Isolation and detection of bacterial species on mitis salivarius bacitracin agar from the plaque samples of caries active children.

BACKGROUND: In cariology studies, mitis-salivarius-bacitracin (MSB) agar has been commonly considered as the selective medium for Streptococcusmutans growth. The present study was the part of a funded project (a noninferiority randomized controlled trial) which compared the efficacy of a plant extract-based mouth rinse with that of a fluoride mouth rinse on the S.mutans counts of the children. AIM: This study aimed to identify the frequency of detection of S.mutans and nonstreptococcal bacterial species from the dental plaque of caries active children using a combined technique of anaerobic culture and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry. SETTINGS AND METHODS: Caries-active children (8-12 years old) were enrolled from a pediatric dental outpatient department at a tertiary care hospital. From each participant, dental plaque samples were collected from carious surfaces under sterilized conditions and then subjected to anaerobic culture. After 48 h of incubation, the bacterial colonies were isolated by sub-culture and identified by the MALDI-TOF. RESULTS: In all, 13 different bacterial species were isolated from the MSB agar medium. Other than S.mutans species, colonies of bacterial species such as Veillonelladispar,Streptococcusanginosus, Veillonellaparvula, and Streptococcusgordonii were also frequently observed from the medium. CONCLUSIONS: The study concluded that several bacterial strains, both streptococcal and nonstreptococcal, could be isolated from the MSB agar medium; hence, this medium should no longer be considered selective medium for the culture of S.mutans in clinical and epidemiological studies.

A Novel Potential Treatment for Diabetic Foot Ulcers and Non-Healing Ulcers - Case Series.

INTRODUCTION: Appropriate care and treatment of a wound is the need of the hour whether it is an infected or a non-infected wound. If wound healing is delayed for some reason, it leads to serious complications and further increases the hospital stay and cost of treatment. Herein, we describe a novel antimicrobial wound dressing formulation (VG111), with an objective to generate the preliminary data showing the distinct advantages in various types of wounds. METHOD: This case series involved the treatment of acute cases of wounds or chronic wounds that did not respond well to conventional wound healing treatments with VG111 in patients with different etiologies. Thirteen cases of patients that included patients with diabetes, pressure ulcers, burns, trauma, and others treated with VG111 showed rapid wound healing in all the cases, even obviating the need for a graft when complete skin regeneration occurred RESULT: This was illustrated by clearing of the wound infections, reduction/disappearance of the exudate, appearance of intense granulation, epithelialization, and anti-biofilm activity followed by complete wound closure. This VG111 precludes the need for systemic antimicrobial agents in localized infections and therefore, this single agent is an attempt to address the limitations and the drawbacks of the available products. CONCLUSION: Despite patients belonging to the old age group and having comorbidities like diabetes, still VG111 showed effective rapid wound healing, and that too without any scar formation in hardto-heal, infected, and non-infected wounds

Case report of SARS-CoV-2 with secondary Clostridioides difficile infection.

There has been an overuse of antibiotics in most patients suffering from COVID-19 which predisposes patients to hospital acquired C. difficile Infection (CDI). Also, COVID infection of the gastrointestinal tract also predisposes the patient to CDI. We here present a fatal case of SARS-CoV-2 coinfection with CDI.

A rare case of complicated pancreatitis due to leptospirosis.

The COVID-19 pandemic and the actions taken to combat it have greatly impacted the health infrastructure of all nations. Here we present a rare case of leptospirosis with severe acute pancreatitis, bilateral peripheral gangrene, disseminated intravascular coagulopathy and multiorgan failure. This is a rare presentation of leptospirosis wherein the patient had no history suggestive of acquisition of leptospires. The patient was started on doxycycline but still could not be saved due to the multisystem involvement.

Scrub typhus in two COVID-19 patients: a diagnostic dilemma.

