Detection of Biofilm Production and Antibiotic Susceptibility Pattern among Clinically Isolated Staphylococcus aureus.

Aim: The increasing antibiotic resistance and the ability to form biofilms in medical devices have become the leading cause of severe infections associated with Staphylococcus aureus (S. aureus). Since the bacteria living in biofilms can exhibit 10- to 1,000-fold increase in antibiotic resistance and implicate chronic infectious diseases, the detection of S. aureus ability to form biofilms is of great importance for managing, minimizing, and effectively treating infections caused by it. This study aimed to compare the tube and tissue culture methods to detect biofilm production and antibiotic susceptibility in MRSA and MSSA. Materials and Methods: The S. aureus isolates were identified by the examination of the colony morphology, Gram staining, and various biochemical tests. Antimicrobial susceptibility testing of all isolates was performed by the modified Kirby-Bauer disc diffusion method as recommended by CLSI guidelines. MRSA screening was performed phenotypically using a cefoxitin disc (30 µg). Isolates were tested for inducible resistance using the D-test, and two phenotypic methods detected biofilm formation. Results: Among 982 nonrepeated clinical specimens, S. aureus was isolated from 103 (10.48%). Among 103 clinical isolates of S. aureus, 54 (52.42%) isolates were MRSA, and 49 (47.57%) were MSSA. Among 54 MRSA isolates, the inducible MLSB phenotype was observed in 23/54 (42.59%) with a positive D-test. By TCP method, 26 (48.1%) MRSA isolates were strong biofilm producers, whereas, among all MSSA isolates, only 6 (12.2%) were strong biofilm producers. Conclusion: MRSA showed strong biofilm production in comparison with MSSA. The TCP method is a recommended reliable method to detect the biofilm among S. aureus isolates, and the TM method could be useful for the screening of biofilm production in S. aureus in the routine clinical laboratory.

Inhibition and Reduction of Biofilm Production along with Their Antibiogram Pattern among Gram-Negative Clinical Isolates.

Background: Bacterial biofilm is a significant virulence factor threatening patients, leading to chronic infections and economic burdens. Therefore, it is crucial to identify biofilm production, its inhibition, and reduction. In this study, we investigated biofilm production among Gram-negative isolates and assessed the inhibitory and reduction potential of ethylene diamine tetra acetic acid (EDTA) and dimethyl sulfoxide (DMSO) towards them. In addition, we studied the antimicrobial resistance pattern of the Gram-negative isolates. Methods: Bacterial isolation and identification was done using standard microbiological techniques, following the Clinical and Laboratory Standards Institute (CLSI) guideline, 28th edition. The Kirby-Bauer disk diffusion method was used to determine the antibiotic susceptibility pattern of the isolates, and ß-lactamase production was tested via the combination disk method. Biofilm formation was detected through the tissue culture plate (TCP) method. Different concentrations of EDTA and DMSO were used to determine their inhibitory and reduction properties against the biofilm. Both inhibition and reduction by the various concentrations of EDTA and DMSO were analyzed using paired t-tests. Results: Among the 110 clinical isolates, 61.8% (68) were found to be multidrug resistant (MDR). 30% (33/110) of the isolates were extended-spectrum ß-lactamase (ESBL) producers, 14.5% (16/110) were metallo-ß-lactamase (MBL), and 8% (9/110) were Klebsiella pneumoniae carbapenemase (KPC) producers. Biofilm formation was detected in 35.4% of the isolates. Biofilm-producing organisms showed the highest resistance to antibiotics such as cephalosporins, chloramphenicol, gentamicin, and carbapenem. The inhibition and reduction of biofilm were significantly lower (p < 0.05) for 1 mM of EDTA and 2% of DMSO. Conclusion: Isolates forming biofilm had a higher resistance rate and ß-lactamase production compared to biofilm nonproducers. EDTA and DMSO were found to be potential antibiofilm agents. Hence, EDTA and DMSO might be an effective antibiofilm agent to control biofilm-associated infections.

Prevalence and antibiogram of bacteria causing urinary tract infection among patients with chronic kidney disease.

Identifying and appropriately managing urinary tract infections (UTIs) among chronic kidney disease (CKD) patients are essential to reduce further disease complications and economic burden. Hence, this study aims to determine the prevalence of UTIs among CKD patients and study the antibiogram of the bacterial isolates. Four hundred eighty-two clean catch midstream urine samples were collected from CKD patients during the study period. The samples were cultured, and bacteria were isolated using standard microbiological techniques. Antibiotic susceptibility testing was performed by the Kirby-Bauer disc diffusion method following the Clinical and Laboratory Standards Institute guidelines. Of the 482 CKD patients, 15.8% were culture positive, and the majority was elderly aged group population. Most bacterial isolates were Escherichia coli 50%, followed by Pseudomonas aeruginosa 15.80%, Enterococcus species 15.80%, and Klebsiella pneumoniae 11.84%. The majority of bacteria were found to be resistant to beta-lactam antibiotics, ampicillin (94.67%), ceftriaxone (89.04%), cefotaxime (87.5%), and ceftazidime (84.0%), while polymyxin, colistin, vancomycin, meropenem, and imipenem were the most sensitive antibiotics. In our study, higher levels of antibiotic resistance were observed among urinary isolates. Therefore, our findings suggest clinicians to choose better antibiotic options to treat UTIs among CKD patients.

