Comparative evaluation of Vitek®2 and broth microdilution method for colistin susceptibility testing of Gram-negative isolates from intensive care unit in a tertiary care hospital.

INTRODUCTION: Colistin is the last resort treatment against resistant Gram-negative bacteria, necessitating reliable and rapid means for sensitivity testing of colistin. Automated systems like VITEK®2 are adopted to determine the minimum inhibitory concentration (MIC) due to easy usage. Broth microdilution (BMD) for colistin MIC was suggested by EUCAST and CLSI. OBJECTIVE: To compare and evaluate colistin MIC by BMD and VITEK®2 against Gram-negative organisms from the ICU in a tertiary care hospital. METHOD: Clinically significant organisms isolated from ICU patients were included. MIC was determined using BMD and VITEK®2. Very major error (VME), major error (ME), essential agreement (EA), categorical agreement (CA), positive predictive value (PPV), negative predictive value (NPV), sensitivity, and specificity were analysed. RESULT: 533 isolates were obtained from blood (435,81.60%), respiratory samples (57,10.70%), pus and exudates (20,3.80%), urine (18,3.40%), and CSF (3,0.60%). The Enterobacterales were K. pneumoniae (185,34.70%) E. coli (73,13.70%) and E. cloacae (26,4.90%) while non-fermenters were A. baumannii (209,39.20%) and P. aeruginosa (40,7.50%). The VITEK®2 sensitivity was >99%; specificity ranged from 14.28 to 52.94%. PPV was 93.81% while NPV was 93.75%. VME ranged from 47 to 100% between isolates. ME was up to 20%. The highest VME was obtained in E. coli (100%). The total EA and CA observed were 68.5% and 99.79% respectively. CONCLUSION: Automated system VITEK®2 failed to detect the resistance in 32 (60%) isolates. The obtained VME and ME values were >3%, which is unacceptable as per the standard guidelines. EA of ≥90% wasn't obtained. Sensitivity for VITEK®2 was >99%, but had low specificity (14.28%). Hence, VITEK®2 is not reliable for colistin susceptibility testing.

MIST (Modified Intubating Sequence for Transmissibility) Bundle for Infectious Diseases with Aerosol Hazard.

The current global severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has magnified the risk to healthcare providers when inititiating airway management, and safe tracheal intubation has become of paramount importance. Mitigation of risk to frontline providers requires airway management to be an orchestrated exercise based on training and purposeful simulation. Role allocation and closed-loop communication form the foundation of this exercise. We describe a methodical, 10-step approach from decision-making and meticulous drug and equipment choices to donning of personal protective equipment, and procedural concerns. This bundled approach will help reduce unplanned actions, which in turn may reduce the risk of aerosol transmission during airway management in resource-limited settings.

Design, synthesis, in silico and in vitro evaluation of novel diphenyl ether derivatives as potential antitubercular agents.

Diphenyl ether derivatives inhibit mycobacterial cell wall synthesis by inhibiting an enzyme, enoyl-acyl carrier protein reductase (InhA), which catalyses the last step in the fatty acid synthesis cycle of genus Mycobacterium. To select and validate a protein crystal structure of enoyl-acyl carrier protein reductase of Mycobacterium tuberculosis for designing inhibitors using molecular modelling, a cross-docking and correlation study was performed. A series of novel 1-(3-(3-hydroxy-4-phenoxyphenyl)-5-phenyl-4,5-dihydro-1H-pyrazol-1-yl) ethan-1-ones were synthesized from this model and screened for their antitubercular activity against M. tuberculosis H37Rv. Compound PYN-8 showed good antitubercular activity on M. tuberculosis H37Rv (MIC = 4-7 µM) and Mycobacterium bovis (% inhibition at 10 µM = 95.91%). Cytotoxicity of all the synthesized derivatives was assessed using various cell lines, and they were found to be safe. Structure of PYN-8 was also confirmed by single-crystal X-ray diffraction. The molecular modelling studies also corroborated the biological activity of the compounds. Further, in silico findings revealed that all these tested compounds exhibited good ADME properties and drug likeness and thus may be considered as potential candidates for further drug development.

Diagnostic utility of in-house loop-mediated isothermal amplification and real-time PCR targeting virB gene for direct detection of Brucella melitensis from clinical specimens.