The authors describe a case series of co-infection with COVID-19 and scrub typhus in two Indian patients. Clinical features like fever, cough, dyspnea and altered sensorium were common in both patients. Case 1 had lymphopenia, elevated IL-6 and history of hypertension, while case 2 had leukocytosis and an increased liver enzymes. Both patients had hypoalbuminemia and required admission to the intensive care unit; one of them succumbed to acute respiratory distress syndrome further complicated by multiple organ dysfunction syndrome. Seasonal tropical infections in COVID-19 patients in endemic settings may lead to significant morbidity and mortality. Therefore, high clinical suspicion and an early diagnosis for co-infections among COVID-19 patients are essential for better patient management.

Anaerobes in cardiac infections: A decade experience from the tertiary care center.

PURPOSE: Anaerobic infections are common yet life-threatening. They are being recovered from all sites of the body, including the cardiovascular system. This study was aimed to determine the retrospective analysis on the isolation of anaerobes in cardiovascular samples received for a decade-long duration. It helps in knowing the frequency of isolation of anaerobic causes of cardiovascular infection. METHODS: All cardiovascular samples from the department of Cardio-thoracic vascular surgery from January 2010 to December 2020 were studied. RESULTS: Of 601 samples received, predominant samples were vegetations and valvular tissues of 258, followed by 98 samples of pericardial tissues, 92 samples of embolus, 90 samples of blood and post-operative collections, and 63 excised aneurysms and vascular grafts. Of the total, 15 samples grew anaerobes where Clostridium species were the predominant isolates. Clostridioides difficile was isolated in 2 samples. CONCLUSIONS: Anaerobes in cardiovascular samples are uncommon yet form a significant cause of morbidity and mortality. Most infections are from the contiguous spread, penetrating trauma, and hematogenous causing endocarditis or valvular infections. These conditions and samples form the seat of infectious focus and clinical suspicion towards the anaerobic cause of these conditions, especially in conventional routine culture-negative samples. Timely diagnosis of anaerobic infections plays a vital role in the good prognostic outcome of patients undergoing cardiothoracic and vascular surgery.

Atypical bacterial co-infections among patients with COVID-19: A study from India.

Emerging evidence shows co-infection with atypical bacteria in coronavirus disease 2019 (COVID-19) patients. Respiratory illness caused by atypical bacteria such as Mycoplasma pneumoniae, Chlamydia pneumoniae, and Legionella pneumophila may show overlapping manifestations and imaging features with COVID-19 causing clinical and laboratory diagnostic issues. We conducted a prospective study to identify co-infections with SARS-CoV-2 and atypical bacteria in an Indian tertiary hospital. From June 2020 to January 2021, a total of 194 patients with laboratory-confirmed COVID-19 were also tested for atypical bacterial pathogens. For diagnosing M. pneumoniae, a real-time polymerase chain reaction (PCR) assay and serology (IgM ELISA) were performed. C. pneumoniae diagnosis was made based on IgM serology. L. pneumophila diagnosis was based on PCR or urinary antigen testing. Clinical and epidemiological features of SARS-CoV-2 and atypical bacteria-positive and -negative patient groups were compared. Of the 194 patients admitted with COVID-19, 17 (8.8%) were also diagnosed with M. pneumoniae (n = 10) or C. pneumoniae infection (n = 7). Confusion, headache, and bilateral infiltrate were found more frequently in the SARS CoV-2 and atypical bacteria co-infection group. Patients in the M. pneumoniae or C. pneumoniae co-infection group were more likely to develop ARDS, required ventilatory support, had a longer hospital length of stay, and higher fatality rate compared to patients with only SARS-CoV-2. Our report highlights co-infection with bacteria causing atypical pneumonia should be considered in patients with SARS-CoV-2 depending on the clinical context. Timely identification of co-existing pathogens can provide pathogen-targeted treatment and prevent fatal outcomes of patients infected with SARS-CoV-2 during the current pandemic.

Coinfections with Other Respiratory Pathogens among Patients with COVID-19.

