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Abstract Background Healthcare-associated bloodstream infections (BSI) threaten patient safety and are the third most common healthcare-associated infection (HAI) in low- and middle-income countries. An intensive-care-unit (ICU) based HAI surveillance network recording BSIs was started in India in 2017. We evaluated this surveillance network’s ability to detect BSI to identify best practices, challenges, and opportunities in its implementation. Methods We conducted a mixed-methods descriptive study from January to May 2022 using the CDC guidelines for evaluation. We focused on hospitals reporting BSI surveillance data to the HAI network from May 2017 to December 2021, and collected data through interviews, surveys, record reviews, and site visits. We integrated quantitative and qualitative results and present mixed methods interpretation. Results The HAI surveillance network included 39 hospitals across 22 states of India. We conducted 13 interviews, four site visits, and one focus-group discussion and collected 50 survey responses. Respondents included network coordinators, surveillance staff, data entry operators, and ICU physicians. Among surveyed staff, 83% rated the case definitions simple to use. Case definitions were correctly applied in 280/284 (98%) case reports. Among 21 site records reviewed, 24% reported using paper-based forms for laboratory reporting. Interviewees reported challenges, including funding, limited human resources, lack of digitalization, variable blood culture practices, and inconsistent information sharing. Conclusion Implementing a standardized HAI surveillance network reporting BSIs in India has been successful, and the case definitions developed were simple. Allocating personnel, digitalizing medical records, improving culturing practices, establishing feedback mechanisms, and funding commitment are crucial for its sustainability.

ABSTRACT The in vitro activity of two novel antibiotics with different modes of action, (i) siderophore cefiderocol and (ii) β-lactam-enhancer mechanism-based cefepime/zidebactam, was tested against New Delhi Metallo-β-lactamase (NDM)-producing Enterobacterales and Pseudomonas aeruginosa collected in India. Minimum inhibitory concentrations of antibiotics against multicentric NDM-producing Escherichia coli (n = 117), Klebsiella pneumoniae (n = 103), and P. aeruginosa (n = 72) were determined by the reference broth microdilution method. Among E. coli, 111 isolates were NDM-alone, and six were NDM + OXA-48-like producers. Among K. pneumoniae, 47 and 56 isolates were NDM-alone and NDM + OXA-48-like producers, respectively. All E. coli isolates harbored four amino acid inserts in their penicillin-binding protein 3. Using the highest susceptible breakpoint among CLSI, FDA, and EUCAST interpretive criteria, cefiderocol susceptibility was 39.3%, ≤80%, and 57%, for NDM ± OXA-48-like-producing E. coli, NDM ± OXA-48-like-producing K. pneumoniae, and NDM-producing P. aeruginosa, respectively. At a cefepime break point of ≤8 mg/L, 100% of Enterobacterales and ≥90% of P. aeruginosa isolates were cefepime/zidebactam-susceptible. NDM being a dominant carbapenemase among Enterobacterales and P. aeruginosa in India, the variable activity of cefiderocol against NDM producers is a concern. Post approval, cefepime/zidebactam could offer a promising treatment option against NDM producers.IMPORTANCEMetallo-β-lactamases are therapeutically challenging due to the limited treatment options. Against such isolates, currently approved newer β-lactam/β-lactamase inhibitor combinations are ineffective. In this study, we tested siderophore cephalosporin, cefiderocol, which utilizes an unconventional iron uptake pathway for efficient cellular penetration, and cefepime/zidebactam that utilizes novel β-lactam enhancer mechanisms for overcoming diverse carbapenemases. Cefiderocol showed limited activity against Escherichia coli isolates co-harboring New Delhi metallo-β-lactamase (NDM) with PBP3 insert, dual carbapenemase (NDM with OXA-48 like)-producing Klebsiella pneumoniae, and NDM-producing Pseudomonas aeruginosa isolates, while cefepime/zidebactam potently inhibited NDM-producing Enterobacterales and P. aeruginosa isolates. NDM being a dominant carbapenemase among Enterobacterales and P. aeruginosa in India, the variable activity of cefiderocol against NDM producers is a concern. Post approval, cefepime/zidebactam could offer a promising treatment option against NDM producers.

Summary: Background: Ventilator-associated pneumonia (VAP) is a major cause of morbidity and mortality in patients receiving mechanical ventilation in India. Surveillance of VAP is essential to implement data-based preventive measures. Implementation of ventilator-associated events (VAE) criteria for surveillance has major constraints for low resource settings, which can lead to significant underreporting. Surveillance of VAP using common protocols in a large network of hospitals would give meaningful estimates of the burden of VAP in low resource settings. This study leverages a previously established healthcare-associated infections (HAI) surveillance network to develop and test a modified VAP definition adjusted for Indian settings. Methods: In this observational pilot study, thirteen hospitals from the existing HAI surveillance network were selected for developing and testing a modified VAP definition between February 2021 and April 2023. The criteria used for diagnosing VAP were adapted from the CDC’s Pediatric VAP definition and modified to cater to the needs of Indian hospitals. Designated nurses recorded each VAP event in a case report form (CRF) and also collected denominator data. The data was entered into an indigenously developed database for validation and analysis. At the time of data analysis, a questionnaire was sent to sites to get feedback on the performance of the modified VAP definitions. Findings: Out of 133,445 patient days and 40,533 ventilator days, 261 VAP events were recorded, with an overall VAP rate of 6.4 per 1000 ventilator days and a device utilization ratio (DUR) of 0.3. A total of 344 organisms were reported from the VAP events. Of these, Acinetobacter spp (29.6%, 102) was the most frequent, followed by Klebsiella spp (26.7%, 92). Isolates of Acinetobacter spp (98%) and Enterobacterales (85.5%) showed very high resistance against Carbapenem. Colistin resistance was observed in 6% of Enterobacterales and 3.2% of Acinetobacter spp. Interpretation: Data from this pilot study needs to validated in the larger Indian HAI surveillance network so that it can help in wider implementation of this protocol in order to assess its applicability p VAP across India. Funding: This work was supported by a grant received from the Indian Council of Medical Research (code I-1203).

ABSTRACT The availability of syndrome-based panels for various ailments has widened the scope of diagnostics in many clinical settings. These panels can detect a multitude of pathogens responsible for a particular condition, which can lead to a timely diagnosis and better treatment outcomes. In contrast to traditional identification methods based on pathogen growth on culture, syndrome-based panels offer a quicker diagnosis, which can be especially beneficial in situations requiring urgent care, such as intensive care units. One such panel is the Biofire Filmarray Pneumonia plus Panel (BFP), which we have compared against microbiological culture and identification. The lower respiratory samples from patients were tested with BFP, culture, and identification with culture considered the gold standard. The phenotypic antibiotic susceptibility results (Vitek 2) were compared with the antimicrobial resistance (AMR) genes detected in BFP. Statistical analysis was carried out using GraphPad 7.0 and MS Excel (Microsoft Inc.). The results showed a positive percent agreement of 100% and a negative percent agreement of 47.8% with an overall agreement of 76.72% compared to culture. BFP was better at identifying fastidious bacteria, and the agreement with culture was higher for high bacterial identification numbers (107 and 106). There was also a correlation between the number of pathogens detected and growth in culture. Carbapenemase genes were detected in around 80% of phenotypically resistant samples and correlated with in-house PCR 60% of the time. Hence, BFP results need to be interpreted with caution especially when multiple pathogens are detected. Similarly, the presence or absence of AMR genes should be used to guide the therapy while being watchful of unusual resistance or susceptibility. The cost constraints and low throughput call for patient selection criteria and prioritization in emergency or resource-limited conditions.IMPORTANCEApplication of syndrome-based panels in clinical microbiology is of huge support in infectious conditions requiring urgent interventions, such as pneumonia. Interpreting the results requires caution; hence, we have compared the results obtained from Biofire Filmarray Pneumonia plus Panel with standard microbiological methods.

