Coexistence of blaNDM and blaOXA-48 Genes in Carbapenem-Resistant Burkholderia pseudomallei Isolated From Pus: A Rare Phenomenon.

Burkholderia pseudomallei causes melioidosis in both humans as well as animals and is classified as a tier 1 pathogen by the US CDC. Melioidosis is a disease that occurs predominantly in subtropical and tropical regions. It is endemic to northern Australia and parts of Southeast Asia, as well as the Indian subcontinent. Diagnosis can be made through history, clinical examination, imaging, and microbiological studies. We report a case where Burkholderia pseudomallei was isolated froma 41-year-old man who complained of pain in the left hip and the left shoulder and swelling in both lower limbs. Chest X-ray showed bilateral consolidation. USG of the left shoulder and bilateral hips showed a mass in the anterior region of the left upper arm and the lateral region of the left thigh. Pus aspirated from left shoulder grew Burkholderia pseudomallei on culture and was carbapenem-resistant. The isolate harbored two carbapenemase genes, blaNDM and blaOXA-48, which is a novel finding.

Evaluation of In-Vitro Activity of Ceftaroline Against Methicillin-Resistant Staphylococcus aureus Clinical Isolates.

Methicillin-resistant Staphylococcus aureus (MRSA) is one of the major causes of hospital and community-acquired infections. Fewer drugs, such as vancomycin, teicoplanin, and daptomycin, are effective against it, but they come with high toxicity. Fifth-generation cephalosporins like ceftaroline and second-generation cefuroxime are effective against MRSA. Limited studies are available on ceftaroline resistance in the literature. This study was undertaken to determine ceftaroline resistance in MRSA in a tertiary care hospital in Eastern India. A cross-sectional, hospital-based study was carried out with MRSA isolates obtained from various clinical samples of patients. Identification of the isolates to the species level was performed by an automated Vitek system, and selected samples were genotypically confirmed by detecting the mecA gene via real-time PCR. Out of a total of 334 Staphylococcus aureus isolates examined in this study, the prevalence of MRSA was seen in 59.3% (198/334), and methicillin-sensitive Staphylococcus aureus was in 40.7% (136/334). Of the total 198 MRSA isolates, ceftaroline intermediate MRSA was seen in 8.6% (17/198), and ceftaroline sensitive MRSA was in 91.4% (181/198), respectively. Among the 17 ceftaroline intermediate MRSA isolates, 88.2% (15/17) showed a minimum inhibitory concentration (MIC) of 2 µg/ml, and 11.8% (2/17) showed an MIC of 3 µg/ml. All the remaining 91.4% (181/198) isolates were sensitive to ceftaroline and showed an MIC ≤1 µg/ml. Real-time PCR confirmed the presence of the mecA gene in MRSA isolates. In this present study, not a single isolate was resistant to ceftaroline, suggesting that it, being a safer drug, can be used in place of glycopeptides such as vancomycin or teicoplanin and linezolid, where resistance has already been detected. The rational use of ceftaroline could be useful in clinical settings, and further studies will confirm the findings.

Colistin Resistance Among Multi-Drug Resistant Gram-Negative Bacterial Isolates From Different Clinical Samples of ICU Patients: Prevalence and Clinical Outcomes.