Emerging evidence indicates that severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-infected individuals are at an increased risk for coinfections; therefore, physicians need to be cognizant about excluding other treatable respiratory pathogens. Here, we report coinfection with SARS-CoV-2 and other respiratory pathogens in patients admitted to the coronavirus disease (COVID) care facilities of an Indian tertiary care hospital. From June 2020 through January 2021, we tested 191 patients with SARS-CoV-2 for 33 other respiratory pathogens using an fast track diagnostics respiratory pathogen 33 (FTD-33) assay. Additionally, information regarding other relevant respiratory pathogens was collected by reviewing their laboratory data. Overall, 13 pathogens were identified among patients infected with SARS-CoV-2, and 46.6% (89/191) of patients had coinfection with one or more additional pathogens. Bacterial coinfections (41.4% [79/191]) were frequent, with Staphylococcus aureus being the most common, followed by Klebsiella pneumoniae. Coinfections with SARS-CoV-2 and Pneumocystis jirovecii or Legionella pneumophila were also identified. The viral coinfection rate was 7.3%, with human adenovirus and human rhinovirus being the most common. Five patients in our cohort had positive cultures for Acinetobacter baumannii and K. pneumoniae, and two patients had active Mycobacterium tuberculosis infection. In total, 47.1% (90/191) of patients with coinfections were identified. The higher proportion of patients with coinfections in our cohort supports the systemic use of antibiotics in patients with severe SARS-CoV-2 pneumonia with rapid de-escalation based on respiratory PCR/culture results. The timely and simultaneous identification of coinfections can contribute to improved health of COVID-19 patients and enhanced antibiotic stewardship during the pandemic. IMPORTANCE Coinfections in COVID-19 patients may worsen disease outcomes and need further investigation. We found that a higher proportion of patients with COVID-19 were coinfected with one or more additional pathogens. A better understanding of the prevalence of coinfection with other respiratory pathogens in COVID-19 patients and the profile of pathogens can contribute to effective patient management and antibiotic stewardship during the current pandemic.

Mycoplasma pneumoniae co-infection with SARS-CoV-2: A case report.

We report co-infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and Mycoplasma pneumoniae in a patient with pneumonia in India. Atypical bacterial pathogens causing community-acquired pneumonia may share similar clinical presentations and radiographic features with SARS-CoV-2 making a thorough differential diagnosis essential. The co-infection of SARS-CoV-2 and M. pneumoniae is infrequently reported in the literature. Broader testing for common respiratory pathogens should be performed in severe COVID-19 cases to rule out other concurrent infections. Early identification of co-existing respiratory pathogens could provide pathogen-directed therapy, and can save patient lives during the ongoing COVID-19 outbreak.

A case of neglected hepatorenal dysfunction during the COVID pandemic.

As the world fights with the Coronavirus, most of the hospitals are gearing up for the care of these patients. As most of the attention these days is being given on Coronavirus, the patients suffering from other clinical infections are being neglected. SARS-CoV-2 is being kept as the top differential in patients presenting with fever and respiratory distress. We hereby present a case of patient returning from Indonesia during the pandemic presenting with a history of hepatic, renal dysfunction with fever and cough. Due to the pandemic, the patient's fever and cough outweighed the hepatic and renal dysfunction, and the patient had to undergo dialysis before the final diagnosis of leptospirosis could be made.

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..

Efficacy of probiotics in the prevention of VAP in critically ill ICU patients: an updated systematic review and meta-analysis of randomized control trials.

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.

Susceptibility pattern of oral antimicrobials in uncomplicated UTI: Does fosfomycin still stand effective?

