Background: Multidrug-resistant (MDR) Gram-negative bacilli that release carbapenemase enzymes ensure that they are not affected by many beta-lactam antibiotics and carbapenems. The methodology of testing susceptibility to polymyxin B can pose a challenge in a clinical scenario. Aims: To assess polymyxin B, E-test susceptibility against Gram-negative bacteria that are MDR and compare the results of it with broth micro dilution (BMD). Settings and Design: This prospective observational study was done at a tertiary care center laboratory for 1 year. Materials and Methods: Polymyxin B susceptibility against 74 Gram-negative clinical isolates were assessed using the E-test and BMD as per the Clinical Laboratory Standards Institute recommendations. Statistical Analysis: Descriptive statistics of frequency, percentage, median, and intra-quartile range were done. Kolmogorov–Smirnov test was performed to assess the normality of the numerical variables. Results: The predominant organisms isolated were Klebsiella pneumoniae (55.4%) followed by Pseudomonas aeruginosa (21.6%), Acinetobacter baumanni complex (16.2%), Escherichia coli (4.1%), and Enterobacter cloacae (2.7%). The polymyxin B susceptibility ranges were much higher by E-test method when cross checked with BMD (97% as against 77%; with P < 0.0001). E-test however correlated poorly with BMD. Conclusion: BMD testing proved to be a more reliable method in situations where the clinical response to infection was inadequate or compromised.

Background Hepatitis B caused by hepatitis B virus HBV is a major global health concern. It can be transmitted through parenteral sexual and vertical routes and may lead to severe liver disease. Screening for hepatitis B is vital for effective patient management thereby reducing the burden of the disease and associated mortality.Aim To assess the role of HBsAg confirmatory neutralization test in screening hepatitis B infection and to determine the probable cut-off value of the quantitative HBsAg enhanced chemiluminescence immunoassay ECLIA test for accurate interpretation.Methods Patientrsquos blood samples were screened for HBsAg using the VITROS qualitative HBsAg test ECLIA. All reactive samples were further evaluated using two rapid diagnostic tests TRUST Line HBsAg rapid lateral flow chromatographic immunoassay and HEPACARD one-step sandwich immunoassay. Reactive samples with cut-off index COI between 1 and 199 were subjected to the VITROS confirmatory HBsAg neutralization test.Results Out of the 636 blood samples that were reactive by the VITROS qualitative HBsAg ECLIA 536 samples were also reactive by the rapid diagnostic tests RDTs. Among the 100 discordant results 61 were confirmed positive by the VITROS HBsAg neutralization test.Conclusion Sequential serological testing should be employed for the screening of HBsAg to minimize diagnostic errors. The VITROS qualitative HBsAg ECLIA can be used to screen HBsAg however for weakly reactive ECLIA samples a confirmatory neutralization test should be performed to avoid any false-positive results. Reliance solely on RDTs may lead to misclassification of weakly reactive cases as non-reactive.

Background and Aim: Non-Typhoidal Salmonellae (NTS) is emerging as a major health problem globally. They cause foodborne infections, especially gastroenteritis as well as extraintestinal manifestations. As this is a zoonotic disease, certain animal sources such as poultry, egg, meat and dairy products are implicated. Fruits and vegetables too contaminated by manure of infected animal excreta are possible vehicles of transmission of this disease. The present report analyzes the spectrum of NTS with seroprofiling, isolated in our institute for the last 12 years. Materials and Methods: Fresh fecal samples from patients with acute gastroenteritis and blood samples from cases of bacteremia were processed according to the standard protocol. Nonlactose fermenting colonies resembling Salmonella were subjected to biochemical characterization and antimicrobial susceptibility testing followed by serotyping. Results: One hundred fifty-two NTS strains were isolated from feces and 13 from blood. Serotyping of these strains revealed that the most common NTS species was Salmonella Typhimurium (44.8%), followed by Salmonella Oslo (15.2%) and Salmonella Weltevreden (11.5%). Many rare strains were also encountered in our study. Conclusion: As many of the NTS species are biochemically similar, serotyping of NTS should be made mandatory to know the burden of the disease and to detect the emergence of new serovars. Emergence of multidrug resistance in NTS strains is a major concern nowadays. It is very essential to adopt strict quality measures in poultry, fish, dairy, and meat processing units as they are widely distributed in animals. In addition, strategies should be formulated by health education and public awareness programs to sensitize the population regarding this menace.