AIMS: In this present study, the utility of a newly developed loop-mediated isothermal amplification (LAMP) and real-time PCR assays designed to amplify the virB gene region of Brucella melitensis was evaluated from human clinical specimens. METHODS AND RESULTS: Fifty-four culture-confirmed cases of brucellosis and 54 culture negative but clinically suspected cases of brucellosis were included in the study. Whole blood, serum and other nonblood specimens were collected and subjected to blood culture using automatic blood culture system, serological tests, LAMP assay and real-time PCR. Overall sensitivities of LAMP and real-time PCR assays were 67·5 and 68·3% respectively. For nonblood clinical specimens, we noticed a marked increase in the sensitivities of LAMP (88·9%) and real-time PCR (100%) assays. CONCLUSIONS: Performance of LAMP and real-time PCR was not satisfactory for whole-blood specimens because of the low abundance of bacteria or DNA. On the other hand, using nonblood specimens, both the assays showed higher sensitivity and specificity which makes them a good alternative for the rapid diagnosis of human brucellosis. SIGNIFICANCE AND IMPACT OF THE STUDY: The developed LAMP and real-time PCR assays are a specific and rapid diagnostic tool for direct and early detection of Brucella in clinical specimens.

Assessment of core capacities for antimicrobial stewardship practices in indian hospitals: Report from a multicentric initiative of global health security agenda.

Introduction: Antimicrobial-resistant HAI (Healthcare associated infection) are a global challenge due to their impact on patient outcome. Implementation of antimicrobial stewardship programmes (AMSP) is needed at institutional and national levels. Assessment of core capacities for AMSP is an important starting point to initiate nationwide AMSP. We conducted an assessment of the core capacities for AMSP in a network of Indian hospitals, which are part of the Global Health Security Agenda-funded work on capacity building for AMR-HAIs. Subjects and Methods: The Centers for Disease Control and Prevention's core assessment checklist was modified as per inputs received from the Indian network. The assessment tool was filled by twenty hospitals as a self-administered questionnaire. The results were entered into a database. The cumulative score for each question was generated as average percentage. The scores generated by the database were then used for analysis. Results and Conclusion: The hospitals included a mix of public and private sector hospitals. The network average of positive responses for leadership support was 45%, for accountability; the score was 53% and for key support for AMSP, 58%. Policies to support optimal antibiotic use were present in 59% of respondents, policies for procurement were present in 79% and broad interventions to improve antibiotic use were scored as 33%. A score of 52% was generated for prescription-specific interventions to improve antibiotic use. Written policies for antibiotic use for hospitalised patients and outpatients were present on an average in 72% and 48% conditions, respectively. Presence of process measures and outcome measures was scored at 40% and 49%, respectively, and feedback and education got a score of 53% and 40%, respectively. Thus, Indian hospitals can start with low-hanging fruits such as developing prescription policies, restricting the usage of high antibiotics, enforcing education and ultimately providing the much-needed leadership support.

Antibiotic prophylaxis in organophosphorus poisoning: A study of health and economic outcomes.

Objective: Organophosphorus poisoning (OPP) is a major concern for developing countries. There are no guidelines for the prophylactic use of antibiotics in the management of OPP which in such critical cases might add to the economic burden of the patients as well as antibiotic resistance. We compared the health and economic outcomes in patients prescribed with prophylactic antibiotics with respect to the patients not prescribed with any antibiotics. Methods: A retrospective observational study was carried out for two years for patients admitted to ICU with OPP. Patients were graded for severity of OPP, and divided into two groups based on prophylactic prescription and no prescription of antibiotics. The length of stay (LOS), hospitalization cost and outcomes were measured and compared between the two groups using statistical tests. Results: Out of the 254 patients observed, 108 were prescribed with prophylactic antibiotics and 94 were not prescribed with any antibiotic. There was a significant difference between LOS, cost of treatment and outcomes in the two groups (p < 0.001). When antibiotics were not prescribed, the odds of improvement was 1.854 times higher compared to those who received prophylactic antibiotics although after adjusting for severity of poisoning, significance was lost. On an average, 2-3 antibiotics were prescribed to every patient in the first group. Conclusion: OPP is an important health concern where issues of antibiotic misuse and overuse are practiced. Our study suggested that systemic antibiotic prophylaxis did not offer any advantage over non-use of any antibiotics in patients with OPP.

Melioidosis: Reinfection going incognito as relapse.