Introduction Intestinal parasitic infections continue to loom in developing countries with low sanitation and socioeconomic conditions. Pandemic times are especially important to study the prevalence of these pathogens since the focus of all healthcare services was coronavirus disease 2019 (COVID-19). This study aimed to evaluate the prevalence and time-trend of intestinal parasitic infections in the capital region of India during the pandemic times.

Invasive Fungal Infections have posed a difficult challenge in the recovery of people infected with COVID-19. COVID-19-associated pulmonary aspergillosis (CAPA) has been described and found in about 30 % of ICU patients worldwide. Here we present an unusual microscopic finding in a case of CAPA in an ICU patient which was helpful in initiating early definite therapy. A 56-year-old gentleman presented with complaints of fever and shortness of breath and subsequently tested positive for COVID. Post admission, his respiratory distress worsened and his condition deteriorated. A provisional diagnosis of COVID pneumonia with acute respiratory distress syndrome (ARDS) was established based on chest radiographic finding of left lower lobe consolidation, increased pulmonary infiltrates in bilateral lung fields with evidence of pleural effusion. Pleural Aspirate obtained via ultrasound guided thoracocentesis revealed branched hyaline septate hyphae along with structures which were composed of elongated vesicle with one layer of phialides covering two-thirds of the vesicle and bearing globose conidia on KOH direct mount. Aspergillus flavus grew on culture, which was later confirmed by MALDI TOF VITEK MS. Patient was diagnosed with Proven Invasive Pulmonary Aspergillosis with COVID-19 and voriconazole was started. Patient successfully recovered and was discharged. Identifying the Aspergillus species directly on the basis of KOH Mount, helped in decreasing the turnaround time, in early initiation of definite therapy and possibly contributed to the favorable outcome. CAPA is a potentially life-threatening complication in patients with severe COVID-19, thus a timely diagnosis and treatment becomes crucial in the management. Keywords: CAPA, COVID-19, Aspergillus

Background Human microsporidiosis presents as an important and rapidly emerging opportunistic infection. However, the exact burden of this infection especially in the pediatric population of Northern India remains unknown. In this study, we investigated the prevalence of microsporidia among human immunodeficiency virus (HIV)-positive and HIV-negative pediatric patients who presented with diarrhea.

Abstract Background Post severe traumatic brain injury (sTBI), axonal alterations lead to myelin loss and its degeneration. In the recovery phase, numerous intermingled biochemical pathways involving complex inflammatory reactions cloud the understanding of this yet undiscerned process that also varies with agonal period. In cases with dubious histories, approximating the survival time can be challenging, and expression levels of characteristic markers may aid forensic experts in the same. Methods This exploratory study recruited 100 samples—68 sTBI, 22 non-TBI and 10 age- and sex-matched control samples. Male:female ratio was 87:13. Histochemical staining using H&E was used to characterize myelination pattern, and IHC of GFAP and CD-68 were performed to assess astroglial and microglial reactions with respect to survival time in specific sites. Result Among sTBI, non-TBI and control recruits, sTBI patients depicted significant myelination abnormalities, astroglial proliferation and microglial reaction and varying with survival time. Non-TBI and control samples depicted nearly similar profiles. Conclusion In order to untangle the complex mesh of biochemical responses, nuanced research on individual factors (both pre- and post mortem) with regard to specific site and survival time are warranted. Standardizing experimental data and converting it into empirical data shall aid forensic experts in suggesting approximate agonal period.

Aim: The increasing burden of resistance in Gram-negative bacteria (GNB) is becoming a major issue for hospital-acquired infections. Therefore, understanding the molecular mechanisms is important. Methodology: Resistance genes of phenotypically colistin-resistant GNB (n = 60) were determined using whole genome sequencing. Antimicrobial susceptibility patterns were detected by Vitek®2 & broth microdilution. Results: Of these phenotypically colistin-resistant isolates, 78% were also genetically resistant to colistin. Activation of efflux pumps, and point-mutations in pmrB, and MgrB genes conferred colistin resistance among GNB. Eight different strains of K. pneumoniae were identified and ST43 was the most prominent strain with capsular type-specific (cps) gene KL30. Discussion: These results, in combination with rapid diagnostic methods, will help us better advice appropriate antimicrobial regimens.

Background: Secondary bacterial infections during COVID-19 hospitalization have been reported in about 6–15% of patients. Aims: To study the secondary bacterial infections that affected the COVID-19 patients during their hospitalisation and to unearth the bacteriological profile of samples obtained after their demise. Settings and Design: This prospective study was carried out at a COVID-19 dedicated, apex tertiary care centre in North India from July 2020 to April 2021. Methods and Materials: Samples of 268 patients were considered for the study. Nasopharyngeal swab specimen, blood, and tissue (lung) were collected from the deceased body as early as possible and processed. Statistical Analysis: Statistical analyses were performed using STATA version 11.1 (Stata Corp., College Station, TX, USA). Results: A total of 170 samples were received from patients before their death, which included blood, urine, respiratory samples, pus, and cerebrospinal fluid. Forty-four pathogens were isolated, which consisted of Acinetobacter baumannii (43.1%), Klebsiella pneumoniae (36.3%), Escherichia coli (11.3%), and Pseudomonas aeruginosa (4.5%), Enterococcus faecium (4.5%). Two hundred fifty-eight samples were collected from the deceased bodies wherein the nasopharyngeal sample was highest, followed by tissue and blood. A total of 43 pathogens were isolated among them which included A. baumannii (44.1%), followed by K. pneumoniae (25.5%), E. coli (20.9%), P. aeruginosa (6.97%) and Enterobacter cloacae (2.3%). All these isolates were highly resistant to antimicrobials. Conclusions: In our study, bacterial profiles in antemortem and postmortem samples were found to be similar, suggesting that resistant pathogens may be the cause of mortality in COVID-19 infected hospitalised patients.

Introduction Stenotrophomonas maltophilia is an emerging environmental, gram-negative, multidrug-resistant organism, associated with risk factors such as prolonged hospitalization, invasive procedures, admission to the intensive care unit, mechanical ventilation, use of indwelling catheters, administration of immunosuppressants or corticosteroids, human immunodeficiency virus infection, underlying malignancy, and organ transplantation. The organism, despite being of low invasiveness in immune-competent individuals, is difficult to treat because of intrinsic resistance to several antimicrobial agents.

Background: There are studies available on the prevalence of coronavirus disease 2019 (COVID-19)-associated mucormycosis (CAM) in hospitalized patients but not on the incidence of CAM in post-discharge patients. The aim of our study was to find the incidence of CAM in the patients discharged from a COVID hospital. Material and Methods: Adult patients with COVID discharged between March 1, 2021 and June 30, 2021 were contacted and enquired about sign and symptoms of CAM. Data of all included patients were collected from electronic records. Results: A total of 850 patients responded, among which 59.4% were males, 66.4% patients had co-morbidities, and 24.2% had diabetes mellitus. Around 73% of patients had moderate to severe disease and were given steroids; however, only two patients developed CAM post discharge. Conclusion: The incidence of CAM post discharge was low in our study, which could be attributed to protocolized therapy and intensive monitoring.

Background Expansion of the testing capacities for severe acute respiratory syndrome-coronavirus-2 is an important issue in the face of ever-increasing case load. So, there is need of point-of-care diagnostic tests in the existing laboratory capacities for early treatment, isolation, and clinical decision making, especially in resource limited settings. Materials and Methods This prospective cohort study was conducted at Jai Prakash Narayan Apex Trauma Center, All India Institute of Medical Sciences, New Delhi. Nasopharyngeal samples and blood samples were collected for antigen and antibody testing. Rapid antigen test was performed as per the kit's instructions. The performance of the kit was compared with the gold standard reverse transcription polymerase chain reaction (RT-PCR) testing. Results Eighty-eight out of 110 patients tested positive by RT-PCR for coronavirus disease 2019 in last 48 to 72 hours were included in the study. Overall, the sensitivity of combined antibody test was 52%, antigen test 26%, and combined sensitivity of both antigen and antibody was 72.7%, respectively. Conclusion The combo kit needs to be used with caution in low prevalence settings, where cases may be missed.

