PREVALENCE AND DRUG RESISTANCE IN ACINETOBACTER SP. ISOLATED FROM INTENSIVE CARE UNITS PATIENTS IN PUNJAB, INDIA

Objective: This study was designed to study the prevalence and antibiotic susceptibility patterns of Acinetobacter sp. as isolated from patients lodged in intensive care units (ICUs) of a tertiary care hospital, Ludhiana, Punjab, India.Methods: The clinical samples were simultaneously streaked on Blood agar and MacConkey agar. The identification of the bacterial isolates was carried out with the aid of Gram stain, motility test and along with a combination of other commonly employed biochemical tests. The antimicrobial susceptibility testing (AST) of all the bacterial isolates was carried out on Muller-Hinton agar through Kirby-Bauer disc diffusion method.Results: Acinetobacter sp. formed a fair allowance contributing at 42% among all ICU culture positive samples. The respiratory tract samples had a major share at 63.15% for all samples attributed to be positive for Acinetobacter sp. nosocomial etiology. The antibiotic sensitivity pattern portrayed that more than 95% of Acinetobacter sp. isolates were multiple drug resistant (MDR) whereas >50% Acinetobacter sp. showed extensive drug resistant (XDR). The last resort for such Acinetobacter sp. nosocomial infections is left to colistin and polymyxin B.Conclusion: Acinetobacter sp. is a highly prevalent microorganism among ICU patients of Ludhiana, Punjab, India, while its potential to acquire resistance toward commonly used antibiotics represents it as a grave threat to the health-care industry, therefore signifying the need for its regular monitoring in the health-care setups

Rationale use of microbiological diagnostic techniques: A module for undergraduate medical students

Introduction: As we all know that 2nd-year professional students are just the beginner to learn the clinical skills. By teaching rational use of various laboratory techniques, one can shortlist the investigations to be ordered, thereby making it cost effective for the patients. This also encourages rapid and precise investigations, decreasing hospital stay and cost and lesser number of nosocomial infections. Aims and Objectives: The aims of this study were as follows: (1) to develop a module for an interactive disease-based teaching session to impart practical knowledge to undergraduate medical students of the microbiological diagnostic techniques, (2) to evaluate the outcome of this intervention on students for better perception and understanding of clinical microbiology, and (3) to encourage implementation of this knowledge imparted to students later on for rapid and precise investigations, diagnosis, and cost-effective hospital stay in a clinical scenario. Materials and Methods: The study was conducted in the Microbiology Department of Christian Medical College and Hospital, Ludhiana. Forty-five 2nd-year professional students participated in the study. Topics were taught in the theory classes as usual followed by preproject assessment by multiple-choice questions (MCQs). The same topics were taught in the practical classes with special emphasis to diagnostic tests (which test when required). Students were again evaluated by the same set of MCQs as given for preproject assessment. Marks obtained by both teaching methods were analyzed statistically. Results: By postproject analysis MCQ test, it was seen that 22.2% of students scored marks in the range 80%–90% (preproject marks 4.44%). Majority of the students (66.6%) scored between 70% and 80% postproject while the students performing in this range preproject were 6.66% only. The postproject performance in the range 60%–70% and 60%/below 60% was seen among 6.66% and 4.44%, respectively. Paired t-test was used for comparison and was statistically significant (P < 0.05). Conclusion: Results of the current study appear to support continued use of the interactive disease-based session. While prospective research is needed to best assess the utility of the session, the current study does provide support for session benefits. This project is still going on in the department and students are taught clinical microbiology at the end of the syllabus

Vibrio isolates from cases of acute diarrhea and their antibiotic susceptibility pattern in a tertiary care hospital of Punjab

Introduction: Vibrio cholerae, the cause of cholera, is one of the leading causes of morbidity and mortality in many developing countries. Especially during monsoon season, cholera outbreaks have been frequently reported from one or the other part of this country. However, occasional sporadic cases have also been reported, majorly due to lack of proper sanitation and impure water supply. Aim: To determine the prevalence of serotypes of Vibrio isolates from cases of acute diarrhea and their antibiotic susceptibility pattern in a tertiary care hospital, North India. Materials and Methods: A retrospective study was conducted over a period of 1 year from January 2013 to December 2013 in the department of microbiology, in a tertiary care hospital in Punjab. The diagnosis was suggested by the clinical picture and confirmed by stool culture. The stool samples received in the laboratory were processed by standard microbiological techniques for identification of V. cholerae. The suspected colonies of Vibrio were identified by standard biochemical tests and serotyping was done by group specific antisera. The susceptibility of all the isolated Vibrio species to different antibiotics were done by Kirby-Bauer′s disk diffusion technique as per the Clinical and Laboratory Standards Institute (CLSI) guidelines. Results: V. cholerae were isolated from 41 (3.8%) of total 1063 stool samples received during the study period. On serotyping, 100% (41) of these positive samples were identified as V. cholerae 01 ogawa serotype. 95% (39) cases were encountered during monsoon season. In the present study analysis of the antibiotic susceptibility data showed highest susceptibility to gentamicin, amikacin, tetracycline and chloramphenicol. There is gradual increase in resistance to ampicillin and high level of resistance was observed for furazolidone and trimethoprim-sulphamethoxazole during the same period. Conclusions: Our study reveals a significant increase in cases of Vibrio infection during monsoon season, with V. cholerae 01ogawa as the predominant serotype. The results also suggest that this organism has begun to develop resistance against ampicillin, trimethoprim-sulphamethoxazole and furazolidone drugs and therefore these should not be used in the first line treatment

