acanthamoeba keratitis (AK), a potentially devastating disease is caused by free-living amoebae of the genus Acanthamoeba . The number of reported cases worldwide is increasing year after year, mostly in contact lens wearers as also in non-contact lens wearers. Interestingly, AK has remained significant due to an incomplete understanding of the pathogenesis and pathophysiology of the disease, diagnostic delays and problems associated with chemotherapeutic interventions. Corneal blindness is a major public health problem worldwide and infectious keratitis is one of the predominant causes. Aim: To find out the Parasitic Etiological Agents in Suppurative Keratitis at a tertiary eye care referral centre in South India. Materials and Methods: A hospital based study was done on total of 350 cases of keratitis, during the period from January 2013 to December 2013 .The samples were examined as wet mount as well as cultured onto Non nutrient agar with E.coli overlay. Result: Acanthamoeba species were isolated from 314 cases. Most of the corneal ulcers responded well to treatment which was started early in their course due to early detection of the parasite by direct microscopy and confirmation by culture.

76-79

Authors: Mahesh Kumar Shankar, Sharon V. A..

Introduction: Worldwide, the most common type of acute viral hepatitis is hepatitis A, transmitted via the fecal-oral route occasionally leading to death due to acute fulminant hepatitis. The present study was undertaken to study the seroprevalence of HAV infection in a tertiary care hospital over a five year period. Materials and Methods: A cross-sectional study was conducted from January 2012 to December 2016 on samples of blood obtained from cases of clinically suspected hepatitis infection by IgM 'capture' assay ELISA. Results: A total of 173 cases were included in the study. The number of positive cases were 35(20.2%). The percentage positivity rate of anti HAV IgM was higher among the children tested 32(30.8%) when compared to adults tested 3(4.3%). Seasonal distribution showed maximum number of seropositive cases in the monsoon months. Conclusion: Increase in the number of samples received during 2014 and 2015 and a zero positive rate in 2015 indicates a rise in the awareness about preventive measures against hepatitis A virus infection. Uniform and well structured protocols which monitor HAV cases and assess disease load are necessary to capture and probe into the sudden rise in the seropositivity, as seen in the last four months of 2016 in this study.

84-86

Authors: Sudheendra Kulkarni, B. V. Peerapur.

Introduction: Urinary tract infection (UTI) due to Escherichia coli (E. coli) is a common health problem among women. Pregnant women are at increased risk from both symptomatic and asymptomatic UTI because of the physiologic changes associated with pregnancy which may cause complications such as pyelonephritis, chronic renal failure, premature delivery and foetal mortality. Our study aimed to check the prevalence of asymptomatic UTI due to E. coli and its antibiotic profile among pregnant women of Bidar district of North Karnataka, India. Materials and Methods: An independent study was done from January to February 2015 to investigate asymptomatic UTI among pregnant women attending Antenatal care clinic (ANC) of Bidar Institute of Medical sciences (BRIMS) Teaching hospital Bidar. 150 mid-stream urine samples were processed for culture and antimicrobial drug susceptibility testing at Department of Microbiology, BRIMS, Bidar as per the routine microbiological techniques. Diagnosis of UTI was made when there were at least 105 colony forming unit (CFU)/ml of urine. Isolates other than E. coli are not considered for this study. Results: Among 150 samples tested, 65 samples grown E. coli with colony count of 105 CFU/ml of urine. (43.33%).The Majority of pregnant women were in their 2nd and 3rd trimester and the mean age of study population was 23.46. The rate of UTI was highest among age group of 20-25 years. E. coli isolates showed highest resistance to antibiotics Ampicillin (96.92%), Cefuroxime (55.38%) and ceftriaxone (53.85%). It was uniformly sensitive to routinely used drugs for treatment of UTI. Discussion: Many Gram negative and Gram positive organisms including Yeast cause UTI in pregnant women. Our study revealed the high prevalence of asymptomatic UTI among pregnant women due to E. coli in Bidar (43.33%) as compared to studies conducted in other parts Karnataka. This study also showed the antibiotic profile of E. coli to different drugs tested and showed highest resistance to Ampicillin. Conclusion: Asymptomatic Bacteriuria in pregnant women due to E. coli is relatively higher in Bidar district and it showed resistance against commonly used antimicrobial agents. Hence it is strongly recommended to undertake routine urine culture in pregnancy and provide appropriate treatment to reduce its complications.

92-95

Authors: Meerah Srinivasagam, Vijayalakshmi Arumugam, Rajeswari Jayakumar.

