Regional variations in antimicrobial susceptibility of community-acquired uropathogenic Escherichia coli in India: Findings of a multicentric study highlighting the importance of local antibiograms.

Objectives: Evidence-based prescribing is essential to optimize patient outcomes in cystitis. This requires knowledge of local antibiotic resistance rates. Diagnostic and Antimicrobial Stewardship (DASH) to Protect Antibiotics (https://dashuti.com/) is a multicentric mentorship program guiding centers in preparing, analyzing and disseminating local antibiograms to promote antimicrobial stewardship in community urinary tract infection. Here, we mapped the susceptibility profile of Escherichia coli from 22 Indian centers. Methods: These centers spanned 10 Indian states and three union territories. Antibiograms for urinary E. coli from the outpatient departments were collated. Standardization was achieved by regional online training; anomalies were resolved via consultation with study experts. Data were collated and analyzed. Results: Nationally, fosfomycin, with 94% susceptibility (inter-center range 83-97%), and nitrofurantoin, with 85% susceptibility (61-97%), retained the widest activity. The susceptibility rates were lower for co-trimoxazole (49%), fluoroquinolones (31%), and oral cephalosporins (26%). The rates for third- and fourth-generation cephalosporins were 46% and 52%, respectively, with 54% (33-58%) extended-spectrum ß-lactamase prevalence. Piperacillin-tazobactam (81%), amikacin (88%), and meropenem (88%) retained better activity; however, one center in Delhi recorded only 42% meropenem susceptibility. Susceptibility rates were mostly higher in South, West, and Northeast India; centers in the heavily populated Gangetic plains, across north and northwest India, had greater resistance. These findings highlight the importance of local antibiograms in guiding appropriate antimicrobial choices. Conclusions: Fosfomycin and nitrofurantoin are the preferred oral empirical choices for uncomplicated E. coli cystitis in India, although elevated resistance in some areas is concerning. Empiric use of fluoroquinolones and third-generation cephalosporins is discouraged, whereas piperacillin/tazobactam and aminoglycosides remain carbapenem-sparing parenteral agents.

Are we missing the silver lining of COVID-19 pandemic: An analytical study to determine effects of three COVID-19 peaks on antimicrobial susceptibility of Staphylococcus aureus isolates.

Background: Given the evolving nature of COVID-19, for better understanding of its effect on antimicrobial resistance of Staphylococcus aureus (S. aureus), it becomes crucial that we follow the resistance patterns across different surges of COVID-19 cases. Methods: This prospective surveillance study extended over two years from January 2020-March 2022 and was conducted in a healthcare center of North India. Susceptibility patterns of Staphylococcus aureus during January-March 2020 were considered as prepandemic patterns. Processing of clinical specimens, identification of S. aureus, and in-vitro antibiotic susceptibility testing were performed in accordance with standard microbiological testing procedures and Clinical Laboratory Standard Institute guidelines. Results: Lowest prevalence (38.9%) of Methicillinresistant S. aureus was reported during January-March 2021 and July-September 2021. More than 50% S. aureus isolates were susceptible to linezolid, cotrimoxazole, tetracycline, and gentamicin in January-March 2020. In January-March 2021, ≥50% of S. aureus isolates from clinical specimens were additionally susceptible to clindamycin and erythromycin. Antibiotic agents of linezolid, tetracycline, clindamycin, and cotrimoxazole were susceptible in ≥50% of S. aureus isolates in January-March 2022. Conclusions: This study reveals a sharp decline in overall resistance to commonly prescribed antibiotic agents for S. aureus isolates after first peak of COVID-19 cases. However, same trend was not observed in subsequent peaks and probably we are approaching the same resistance levels that were seen prior to COVID-19 pandemic.

Predictors of Candidemia during Febrile Episode in Lymphoreticular Malignancy Affecting Paediatric Population.

