Molecular mechanisms of colistin- and multidrug-resistance in bacteria among patients with hospital-acquired infections.

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.

Characteristics of Gut Microbiome After Traumatic Brain Injury.

BACKGROUND: Preclinical studies have reported significant changes in the gut microbiome after traumatic brain injury (TBI). We hypothesized that TBI induces the growth of Proteobacteria in the human gut. Our primary outcome was to study the profile of the human fecal microbiome after TBI and the secondary outcome was to identify colonization with colistin-resistant and multidrug-resistant pathogens. METHODS: Consecutive patients with moderate-severe TBI admitted to the neurotrauma-intensive care unit within 48 hours of injury were enrolled into this observational study. Samples from rectal swabs obtained on days 0, 3, and 7 after admission were assessed for microbial growth and antibiotic resistance. Demographic data and variables such as hypotension, blood transfusion, surgery, start of nasogastric feeding, use of antibiotics, length of hospital stay and mortality were noted. RESULTS: One hundred one patients were enrolled into this study; 57 (56.4%) underwent surgery, 80 (79.2%) required blood transfusion, 15 (14.9%) had an episode of hypotension, 37 (36.6%) received enteral feed within the first 3 days, and 79 (78.2%) received antibiotics. Rectal microbiological samples were collected from 101, 95, and 85 patients on days 0, 3, and 7, respectively. All organisms isolated at the 3 time-points belonged to the Proteobacteria phylum, with Enterobacteriaceae forming the largest group. Colistin-resistant organisms were found in 17 (16.8%) of 101 patients and multidrug-resistant organisms in 25 (64.1%) of the 39 patients in whom isolates were tested against the entire panel of antimicrobials. CONCLUSION: TBI is associated with widespread colonization with Proteobacteria as early as 48 hours after injury. Colonization with colistin and multidrug-resistant organisms highlights the importance of the judicious use of antibiotics.

Surveillance for surgical site infections in orthopedic trauma surgeries at an Indian hospital.

PURPOSE: Surgical site infections (SSIs) are one of the most common, causing substantial morbidity, mortality and are highly cost-effective means of reducing healthcare associated infections rates in health care set-ups. In India, there is no existing system for systematic surveillance of SSIs, encompassing post-discharge period. METHODS: An indigenous SSI e-surveillance software was developed. Patients developing SSI as per standard definitions were included in the study. A denominator form and a case report form were filled for each case of SSI detected. The microbiological diagnosis was done as per standard methods. Logistic regression analysis was used to test for association of SSI and risk factors and determining the prevalence odds ratios. RESULTS: Of the total of 850 patients enrolled in the SSI surveillance, 47 (5.5%) developed SSI. Most patients (490/850, 58%) underwent the open reduction internal fixation (ORIF) and also developed an SSI (33/490, 6.7%). Clean contaminated wound class and Dressing were found to be associated with increased risk of SSI significantly, Also increase in the length of stay was found to be associated with increased risk of SSI significantly. High antimicrobial resistance was observed in the microbial isolates recovered from SSIs. Patients who developed SSI had longer hospital stays. CONCLUSIONS: Our study has been the first systematic surveillance effort in India, where patients were followed up till six months post surgeries. This pilot study was later expanded to other Indian hospitals. This network of SSI-Surveillance will lay the foundation for initiation of SSI-surveillance across the country.

Association of screen time, quality of sleep and dry eye in college-going women of Northern India.

PURPOSE: To evaluate the association of daily screen time and quality of sleep with the prevalence of dry eye among college-going women. METHODS: This study was a cross-sectional, comparative questionnaire-based study of 547 college-going women in northern India. A 10-item Mini Sleep Questionnaire was used to check the quality of sleep, and the Standard Patient Evaluation of Eye Dryness (SPEED) scale was used to examine the prevalence of dry eye among college-going women. RESULTS: Multinomial logistic regression showed a significant association between dry eye with daily screen time spent (P < 0.05) and the quality of sleep (P < 0.05) among college-going girls. Using Latent Class Analysis, two latent classes were selected based on the Bayesian Information Criteria. It was found that the majority population falls in class two and was having Severe Sleep-Wake difficulty. It was seen that the participants in class two belonged to the age bracket of 18-21 years, were from stream Humanities, education of father and mother equal to graduation, father working only, belonging to the nuclear family, having one sibling, hailing from the urban locality, spending more than 6 h daily on-screen, a majority of them using mobile phones, not using eye lubricants, and reported an increase in screen time during COVID-19. CONCLUSION: Dry eye and sleep quality are essential global health issues, and coupled with increased screen time, may pose a challenge in the present era. Preventive strategies need to be incorporated in school and college curriculums to promote physical, social, and psychological well-being and quality of life.