INTRODUCTION: Urinary tract infections (UTIs) are a leading cause of morbidity amongst all age groups and most patients with uncomplicated UTI who visit the OPD are treated with empirical antibiotics without doing any culture analysis. Of all the oral antibiotics used in UTI, fosfomycin is well excreted in the urine and is being used for the treatment of UTIs with a single oral dose. METHODOLOGY: The current study was planned to determine the resistance amongst uropathogens to various oral antibiotics including fosfomycin isolated over a 4-year period (April 2015 to March 2019). RESULTS: A total of 22,546 urine samples were received from OPD patients over 4 years and of these, 7,295 isolates were obtained from patients with uncomplicated UTI. About 82% of the isolates were gram-negative bacilli (GNBs). The most common isolate was E. coli 1023 (67.2%) followed by Klebsiella spp 254 (16.7%), and Pseudomonas spp 63 (4.1%). Of all the antibiotics tested, maximum sensitivity in the year 2018 was found to fosfomycin followed by nitrofurantoin for all the isolates tested. There was a statistically significant increase in the resistance pattern for almost all antibiotics tested in gram-negative bacteria (both Enterobacteriaceae and nil fermenters) whereas the increase in the resistance was not statistically significant in gram-positive cocci. CONCLUSION: It can be easily seen that the level of antibiotic resistance has been increasing even in the community with the rampant injudicious use of antibiotics in humans as well as livestock. As the level of resistance to fosfomycin is still low in the community, it can play a promising role in the treatment of infection in patients with uncomplicated UTI.

Prevalence and Characterization of Carbapenemase-producing Escherichia coli from a Tertiary Care Hospital in India.

The purpose of this study was to estimate the prevalence and to characterize the carbapenemase-producing Escherichia coli by various phenotypic antimicrobial susceptibility testing methods, and its performance was compared to the gold standard genotypic method. The prevalence of carbapenemase-resistant E. coli was found to be 65%. The phenotypic methods evaluated are cost-effective and can be used in resource-limited laboratories to rule out carbapenem resistance.

Prevalence and characterization of beta-lactamase-producing Escherichia coli isolates from a tertiary care hospital in India.

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-ß 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 ß-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, blaFOX gene (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.

Antibiotic resistance profile and co-production of extended spectrum beta lactamases and AmpC in Acinetobacter spp. in a level 1 trauma center from India.

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.

Phenotypic & genotypic profile of antimicrobial resistance in Pseudomonas species in hospitalized patients.

Background & objectives: Nosocomial infections caused by multidrug-resistant, Pseudomonas species have become a major clinical and public health concern. The aim of this study was to characterize phenotypic and genotypic profile of antimicrobial resistance (AMR) in Pseudomonas spp. isolated from hospitalized patients. Methods: A total of 126 consecutive, non-duplicate isolates of Pseudomonas spp. isolated from various clinical samples were included in the study over a period of two years. Identification and antimicrobial sensitivity was performed using automated culture system according to the Clinical and Laboratory Standards Institute (CLSI) recommendations. Phenotypic detection of extended-spectrum ß-lactamases (ESBLs), Amp-C ß-lactamase (AmpC) and metallo-ß-lactamases (MBLs) were done by various combinations of disc-diffusion and E-test methods, followed by polymerase chain reaction-based detection of ß-lactamase-encoding genes. Results: Among 126 clinical isolates, 121 (96.1%) isolates were identified as Pseudomonas aeruginosa. Most of the isolates were recovered from pus sample, 35 (27.8%) followed by urine, 25 (19.84%); endotracheal aspirate, 24 (19.04%); blood, 14 (11.11%) and sputum, four (3.17%). The highest rate of resistance was against ticarcillin-clavulanic acid, 113 (89.7%) followed by meropenem, 92 (72.5%) and ceftazidime, 91 (72.3%). Overall, ESBLs, AmpC and carbapenemase production was detected in 109 (96.4%), 64 (50.8%) and 105 (94.6%) isolates by phenotypic methods. The most prevalent ESBL gene was blaTEMin 72 (57.1%) and the least prevalent was blaSHVin 19 (15.1%) isolates. AmpC gene was seen less compared to ESBL gene. The most prevalent carbapenemases gene was blaNDM-141 (46.06%) followed by blaVIM and blaOXA-1. Interpretation & conclusions: Our findings suggested that a high rate of ESBLs and carbapenemases production was observed in Pseudomonas spp. Therefore, phenotypic and genotypic detection of AMR needs to be combined for better characterization of resistance patterns in Pseudomonas spp.