Background: At what rate does the RNA of SARS CoV-2 shed from cadavers? Although, there have been numerous studies which have demonstrated the persistence of the virus on dead bodies, there is a lack of conclusive evidence regarding the variation of viral RNA content in cadavers. This has led to a knowledge gap regarding the safe handling/management of COVID-19 decedents, posing a barrier in forensic investigations. Methods: In this study, we report the presence of RNA of SARS CoV-2 by real time RT-PCR, in nasopharyngeal swabs collected after death from two groups of bodies – one who died due to COVID-19 and the other who died due to other diagnoses. A prospective study on 199 corpses, who had tested positive for COVID-19 ante-mortem, was conducted at a tertiary care center. RNA testing was conducted at different time intervals (T1-T5). Results: 112(56.3%) died primarily due to COVID-19 and 87(43.7%) died due to other diagnoses. 144(72.4%) were male and 55(27.6%) were female. A total of 115 (57.8%) tested positive for COVID-19 after death at different time points. The mean age was 50.7 ± 18.9 years and the length of hospitalization ranged from 1 to 50 days with a mean of 9.2 ± 7.6 days. Realtime RT-PCR positivity of SARS CoV-2 RNA decreases with time. Conclusion: We observed that real time RT-PCR positivity, indicating viral RNA detection, decreases with time. Therefore, it is advisable to follow appropriate COVID-19 precautions to carry out scientific studies, medico-legal investigations and mortuary services on suspected/confirmed COVID-19 corpses.

Rapid antigen testing for coronavirus disease 2019 (COVID-19) available at present provides immediate results at low cost with less expertise and without any need of sophisticated infrastructure. Most of these test kits available are for nasopharyngeal samples. This is a novel study to detect the presence of COVID antigen in samples other than throat and oropharyngeal. Various samples received from patients admitted in the COVID-19 dedicated center were tested for the presence of antigen. Same procedure was followed as done for the nasopharyngeal sample. A total of 150 samples were tested, which included ascitic fluid, pleural fluid, drain fluid, bile, bronchoalveolar lavage, cerebrospinal fluid, endotracheal tube aspirate, sputum, tissue, and urine. Out of 150, 11 (7.33%) were positive and 138 (92.66%) were negative for the antigen test. The COVID-19 antigen test kit, though designed for nasopharyngeal samples, was able to detect the presence of antigen in other clinical samples.

ABSTRACT: Objectives: The genetic basis for the spread of vancomycin resistance in Enterococcus faecium is largely unexplored in India. The present study aimed to investigate the plasmid diversity and variation of Tn1546 associated with vanA harbouring VREfm isolates. Methods: A total of 122 VREfm isolates collected from blood cultures were included in this study. MLST analysis was performed on all isolates, and they were also screened for the presence of vanA and vanB genes. Whole genome sequencing was performed for a subset of fifteen VREfm isolates belonging to ST1643. Results: All of the 122 VREfm isolates carried the vanA gene. Twenty-four different sequence types were seen; of these, ST1643, ST80 and ST17 were predominant. Whole genome sequencing was performed on 15 VREfm isolates belonging to ST1643. For eight isolates the vanA gene was found on pRUM-like circular plasmids, and for the remaining seven isolates, the vanA gene was found on the linear plasmids. Novel Tn1546 variants carrying vanA were found on both circular and linear plasmids. Interestingly, co-presence of vanA and optrA were seen in the backbone of three linear plasmids. Conclusion: Multiple vanA-carrying plasmids and Tn1546-like elements were involved in the dissemination of vancomycin resistance in VREfm. The co-occurrence of Tn1546 carrying vanA and Tn554 family transposon carrying optrA on the backbone of plasmids is worrisome. The dissemination of such plasmids may pose treatment and infection control challenges.

Background The burden of hospital-acquired infections (HAIs) is all assumption based, and the true burden remains unknown in most countries, particularly in the developing countries where healthcare facilities are suboptimal and knowledge is limited. Methodology This cross-sectional study was conducted at the trauma center of a tertiary care institute from August to September 2019, to assess the burden of HAI and antibiotic resistance pattern of HAI. The total sample size in our study was 105. Our objective was to estimate the point prevalence of HAI and study the associated factors in a tertiary care hospital. Result In this study, the point prevalence of HAI was five to six times higher when compared with that of developed countries. Gram-negative organisms were the predominant bacteria; with Acinetobacter baumannii the most common among them. Conclusion Point-prevalence survey is an important objective of the antimicrobial stewardship program; it will be helpful in controlling antimicrobial resistance and this tool plays a significant role in hospital settings. Our study is quite pertinent to assess the point prevalence of HAI. It will help in knowing the current prevalence and pattern of the HAI. Therefore, as healthcare administrators, we can further decrease the HAI for better patient outcomes in the future.

Abstract Angiotensin-converting enzyme 2 (ACE2) is a key host protein by which severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) enters and multiplies within cells. The level of ACE2 expression in the lung is hypothesised to correlate with an increased risk of severe infection and complications in COrona VIrus Disease 2019 (COVID-19). To test this hypothesis, we compared the protein expression status of ACE2 by immunohistochemistry (IHC) in post-mortem lung samples of patients who died of severe COVID-19 and lung samples obtained from non-COVID-19 patients for other indications. IHC for CD61 and CD163 was performed for the assessment of platelet-rich microthrombi and macrophages, respectively. IHC for SARS-CoV-2 viral antigen was also performed. In a total of 55, 44 COVID-19 post-mortem lung samples were tested for ACE2, 36 for CD163, and 26 for CD61, compared to 15 non-covid 19 control lung sections. Quantification of immunostaining, random sampling, and correlation analysis were used to substantiate the morphologic findings. Our results show that ACE2 protein expression was significantly higher in COVID-19 post-mortem lung tissues than in controls, regardless of sample size. Histomorphology in COVID-19 lungs showed diffuse alveolar damage (DAD), acute bronchopneumonia, and acute lung injury with SARS-CoV-2 viral protein detected in a subset of cases. ACE2 expression levels were positively correlated with increased expression levels of CD61 and CD163. In conclusion, our results show significantly higher ACE2 protein expression in severe COVID-19 disease, correlating with increased macrophage infiltration and microthrombi, suggesting a pathobiological role in disease severity.

Background Pediatric traumatic brain injury (TBI) has distinctive pathophysiology and characteristics that differ from adults. These can be attributed to age-related anatomical and physiological differences and distinct patterns of injuries seen in children. Our aim was to identify the patient characteristics, clinical variables during intensive care and intraoperative management associated with poor functional outcome in a cohort of pediatric TBI patients. Methods Retrospective chart review of pediatric TBI patients admitted to neurotrauma intensive care unit (NICU) over a period of 1 year. Results A total of 105 children (< 12 years) with head injury were admitted in the NICU during the study period. The most common mechanism of injury was fall in 78% cases. Fifty-four patients (51.4%) presented with a severe head injury (Glasgow coma scale [GCS] ≤ 8), while 31 (29.5%) and 20 (19.1%) had a mild and moderate head injury. The most common finding was skull fractures (59%), contusions (36.2%), and subdural hematoma (SDH) (30.4%). Forty nine patients (46.7%) required surgical management. The median duration of anesthesia was 205 (interquartile range [IQR] 65, 375) minutes, and median blood loss during the surgery was 16.7 mL/kg body weight with 41% requiring intraoperative blood transfusions. Median duration of ICU and hospital stay was 5 (IQR 1, 47) and 8 (IQR 1, 123) days, respectively. GOS at discharge ≤ 3 representing poor outcome was present in 35 patients (33.3%). Mortality was seen in 15 (14.3%) patients. Multivariate analysis identified postresuscitation GCS ≤ 8 on admission as independent predictor of mortality, and postresuscitation GCS ≤ 8 on admission and NICU stay of > 7 days as independent predictor of poor outcome. Conclusion Despite advances in neurointensive care, mortality and morbidity remains high in pediatric head trauma and is mainly dependent on postresuscitation GCS and NICU stay of more than 7 days. Multidimensional approach is required for its prevention and management.