INTRODUCTION: Colistin is considered to be the last resort for the management of infections caused by multi-drug resistant (MDR) gram-negative bacilli (GNB). However, in the recent past, there has been a rise in colistin resistance among MDR isolates in clinical settings with no profound data on the incidences and causes. The purpose of this study was to estimate the prevalence of colistin-resistance (CLR) in MDR isolates collected from different intensive care units (ICUs) and to determine the clinical outcomes of the patients.  Materials and methods: A prospective study was conducted in the ICU of a tertiary care hospital in Eastern Odisha, India from March 2019 to February 2020. MDR GNB isolates from different clinical samples of ICU patients, not intrinsically resistant to colistin, were included in this study. Samples collected for culture and sensitivity testing were processed as per standard guidelines in the microbiology laboratory. MDR organisms were examined for colistin susceptibility by the broth dilution method. Clinical data was collected from hospital electronic medical records and presented as percentage, number (N), and median (range). RESULTS:  The prevalence of colistin resistance MDR GNB was found to be 19.6% in the present study. Colistin resistance among the MDR isolates was found to be the highest (9.2% for Klebsiella pneumonia followed by 5% for Escherichia coli). CLR drug-resistant isolates were commonly (28.8%) isolated from samples of respiratory tract infections and the majority (54.1%) were from neurology ICU. In this study, co-morbidity was not found among 57.9% of the ICU patients and recovery was maximum i.e., 74.2%. CONCLUSION: This study found the prevalence of colistin resistance to be high (19.6%) among all MDR GNB isolates from samples of ICU patients, Klebsiella pneumonia and Escherichia coli commonly acquire colistin resistance. Patients in the neurology ICU were frequently infected with CLR MDR strains. Most of the patients who recovered were without any underlying comorbidities. Prolonged hospital stay and direct antibiotic pressure in the hospital can lead to the development of CLR variants.

A Clinical Significance of Fungal Infections in Diabetic Foot Ulcers.

BACKGROUND: Diabetic foot ulcers (DFUs) are the most common and serious complications in uncontrolled diabetes. Infections are predominantly polymicrobial, with aerobic Gram-positive, anerobic, and fungal infections. Early detection of fungal infection and initiation of appropriate treatment in DFUs may lead to better healing and avoid amputations. The primary objective was to find out the prevalence of DFUs getting infected with fungus and the secondary objective was to identify the appropriate methodology for the detection of the fungus in DFUs. MATERIALS AND METHODS:  This was a cross-sectional observational study carried out in a tertiary care hospital with a sample size of 60 DFUs. Microbiological analysis was done by swab culture and deep tissue culture. Observational data were collected and the significance level was statistically analyzed. RESULTS:  In the present study, the prevalence of fungal infections in DFUs was 31.7%. Only fungal tissue was positive in 15%, the fungal swab was positive in 8.33%, and both tissue and swab were positive in 8.33%. All these patients were treated with antifungal treatment as per the culture report in addition to appropriate antimicrobial therapy. CONCLUSION:  A fungal culture should be done in all patients with non-healing DFUs. Both fungal swab and tissue culture testing should be advocated in patients with DFUs for better mycological evaluation. The addition of antifungal medications may provide better outcomes in selected cases.

Dynamicity and persistence of severe acute respiratory syndrome coronavirus-2 antibody response after double dose and the third dose with BBV-152 and AZD1222 vaccines: A prospective, longitudinal cohort study.

Introduction: Vaccines are available worldwide to combat coronavirus disease-19 (COVID-19). However, the long-term kinetics of the vaccine-induced antibodies against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) have not been sufficiently evaluated. This study was performed to investigate the persistence and dynamicity of BBV-152 (Covaxin)- and AZD1222 (Covishield)-induced immunoglobulin-G (IgG) antibodies over the year and neutralizing antibodies' status after 1-month of booster dose. Materials and methods: This 52-week longitudinal cohort study documented antibody persistence and neutralizing antibodies status among 304 healthcare workers (HCWs) from six hospitals and research facilities in Odisha, enrolled during January 2021 and continued till March 2022. IgG antibodies against spike receptor-binding domain (RBD) of SARS-CoV-2 were quantified in an automated chemiluminescence immune assay-based (CLIA) platform and a surrogate virus neutralization test (sVNT) was performed by enzyme-linked immunosorbent assay (ELISA). Results: Among these 304 HCWs vaccinated with double doses, 154 HCWs (50.66%) were Covaxin recipients and the remaining 150 (49.34%) were Covishield recipients. During the follow-ups for seven times, a total of 114 participants were identified as vaccine breakthrough cases. In 190 non-infected HCWs, the median antibody titer was significantly waned from DD2 to DD10, both for Covaxin (231.8 vs. 42.7 AU/ml) and Covishield (1,884.6 vs. 369.2 AU/ml). No statistically significant differences in antibody titers were observed based on age, gender, comorbidities, and blood groups. The median inhibition activity of sVNT increased from 23.8 to 91.3% for Covaxin booster recipients and from 41.2 to 96.0% for Covishield booster recipients. Among 146 booster dose recipients, 48 were breakthrough cases after booster and all were contracted by the omicron variant. Conclusion: This year-long follow-up study found a 7- and 5-fold antibody waning in Covaxin and Covishield recipients, respectively, without any breakthrough infection history. However, individuals with booster breakthrough had mild symptoms and did not require hospital admission. The data also indicate the possible escape of omicron variants despite the presence of vaccine-induced neutralizing antibodies.