Limited studies on candidemia in malignancy in the paediatric population from developing countries show a high incidence, high morbidity and a unique epidemiology as compared to developed nations. Our prospective observational study aimed to explore the prevalence of invasive candidiasis, especially candidemia, in febrile paediatric patients with lymphoreticular malignancy. A sample size of 49 children, with 100 recorded febrile episodes was studied. The relevance of candida colonization and mannan antigen detection as indicators of impending candidemia was evaluated. Genotypic identification of the yeast isolates was followed by sequence analysis using the NCBI-BLAST program, and the generation of the phylogenetic tree using MEGA 6.0 software. We observed a 5% prevalence of candidemia among febrile paediatric patients with lymphoreticular malignancy, predominantly caused by non-albicans candida. Colonization at multiple anatomical sites decreased from day 1 to day 8 of febrile episodes. Significant candida colonization (colonization index ≥0.5) was seen in a larger proportion of candidemia patients on day 1 and day 4 (p < 0.001) displaying a definite association between the two. The receiver operator characteristic (ROC) curve analysis for mannan antigen level revealed a cut-off of ≥104.667 pg/mL, suitable for predicting candidemia with a sensitivity of 100%, specificity of 92% and area under ROC value of 0.958 (95% CI: 0.915-1; p < 0.001). A phylogenetic tree with three population groups, clade 1, 2 and 3, consisting of Candida auris (1), Candida tropicalis (2) and Candida parapsilosis (2), respectively, was generated. The diagnosis of candidemia based on mannan antigen detection gives early results and has high negative predictive values. It can be combined with other biomarkers to increase sensitivity, specificity and positive predictive value.

Colistin Susceptibility Testing by Colistin Broth Disk Elution MIC Method among Carbapenem-resistant Gram-negative Blood Culture Clinical Isolates in a Tertiary Care Setting, East Delhi, India.

Background: Antibiotic resistance is a growing concern for bloodstream infections (BSIs), especially with the emergence of multidrug-resistant (MDR) gram-negative bacteria. In this study, we aimed to assess the pattern of colistin susceptibility using the colistin broth disc elution (CBDE) method among carbapenem-resistant gram-negative clinical isolates from blood cultures in a high burden tertiary healthcare setting in East Delhi. Methods: A total of 106 carbapenem-resistant gram-negative clinical isolates were tested. The most common isolates were Klebsiella pneumoniae, Escherichia coli, Enterobacter species, and Klebsiella oxytoca by CBDE method. Result: All the carbapenem resistant gram-negative bacterial blood culture isolates showed intermediate colistin susceptibility. This was statistically significant by chi-square test (p<0.5). Conclusion: This study highlights the need to monitor colistin resistance trends in the face of increasing antimicrobial resistance. Accurate surveillance of emerging colistin resistance is crucial for effective management of BSIs caused by carbapenem-resistant gram-negative bacteria.

First study on detection of cryptic resistance to linezolid among clinical isolates of methicillin resistant Staphylococcus aureus from India.

PURPOSE: Linezolid is an oral antibiotic which is widely used for serious infections caused by Methicillin Resistant Staphylococcus aureus (MRSA). With emergence of vancomycin MIC creep among clinical strains of MRSA, it is essential to know the possible emergence of subclinical resistance against linezolid as well. With this background, we aimed to detect evident (phenotypic) and cryptic (hidden or genotypic) linezolid resistance among MRSA isolates. METHODS: 250 clinical isolates of MRSA were collected and their susceptibility patterns were determined. Every third MRSA isolate was subjected to PCR for domain V of the 23S rRNA for the mutation hotspot in the 746bp segment which harbors the classical mutation for linezolid resistance. Restriction Fragment Length Polymorphism was done to confirm presence of the G2576U mutation. RESULTS: Six isolates (2.4%) were phenotypically resistant to linezolid. Among these six LRSA isolates, 5 demonstrated the G2576U mutation by PCR - RFLP. Cryptic resistance to Linezolid was identified in two isolates among linezolid susceptible isolates. CONCLUSIONS: In the present study, hidden resistance to linezolid was observed in linezolid susceptible clinical isolates. Emergence of resistance against over-the-counter drugs like linezolid is major challenge. Identification of cryptic resistance among patients implies impending resistance to linezolid. Judicious use of antimicrobials, application of strict infection control practices and prescription audit needs to be made mandatory to preserve such drugs.