Online) An Open Access

ABSTRACT Tuberculosis is a disease associated with significant morbidity and mortality, more so in a developing nation like India. Most often it remains undiagnosed and therefore untreated. The gold standard for diagnosis of tuberculosis is by microscopic detection of Mycobacterium or its isolation from the specimen. But because of the poor performance of these conventional methods on extrapulmonary specimens, one has to opt for molecular methods like PCR. However, the cost per test and lack of treatment monitoring by PCR are the disadvantages in a developing country like ours. Therefore, a study was done comparing the conventional methods with PCR. It is a retrospective study conducted over a period of one year, in which a total of 114 samples sent for smear, culture &amp; PCR to the microbiology laboratory of a tertiary care hospital in northern India were included. Sample was first concentrated by Petroff&apos;s method and then with the concentrate obtained was inoculated on Lowenstein Jensen medium for culture and rest was used to make smear which was stained by Ziehl Neelsen stain. Another sample was sent to Dr. Lal Path Lab for Real Time PCR. Out of 114 samples, 88 (77.2%) were negative for Mycobacterium by all three methods. 26 (22.8%) samples were positive by PCR, out of which 24(92.3%) were extrapulmonary samples and 2 (7.7%) were pulmonary samples. Out of the 2 pulmonary samples, 1(50%) which was positive by PCR was also positive by conventional techniques; rest 25 samples (mainly extrapulmonary) were negative by conventional techniques. Conventional methods of microscopy &amp; culture remain gold standard for diagnosis of pulmonary tuberculosis &amp; monitoring of treatment. However, due to paucity of bacillary load &amp; uneven distribution of bacilli in extrapulmonary samples, one has to count on PCR. In conclusion, molecular methods like PCR work complementary to conventional methods rather than stand alone superior in the field of tuberculosis diagnosis

Clostridium difficile associated diarrhea - ′suspect, inspect, treat, and prevent′

Context: Clostridium difficile is a fastidious, gram-positive, spore-forming bacterium responsible for infectious diarrhea and pseudomembranous colitis associated with significant morbidity and mortality. Only toxigenic strains produce disease in humans. Pathogenicity is dependent on the presence of diarrhea-producing toxins, named toxin A (TcdA) and toxin B (TcdB). Risk factors include depletion of protective gut flora by antibiotics and diminished immune response to C. difficile due to age and medical co morbidities; and increased use of proton-pump inhibitors (PPI). Treatment includes the stoppage of inciting antibiotics if possible, to allow regeneration of the normal gut microflora, and starting an antibiotic with activity against C. difficile. A good clinical suspicion in patient with co morbidities developing diarrhea during hospital stay can help reduce the burden of this treatable morbid infection. Our aim of doing this study was to determine the prevalence of this infection in our tertiary care hospital, so as to monitor its burden in future. Aims: To determine the prevalence of this infection in our tertiary care hospital. Settings and Design: A retrospective study was conducted in the department of Microbiology in a tertiary care hospital in North India. Materials and Methods: A total of 195 stool samples received over a period of 2 years were included in the study. An enzyme immunoassay was performed for the qualitative determination of toxins A and B from Clostridium difficile in stool samples. Results: A total of 13 (6.67%) stool samples out of 195 samples processed were positive for the presence of Clostridium difficile toxins A/B. Conclusions: CDI has become a global public health challenge today. Various studies show a prevalence rate between 11% - 22%. Lower prevalence rate revealed from our study (6.7%), makes it imperative to maintain a strict surveillance in our patients to ensure opportune detection and treatmen