A medical postgraduate course in the field of Laboratory Medicine for the Bachelor of Medicine and Bachelor of Surgery (MBBS) degree holders has existed for more than two decades in India, initiated and offered by the All India Institute of Medical Sciences, New Delhi, which was created under the special Act of Parliament of India 1956. This course has recently been included in the draft of National Medical Commission's Post Graduate Regulation 2021 list of medical courses, and the foundation guidelines have been laid for other medical colleges and teaching hospitals across the country to start this course. This article, written purely in academic interest, describes the past, present and future of this postgraduate training program in India with an aim to answer several doubts regarding this unique and holistic course with a view to providing a direction to those who are willing to become a laboratory physician through this post-graduation.

Introduction: During the second wave of coronavirus disease 2019 (COVID-19), superinfection caused by fungus and multidrug-resistant bacteria worsened the severity of illness in COVID-19 patients. Limited studies from India reported the antimicrobial resistance pattern of secondary infections. In this study, we aim to study the epidemiology of pathogens causing superinfections and genotyping of Gram-negative isolates in COVID-19 patients. Methods: This retrospective study was conducted at a dedicated COVID-19 center, India. The identification of bacteria/fungi was done by Vitek2® and matrix-assisted laser desorption/ionization-time of flight mass spectrometry system. Identification of beta-lactamase genes was done using thermal cycler. The diagnosis of mucormycosis was based on 10% potassium hydroxide direct microscopy. Statistical analyses were performed using STATA version 15.1 (StataCorp., College Station, TX, USA). For continuous variables, mean and standard deviation were computed. For comparing proportions of secondary infections across admission location and outcomes, the Chi-squared test of independence was used. To compare the mean and median between intensive care units and outcomes, an independent t-test and a Mann–Whitney test were used. Results: Of all the clinical samples, 45.4% of samples were cultured positive for secondary infections. Acinetobacter baumannii (35%) was the most common Gram-negative pathogen, while among Gram positive, it was Enterococcus faecium (40%). Among fungus, Candida spp. (61%) predominates followed by molds. Colistin and tigecycline proved effective against these pathogens. blaNDM was the most prevalent gene followed by the blaOX among the carbapenemase genes. Conclusions: The mortality rate among COVID-19 patients with secondary infection was significantly higher compared to the overall mortality rate in COVID-19 patients.

ABSTRACT In the second wave of COVID-19 in India, there was a new challenge in the form of mucormycosis. Coinfection with mucormycosis was perilous as both conditions required a prolonged hospital stay, thus serving as an ideal platform for secondary infections. Using a retrospective observational study, we studied secondary infections and their impact on the outcome in COVID-19 patients with mucormycosis. The outcome in these patients was evaluated and compared with COVID-19 patients with mucormycosis but without any secondary infection. SPSS V-20 was used for data analysis. Fifty-five patients tested positive for mucormycosis (55/140; 39.28). Twelve out of these 55 (21.8%) developed secondary infections during their hospital stay. Bloodstream infection was the most common (42.86%) secondary infection. The Gram-negative (GN) organisms were more common (11/16; 68.75%) compared with the Gram-positives (GP) (5/16; 31.25%). But the most common isolate was Enterococcus faecium (5/16; 31.25%). A high percentage of microorganisms isolated were multidrug-resistant (15/16; 93.75%). Two out of five (40%) isolates of Enterococcus faecium were vancomycin-resistant (VRE). High resistance to carbapenems was noted in the GN isolates (9/11; 81.81%). The comparison of length of stay in both subgroups was statistically significant (P value

Abstract SARS-CoV-2 has posed an unprecedented challenge to the world. Pandemics have been caused previously by viruses of this family like Middle East Respiratory Corona Virus (MERS CoV), Severe Acute Respiratory Syndrome Corona Virus (SARS CoV). Although these viruses are primarily respiratory viruses, but they have been isolated from non-respiratory samples as well. Presently, the detection rate of SARS‐CoV‐2 RNA from different clinical specimens using Real Time Reverse Transcriptase Polymerized Chain Reaction (qRT‐PCR) after onset of symptoms is not yet well established. Therefore, the aim of this systematic review was to establish the profile of detecting SARS‐CoV‐2, MERS CoV, SARS CoV from different types of clinical specimens other than the respiratory using a standard diagnostic test (qRT‐PCR). A total of 3429 non-respiratory specimens were recorded: SARS CoV (total sample—802), MERS CoV (total sample—155), SARS CoV-2 (total sample—2347). Out of all the samples studied high positive rate was seen for saliva with 96.7% (14/14; 95% CI 87.6–100.0%) for SARS CoV and 57.5% (58/250; 95% CI − 1.2 to 116.2%) for SARS CoV-2, while low detection rate in urine samples for SARS CoV-2 with 2.2% (8/318; 95% CI 0.6–3.7%) and 9.6% (12/61; 95% CI − 0.9 to 20.1%) for SARS CoV but there was relatively higher positivity in urine samples for MERS CoV with detection rate of 32.4% (2/38; 95% CI − 37.3 to 102.1%). In Stool sample positivity was 54.9% (396/779; 95% CI 41.0–68.8%), 45.2% (180/430; 95% CI 28.1–62.3%) and 34.7% (4/38; 95% CI − 29.5 to 98.9%) for SARS CoV-2, MERS CoV, and SARS CoV, respectively. In blood sample the positivity was 33.3% (7/21; 95% CI 13.2–53.5%), 23.7% (42/277; 95% CI 10.5–36.9%) and 2.5% (2/81; 95% CI 0.00–5.8%) for MERS CoV, SARS CoV-2 and SARS CoV respectively. SARS‐CoV‐2 along with previous two pandemic causing viruses from this family, were highly detected stool and saliva. A low positive rate was recorded in blood samples. Viruses were also detected in fluids along with unusual samples like semen and vaginal secretions thus highlighting unique pathogenic potential of SARS‐CoV‐2.

Introduction Multiple drug resistance emergences among bacteria at an alarming rate worldwide are posing a serious threat to the treatment benefits that have been achieved with antibiotics. This crisis is due to the inappropriate and overuse of existing antibiotics. We evaluated the antimicrobial resistance pattern of Enterobacteriaceae pathogens isolated from intensive care units (ICUs), wards, and outpatient department (OPD) patients. Objectives The aim of the study is to determine the antimicrobial resistance pattern in bacteria of Enterobacteriaceae family. Material and Methods This is a retrospective study conducted at a tertiary care level-1 trauma center in the capital city of India. We collected all the retrospective data of 5 years from the laboratory information system software of the microbiology laboratory. The retrospective data included patients’ details, samples detail, organism’s identification, and their antimicrobial susceptibility testing, done by Vitek2 compact system and disk diffusion test according to each year’s Clinical and Laboratory Standards Institute (CLSI) guidelines. This study included the interpretation of zone diameters and minimum inhibitory concentrations of all isolates according to CLSI guidelines, 2018. Results Among all the Enterobacteriaceae, Klebsiella spp. was the most commonly isolated pathogen, followed by Escherichia coli and Enterobacter spp. in ICUs and wards, while in OPD patients E. coli was the most commonly isolated pathogen, followed by Klebsiella spp. and Enterobacter spp. Enterobacteriaceae isolates remained resistant to all classes of cephalosporins in all settings. In addition, β lactam and β-lactamase inhibitor remained less effective. Carbapenems showed less resistance than quinolones and aminoglycosides. Among the different antimicrobial agents, tigecycline proved most effective in all settings; however, it showed more resistance than other studies. Conclusion Tigecycline proved effective among different multidrug resistance bacteria. Multidrug resistance in bacteria leads to prolonged hospital stays as well as makes the treatment less cost effective. Proper and judicious use of antimicrobials is the need of the hour.