Utility of Cerebrospinal Fluid and Serum Procalcitonin for the Diagnosis of Neonatal Meningitis.

OBJECTIVE: Positive CSF culture is the gold standard for the diagnosis of meningitis but it carries poor sensitivity. CSF procalcitonin (PCT) is shown to have some utility for the diagnosis of meningitis though there are limited studies in neonatal age group. We planned this study to compare CSF, serum, and CSF to serum PCT levels in neonates with confirmed, probable, and nonmeningitis groups to determine its optimal cut-off in CSF and serum for diagnosing meningitis. STUDY DESIGN: Sixty-seven neonates who qualified for lumbar puncture were enrolled in the study. Neonates were categorized into confirmed meningitis, i.e., CSF cytochemistry and culture positive (N = 17), probable meningitis, i.e., CSF cytochemistry positive but culture negative (N = 25) and nonmeningitis, i.e., both cytochemistry and culture negative (N = 25). CSF and serum samples were stored at -80°C for PCT assay. RESULTS: Significant difference was seen in mean of CSF PCT in neonates with confirmed (0.31 ng/mL), probable (0.22 ng/mL), and nonmeningitis (0.11 ng/mL) groups. Similarly, significant difference was observed in serum PCT levels also, though the ratio of serum to CSF PCT was comparable. At cut-off of 0.2 ng/mL, CSF PCT had sensitivity of 95.2% and specificity of 96% in the diagnosis of meningitis. CONCLUSION: CSF PCT is more specific marker for the diagnosis of neonatal meningitis as compared with serum PCT and CSF to serum PCT ratio. KEY POINTS: · CSF procalcitonin is a better marker than serum procalcitonin for diagnosing neonatal meningitis.. · It is better than serum procalcitonin and CSF to serum procalcitonin ratio.. · At cut-off of >0.2 ng/mL CSF procalcitonin can diagnose neonatal meningitis with 96% specificity..

Paradigm Shift in Antimicrobial Resistance Pattern of Bacterial Isolates during the COVID-19 Pandemic.

Antimicrobial resistance (AMR) is an emerging public health problem in modern times and the current COVID-19 pandemic has further exaggerated this problem. Due to bacterial co-infection in COVID-19 cases, an irrational consumption of antibiotics has occurred during the pandemic. This study aimed to observe the COVID-19 patients hospitalized from 1 March 2019 to 31 December 2020 and to evaluate the AMR pattern of bacterial agents isolated. This was a single-center study comprising 494 bacterial isolates (blood and urine) that were obtained from patients with SARS-CoV-2 admitted to the ICU and investigated in the Department of Microbiology of a tertiary care hospital in Delhi, India. Out of the total bacterial isolates, 55.46% were gram negative and 44.53% were gram positive pathogens. Of the blood samples processed, the most common isolates were CoNS (Coagulase Negative Staphylococcus) and Staphylococcus aureus. Amongst the urinary isolates, most common pathogens were Escherichia coli and Staphylococcus aureus. A total of 60% MRSA was observed in urine and blood isolates. Up to 40% increase in AMR was observed amongst these isolates obtained during COVID-19 period compared to pre-COVID-19 times. The overuse of antibiotics gave abundant opportunity for the bacterial pathogens to gradually develop mechanisms and to acquire resistance. Since the dynamics of SARS-COV-2 are unpredictable, a compromise on hospital antibiotic policy may ultimately escalate the burden of drug resistant pathogens in hospitals. A shortage of trained staff during COVID-19 pandemic renders it impossible to maintain these records in places where the entire hospital staff is struggling to save lives. This study highlights the extensive rise in the use of antibiotics for respiratory illness due to COVID-19 compared to antibiotic use prior to COVID-19 in ICUs. The regular prescription audit followed by a constant surveillance of hospital infection control practices by the dedicated teams and training of clinicians can improve the quality of medications in the long run and help to fight the menace of AMR.

Clinico-immunological profile in a case of hepatic tubercular abscess: A rare manifestation of extrapulmonary TB in an immunocompetent child.