Evaluation of COVID-19 Antigen Fluorescence Immunoassay Test for Rapid Detection of SARS-CoV-2.

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.

Reworking protocols of ophthalmic resident surgical training in the COVID-19 era - Experiences of a tertiary care institute in northern India.

PURPOSE: Coronavirus Disease 2019 (COVID-19) pandemic has negatively impacted medical professionals in all fields of medicine and surgery in their academic, clinical and surgical training. The impact of surgical training has been described as 'severe' by most ophthalmology residents worldwide due to their duties in COVID-19 wards, disruption of outpatient and camp services. METHODS: Ophthalmic surgery demands utmost accuracy and meticulousness. Fine motor proficiencies, stereoscopic skills and hand-eye coordination required can only be achieved by practice. So, a multileveled structured wet-lab teaching schedule was prepared for the residents and implemented to bridge this gap between theory and practice at our tertiary care institute. A semester-wise training schedule was made with the proper distribution of wet-lab and simulator training. Surgeries like phacoemulsification, scleral buckling, pars plana lensectomy and vitrectomy, trabeculectomy and intravitreal injections were practised by the residents on the goat eyes. Simulator training was provided for phacoemulsification and vitrectomy to increase the hand-eye coordination of the residents. RESULTS: Residents noticed improvement in their surgical skills and ambidexterity post wet-lab and simulator training . It also increased their confidence and provided essential surgical skills required to be used in the operation theater later. CONCLUSION: It is imperative that wet-lab training be included in the residency training programme in this COVID-19 era.

Profile of co-infections & secondary infections in COVID-19 patients at a dedicated COVID-19 facility of a tertiary care Indian hospital: Implication on antimicrobial resistance.

BACKGROUND: The COVID-19 pandemic has raised concerns over secondary infections because it has limited treatment options and empiric antimicrobial treatment poses serious risks of aggravating antimicrobial resistance (AMR). Studies have shown that COVID-19 patients are predisposed to develop secondary infections. This study was conducted to ascertain the prevalence and profiles of co- & secondary infections in patients at the COVID-19 facility in North India. METHODS: We studied the profile of pathogens isolated from 290 clinical samples. Bacterial and fungal pathogens were identified, and antimicrobial susceptibility was determined by the Vitek2® system. Additionally, respiratory samples were tested for any viral/atypical bacterial co-infections and the presence of AMR genes by FilmArray test. The clinical and outcome data of these patients were also recorded for demographic and outcome measures analyses. RESULTS: A total of 151 (13%) patients had secondary infections, and most got infected within the first 14 days of hospital admission. Patients aged >50 years developed severe symptoms (p = 0.0004) and/or had a fatal outcome (p = 0.0005). In-hospital mortality was 33%.K.pneumoniae (33.3%) was the predominant pathogen, followed by A. baumannii (27.1%). The overall resistance was up to 84%.Majority of the organisms were multidrug-resistant (MDR) harbouring MDR genes. CONCLUSION: A high rate of secondary infections with resistant pathogens in COVID-19 patients highlights the importance of antimicrobial stewardship programs focussing on supporting the optimal selection of empiric treatment and rapid-de-escalation, based on culture reports.

Pediatric cataract surgery practices in the COVID-19 era: Perspectives of a tertiary care institute in Northern India.