Introduction: Scrub typhus is a disease caused by Orientia (Rickettsia) tsutsugamushi which belongs to Rickettsiaeceae family. Early diagnosis of Scrub typhus is important because timely anti-microbial therapy may help an excellent response and prevent complications. Serology is widely used for diagnosis of scrub typhus. The Aim of this study isto detect the IgM antibodies by Capture Enzyme linked immuno sorbent assay (ELISA) and Weil –Felix agglutination test against Scrub typhus and to compare the results of IgM ELISA with Weil – Felix test. Materials and Methods:Total of 723 blood samples were collected from clinically suspected Scrub typhus patients. IgM ELISA (commercial) and Weil –Felix agglutination test for Scrub typhus were performed on all serum samples. Results: Among 723 patients, 31(4.2%) showed positive by Weil-Felix test against OX K antigen and 43 (6%) showed positive by IgM ELISA. Among 43samples which were positive by IgM ELISA, 13(1.8 %) showed negative by Weil-Felix test. Out of 31samples positive by Weil-Felix test, one sample showed negative by IgM ELISA. The sensitivity of IgM ELISA and Weil –Felix agglutination test is 96.77% and 69.76 % respectively. Conclusion:Early diagnosis and treatment will prevent complications of Scrub typhus. This study has found out the IgM ELISA is highly sensitive and dependable for diagnosis of scrub typhus than Weil –Felix agglutination test.

100-105

Authors: Sodani Sadhna, Hawaldar Ranjana.

Introduction: Respiratory tract infections (RTI) are one of the most common cause of infections and are responsible for loss of working hours in affected individuals. Many newer antibiotics have been introduced to treat these infections whose sensitivity pattern is still not clearly understood due to paucity of published data in this regard.Several studies have been published in different parts of the world but still there are limited data from Indian subcontinent. The present retrospective study was carried out in microbiology department of our diagnostic centre with the aim of analyzing the various causative bacteria of CAP and to study their antibiotic sensitivity pattern. Materials and Methods: The present retrospective study of sputum cultures was carried out between July to September 2017 in the microbiology department of our diagnostic centre. A total of 100 sputum samples collected from patients of all ages and both sexes were included in the study. Sputum samples were collected in wide mouth, sterile containers by standard protocols. Gram’s staining and Zheil Neelsen staining for acid fast bacilli was done after inoculating the specimen on Blood and MacConkey Agar plates. Results: There were 68 males and 32 females with a male to female ratio of 2.1:1. Maximum 23% patients were in 61-70 years age group followed by 15% each in 41-50 and 51-60 years age group. 3% patients were above 80 years of age. Normal bacterial flora was grown in 71% cases. Growth of pathogenic bacteria was observed in 29% cases. Klebsiella pneumoniae was the most commonly isolated micro-organism accounting for 13% cases. Staphylococcus aureus was isolated in 6% cases followed by Pseudomonas aeruginosa in 3% and 2% each of Enterococcus, E.coli and Candida species. Conclusion: The role of sputum culture has for long been debatable and is limited by the fact that it is difficult to obtain a deep cough specimen in paediatric and geriatric patients .However, epidemiological value of sputum cultures as a tool in providing information about microbiological profile and antibiotic sensitivity pattern in different geographical areas of the world cannot be underestimated.

110-114

Authors: Prakash H. Waghmare, Shalaka S. Rudrakshi, Kishor V. Ingole, Rashmi S. Bawane, Sadiya V. Shaikh.

Asymptomatic bacteriuria leading to urinary tract infection is the most frequently encountered condition in pregnancy, which has been associated with an increased risk of pre-term delivery and low birth weight. Clinically the diagnosis of UTI can be difficult as symptoms are non-specific. Gold standard method for diagnosis of UTI is urine culture. Up to 18 hours are required for bacterial growth on culture media by standard laboratory techniques, which leads to delay in the treatment. In the present study 100 midstream urine samples from asymptomatic pregnant women were collected. Gram staining, Griess Nitrite Test, Triphenyl tetrazolium chloride test (TTC), Catalase test, these four screening tests were performed on the samples and their results were compared with the results of urine culture. Among these 100 samples, the study revealed that 14 pregnant women had significant bacteriuria (1,00,000 or more bacteria/ ml of urine) giving a prevalence of 14%. This study showed that Gram staining has highest sensitivity i.e. 92.85% and TTC has highest specificity i.e. 98.83% for the detection of significant bacteriuria. To conclude if we perform these two tests in combination then we get a sensitivity of 98.97% and specificity of 94.22% which is not 100% but very close to it. Use of these screening tests will help in early diagnosis and initiation of treatment of UTI in pregnant women.