Tubercular liver abscess is a rare entity even in an endemic area for TB. We report here a rare case of pediatric tuberculous liver abscess, the etiology of which was established using recently introduced Cartridge based nucleic acid amplification test (CBNAAT). A 7 years old male child presented with vomiting, pain abdomen and fever. Hepatomegaly was found on examination. Ultrasound of abdomen revealed two liver abscesses in the right lobe. Patient remained symptomatic even after empirical antimicrobial therapy. On diagnostic tap Gram stained smear of the pus showed polymorphs with negative culture. CBNAAT was positive for Mycobacterial tuberculosis and sensitive to rifampicin. Subjecting difficult extrapulmonary specimens to relevant microbiological investigations along with CBNAAT and other newer methods may improve diagnosis of tuberculosis in such rare cases thus leading to an early management and decrease in morbidity.

Development, optimization, standardization, and validation of a simple in-house agar gradient method to determine minimum inhibitory concentration of vancomycin for Staphylococcus aureus.

BACKGROUND: The Clinical and Laboratory Standards Institute recommends reporting minimum inhibitory concentration (MIC) values of vancomycin for Staphylococcus aureus. Commercial MIC strips are expensive, and the traditional broth microdilution method is cumbersome. With this background, we attempted to develop and standardize an in-house agar gradient method to determine MIC values of vancomycin for S. aureus. OBJECTIVES: To develop and validate an in-house vancomycin MIC strip, based on simple agar gradient method for S. aureus as per bioassay development guidelines. MATERIALS AND METHODS: Filter paper gradient strips were made in house and impregnated with varying concentrations of vancomycin to create an antibiotic gradient. During standardization, MICs of ninety clinical strains of S. aureus and ATCC 29213 were tested by the broth microdilution and commercial strip followed by the in-house strip. During the validation stage, MICs of ninety different clinical strains of S. aureus and ATCC 29213 were determined by the in-house strip followed by MIC detection by broth microdilution and commercial strips. A reading of more than ± 1log2 dilution compared with broth microdilution was considered as an outlier. RESULTS: During the initial stage, there were 7/90 outliers in the clinical strains, and no outliers were seen with the ATCC 29213 control strain. Corrective action included increasing precaution during the antibiotic impregnation on the strip. During validation stage, only 4/90 outliers were observed in the clinical strains. The commercial strips had 29/90 among clinical and 15/30 outliers in the control strain during the prevalidation phase. Despite maintaining cold chain during the validation phase, the outliers for commercial strip were 18/90 and 4/30 for clinical and control strains, respectively. CONCLUSION: Reporting vancomycin MIC for S. aureus may be attempted using the in-house method after validating it with a gold standard broth microdilution method and quality control as per protocol.

Candida auris colonization in an immunocompetent patient: A new threat in medical ICU.

Candida auris has become a great challenge in diagnostic, therapeutic and hospital environmental adaptation. With a prevalence of 5.3% in intensive care unit (ICU) acquired candidemia in India, its colonization is very rapid which hastens hospital transmission. Strict surveillance and preventive measures need to be adopted in ICU as it can persist on dry, inanimate object, prompt adaptation and antifungal resistance can pose a future threat of a new drug hospital acquired pathogen.

Predictors for gut colonization of carbapenem-resistant Enterobacteriaceae in neonates in a neonatal intensive care unit.

BACKGROUND: With the emergence of carbapenem-resistant isolates, the therapeutic alternatives have become limited. Various factors are responsible for carbapenem-resistant Enterobacteriaceae (CRE) gut colonization. This study was conducted to determine predictors for CRE gut colonization in neonates who were hospital delivered and admitted in a neonatal intensive care unit (NICU). METHODS: Three rectal swabs were collected from 300 hospital-delivered and NICU-admitted neonates (likely to stay for >3 days). The data collected for the possible risk factors for CRE gut colonization were namely mode of delivery, prolonged rupture of membrane >18 hours, period of gestation, birth weight, meconium-stained liquor, ventilation, intravenous catheter, nasogastric (NG) tube, NG feeding, breastfeeding, katori spoon feeding, top feeding, expressed breastmilk, antibiotics administration, and duration of hospitalization. P < .05 was considered as statistically significant. RESULTS: A total of 26 cases of CRE were isolated from 300 neonates. Statistically significant risk factors were found to be NG tube, breastfeeding, NG feeding, top feeding, expressed breastmilk, ventilation, antibiotic administration, and duration of hospitalization. Top feeding and antibiotics administration were identified as 2 independent risk factors by multiple logistic regression. CONCLUSIONS: Active surveillance of cultures from hospitalized patients and implementation of preventive efforts can reduce the risk of CRE.