Abstract Background: A good awareness and application of hand hygiene (HH) among health-care personnel form a solid foundation for any health-care center to prevent nosocomial infections. The objective of this study was to gauge the expertise and execution of HH among various health-care professionals in a tertiary care teaching hospital. Methodology: This analytical cross-sectional study was conducted over 2 months. A self-administered online questionnaire adapted from the WHO’s “Hand Hygiene Knowledge Questionnaire for Healthcare Workers” was sent to 217 participants. Their answers were scored based on the level of knowledge and practice of HH among them. Results: All age groups practiced HH to a fair extent, although respondents over the age of 31 (94.1%) showed significantly higher levels of understanding of HH in comparison to younger adults. There was no statistical difference in the knowledge level between male and female gender; however, females (91.2%) showed a better practice of HH than males. Although doctors (87.8%) had a higher degree of knowledge than nurses (70%), nurses (98%) had better HH practice than doctors. Overall, knowledge of HH was adequate with good practice. Conclusion: The adoption of excellent HH habits is thought to be predicted by greater knowledge about HH. However, to foster better HH habits that can significantly contribute to maintaining compliance with good practices, it is crucial to convey lacunae in knowledge during routine training sessions.

Background: The increasing cases of carbapenemase resistant Enterobacteriaceae (CRE) across the world is a cause of concern. Asymptomatic carriage of CRE in critical care units is a menace to infection control. Aims: This study determines the carriage rate of CRE in patients admitted to the intensive care units (ICU's) and evaluates the potential risk factors, leading to colonization in patients with CRE. Materials and Methods: Sixty rectal swabs from patients in the ICU's were screened for carriage of CRE. The samples were inoculated onto ChromID CARBA SMART bi-plate. The organisms showing color appearances as per the manufacturer's instructions were considered as CRE. Routine disk diffusion technique was also employed and CRE was defined as an organism belonging to the Enterobacteriaceae family which was resistant to either imipenem or meropenem. Results: The organisms isolated were identified and the percentage of carriage of carbapenem-resistant organisms was 12 (20%), of which Klebsiella pneumoniae was 4 (33.3%), Escherichia coli 6 (50%), Citrobacter freundii 1 (8.3%), and Enterobacter spp. 1 (8.3%). Out of these, 2 (3.3%) showed OXA 48 type resistance seen with K. pneumoniae and E. coli. Prior hospitalization, the use of high-end antibiotics and patients who have undergone surgeries were the most common potential risk factors for colonization with CRE. Conclusion: The prompt detection of CRE by routine screening using cost-effective methods and reduction of potential risk factors for gut colonization reduce the transmission of drug resistance in any hospital setting and pave the way for better antibiotic stewardship and appropriate contact isolation precautions.

Background: A good awareness and application of hand hygiene (HH) among health-care personnel form a solid foundation for any health-care center to prevent nosocomial infections. The objective of this study was to gauge the expertise and execution of HH among various health-care professionals in a tertiary care teaching hospital. Methodology: This analytical cross-sectional study was conducted over 2 months. A self-administered online questionnaire adapted from the WHO’s “Hand Hygiene Knowledge Questionnaire for Healthcare Workers” was sent to 217 participants. Their answers were scored based on the level of knowledge and practice of HH among them. Results: All age groups practiced HH to a fair extent, although respondents over the age of 31 (94.1%) showed significantly higher levels of understanding of HH in comparison to younger adults. There was no statistical difference in the knowledge level between male and female gender; however, females (91.2%) showed a better practice of HH than males. Although doctors (87.8%) had a higher degree of knowledge than nurses (70%), nurses (98%) had better HH practice than doctors. Overall, knowledge of HH was adequate with good practice. Conclusion: The adoption of excellent HH habits is thought to be predicted by greater knowledge about HH. However, to foster better HH habits that can significantly contribute to maintaining compliance with good practices, it is crucial to convey lacunae in knowledge during routine training sessions.