Microbiome Diversity in Vaginal Fluid and Sensitivity Patterns in Preterm Premature Rupture of Membrane Cases.

INTRODUCTION: Preterm premature rupture of membranes (PPROM) is spontaneous rupture of the fetal membranes before 37 completed weeks and before the onset of labor. PPROM occurs in 3% of all pregnancies and is responsible for approximately one-third of all preterm deliveries. It leads to increase in perinatal morbidity and mortality. AIM: The present study aimed to characterize the microbiome of vaginal fluid, which will be helpful in the selection of empiric antimicrobial therapy. MATERIALS AND METHOD: A prospective observational study was conducted in the Department of Obstetrics and Gynaecology, Pradyumna Bal Memorial Hospital, Kalinga Institute of Medical Sciences (KIMS), Bhubaneswar during the period of October 2019 to June 2021 to characterize the microorganisms in the vaginal fluid and their antimicrobial sensitivity patterns found in antenatal women presenting with PPROM. A total of 160 antenatal women diagnosed with PPROM, gestational age between 28 weeks to 36 weeks and 6 days were included in the study. High vaginal swabs were collected for microbial culture and sensitivity. RESULTS: Out of 160 samples, the growth of organisms was observed in 134 (85.09%) samples. Out of them, 133 were monomicrobial, one was polymicrobial. Common isolated infections included Enterococcus faecalis (17.39%), followed by Staphylococcus aureus (14.29%), Escherichia coli (11.18%), and Staphylococcus haemolyticus (6.21%). Most of them were sensitive to ampicillin followed by linezolid and vancomycin. S. aureus was most sensitive to linezolid followed by gentamicin and vancomycin. Most isolates were multidrug-resistant. CONCLUSION: The empirical antimicrobial treatment started for PPROM management should be based on the established changing microbiological pattern and sensitivities with due consideration of geographical and demographic variations.

Comparative Evaluation of Cartridge-Based Abbott ID NOW Test With Probe-Based Real-Time Reverse Transcription Polymerase Chain Reaction Assay for Detection of SARS-CoV-2.