Changing trends in antimicrobial susceptibility pattern of bacterial isolates in a burn unit.

INTRODUCTION: Emerging antimicrobial resistance in nosocomial bacterial isolates, limits the available treatment options for burn wound infections, among them multi-drug resistant Gram negative bacteria and methicillin-resistant Staphylococcus aureus (MRSA) are major contributors to the increase in morbidity and mortality rates. MATERIAL AND METHODS: A retrospective cross-sectional study was done in the Department of Microbiology, University College of Medical Sciences & Guru Teg Bahadur Hospital, Delhi. A total of 818 wound samples from patients admitted in the burn wards and Intensive Care Units (ICUs) examined between 2010-2014 (5 years period). Pseudomonas aeruginosa was found as the most common isolate (37%) followed by Klebsiella pneumoniae (15%) and Acinetobacter baumanii (12%) among Gram negative organisms while S. aureus (12%) remained the major isolates among Gram positive organisms. A significant decrease in incidence of Gram positive organisms was observed in comparison with previous study. However, resistance to ceftazidime and aminoglycosides were increased significantly in Gram negative organisms. Multi-drug resistant P. aeruginosa (MDR PA) accounted for 15.2%, multi-drug resistant A. baumanii (MDR AB) was prevalent in 13.8% and MRSA in 77.4% of burn wound infections. DISCUSSION AND CONCLUSION: Emerging bacterial drug resistance has both clinical and financial implications for the therapy of infected burn patients. Spectrum of bacterial drug resistance in an institution is important for epidemiological as well as clinical purposes. Rising frequency of MDR strains in burn patients is alarming for clinicians as it downgrades the treatment efficacy.

Carriage prevalence of carbapenem-resistant Enterobacteriaceae in stool samples: A surveillance study.

BACKGROUND: With more people being exposed to antibiotics, intestinal microflora faces constant pressure of antibiotic selection, which has resulted in the emergence of multidrug resistant strains. This may pose a severe problem as intestinal Enterobacteriaceae members are commonly implicated in human infections. AIMS: This surveillance study was undertaken to investigate the carriage of carbapenem-resistant Enterobacteriaceae (CRE) in the gastrointestinal tract among patients attending the outpatient clinic in a tertiary care center of East Delhi, India. METHOD: We performed a prospective surveillance study to screen 242 Enterobacteriaceae isolates for carbapenemase production from the stool samples of 123 outpatients attending a tertiary care hospital in East Delhi over a four-month period. RESULTS: Twenty-four (9.9 per cent) isolates demonstrated carbapenemase activity among 242 screened Enterobacteriaceae isolates. Four stool samples had two isolates of different species, both eliciting this feature and therefore indicating presence of multiple carbapenem-resistant Enterobacteriaceae (CRE) isolates in a single sample. CONCLUSION: Screening for carriage of CRE in stools of patients undergoing elective or emergency gastrointestinal surgical procedures, with haematological malignancies taking chemotherapy, or those planned for bone marrow transplantation can guide clinicians about gut colonisation of multidrug-resistant Enterobacteriaceae as these groups of patients are at risk of possible endogenous infection.

Antibiotic pressure mediated selection of non-biofilm forming strain of Elizabethkingia meningosepticum causing fatal nosocomial meningitis in a term infant.

We describe a fatal case of hospital acquired meningitis in a term infant due to the antibiotic pressure mediated selection of Elizabethkingia meningosepticum. The antibiotics were administered for multi-drug resistant Acinetobacter baumannii infection. The strain was also phenotypically characterized for beta lactamase production, biofilm forming capability and resistance to in use disinfectants.