With the excessive genome plasticity, Acinetobacter baumannii can acquire and disseminate antimicrobial resistance (AMR) genes often associated with mobile genetic elements (MGEs). Analyzing the genetic environment of resistance genes often provides valuable information on the origin, emergence, evolution, and spread of resistance. Thus, we characterized the genomic features of some clinical isolates of carbapenem-resistant A. baumannii (CRAb) to understand the role of diverse MGEs and their genetic context responsible for disseminating carbapenem resistance genes. For this, 17 clinical isolates of A. baumannii obtained from multiple hospitals in India between 2018 and 2019 were analyzed. AMR determinants, the genetic context of resistance genes, and molecular epidemiology were studied using whole-genome sequencing. This study observed an increased prevalence of blaOXA–23 followed by dual carbapenemases, blaOXA–23, and blaNDM. This study identified three novel Oxford MLST sequence types. The majority of the isolates belonged to the dominant clone, IC2, followed by less prevalent clones such as IC7 and IC8. This study identified variations of AbaR4 and AbGRI belonging to the IC2 lineage. To the best of our knowledge, this is the first study that provides comprehensive profiling of resistance islands, their related MGEs, acquired AMR genes, and the distribution of clonal lineages of CRAb from India.

To elucidate the molecular mechanisms that manifest lung abnormalities during severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections, we performed whole-transcriptome sequencing of lung autopsies from 31 patients with severe COVID-19 and ten uninfected controls. Using metatranscriptomics, we identified the existence of two distinct molecular signatures of lethal COVID-19. The dominant ‘classical’ signature (n=23) showed upregulation of the unfolded protein response, steroid biosynthesis and complement activation, supported by massive metabolic reprogramming leading to characteristic lung damage. The rarer signature (n=8) that potentially represents ‘cytokine release syndrome’ (CRS) showed upregulation of cytokines such as IL1 and CCL19, but absence of complement activation. We found that a majority of patients cleared SARS-CoV-2 infection, but they suffered from acute dysbiosis with characteristic enrichment of opportunistic pathogens such as Staphylococcus cohnii in ‘classical’ patients and Pasteurella multocida in CRS patients. Our results suggest two distinct models of lung pathology in severe COVID-19 patients, which can be identified through complement activation, presence of specific cytokines and characteristic microbiome. These findings can be used to design personalized therapy using in silico identified drug molecules or in mitigating specific secondary infections.

Abstract Introduction Ventilator-associated pneumonia (VAP) is reported as the second most common nosocomial infection among critically ill patients with the incidence ranging from 2 to 16 episodes per 1000 ventilator days. The use of probiotics has been shown to have a promising effect in many RCTs. Our systematic review and meta-analysis were thus planned to determine the effect of probiotic use in critically ill ventilated adult patients on the incidence of VAP, length of hospital stay, length of ICU stay, duration of mechanical ventilation, the incidence of diarrhea, and the incidence of oropharyngeal colonization and in-hospital mortality. Methodology Systematic search of various databases (such as Embase, Cochrane, and Pubmed), published journals, clinical trials, and abstracts of the various major conferences were made to obtain the RCTs which compare probiotics with placebo for VAP prevention. The results were expressed as risk ratios or mean differences. Data synthesis was done using statistical software - Review Manager (RevMan) Version 5.4 (The Cochrane Collaboration, 2020). Results Nine studies met our inclusion criterion and were included in the meta-analysis. The incidence of VAP (risk ratio: 0.70, CI 0.56, 0.88; P = 0.002; I 2 = 37%), duration of mechanical ventilation (mean difference −3.75, CI −6.93, −0.58; P 0.02; I 2 = 96%), length of ICU stay (mean difference −4.20, CI −6.73, −1.66; P = 0.001; I 2 = 84%) and in-hospital mortality (OR 0.73, CI 0.54, 0.98; P = 0.04; I 2 = 0%) in the probiotic group was significantly lower than that in the control group. Probiotic administration was not associated with a statistically significant reduction in length of hospital stay (MD −1.94, CI −7.17, 3.28; P = 0.47; I 2 = 88%), incidence of oro-pharyngeal colonization (OR 0.59, CI 0.33, 1.04; P = 0.07; I 2 = 69%), and incidence of diarrhea (OR 0.59, CI 0.34, 1.03; P = 0.06; I 2 = 38%). Discussion Our meta-analysis shows that probiotic administration has a promising role in lowering the incidence of VAP, the duration of mechanical ventilation, length of ICU stay, and in-hospital mortality.

Objectives: Acinetobacter baumannii emerged as a major nosocomial pathogen responsible for infections. In this study, we report the molecular characterization, association of insertion sequences and sequence types of clinical isolates of carbapenem resistant A. baumannii. Materials and Methods: A total of 763 non-duplicate isolates of A. baumannii received from 8 centres across India during January 2014 to December 2017 were studied. Susceptibility testing was done by Kirby-Bauer method. PCR was performed for detection of extended spectrum β-lactamases, metallo β-lactamases, oxacillinases and ISAba1. Mapping PCR was performed to identify the position of ISAba1 with respect to blaOXA-23 like and blaOXA-51 like gene. MLST was performed to identify the sequence type. Whole genome sequencing was done to decipher the genetic arrangement of ISAba1 with blaOXA-23 like and with blaOXA-51 like. Results: All the isolates were resistant to imipenem and meropenem. blaOXA-23 like was the predominant carbapenemase. All isolates were positive for ISAba1. The common sequence types were ST848, ST451 and ST1305 which belongs to International clone II. Whole genome sequencing showed considerable variation in the insertion site location. Conclusions: In conclusion, high prevalence of blaOXA-23 like in A. baumannii and its association with ISAba1 and sequence types belonging to IC-II facilitates the successful dissemination of these extremely drug resistant strains.

Introduction Compliance to hand hygiene (HH) is an important measure in preventing infections to patients in health-care settings. Wellness and safety of patients and health-care workers (HCWs) can be achieved by promoting best practices in infection control through education and advocacy. Aims and Objectives To assess the compliance to HH among all cadres of HCWs and its association with hospital-acquired infection (HAI) in patients. Materials and Methods A prospective, observational study was conducted for a period of 5 years (January 2014 to December 2018) in Neuro Trauma intensive care unit. A standard checklist based on World Health Organization’s 5 Moments for Hand Hygiene was used as a tool to measure the HH compliance. Results HAI rate was found to be directly proportional to the compliance to HH. Reduction in HAI rates was reported when there was an increase in HH compliance. HAI of 4.25% was found to be lowest in the year 2015 with the compliance to HH of 63.65%. The HH compliance was also found to be highest (64.63%) in the year 2016 followed by 64.12% in the year 2017. During this period HAI rates were 4.35% and 4.8%, respectively. When the HH compliance declined in the year 2018 to 53.95%, there was an increase in the rate of HAI to 6.9%. Conclusion It could be concluded that HH compliance was associated with the decrease in HAIs. HH could be a simple and cost-effective method in the prevention of HAIs.

We evaluated the appropriateness of antibiotic prescriptions at discharge from a tertiary-care hospital in India. Of the 790 adult patients included, 84.4% received antibiotics. Microbiological specimens were taken from 67.3% of these patients, and pathogens were identified in 28.8% of cases. Overuse of antimicrobials at hospital discharge should be curtailed.