Healthcare associated infections (HAI’s) or nosocomial infections are frequently encountered in neonatal intensive care units (NICU’s), resulting in morbidity as well as mortality of new born babies, with varying predisposing factors and microbiological profile. This study was conducted to identify the most common HAI among the neonates admitted in the NICU, to identify the risk factors of HAI’s in neonates and to compare the bacteriological profile of culture positive cases with the results of environmental survey. This was a prospective study, conducted for a duration of one and a half years from December 2018 to May 2020. Environmental samples from the NICU as well as clinical samples from neonates admitted to the NICU of a tertiary care centre for a duration of more than 2 calendar days were collected. The most common HAI in the NICU was VAP (41.6%), followed by CLABSI (21.8%). Statistically significant risk factors for neonatal nosocomial infection were preterm birth, LSCS, presence of comorbidities, Low Birth Weight, absence of breast feeding and Apgar score. The most common Gram negative organism isolated was Acinetobacter baumannii (18.8%) and Gram positive organism as Methicillin Resistant Coagulase negative Staphylococcus (MRCoNS) (13.9%). Acinetobacter baumanni above average isolation rate was matching with the growth of the similar organism each month, according to the surveillance report. The most common HAI among neonates in our study was VAP, majority of which were caused by Gram negative bacteria. Acinetobacter baumanni was found to have an isolation rate above average on environmental surveillance, indicating its role in causing HAI’s in neonates admitted to the NICU. However, this study did not show a definitive association between HAI and the environmental surveillance report.

Background: Among Gram-negative bacteria, the presence of beta-lactamases is a defined modality of resistance to antibiotics of the beta-lactam group. Proteus species also has been showing various degrees of resistance to these groups of antibiotics and thereby appearing multidrug resistant. The aim of our study was to know the occurrence of Proteus species from different clinical samples their antibiotic susceptibility, extended-spectrum β-lactamase (ESBL) production, and the associated patient risk factors. Materials and Methods: This observational descriptive study was done from December 2020 to December 2021. All Proteus isolates obtained from various clinical samples, were biochemically identified, antibiogram, and screening for ESBL production was done using a combination of cefotaxime and cefotaxime clavulanic acid antibiotic discs. Results: Seventy-nine Proteus isolates were obtained from hospitalized patients. In that, Proteus mirabilis was the most common isolate followed by Proteus vulgaris. Proteus infections were common in the age groups of 41–60 years and males were affected more than females. Out of the 79 Proteus isolates 12 were ESBL producers. Among these, 25% of ESBL producers were P. mirabilis and 75% of them were P. vulgaris. Conclusion: The most common isolate was P. mirabilis among hospitalized patients and P. vulgaris was the main ESBL producers when compared to P. mirabilis. These pathogens mainly caused wound and urinary tract infections. The patient age groups ranged mainly from 49–60 years. Diabetes mellitus was the most significant risk factor in these Proteus infections.