Melioidosis has recently gained importance as an emerging disease in India. Recurrent melioidosis has been reported from different parts of the world and can be due to relapse or reinfection. Distinction between relapse and reinfection is important for epidemiology, investigation and management. Here, we present the data regarding rate of recurrence and utility of multilocus sequence typing (MLST) in differentiating relapse form reinfection amongst melioidosis patients from a tertiary care hospital in South India. Amongst the 31 patients who survived and underwent follow-up, 4 (13%) presented with recurrence. Three cases (75%) were identified as reinfection and one (25%) as relapse based on MLST. Re-exposure to environmental Burkholderia pseudomallei amongst patients with melioidosis in endemic areas is likely. In such a scenario, more often than not, recurrence of melioidosis can be attributed to reinfection.

Genome Sequence of a Burkholderia pseudomallei Clinical Isolate from a Patient with Community-Acquired Pneumonia and Septicemia.

Here, we report the draft genome sequence of Burkholderia pseudomallei CM_Manipal, the causative agent of melioidosis isolated from a diabetic patient in Manipal, southern India. The draft genome consists of 107 contigs and is 7,209,157 bp long. A total of 5,600 coding sequences (CDSs), 60 tRNAs, 12 rRNAs, and one noncoding RNA (ncRNA) were predicted from this assembly.

Spontaneous ascitic fluid infection in liver cirrhosis: bacteriological profile and response to antibiotic therapy.

BACKGROUND: Spontaneous ascitic fluid infection (SAI) is common in cirrhotic patients leading to significant morbidity and mortality. Third-generation cephalosporins are currently recommended as first-line therapy. We conducted a prospective study to determine bacterial etiology, susceptibility patterns, and clinical epidemiology including 1-month mortality of SAIs among patients with cirrhosis. METHODS: Records of 600 patients with suspected SAI over a 4-year period were analyzed. Empirical cefotaxime/ceftriaxone was initiated in patients who had a neutrophil count >250/mm(3). Treatment failure was defined by absence of clinical improvement and/or significant decrease in neutrophil count of ascites (<25 % of base line value) by 72 h of therapy. RESULTS: Seventy patients (11.6 %) had SAI, including 40 (57.1 %) culture-negative neutrocytic ascites (CNNA), 25 (35.8 %) spontaneous bacterial peritonitis (SBP), and five (7 %) monomicrobial non-neutrocytic bacterascites (MNB). Gram-negative bacilli (Klebsiella and E. coli) were the commonest organisms. The overall response rate to ceftriaxone was 62.8 % (44/70). Among culture-positive patients (SBP and MNB), sensitivity rates to ceftriaxone was 50 %, while it was 53.3 % for quinolones, 70 % for piperacillin-tazobactam, and 93.3 % for cefoperazone-sulbactam combination. Thirty-day mortality was lower for CNNA compared to SBP (20 % vs. 40 %, p < 0.001) and for patients with response compared to no response to first antibiotic (11.3 % vs. 53.8 %, p < 0.001). CONCLUSION: The response of SAI to third-generation cephalosporins was low at our center. Cefoperazone-sulbactam could be a better alternative choice.

Prevalence of measles, mumps, rubella, and varicella susceptibility among health science students in a University in India.

BACKGROUND: Health science students (HSS) are at increased risk of contracting and transmitting viral diseases such as measles, mumps, rubella, and varicella (chickenpox). This study was undertaken to determine the prevalence of susceptibility of HSS to these infections. METHODS: Using a cross-sectional design, 790 HSS of Manipal University, Manipal, India, answered a questionnaire and provided a blood sample which was tested for specific IgG antibodies to measles, mumps, rubella and varicella by ELISA (Enzygnost(®)). RESULTS: The study group was comprised of medical (53.9%), nursing (16.6%), and allied health (29.5%) students. Among the overall group (n = 790), the prevalence of serological susceptibility to measles, mumps, rubella, and varicella were 9.5%, 32.0%, 16.6%, and 25.8%, respectively. Among the subgroup of vaccinated subjects, susceptibility to measles, mumps, rubella, and varicella were 7.9%, 34.7%, 10.7%, and 35.2%, respectively. CONCLUSION: HSS susceptible to measles, mumps, rubella, and/or varicella are at risk of acquiring these diseases during their training period. In addition, they may be a potential source for nosocomial transmission posing a risk to immunocompromised patients. Hence, in the Indian setting, HSS should be immunized against measles, mumps, rubella, and varicella at the time of joining the medical school.

A child with Erysipelothrix arthritis-beware of the little known.

Erysipelothrix rhusiopathiae is an established animal pathogen while the zoonotic infections in humans are rarely reported. Infections occur after exposure to animals or animal products that are mostly occupational in adults. Here we report in a child for the first time septic arthritis and osteomyelitis without an identifiable risk factor. A 5-year-old male child was admitted with pain in the left hip joint and inability to bear weight on the limb. Clinical examination followed by radiological and magnetic resonance imaging was suggestive of septic arthritis. Erysipelothrix rhusiopathiae grew from peroperative joint specimen. The infection was resolved following arthrotomy, joint lavage and antibiotic therapy.