Background The role of enhanced recovery after surgery (ERAS) has been established in elective operations. However, its role in emergency operations especially in trauma is under-recognized. The aim of this study was to explore the safety and efficacy of ERAS program in patients undergoing emergency laparotomy for trauma.Methods In this single-center study, patients who underwent emergency laparotomy after trauma were randomized to the ERAS protocol or conventional care. The ERAS protocol included early removal of catheters, early initiation of diet, use of postoperative prophylaxis and optimal usage of analgesia. The primary endpoint was duration of hospital stay. The secondary endpoints were recovery of bowel function, pain scores, complications and readmission rate.Results Thirty patients were enrolled in each arm. The ERAS group had significant reduction in duration of hospital stay (3.3±1.3 vs. 5.0±1.7; p

BackgroundThe incidence of hypervirulent (hv) carbapenem-resistant (CR) Klebsiella pneumoniae (Kp) is increasing globally among various clones and is also responsible for nosocomial infections. The CR-hvKp is formed by the uptake of a virulence plasmid by endemic high-risk clones or by the uptake of plasmids carrying antimicrobial resistance genes by the virulent clones. Here, we describe CR-hvKp from India belonging to high-risk clones that have acquired a virulence plasmid and are phenotypically unidentified due to lack of hypermucoviscosity.MethodsTwenty-seven CRKp isolates were identified to possess rmpA2 by whole-genome sequencing; and resistance and virulence determinants were characterized. By in silico protein modeling (and validation), protein backbone stability analysis, and coarse dynamics study, the fitness of RmpA, RmpA2, and aerobactin-associated proteins-IucA and IutA, were determined to establish a reliable marker for clinical identification of CR-hvKp.ResultsThe CR-hvKp belonged to multidrug-resistant (MDR) high-risk clones such as CG11, CG43, ST15, and ST231 and carried OXA-232 as the predominant carbapenemase followed by NDM. The virulence plasmid belonged to IncHI1B replicon type and carried frameshifted and truncated rmpA and rmpA2. This resulted in a lack of hypermucoviscous phenotype. However, functional aerobactin was expressed in all high-risk clones. In silico analysis portrayed that IucA and IutA were more stable than classical RmpA. Furthermore, IucA and IutA had lower conformational fluctuations in the functional domains than the non-functional RmpA2, which increases the fitness cost of the latter for its maintenance and expression among CR-hvKp. Hence, RmpA and RmpA2 are likely to be lost among CR-hvKp owing to the increased fitness cost while coding for essential antimicrobial resistance and virulence factors.ConclusionIncreasing incidence of convergence of AMR and virulence is observed among K. pneumoniae globally, which warrants the need for reliable markers for identifying CR-hvKp. The presence of non-functional RmpA2 among high-risk clones highlights the significance of molecular identification of CR-hvKp. The negative string test due to non-functional RmpA2 among CR-hvKp isolates challenges phenotypic screening and faster identification of this pathotype. This can potentially be counteracted by projecting aerobactin as a stable, constitutively expressed, and functional marker for rapidly evolving CR-hvKp.

Introduction: Tests detecting SARS-CoV-2-specific antigen have recently been developed, and many of them are now commercially available. However, the real-world performance of these assays is uncertain; therefore, their validation is important. In this study, we have evaluated the performance of STANDARD F COVID-19 antigen fluorescence immunoassay (FIA) kit. Methods: Nasopharyngeal samples collected from patients were subjected to the test as per manufacturer's instructions. The performance of the kit was compared with the gold standard real-time polymerase chain reaction. Results: A total of 354 patients were tested with STANDARD F COVID-19 antigen FIA test kit. The overall sensitivity, specificity, positive predictive value, and negative predictive value of this test were found to be 38%, 99%, 96.2%, and 72%, respectively, with a diagnostic accuracy of 75.7%. Conclusion: STANDARD F COVID-19 antigen FIA showed high specificity and positive predictive value but low sensitivity and negative predictive value.

Background: The management of geriatric trauma patients is challenging because of the altered physiology and co-existent medical conditions. To study the in-hospital mortality profile of geriatric trauma victims and the parameters associated with the mortality, we conducted this retrospective analysis. Methods: In a retrospective review of geriatric trauma admissions (above 60 years) over a 3-year period, we studied the association of age, gender, comorbidities, mechanism of injury (MOI), Glasgow coma score (GCS), injury severity score (ISS), systolic blood pressure, and hemoglobin (Hb) level on admission with hospital mortality. Univariate and Multivariable logistic regression was used to estimate odds and find independent associated parameters. P < 0.05 was considered as statistically significant. Results: Out of 881 patients, 208 (23.6%) patients died in hospital. The most common MOI was fall (53.3%) followed by motor vehicle collision (31.1%) and other mechanisms (14.5%). The in-hospital mortality was significantly higher and adjusted odds ratio (OR) for mortality were higher for male gender (2.11 [1.04–4.26]), higher ISS (6.75 [2.07–21.95] for ISS >30), low GCS (

Streptococcus pneumoniae is a rare cause of appendicitis, skin soft tissue, and bloodstream infections. The clinical significance of its isolation from samples of skin or soft tissues and pus from the appendix is poorly understood. Invasive pneumococcal disease (IPD) continues to be a problem in India, associated with a high case fatality rate despite treatment facilities available in the hospital settings. In the present study, we report three adult cases, one presented as acute appendicitis, the other had skin and soft tissue infection, and third presented with bloodstream infection caused by Streptococcus pneumoniae from our level 1 trauma center. The patients with acute appendicitis and soft tissue infection recovered when treated with appropriate antimicrobial therapy, however, the one with pneumococcal sepsis could not be revived.

Abstract Background Recently, in India, there has been a shift from NDM to OXA48-like carbapenemases. OXA-181 and OXA-232 are the frequently produced variants of OXA48-like carbapenemases. OXA48-like carbapenemases are also known to be carried on transposons such as Tn1999, Tn1999.2 and it is also associated with IS1R carried on Tn1999. In India, there are no previous reports studying the association of mobile genetic elements (MGEs) with OXA48-like carbapenemases. The present study was aimed at determining the genetic backbone of OXA48-like carbapenemases to determine the role of MGEs in its transfer and to investigate the Inc plasmid type carrying bla OXA48-like. Results A total of 49 carbapenem resistant K. pneumoniae which included 25 isolates from South India and 24 isolates from North India, were included in the study. Whole genome sequencing using Ion Torrent PGM was performed to study the isolates. OXA-232 was present in 35 isolates (71%). In 19 isolates (39%), bla OXA48-like was associated with MGEs. Insertion sequences such as ISX4, IS1, IS3, ISKpn1, ISKpn26, ISKpn25, ISSpu2, ISKox1, IS4321R, ISEc36, and ISPa38; and transposons such as TnAs3 and Tn2, were present. Isolates from northern and southern India belonging to same sequence type (ST) had diverse genetic backbone for bla OXA48-like. ST14 isolates from north had IS5 and Tn3 families while from south they had IS1, IS5 and IS630 families. ST231 from north had IS5, IS6 and Tn3 families with bla OXA-232 while from south, IS1, IS3 and IS5 families were observed; with ISKpn26 being present among isolates from both the regions. bla OXA48-like was predominantly found on ColKP3 plasmid. ST231 was the predominant ST in 22 isolates (45%). Conclusion OXA-232 is the predominant variant of OXA48-like carbapenemase with ST231 being the commonest ST of OXA48-like carbapenemase producing K. pneumoniae in India. Diverse MGEs have been associated with both bla OXA-232 and bla OXA-181 which contribute to their spread. The MGEs in the present study are different from those reported earlier. There is no clonal expansion of bla OXA48-like producing K. pneumoniae since diverse STs were observed. Monitoring the genetic backbone of OXA48-like carbapenemase is essential to better understand the transmission dynamics of XDR K. pneumoniae.