Introduction: Multiple drug resistance (MDR) of the bacteria is an issue across any health-care setting worldwide. This leads to prolonged hospital stay and high cost of treatment. One must be cautious to use reserve drugs such as imipenem, meropenem, colistin, polymyxin, and tigecycline. Tigecycline is a mainstay to treat emerging single or MDR pathogens. The study was conducted to assess the in vitro sensitivity of tigecycline to MDR isolates and to compare the sensitivity of Kirby–Bauer (KB) disc-diffusion method with the Epsilometer (E-test) against various MDR clinical isolates. Materials and Methods: This prospective study was done from January 2019 to 2020 on MDR isolates from clinical specimens. The isolates were tested for their antibiotic sensitivity to tigecycline by KB and E-test method to know the minimum inhibitory concentration (MIC) of tigecycline. Statistical data were analyzed by the percentage method. Results: Tigecycline showed a higher in vitro sensitivity against multidrug-resistant isolates in our study. Among the 89 MDR isolates obtained, 38 (42.7) were found to be sensitive to tigecycline by the KB diffusion and 67 (75.3) were sensitive by the E-test method. Conclusions: Tigecycline is a reserve antibiotic against MDR organisms, especially useful to treat coinfections of Gram-positive and Gram-negative superbugs. The E-test was found to be far superior to the KB method to detect in vitro activity and the results of this test will guide the clinician to judiciously use this antibiotic in their day-to-day practice.

A study to analyze the differences between GeneXpert MTB/RIF assay and LED fluorescent microscopy for the diagnosis of pulmonary tuberculosis in a tertiary care centre - IJMR- Print ISSN No: - 2394-546X Online ISSN No:- 2394-5478 Article DOI No:- 10.18231/j.ijmr.2020.009, Indian Journal of Microbiology Research-Indian

Tuberculosis remains world's leading cause of death from a single infectious agent. Pulmonary tuberculosis (PTB) is the most common presentation of tuberculosis. Here we compare Ziehl-Neelsen (ZN), Gabbet's and Fluorescent staining (FS) methods to determine efficiency and cost effectiveness of diagnosing pulmonary tuberculosis from sputum samples. Three smears from 148 sputum samples of PTB cases were prepared and stained by the above methods. Of the samples examined, 71.6% and 53.4% and 85.8% cases were detected by Ziehl-Neelsen, Gabbet's and Fluorescence staining respectively. Fluorescent staining was found to be more reliable and cost effective method particularly when dealing with large sample loads than Ziehl-Neelsen and Gabbet's staining technique

Context: Anaerobes are significant pathogens as well as normal flora in a variety of body sites. As the conventional anaerobic culture techniques are laborious, time-consuming and relatively expensive, the interest in anaerobes started narrowing down in the recent past. Objective: This study is aimed at the isolation of bacteria encountered in the surgical infections with an emphasis on anaerobes and to create an awareness among the clinicians regarding the potential role of these silent pathogens Materials and Methods: The present prospective study that was conducted for a period of 1 year, employed 393 specimens collected from various surgical infections. All the samples were subjected for the isolation of both aerobes and anaerobes. Results: Out of 393 specimens, 193 anaerobes and 311 aerobes were obtained. Majority of the cases (226) exhibited polymicrobial etiology. Among the anaerobes, more frequently isolated organisms were nonsporing gram-negative bacilli, such as Bacteroides fragilis group, Prevotella-Porphyromonas group and Fusobacterium, which constitute 68.91% of the cases. Conclusions: This study emphasizes the need to employ anaerobic culture techniques routinely in microbiology laboratories and to create an awareness among the clinicians regarding the potential role of anaerobes.

Streptococcus pneumoniae are Gram positive lanceolate diplococci. In some individuals they are present as normal commensals of the nasopharynx. They cause pneumonia, otitis media, sinusitis, bronchitis and bacterial meningitis. Infection may be exogenous or endogenous in origin. S.pneumoniae however are not a part of the normal vaginal flora. We present a case of a 19 year old primigravida of 25 weeks amenorrhea, who presented with a history of cough since 3 days, abdominal pain since a day and spotting per vagina. She had a previous history of IgA nephropathy, and underwent dialysis and was on oral steroids. S. pneumoniae was isolated from the high vaginal swab culture. As it may predispose to neonatal sepsis, she was started on IV Cefazolin. Due to severe pregnancy induced hypertension and premature rupture of membrane an elective caesarean section was done and a preterm low birth weight live female child was delivered. As the baby had an acute respiratory distress syndrome, feed intolerance, and a raised C- Reactive protein, she was put on a ventilator and given antibiotics. This signifies that S. pneumoniae can be a resident vaginal flora especially in patients with predisposing factors. Patients with high risk of S.pneumoniae infection should be screened and vaccinated to reduce morbidity and mortality outcome for both mother and child.