PURPOSE: To discuss the impact of COVID-19 pandemic on the pediatric cataract surgery services in a tertiary care institute in India, as well as the protocol followed for these surgeries. METHODS: COVID-19 has hampered outpatient and elective services and surgeries throughout the world. During the national lockdown imposed in March in India, outpatient services were suspended in our institute, leading to a tremendous backlog of pediatric patients with cataract. Since the delay in surgery in pediatric cataract can cause amblyopia, our institute had resumed pediatric cataract surgeries in June 2020 at the time of Unlock-1 in the country. RESULTS: We have discussed the percentage of reduction in pediatric cataract surgeries in 2020 during the Unlock 1, 2, 3, and 4, as compared to the number of surgeries done by the pediatric ophthalmology unit in the same months last year. We had introduced triage and telemedicine in our department. We have discussed the preoperative, intraoperative, and postoperative protocol followed in our institute for children with pediatric cataract, and also the measures which can be taken for the safety of patients and staff. CONCLUSION: It is essential to ensure COVID-19 protocol, i.e., wearing a mask, social distancing, and frequent hand hygiene, among the patients and health care personnel. Redesigning pediatric cataract surgery practices is essential to ensure the safety of the health care workers and the patients.

Rapid chromatographic immunoassay-based evaluation of COVID-19: A cross-sectional, diagnostic test accuracy study & its implications for COVID-19 management in India.

Background & objectives: Coronavirus disease 2019 (COVID-19) has so far affected over 41 million people globally. The limited supply of real-time reverse transcription-polymerase chain reaction (rRT-PCR) kits and reagents has made meeting the rising demand for increased testing incompetent, worldwide. A highly sensitive and specific antigen-based rapid diagnostic test (RDT) is the need of the hour. The objective of this study was to evaluate the performance of a rapid chromatographic immunoassay-based test (index test) compared with a clinical reference standard (rRT-PCR). Methods: A cross-sectional, single-blinded study was conducted at a tertiary care teaching hospital in north India. Paired samples were taken for RDT and rRT-PCR (reference standard) from consecutive participants screened for COVID-19 to calculate the sensitivity and specificity of the RDT. Further subgroup analysis was done based on the duration of illness and cycle threshold values. Cohen's kappa coefficient was used to measure the level of agreement between the two tests. Results: Of the 330 participants, 77 were rRT-PCR positive for SARS-CoV-2. Sixty four of these patients also tested positive for SARS-CoV-2 by RDT. The overall sensitivity and specificity were 81.8 and 99.6 per cent, respectively. The sensitivity of RDT was higher (85.9%) in participants with a duration of illness ≤5 days. Interpretation & conclusions: With an excellent specificity and moderate sensitivity, this RDT may be used to rule in COVID-19 in patients with a duration of illness ≤5 days. Large-scale testing based on this RDT across the country would result in quick detection, isolation and treatment of COVID-19 patients.

Changes in extracellular cytokines in predicting disease severity and final clinical outcome of patients with blunt chest trauma.

BACKGROUND: Chest trauma causes substantial morbidity and mortality and its severity is assessed using clinical diagnosis or scoring systems like Injury severity score (ISS) and thoracic trauma severity score (TTSS). Association of inflammatory cytokines with severity of disease and final clinical outcome is not clearly defined in patients with chest trauma. In this study, we thought to evaluate the inflammatory response in serum and bronchoalveolar lavage fluid (BALF) in chest trauma patients and correlate the level of extracellular cytokines with diseases severity and final outcome. METHODS: A total of 65 patients with blunt chest trauma and 30 healthy controls were enrolled in this prospective observational study. Assessment of inflammatory cytokines such as Interleukin (s) - IL-5, IL-13, IL-2, IL-6, IL-9, IL-1ß, IFN-γ, TNF-α, IL-17A, IL-17F,IL-4, IL-21 and IL-22 was performed in both serum and bronchoalveolar lavage fluid using 13-plex multiplex kit using fluorescence-encoded bead based immunoassays. RESULTS: A significantly higher level of IL-13, IL-2, IL-6, IL-9, IL-1ß, IFN-γ, TNF-α, IL-17A, IL-17F, IL-21 and IL-22 cytokines were observed in patients with blunt chest trauma compared to healthy controls. Level of IL-2, IL-6, IL-1ß and IL-17A was significantly raised in the patients with blunt chest trauma who had a fatal outcome during the hospital stay. An elevated cytokine response of IL-13, IL-4, and IL-21 was noted in the group of patients with high (>5) thoracic trauma severity score. CONCLUSION: Routine monitoring of the inflammatory cytokine level in patients with chest trauma may be used routinely. Longer prospective studies should be encouraged to determine the role of cytokines in patients with chest trauma in predicting the patient final clinical outcome.