INTRODUCTION: Acinetobacter baumannii has now emerged as a significant nosocomial pathogen in health-care setting ESP in intensive care units. Rapidly growing resistance among clinical isolates suggests a need to detect resistance mechanisms in this organism. The present study was designed to compare the various phenotypic tests available with the gold standard of genotype. METHODOLOGY: The present study was conducted to include all isolates of Acinetobacter spp. isolated over 3 years. Their resistance to various antibiotics was determined and extended spectrum beta-lactamases (ESBL) and AmpC production in the isolates showing resistance to ceftazidime/ceftriaxone/cefotaxime (CAZ/CTR/CTX) was determined. ESBL and AmpC production was confirmed using polymerase chain reaction (PCR). RESULTS: A total of 154 strains were isolated, and all the strains were tested for ESBL and AmpC detection. Of the strains tested, 15 (9.7%), 17 (11%), 24 (15.6%), 27 (17.5%), 54 (35%), 67 (43.5%), and 72 (46.7%) strains showed ESBL production using CTX/CTX-clavulanate double-disc synergy test (DDST), CTX/CTX-clavulanate E-test, CAZ/CAZ-clavulanate DDST, CAZ/CAZ-clavulanate E-test, Piperacillin/Piperacillin-tazobactam (TZ) DDST, CTR/CTR-Sulbactum DDST, and Piperacillin/Piperacillin-TZ E-test, respectively. 20 (12.9%) and 19 (12.3%) of strains were positive for AmpC production using AmpC disc test and Boronic acid inhibition test, respectively. Genotype analysis using PCR for TEM, SHV, CTXM, PER, and VEB genes was done and 69 (51.5%) strains were positive for TEM gene. DISCUSSION: ESBL detection in Acinetobacter spp. is difficult as standard guidelines for the same are not available unlike in enterobacteriaceae, and there are no zone diameter breakpoints for aztreonam and cefpodoxime. In comparison, piperacillin/piperacillin-TZ E-test had the best sensitivity and specificity for ESBL detection. CONCLUSION: Standard guidelines for ESBL detection in nil fermeners like Acinetobacter spp. must be laid down for ease of detection. Use of piperacillin/piperacillin-tazobactam E-test could be used as one of the standard methods.

BACKGROUND: The purpose of the study was to determine the prevalence and characterize the resistance profiles of Escherichia coli isolated from various clinical specimens by various phenotypic and genotypic methods. MATERIALS AND METHODS: A total of 196 consecutive, nonduplicate strains of clinically significant E. coli isolated from various clinical specimens were included in the study. Identification and antimicrobial susceptibility testing was performed by using Vitek-2 system (Biomerieux, France). Phenotypic detection of extended spectrum beta-lactamase (ESBLs), Amp-C-β lactamase (Amp C), and carbapenemase production was done by various combination of disc diffusion methods, minimum inhibitory concentration determination by E-test, followed by polymerase-chain-reaction for the detection of β-lactamase-encoding genes. RESULTS: Overall prevalence of ESBLs, Amp C, and carbapenemase production was found to be 88.3%, 42.2%, and 65.1% by the phenotypic detection methods. Our study also revealed high resistance rates against other antibiotics such as cefepime (89%), cefotaxime (95.4%), ceftazidime (85.4%), ceftriaxone (91.8%), cefpodoxime (92.7%), aztreonam (56.3%), piperacillin/tazobactam (89.2%), and ticarcillin/clavulanic acid (76.3%). The most prevalent ESBL gene was blaTEM(67.30%), and least prevalent ESBL gene was blaVEB(2.61%). In case of Amp C, blaFOXgene (21.9%) was predominant. Among the genes encoding for carbapenemases, the most common gene was blaNDM(61.7%) followed by blaVIM(30.8%), blaKPC(10.6%), blaOXA-48 (5.3%), and blaIMP(2.1%). CONCLUSION: Our findings suggest a high rate of ESBLs, Amp C, and carbapenemase production among the E. coli isolates. A combination of both phenotypic and genotypic methods would be ideal for better characterization of resistance patterns among the E. coli isolates.

Objective: The detailed epidemiological and molecular characterization of an outbreak of Burkholderia cepacia at a neurotrauma intensive care unit of a level 1 trauma centre is described. The stringent infection control interventions taken to successfully curb this outbreak are emphasized. Methods: The clinical and microbiological data for those patients who had more than one blood culture that grew B. cepacia were reviewed. Bacterial identification and antimicrobial susceptibility testing was done using automated Vitek 2 systems. Prospective surveillance, environmental sampling, and multilocus sequence typing (MLST) were performed for extensive source tracking. Intensive infection control measures were taken to further control the hospital spread. Results: Out of a total 48 patients with B. cepacia bacteraemia, 15 (31%) had central line-associated blood stream infections. Two hundred and thirty-one environmental samples were collected and screened, and only two water samples grew B. cepacia with similar phenotypic characteristics. The clinical strains characterized by MLST typing were clonal. However, isolates from the water represented a novel strain type (ST-1289). Intensive terminal cleaning, disinfection of the water supply, and the augmentation of infection control activities were done to curb the outbreak. A subsequent reduction in bacteraemia cases was observed. Conclusion: Early diagnosis and appropriate therapy, along with the rigorous implementation of essential hospital infection control practices is required for successful containment of this pathogen and to curb such an outbreak. Keywords: Burkholderia cepacia, Outbreak, Neurotrauma unit, MLST typing, Novel ST type

INTRODUCTION: Acinetobacter spp. has emerged as a major cause of nosocomial outbreaks. Multiple antibiotic resistance is an important problem in Acinetobacter isolates in recent years. The aim of this study was to evaluate the rate of antimicrobial resistance and changes in resistance pattern over a period of 5 years (2012–2016) in Acinetobacter spp. isolated from trauma patients. MATERIALS AND METHODS: Acinetobacter spp. was identified by VITEK 2 and antibiotic susceptibility of isolates was investigated by disc-diffusion method and VITEK 2 automated system. Interpretation of susceptibility results was based on the Clinical and Laboratory Standards Institute guidelines. RESULTS: Out of the total 16,210 isolates obtained throughout the period of 5 years, Acinetobacter spp. accounted for 3744 (28.9%). Out of which, the species which was maximally isolated was Acinetobacter baumannii (98.5%), followed by Acinetobacter lwoffii (1.4%) and Acinetobacter hemolyticus (0.1%). The highest number of clinical isolates of Acinetobacter were recovered from neurosurgical ward (n = 1210), followed by the neurosurgical intensive care unit (ICU) (n = 1000) and surgical ICU (n = 948) and the most common sample of Acinetobacter isolation was from tracheal aspirate (37.1%), followed by wound swab (18.8%). The highest level of resistance was observed against ciprofloxacin (96%), followed by cefepime (95%), ceftazidime (95%), piperacillin (95%), and amikacin (92%). The trend of antibiotic resistance was found to be statistically significant (P < 0.001) for most of the antibiotics being tested such as amikacin and carbapenems. CONCLUSION: The high rate of antibiotic resistance of the Acinetobacter strains indicated that there is an urgent need for controlled antibiotic usage and appliance of hospital infection control measures.

Introduction: MBL containing genes have been reported in all GNBs including Acinetobacter spp since 1990s which are worrisome as they are transmitted by mobile genetic elements. Thus, early detection of MBL encoding organisms is necessary. The current study was designed to identify the most sensitive cost-effective test which could be used as a screening test for detection of cabapenamase producing Acinetobacter isolates. Methodology: All consecutive strains of Acinetobacter spp isolated from various clinical samples were included. All isolates found resistant to any of the carbapenems were tested for MBL production using MHT (on MacConkey Agar and Mueller Hinton Agar), Etest (using Imipenem/Meropenem-EDTA) and Combined Disc Test (using EDTA and 2 MPA as inhibitors and Ceftazidime/Imipenem/Meropenem as substrate discs). PCR was performed for representative strains for IMP, VIM, KPC, OXA and NDM-1 gene. Results: Total of 154 non-duplicate strains of Acinetobacter spp were isolated and identified, of which, 134 (88%) and 126 (82%) were resistant to meropenem and imipenem respectively. All 134 meropenem resistant strains were tested for MBL production and PCR was performed on 100 strains. 3(3%), 5(5%), 7(7%), 26(26%), and 51(51%) strains had IMP gene, VIM gene, KPC gene, OXA gene and NDM-1 gene. MHT on MAC had better performance than on MHA and dilution to 0.05 McFarland was not required. Conclusion: MHT on MAC had best sensitivity when compared with gold standard PCR and was also cost effective. With ROC curve, we found that 2MPA was not a good MBL inhibitor when compared with EDTA..