Background: Emerging trends in non ICUs (Intensive care units) reveal an increase in morbidity and mortality directly proportional to risk factors, virulence capabilities and resistant antibiograms in contrast to the ICU settings. Aims & Objectives: To determine the bacterial blood pathogens isolated from adult ICUs and compare it with isolates from non ICU ( wards ) settings in regard to antimicrobial resistance and analyze the role of carbapenems and colistin with special reference to non fermenters like Acinetobacter and Pseudomonas species. Methods: A one year prospective cohort study of patients in adult ICUs and non ICUs. Positive bacterial Bactec blood cultures from patients admitted in both these settings and possible risk factors and antibiogram patterns were noted. Analysis was done using SPSS software. Results: The most significant bacterial blood isolates in both settings was Staphylococcus aureus, Acinetobacter and Pseudomonas species from non ICUs were predominant and demonstrated a higher degree of drug resistance in comparison with ICU strains. The major risk factors in the ICUs were intubations and mechanical ventilations and in non ICU settings were elderly patients with a history of recent hospital admissions requiring central line interventions and third line therapies. Conclusion: Growing incidence of community acquired multidrug resistant (MDR) non fermenters in non ICUs is a reminder to adopt systematic surveillance and minimize associated risk factors.

Massive ovarian oedema is a rare pseudotumor often mistaken for a malignant tumour. Our article describes a 47-year-old patient, who presented with acute abdomen suspected to be due to ovarian torsion. The patient was posted for staging laparotomy suspecting ovarian malignancy but the histological examination confirmed it to be massive ovarian oedema.

Background: Asymptomatic bacteriuria (AB) is common in-women and increases in prevalence with age or sexual activity. Prompt detection and treatment of this condition and associated factors decreases complications like acute pyleonephritis, intrauterine growth retardation and preterm labour. Chromogenic media is a versatile tool in rapid primary screening of the causative organisms considerably reducing daily routine workload.Aim: To determine the prevalence of AB among pregnant women in a tertiary care set-up and analyse the contributory risk factors, its effects on pregnancy and the role of chromogenic media in the laboratory diagnosis of these cases.Materials & Methods: Urine samples of all pregnant women attending pre-natal check-ups with no genitourinary complaints, history of fever or antibiotic intake were collected for Gram stain, culture and antibiotic sensitivity tests. A second urine specimen for culture and sensitivity testing was obtained from those with significant bacteriuria. The results were compared with patients showing negative urine cultures.Results: The overall prevalence of this clinical condition in our study was 13.2%. The significant isolates were Klebsiella pneumonia and E. coli and the most common risk factor was a previous history of urinary tract infection. The isolates were easily identified by using chromogenic agar (HiCrome) but colonies of uncommon pathogens like Acinetobacter and Streptococcus species appeared white and needed further identification.Conclusion: Screening of pregnant women for AB at first prenatal checkup helps analyse the associated factors and prevents its effects on pregnancy. The use of a chromogenic media can enhance reporting accuracy and will be an effective tool to monitor these cases routinely.

Aspergillosis, an opportunistic infection, is a major cause of morbidity and mortality in immunocompromised patients. But, Aspergillus is also known to cause Pulmonary aspergillosis in immunocompetent host too. Only few cases have been reported in literature of Aspergillosis in Immunocompetent individuals so far. Here, we report 4 different cases of Pulmonary Aspergillosis , who have presented with acute symptoms.The possibility of pulmonary mycosis needs to be considered in patients with acute pulmonary symptoms like breathlessness , hemoptysis, etc with signs of infection even in immunocompetent individuals, without underlying pulmonary disease.