Multiple cutaneous swellings in an immunocompetent host--cryptococcosis overlooked.

Recently, there has been an increase in the incidence of cryptococcosis even among immunocompetent patients. We present a case of multiple subcutaneous swellings later developing into a non-healing ulcer caused by Cryptococcus in a 60 year-old immunocompetent male along with a brief review of clinical presentations of cutaneous cryptococcosis in immunocompetent and immunocompromised individuals. A high index of suspicion is desired in view of possible dissemination and variable non-specific clinical presentations.

Clinical profile of brucellosis from a tertiary care center in southern India.

OBJECTIVE: To highlight the spectrum of clinical manifestations, labs, complications, treatment and outcome of brucellosis. METHODS: Retrospective study was conducted in Kasturba Medical College, Manipal University, Karnataka, India which included 68 confirmed cases of brucellosis from January 2006- April 2010. Diagnosis of brucellosis was made by culturing the sera/body fluids by standard BACTEC method (or) by testing the sera for Brucella agglutinins using the standard agglutination test (SAT). A titer of 1:320 or more was considered as significant. SPSS 16 was used for statistical analysis and Microsoft Excel for graphical representation. RESULTS: Of the 68 patients, 46 (68%) were male and 22 (32%) were female patients with age distribution of 9-75 years. Forty four (64.7%) had history of contact with unpasteurized dairy products or infected animals. Symptoms included fever (68, 100%), myalgia (21, 31%), musculoskeletal symptoms (23, 34%), headache (16, 24%), gastrointestinal symptoms (19, 28%) and altered sensorium (3, 4%). Co-morbidities and associations included HIV positivity (2, 3%), type 2 diabetes mellitus (13, 19%), steroid therapy (3, 4%) and HBsAg positivity (8, 12%). Ten (15%) patients had cervical lymphadenopathy, 4(6%) had splenic enlargement, 6 (7%) had hepatomegaly, 19 (28%) had hepatosplenomegaly and 2(3%) got meningeal signs. Anaemia was observed in 39 (57.3%) cases, high erythrocyte sedimentation rate (ESR) was present in 55 (80.8%) cases, leucocytosis in 10(14.7%), leucopenia in 10(14.7%), thrombocytopenia in 23 (33.82%) and thrombocytosis in 2 (2.94%) cases. CONCLUSIONS: In countries like India, where brucellosis and tuberculosis are endemic; rapid, sensitive and highly specific diagnostic methods are required to make early diagnosis and prevent resistance as there is an overlap in therapy.

Melioidosis in southern India: epidemiological and clinical profile.

Melioidosis, which is mainly prevalent in Thailand and Australia, has shown an increasing trend in India in the last few years. We carried out a retrospective study of 25 culture-proven adult cases of melioidosis who were admitted to a tertiary care hospital in southern India during June 2001 to September 2007. There was a six-fold increase in the number of cases in 2006 and 2007 as compared to 2001. Diabetes mellitus was the predisposing factor in 68% of cases, followed by alcoholism (28%). The clinical presentations were fever (80%), pneumonia and/or pleural effusion (48%), hepatomegaly (56%), joint involvement, and/or osteomyelitis (48%), splenomegaly (40%), splenic abscess (24%) and septicemia (28%). The organism, Burkholderia pseudomallei, was sensitive to co-amoxiclav, cotrimoxazole, ceftazidime, and carbapenem. The study suggests that melioidosis is an emerging infectious disease in the southwestern coastal belt of India, and it is likely to happen at much higher incidence.

Role of laboratory in rapid diagnosis of atypical mumps.

Fairly large number of mumps virus infections present atypically without parotitis leading to delay in diagnosis and increased morbidity. Awareness of such presentations and inclusion of serological test for detecting IgM-specific antibodies could help in solving diagnostic dilemma, especially in unvaccinated individuals from developing countries.

Pulmonary melioidosis in febrile neutropenia: the rare and deadly duet.

We present the first two fulminant cases of pulmonary melioidosis in febrile neutropenic patients with acute and varied presentations seen in our institution and their fatal outcome. A high index of suspicion coupled with microbiological confirmation can facilitate the administration of the appropriate therapy in cases of melioidosis that differ from other bacterial infections in terms of presentation and the response to antimicrobials.