BACKGROUND: The gold standard test for detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) recommended by WHO is real-time reverse transcription polymerase chain reaction (RT-PCR), which has a turnaround time of five to six hours. Abbott ID NOW (Abbott Diagnostics Scarborough, Inc., Scarborough, ME, USA), the cartridge-based loop-mediated isothermal amplification (LAMP) assay, was approved by FDA for Emergency Use Authorization as rapid point of care testing. The present study was planned to evaluate the performance of the cartridge-based Abbott ID NOW test by comparing it to the currently used standard probe-based real-time RT-PCR method for detection of SARS-CoV-2. METHODOLOGY: A cross-sectional study was conducted in a tertiary care hospital in the eastern part of India after getting institutional ethics committee (IEC) approval. Two hundred fifty-nine cases of various age groups of both sexes who were advised for testing for SARS-CoV-2 were included in the study. Nasopharyngeal swabs were collected according to protocol advisory by the Indian Council of Medical Research (ICMR), India. Dry swabs were sent for Abbott ID NOW testing and swabs in viral transport medium were sent for probe-based RT-PCR assay using the CoviPath kit (Thermo Fisher Scientific, Bangalore, India). The data were collected and statistical analysis was performed using Statistical Package for Social Sciences (SPSS) (IBM Corp., Armonk, NY, USA). Sensitivity, specificity, positive and negative predictive values for ID NOW were calculated taking RT-PCR as the gold standard.  Results: Out of 259 patients enrolled in the study, 49% were symptomatic for coronavirus disease 2019 (COVID-19). The prevalence rate of SARS-CoV-2 was 20.84% among the study population. Sensitivity and specificity, positive and negative predictive values of ID NOW test in comparison to RT-PCR assay was found to be 87%, 98%, 92.1% and 96.8% respectively. ID NOW detected seven out of 54 (12.9%) cases as false negative who were found to be positive with RT-PCR, with mean Ct value of the target genes >34. CONCLUSIONS: In this study the overall sensitivity for ID NOW assay was found to be lower, but specificity, positive and negative predictive values were found to be higher. It had the highest correlation to RT-PCR among symptomatic patients and at higher viral loads. Due to the ease of use and shortest result time for detecting COVID-19, ID NOW test could be used as a point-of-care test. But for all tests, the results should be interpreted according to the clinical and epidemiological context.

Prevalence of uropathogens and their antimicrobial resistance pattern among adult diabetic patients.

BACKGROUND: The high prevalence of diabetes, urinary tract infection (UTI) in persons with diabetes, and antibiotic resistance throughout the world including India is a cause of concern for health-care providers. OBJECTIVES: The study aimed to assess the prevalence of uropathogens, their resistance pattern, and associated factors of UTI among adults with diabetes. METHODS: A cross-sectional study was conducted during the year 2018-2019 among 504 adult diabetic patients attending a tertiary care hospital of Bhubaneswar, Eastern India. Using recommended culture methods, clean-catch midstream urine samples were examined for the presence of pathogenic bacteria and their antimicrobial susceptibility pattern. RESULTS: The prevalence of UTI was 75.4% and the predominant isolates were Escherichia coli (25.6%), Enterococcus spp. (18.7%), and Klebsiella spp. (8.1%). Most of the bacteria isolates were sensitive to nitrofurantoin (80.8%), gentamicin (76.8%), and amikacin (72.1%) whereas resistant to cefpodoxime (77.6%), cefixime (70.8%), and cefadroxil (65.0%). The overall multidrug resistance (MDR) to two or more antimicrobial agents was observed in 87.4% bacterial isolates. The odds of developing UTIs were significantly higher in females with diabetes (adjusted odds ratio [AOR]: 2.04; 95% confidence interval CI: 1.33-3.13), those with Stage I/II hypertension (AOR: 1.77; 95% CI: 1.04-3.008), and those having glycated hemoglobin level >9% (75 mmol) (AOR: 2.15; 95% CI: 1.13-4.10) compared with their counterparts. CONCLUSION: The prevalence of UTI and MDR to commonly used antibiotics among diabetic patients is alarming, and thus, isolation of uropathogenic bacteria and antimicrobial susceptibility testing is crucial for the treatment of UTI in persons with diabetes.

Persistence of Antibodies Against Spike Glycoprotein of SARS-CoV-2 in Healthcare Workers Post Double Dose of BBV-152 and AZD1222 Vaccines.