119-125

Authors: Sumaira Qayoom, Mariraj Jeer, Muzaffar Amin, Talat Masoodi, Syed Khurshid.

Objective: To find out the bacteriological profile of post burn infections in wound. Place and duration of study: Vijayanagar institute of medical sciences, ballari, over a period of one year from 1st January 2014-31st December 2014. Study design: Prospective Materials and Methods: Swabs were collected from the burn wounds using standard methods and cultured for the growth of the bacteria.The organisms isolated were identified by standard microbiological methods following which antimicrobial susceptibility test was done by Kirby-baeur disk diffusion test. Results: The predominant organism isolated from swab culture was Pseudomonas aeruginosa(23%), followed by Staphylococcus aureus(22%), E.coli(14%), K.pneumoniae(9%), K.oxytoca(8%), CONS(7%), Proteus mirabilis(5%), C.freundii(4%) and C.koseri(3%),others(3%) and sterile culture(3%).The antibiogram of Pseudomonas aeruginosa showed high susceptibility to Imipenem( 81%) Piperacillin/Tazobactam (78%), Amikacin(76%), Ceftazidime(60%) and low susceptibility to Gentamicin(57%), Tobramycin(57%), Cefotaxime(47%), and Ceftriaxone(39%).Total resistance pattern was noted in Ampicillin, Cotrimoxazole, Ciprofloxacin.

132-137

Authors: Vinoba S., Rashmi K. S..

Introduction: Metallobetalactamase producing Acinetobacter species has been reported to be an important cause of nosocomial infection and is a critical therapeutic problem worldwide, especially in the intensive care unit. Objectives: To determine the frequency of metallo‐β‐lactamases among imipenem‐resistant Acinetobacter species and to compare different phenotypic methods. Materials and Methods: 59 imipenem‐resistant Acinetobacter species isolated from various clinical samples were tested for metallo‐β‐lactamase production using different phenotypic methods. Minimal Inhibitory Concentratrion (MIC) to meropenem was determined by E test. Results: Of all the imipenem resistant isolates, 50.8% of Acinetobacter species were MBL producers. MIC of all those MBL producing isolates were ≥ 16μg/ml. Among the Acinetobacter species MBL production were detected in 30 isolates (50.8%) by E test, 29 isolates (49.2%) by CDT and in 15 isolates (25.4%) by DDST and DPT. The sensitivity and specificity of CDT, DDST and DPT compared to E test was 98%, 48.1%, 48.1% and 100%, respectively. Conclusion: Prevalence of MBL producing Acinetobacter species is being increasingly reported in ICU patients. The MIC of all the MBL producing isolates for meropenem were >16μg/ml (Resistant). E test and CDT were more reliable for MBL detection. CDT was cost effective, easy to perform and interpretation also straightforward. MBL producing isolates were multidrug resistant making therapeutic choices limited. Continuous antibiotic surveillance, infection control practices and an effective antibiotic policy are required to address the problem of MBL – associated infections.

143-146

Authors: Sunayana M. Jangla, Susan Cherian, Raji Naidu.

Introduction: Tuberculosis (TB) causes severe morbidity and mortality in millions of people every year, worldwide and India in particular. Hence this study was undertaken to find out the role of the widely used GeneXpertMTB/Rif test in tuberculosis detection. Materials and Methods: A total of 155 samples (pulmonary and extrapulmonary) from patients clinically suspected to have tuberculosis were received for testing by ZN (Ziehl-Neelsen) stain and GeneXpert/MTB Rif. These samples were also processed for histopathology or cytology and mycobacterial culture on request of clinician. The results were studied and compared. Result: Out of the total samples received, lymph node aspirates were 23%,pleural fluid 20%,sputum 19%,pus 10%,tissue 8%,ascitic fluid 8%,cerebrospinal fluid 7%,endometrial tissue 3%,synovial fluid 1% and urine 1%.Nine sputum samples (31%) were positive by GeneXpertMTB/Rif of which two were negative on smear, two positive on culture and there was no culture request for the remaining seven samples.29 (23%) extrapulmonary samples were positive by GeneXpertMTB/Rif out of which there was no culture request for 16 samples. One sample which was negative by GeneXpert MTB/Rif grew Atypical Mycobacteria on culture. Four isolates were detected as rifampicin resistant by GeneXpertMTB/Rif but none of them were sent for Drug Susceptibility Test (DST). Conclusion: GeneXpertMTB/Rif should be used in combination with other tests for diagnosing tuberculosis.