Online) An Open Access

ABSTRACT Shigella is an important cause of invasive dysentery in children and others. The present study was done to study the prevalence, any seasonal variation, distribution among children and adults, and antimicrobial susceptibility pattern of Shigella isolates in stools. A retrospective study was conducted over a period of 5 year from 1 January 2009 to 31th December 2013 in the department of microbiology, in a tertiary care hospital in Punjab. Stool specimens from diarrhea/dysentery cases in different age-groups were processed. The specimens were processed by standard microbiological techniques. The suspected colonies of Shigella were identified by standard biochemical tests and serotyping was done by group specific antisera. The susceptibility of all the isolated Shigella species to different antibiotics were done by KirbyBauer&apos;s disk diffusion technique as per the Clinical and Laboratory Standards Institute (CLSI) guidelines. Shigella species were isolated from 146 (2.42%) of the total 6025 stool samples received during the study period. Among these, S.flexneri was the predominant species accounting for 67.1% of isolates, followed by S.sonnei 25.3%, S.boydii 5.4% and S. dysenteriae in 2.1% of the total number of isolates. Of these, 53% were from children from 0-18 years and the rest 47% were from adults. Over 70% of Shigella isolates were resistant to two or more drugs including ampicillin, and co-trimoxazole and approximately, 3% Shigella isolates were resistant to ceftriaxone, during the study period. Analysis of the antibiotic susceptibility data showed a gradual increase in resistance to ampicillin, fluctuating resistance to norfloxacin and persistent resistance to co-trimoxazole. No resistance was observed to cefoperazone/sulbactam, piperacillin/tazobactam, and imipenem during the study period. The results of the study revealed the high prevalence of shigellosis with S. flexneri as the predominant species. The result also suggests that ampicillin, co-trimoxazole and norfloxacin should not be used empirically as the first line drugs in the treatment of shigellosis. Periodic analysis and reporting of antibiotic susceptibility is an important measure to check for antibiotic resistance and form appropriate treatment protocols

Opportunistic infections in relation to CD4 counts in human immunodeficiency virus seropositive patients in a tertiary care hospital in North India

Context: The human immunodeficiency virus (HIV) seropositive patients have decreased immunity predisposing them to various opportunistic infections (OI). The CD4 counts in the patient can be taken as a marker to evaluate the immune status. Aim: The aim of the study is to screen the HIV seropositive patients for OI in relation to their CD4 counts in a tertiary care hospital. Settings and Design: The present study was prospective in design conducted in the Department of Microbiology in a tertiary care hospital. Materials and Methods: A total of 80 HIV-seropositive patients, admitted from a period of 1΍ years were screened for OI. CD4 counts were done in all patients and measured by flow cytometry. Sputum and bronchoalveolar lavage samples were tested for tuberculosis and pneumocystis carinii pneumonia (PCP); oral candidiasis was detected by potassium hydroxide preparation; stool samples were subjected to direct and ether concentration method and stained for opportunistic stool infections; serum samples were tested for toxoplasmosis and cytomegalovirus (CMV) infection by enzyme-linked immunosorbent assay. Cerebrospinal fluid samples were tested for cryptococcosis. Results: A total of 38 (47.5%) patients were positive for OI. There were 13.6% patients presenting with infections having a CD4 count of below 200 cells/μL of whom, oral candidiasis (40.8%), cryptosporidiosis (23.68%), tuberculosis (5.92%), and CMV immunoglobulin M (IgM), PCP, isosporiasis and cryptococcosis were (2.96%). Conclusions: Majority of OI are present in patients with HIV with a low CD4 count, which emphasizes the need for monitoring such patients

Species distribution and antifungal susceptibility of candidemia at a multispecialty center in North India

Introduction: Fungi have emerged as major opportunistic pathogens. Candida species account for nearly 96% of all opportunistic mycoses and is an important cause of bloodstream infections. There has been a progressive shift from the predominance of Candida albicans to nonalbicans Candida species as the major cause of candidemia all over the world. Resistance to antifungal drugs is more in nonalbicans Candida species. Hence, speciation and antifungal susceptibility testing is the need of the hour. Materials and Methods: This retrospective study was conducted in a multispecialty center in North India from January 1, 2014 to March 31, 2015. The blood culture samples that were positive for Candida species were processed further. Species identification was done by standard microbiological techniques. Antifungal drug susceptibility was done by disk diffusion method (Clinical and Laboratory Standards Institute M44-A2). Results: A total of 10893 samples were processed, 1440 (13.2%) blood cultures were positive. Candida species was isolated from 105 (7.3%) samples, of which 15 (14.3%) were C. albicans and 90 (85.7%) were nonalbicans Candida. Nonalbicans Candida included Candida tropicalis (50.5%), Candida glabrata (19.0%), Candida parapsilosis (14.3%) and one isolate each of Candida guillermondi and Candida krusei. Majority of the Candida spp. isolates were resistant to clotrimazole (55.5%) fluconazole (42%) and itraconazole (69%) and and ketoconazole (38%). All isolates were uniformly sensitive to amphotericin B. Conclusion: There is predominance of nonalbicans Candida species in hospital setting. A high index of suspicion, early diagnosis, and a prompt and appropriate therapy remains the cornerstone of treatment