Purpose: We investigated the persistence of the vaccine-induced immunoglobulin G (IgG) antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) among healthcare workers (HCWs) in Odisha who received a complete dose of either Covaxin or Covishield vaccine. Methods: This 24-week longitudinal cohort study was conducted from January to July 2021 with participants from 6 healthcare and research facilities of Odisha to understand the dynamicity of the vaccine-induced IgG antibodies against SARS-CoV-2 after the complete dose of vaccines. Results: Serum samples were collected from 614 participants during each follow-up and were tested in two chemiluminescent microparticle immunoassay (CLIA)-based platforms to detect SARS-CoV-2 antibodies both qualitatively and quantitatively. Among these participants, 308 (50.2%) participants were Covishield recipients and the rest 306 (49.8%) participants took Covaxin. A total of 81 breakthrough cases were recorded and the rest 533 HCWs without any history of postvaccination infection showed significant antibody waning either from T3 (Covaxin recipient) or T4 (Covishield recipient). The production of vaccine-induced IgG antibodies is significantly higher (p < 0.001) in Covishield compared with Covaxin. Covishield recipients produced higher median anti-S IgG titer than Covaxin. No statistically significant differences in antibody titers were observed based on age, gender, comorbidities, and blood groups. Conclusion: This 6-month follow-up study documents a 2-fold and 4-fold decrease in spike antibody titer among Covishield and Covaxin recipients, respectively. The clinical implications of antibody waning after vaccination are not well understood. It also highlights the need for further data to understand the long-term persistence of vaccine-induced antibody and threshold antibody titer required for protection against reinfection.

Prevalence of ESBL in Escherichia coli Isolates Among ICU Patients in a Tertiary Care Hospital.

INTRODUCTION: Resistance to common antibiotics is a matter of grave concern in treating infections in hospital settings especially in Intensive Care Units (ICUs). One of the most commonly used and effective group of antibiotics, cephalosporins, exhibit resistance due to production of Extended Spectrum Beta- Lactamases (ESBLs). The prevalence of ESBL producing Escherichia coli (E.coli) has increased throughout the world and is a major cause of treatment failure in ICUs. As per our knowledge studies were not available on the prevalence of ESBL producing E.coli in ICUs of this region. AIM: To determine the prevalence of ESBLs among Escherichia coli isolates in ICUs of a tertiary care hospital. MATERIALS AND METHODS: A cross sectional study was conducted over a period of 4 years (Sept 2011 to Sept 2015) in the Department of Microbiology, Kalinga Institute of Medical Sciences (KIMS), Bhubaneswar. Consecutive non-duplicate isolates of E.coli recovered from 6800 clinical samples of patients admitted to different Intensive Care Units (ICUs) were subjected to ESBL screening test and then to CLSI recommended Phenotypic Confirmatory Disc Diffusion Tests (PCDDT) for ESBL production determination. RESULTS: Out of 6800 samples, 1038 were E.coli isolates and 452(44%) were resistant to third generation cephalosporins. ESBL producing Escherichia coli among them were 276 (61.1%). Paediatric ICU showed the highest prevalence of ESBL E.coli at 80.9%. The highest prevalence of ESBL E.coli was in urine samples (82.6%) followed by pus (9.8%). The most effective antibiotic for ESBL producers was imipenem (96.7% sensitive), followed by amikacin (88.4%) and piperacillin- tazobactum (87%). CONCLUSION: This study has highlighted the high prevalence of ESBL producing E.coli in the ICUs of our hospital. An in depth analysis of their antibiogram will be helpful in formulating the antibiotic policy and prevent spread of ESBL strains. It is recommended that ESBL testing should be done routinely to curtail antibiotic resistance and to effectively implement infection control measures.

Antimicrobial susceptibility pattern of clinical isolates of non-fermentative bacteria.

152 nonfermentative bacteria were isolated from a total number of 965 clinical samples processed routinely in the laboratory of Microbiology Department, M.K.C.G Medical College in South Orissa accounting to a prevalence rate of 15.75%. Pseudomonas spp. (both pigmented and non-pigmented strains) were isolated in maximum percentage (73.6%) followed by Acinetobacter spp. (19.7%) and Alkaligenes faecalis (4.6%). Rarely encountered species were Eikenella corrodens (1.3%) and Stenotrophomonas maltophila (0.6%). Pus from various sites was the major source (116; 76%). 81% of all isolates were sensitive to amikacin and 74% to ofloxacin. Sensitivity to cefotaxime, ciprofloxacin, tobramycin, gentamicin and netlimycin ranged from 53% to 68%. Least effective drugs were carbenicillin and ceftriaxone (48% each).