Device associated infections at a trauma surgical center of India: Trend over eight years.

BACKGROUND: Device-associated infections (DAIs) are an important cause of excessive stay and mortality in ICUs. Trauma patients are predisposed to acquire such infections due to various factors. The prevalence of HAIs is underreported from developing nations due to a lack of systematic surveillance. This study reports the rates and outcomes of DAIs at a dedicated Trauma Center in trauma patients and compares the rates with a previous pilot observation. METHODS: The study reports the finding of ongoing surveillance and the use of an indigenous software at a level-1 trauma center in India. Surveillance for ventilator-associated pneumonia, central line-associated bloodstream infections, and catheter-associated urinary tract infections was done based on standard definitions. The rates of HAIs and the profile of pathogens isolated from June 2010 to December 2018 were analyzed. RESULTS: A total of 7485 patients were included in the analysis, amounting to 68,715 patient days. The rates of VAP, CLABSI, and CA-UTI were respectively 12, 9.8 1st 8.5/1000 device days. There was a significant correlation between device days and the propensity to develop infections. Of the 1449 isolates recovered from cases of DAIs, Acinetobacter sp (28.2%) was the most common isolate, followed by Candida sp. A high rate of multi-resistance was observed. CONCLUSION: Automated surveillance was easy and useful for data entry and analysis. Surveillance data should be used for implementing preventive programs.

Quantification of Inflammation Following Nd:YAG Laser Capsulotomy and Assessing the Anti-inflammatory Effects of Nepafenac 0.1% and Betamethasone 0.1.

Purpose: To quantify the intraocular inflammation, measured using laser flare photometry (LFP), after Nd:YAG capsulotomy and compare the effects of different topical anti-inflammatory regimes.Methods: A single-center, prospective, randomized controlled study, which included 90 eyes of 90 patients with visually significant posterior capsular opacification. In all patients, Nd:YAG capsulotomy was done and was then randomized into three groups: (A) Topical betamethasone 0.1%; (B) Topical nepafenac 0.1%; and (C) Control group: did not receive any anti-inflammatory agent post capsulotomy.Results: There was a significant increase in the mean LFP values 2 h following laser capsulotomy (p < 0.001) and then declined gradually and came back to pre-laser values by 2 weeks in all three groups. The mean LFP values, central macular thickness, and best corrected visual acuity were comparable among groups. None developed central macular edema.Conclusion: Intraocular inflammation following Nd:YAG capsulotomies is minimal and transient. Routine use of prophylactic anti-inflammatory agents may be avoided.

Antimicrobial Resistance in Enterobacteriaceae Bacteria Causing Infection in Trauma Patients: A 5-Year Experience from a Tertiary Trauma Center.

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, ß lactam and ß-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.

Surgical outcomes of cataract surgery in anterior and combined persistent fetal vasculature using a novel surgical technique: a single center, prospective study.

PURPOSE: To study the complications and surgical outcomes of cataract surgery in patients of persistent fetal vasculature (PFV) with cataract. METHODS: In this prospective study, phacoaspiration with/without intraocular lens implantation (IOL) was done in 20 children (mean age 14.2 months) with unilateral cataract with anterior (n = 6) or combined (n = 14) PFV. The rentrolental vascularized membrane was cauterized and dissected circumferentially, followed by cauterization and resection of the PFV stalk. The outcome measures included fixation preference using the CSM (central, steady, maintained) method and intraoperative and postoperative complications in an 18-month follow-up. The difference in outcomes of anterior and combined PFV, as well as aphakic and pseudophakic eyes, was studied. RESULTS: CSM fixation was seen in 16 patients after 18 months. The intraocular lens was implanted in 16 eyes and 4 eyes with combined PFV were left aphakic. None of our patients had intraoperative bleeding. Visual axis obscuration was the major complication seen, requiring membranectomy in 8 children. Pupilloplasty was required with membranectomy in one eye. None of our patients developed glaucoma or retinal detachment. CONCLUSION: Timely surgical intervention and aggressive amblyopia therapy led to good visual results in our study. Poor prognosis was seen in combined PFV, aphakia, and microphthalmia.

Comparison of Genotypic and Phenotypic Methods of Metallo-ß- lactamase Detection in Acinetobacter spp.

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..