INTRODUCTION: Bloodstream infections (BSIs) can lead to life-threatening sepsis and are globally associated with high morbidity and mortality. Although BSIs require immediate antimicrobial treatment, their prevalence, etiology, and antimicrobial susceptibilities differ from one country to other. There is a dearth of such data from India. Here, we report the 4-year etiologic data on BSI in trauma patients admitted to a tertiary care referral hospital in New Delhi, India. MATERIALS AND METHODS: A retrospective study was conducted at the trauma center between January 2013 and December 2016. The routine microbiological data on bacterial BSI were recorded and determined retrospectively from the laboratory records. Antimicrobial susceptibility profiles were statistically analyzed. RESULTS: A total of 2017 bacterial strains isolated from blood culture samples were included for microbiological analysis. During the study, the median age of the patients varied from 30 to 35 years, with the percentage of females in the study population varying from 17% to 19%. The predominant pathogens were Gram-negative bacteria, with Acinetobacter species, followed by Klebsiella species being the most commonly isolated organisms throughout the 4 years of study. Among Gram-positive isolates, Staphylococcus species were the leading pathogens (11%–15%). CONCLUSIONS: A detailed analysis of prevalence, etiology of BSIs in India and its resistance profile is crucial for appropriate antibiotic use, clinical management, and formulation of antibiotic policies and preventive measures.

INTRODUCTION: Antimicrobial resistance is an increasing problem worldwide especially among the surgical site infections (SSIs). SSI is becoming more serious due to hospital-acquired infections/nosocomial infections, which further leads to the overuse of broad-spectrum antibiotics. To investigate the antimicrobial resistance patterns among Gram-negative bacteria in SSI in in- and out-patients the present study was designed. METHODOLOGY: During the 4 years (January 2013–December 2016), the antimicrobial resistant pattern was studied in the admitted patients and in the patients who were followed up to the outpatients department (OPD) after discharge. Antimicrobial resistance pattern testing was done by the disk diffusion method on Mueller-Hinton agar and by E-test for ten antibiotics according to The Clinical and Laboratory Standards Institute guidelines for Gram-negative bacilli. RESULTS: A total of 2,447 strains were isolated from the studied population on over the period of 4 years. Of 2447, 1996 (81%) were isolated from patients who had SSI during the hospital stay, and 451 (18%) were from patients who attended the OPD after discharge. In the outpatients, who followed up in the OPD for the SSI, Escherichia coli (148), and Pseudomonas aeruginosa (93), whereas in the patients who develop SSI during their hospital stay, Acinetobacter baumannii (622), E. coli (424), and Klebsiella pneumoniae (315) were found to be common. A very high resistance pattern was observed in both the studied groups; however, a higher resistance pattern was seen in in-patients as compared to outpatients. CONCLUSION: In our study, we have reported resistance pattern in Gram-negative bacteria isolated from the patients who were came for the follow as well as in the inpatients. For the outpatients, it can be concluded that it could be a community-acquired infection which is also an alarming condition for our society.

INTRODUCTION: Device-associated infections constitute the majority of health-care infections in Intensive Care Units (ICUs). Trauma patients are more prone to acquire such infections; ventilator-associated pneumonia (VAP) being the most common Health care associated infections (HAI) in ICU has serious implications such as increased morbidity, prolonged hospital stay, and mortality. This study aims to compare the clinicomicrobiological profile of VAP and non-VAP trauma patients at Level I trauma center. MATERIALS AND METHODS: A 4-year retrospective study of prospectively maintained database was conducted at Level 1 trauma center from January 2013 to December 2016. The patients were classified into two groups named VAP and non-VAP patients. VAP patients were defined according to the criteria of the Centers for Disease Control and Prevention. The data were compiled and analyzed. Statistical data were analyzed using SPSS version 21 software. RESULTS: During the study period, 134 (13%) cases of VAP and 909 (87%) non-VAP cases were observed in our study. The total number of ventilator days for VAP patients was 5128 days, which ranged from 2 to 82 days (median 42 days). The length of hospital stay in non-VAP category ranged from 1 to 390 days (median 195.5 days). Inhospital mortality was observed in 62 (46%) patients with VAP. Three hundred and eighteen (35%) non-VAP patients had also had a fatal outcome. Gram-negative organisms, most commonly Acinetobacter spp. (13, 21%), were reported in the fatal VAP patients. CONCLUSION AND DISCUSSION: Higher rate of mortality was observed in patients with VAP in comparison to non-VAP patients, both being on mechanical ventilation. Early recognition of VAP, implementation of proper VAP preventive bundle strategies, and stringent infection control practices are essential mandates to prevent VAP.

Antimicrobial resistance is one of the greatest threats to human health worldwide. The rate of development of newer antibiotics is much slower than the rate of development of antibiotic resistance. A survey reported that it takes 15 years and US$800 million (including preclinical and clinical costs) to bring a single drug to the market, whereas the reuse of the older drugs for antimicrobial use takes $17 million, thereby circumventing 40% of the overall cost. The first case is a patient with nosocomial pyrexia of unknown origin who was given treatment with tigecycline and cefepime/tazobactam but failed to respond to the same. However, the patient responded to the treatment with cotrimoxazole. The second case is a patient with meningitis caused by an atypical zoonotic pathogen, Staphylococcus chromogenes. This is the first report of human infection with S. chromogenes, this being a common cause of bovine mastitis. The isolate was obtained from a patient of neurotrauma who developed meningitis after decompressive craniotomy. The strain was obtained from cerebrospinal fluid, blood, and shunt chamber pus. Cotrimoxazole was given for the treatment, and the patient improved after the treatment. Although the newer antibiotics have replaced sulfonamides in the treatment of many infections, they are still of great value and are the agents of choice in many infections. Sulfonamides have wide antimicrobial activity against both Gram-positive and Gram-negative bacteria, but their usefulness has diminished with the emergence of resistant strains. This paper reports cases of two different kinds of infections from a level 1 trauma center, who failed to respond to the newer antibiotics but showed a response to administration of cotrimoxazole.

INTRODUCTION: Stenotrophomonas maltophilia earlier had limited pathogenic potential, but now with growing degree of immunosuppression in general population, it is being recognized as an important nosocomial pathogen. METHODOLOGY: A retrospective 7 years study was carried out to determine the clinical characteristics of all patients with Stenotrophomonas infections, antibiotic resistance pattern, and risk factors associated with hospital mortality. All patients with Stenotrophomonas culture positivity were identified and their medical records were reviewed. Risk factor associated with hospital mortality was analyzed. RESULTS: A total of 123 samples obtained from 88 patients were culture positive. Most patients presented with bacteremia (45, 51%) followed by pneumonia (37, 42%) and skin and soft tissue infections (6, 7%). About 23 of 88 Stenotrophomonas infected patients had co-infection. Percentage resistance to cotrimoxazole; 8 (5.4%) was lower than that for levofloxacin; 18 (12%). Twenty-eight patients died during hospital stay. Intensive Care Unit admission (P = 0.0002), mechanical ventilation (P = 0.0004), central venous catheterization (P = 0.0227), urethral catheterization (P = 0.0484), and previous antibiotic intake (P = 0.0026) were independent risk factors associated with mortality. CONCLUSION: Our findings suggest that Stenotrophomonas can cause various infections irrespective of patient’s immune status and irrespective of potential source. Thus, Stenotrophomonas should be thought of as potential pathogen and its isolation should be looked with clinical suspicion.

Gas gangrene is a necrotic infection of the skin and soft tissue that is associated with high mortality and often necessitating amputation to control the infection. Clostridial myonecrosis is most often cause of gas gangrene and usually present in settings of trauma, surgery, malignancy, and other underlying immunocompromised conditions. The most common causative organism of clostridial myonecrosis is Clostridium perfringens followed by Clostridium septicum. Here, we are reporting an unusual case report of posttraumatic gas gangrene caused by Clostridium sordelli.