Residual solvent determination by head space gas chromatography with flame ionization detector in omeprazole API / Determinação solvente residual, por cabeça de cromatografia gasosa com detector de ionização espaço chama em omeprazol API

Residual solvents in pharmaceutical samples are monitored using gas chromatography with head space. Based on good manufacturing practices, measuring residual solvents is mandatory for the release testing of all active pharmaceutical ingredients (API). The analysis of residual organic solvents (methanol, acetone, cyclohexane, dichloromethane, toluene) in Omeprazole, an active pharmaceutical ingredient was investigated. Omeprazole is a potent reversible inhibitor of the gastric proton pump H+/K+-ATPase. The Head space gas chromatography (HSGC) method described in this investigation utilized a SPB TM-624, Supelco, 30 m long x 0.25 mm internal diameter, 1.4µm-thick column. Since Omeprazole is a thermally labile compound, the selection of the proper injector temperature is critical to the success of the analysis. The injector temperature was set at 170ºC to prevent degradation. The initial oven temperature was set at 40ºC for 12 min and programmed at a rate of 10ºC min-1 to a final temperature of 220ºC for 5 min. Nitrogen was used as a carrier gas. The sample solvent selected was N,N-dimethylacetamide. The method was validated to be specific, linear, precise, sensitive, rugged and showed excellent recovery.

Solventes residuais em amostras farmacêuticas são monitoradas utilizando-se cromatografia a gás "headspace". Com base nas boas práticas de fabricação, a medida de solventes residuais é obrigatória para o teste de liberação de todos os ingredientes farmacêuticos (API). Efetuou-se a análise de solventes orgânicos residuais (metanol, acetona, cicloexano, diclorometano, tolueno) em omeprazol, ingrediente farmacêutico ativo. O omeprazol é potente inibidor reversível da bomba de prótons H+/K+-ATPase. A cromatografia a gás "headspace" (HSGC) descrita nessa pesquisa utilizou um SPB TM-624, Supelco, de 30 m de comprimento x 0,25 mm de diâmetro interno, e coluna de 1,4 µm de espessura. Considerando-se que o omeprazol é termicamente lábil, a seleção da temperatura apropriada do injetor é crítica para impedir a degradação. A temperatura inicial do forno foi de 40 ºC, por 12 minutos, e programada à taxa de acréscimo de 10 ºC min-1 até a temperatura final de 220 ºC, por 5 minutos. Nitrogênio foi utilizado como gás de transporte. Selecionou-se como solvente a N,N-dimetilacetamida. O método foi validado mostrando-se específico, linear, preciso, sensível, robusto e com excelente recuperação.

Cardiorenal syndrome.

Very often, physicians confront with patients who have concomitant heart and kidney failure. The coexistence of kidney and heart failure carries an extremely bad prognosis. The exact cause of deterioration of kidney function and the mechanism underlying this interaction are complex, multifactorial in nature, and still not completely understood. Both the heart and the kidney act in tandem to regulate blood pressure, vascular tone, diuresis, natriuresis, etc. An extension to the Guytonian model of volume and blood pressure control is proposed called cardiorenal connection. Regulating actions of Guyton's model were coupled to their extended actions on structure and function of the heart and the kidney changes in the rennin-angiotensin-aldosterone system, the imbalance between nitric oxide and reactive oxygen species, the sympathetic nervous system, and inflammation are the cardiorenal connectors to develop cardiorenal syndrome. Imbalance in this closed complex will often lead to deterioration of both cardiac and kidney function. The World Congress of Nephrology emphasized vast interrelated derangements that can occur in cardiorenal syndrome and proposed that the recent definition of cardiorenal syndrome be modified into categories whose labels reflect the likely primary and secondary pathology and time frame. For management, drugs that impair kidney function are undesirable, particularly in a population with already compromised or at risk of kidney function. In severe volume-loaded patients who are refractory to diuretics, management of cardiorenal dysfunction is challenging. In the absence of definitive clinical trials, treatment decision must be based on a combination of patient's condition and understanding of individual treatment options.