Associated Biochemical and Hematological Markers in COVID-19 Severity Prediction.

Background: The global threat of COVID-19 has created the need for researchers to investigate the disease's progression, especially through the use of biomarkers to inform interventions. This study aims to assess the correlations of laboratory parameters to determine the severity of COVID-19 infection. Methods: This study was conducted among 191 COVID-19 patients in Sumeru Hospital, Lalitpur, Nepal. According to their clinical outcomes, these patients were divided into severe and nonsevere groups. Inflammatory markers such as LDH, D-dimer, CRP, ferritin, complete blood cell count, liver function tests, and renal function tests were performed. Binary logistic regression analysis determined relative risk factors associated with severe COVID-19. The area under the curve (AUC) was calculated with ROC curves to assess the potential predictive value of risk factors. Results: Out of 191 patients, 38 (19.8%) subjects died due to COVID-19 complications, while 156 (81.7%) survived and were discharged from hospital. The COVID-19 severity was found in patients with older age and comorbidities such as CKD, HTN, DM, COPD, and pneumonia. Parameters such as d-dimer, CRP, LDH, SGPT, neutrophil, lymphocyte count, and LMR were significant independent risk factors for the severity of the disease. The AUC was highest for d-dimer (AUC = 0.874) with a sensitivity of 82.2% and specificity of 81.2%. Similarly, the cut-off values for other factors were age >54.5 years, D-dimer >0.91 ng/ml, CRP >82.4 mg/dl, neutrophil >78.5%, LDH >600 U/L, and SGPT >35.5 U/L, respectively. Conclusion: Endorsement of biochemical and hematological parameters with their cut-off values also aids in predicting COVID-19 severity. The biomarkers such as D-dimer, CRP levels, LDH, ALT, and neutrophil count could be used to predict disease severity. So, timely analysis of these markers might allow early prediction of disease progression.

Co-infection of dengue, scrub typhus, and typhoid during dengue outbreak in Nepal, 2022: A case report.

In midst of the recent dengue outbreak in Nepal, in 2022, the risk of co-infection increases and may lead to fatal outcomes if the diagnosis of multiple infections is delayed. Thus, all available diagnostic approaches must be taken to decrease the burden of illness and lessen mortality.

Implication of Salivary Biochemical Parameters for Diagnosis and Prognosis of Type 2 Diabetes Mellitus.

Background: Clinical laboratory diagnosis and prognosis for diabetes mellitus is performed using blood as a major specimen; however, saliva may represent as an alternative noninvasive specimen of choice. This study aims to evaluate salivary biochemical parameters in diabetic and healthy individuals to substantiate saliva's role in the diagnosis and prognosis of type 2 diabetes mellitus (T2DM). Methods: This case-control study included 150 T2DM patients and 150 apparently healthy individuals. Socio-demographic data and anthropometric measurements were recorded using a standard questionnaire. Correlation between salivary and blood levels for each parameter was determined using Pearson correlation. Linear regression was performed to estimate the blood levels of the parameters from their salivary levels. Receiver operating characteristics (ROC) analysis was done to determine the diagnostic ability of salivary glucose and establish a sensitivity, specificity, and cut-off value. Results: Salivary glucose, TC, LDL-C, urea, and creatinine were significantly higher in people with diabetes than in the control population (p < 0.05). A significant positive correlation was found between salivary and blood parameters including glucose, TC, TG, LDL-C, urea, and creatinine except for HDL-C in both case and control groups. The linear relationship for each parameter, except glucose in case population and HDL-C in case, control, and the total population was observed between blood and saliva. ROC analysis gave a cut-off value of 1.9 mg/dl for salivary glucose with 71.4% sensitivity and 72.3% specificity. Conclusion: Salivary estimation significantly reflects the blood parameters in this study, indicating that saliva can be a noninvasive specimen for the diagnosis and prognosis of T2DM.

Diagnostic Performance of SARS-CoV-2 Rapid Antigen Test in relation to RT-PCR Cq Value.

Background: Early detection of the SARS-CoV-2 is crucial for both the improvement of turnaround time and limiting the spread of the virus in the community. Thus, this study aims to establish rapid antigen tests as an effective diagnostic tool to improve the testing strategies of COVID-19 diagnosis. Methods: A laboratory based cross-sectional study was performed on the patients that visited Sukraraj Tropical and Infectious Disease Hospital (STIDH) in Kathmandu, Nepal, from November 2020 to January 2021. A total of 213 nasopharyngeal swabs were collected from both symptomatic and asymptomatic patients for rapid antigen test, followed by RT-PCR assay as reference test for confirmation of COVID-19. A standard questionnaire was administered to collect other information from patients. Data were collected and analyzed using SPSS version 20. Results: Out of 213 individuals, 75 tested positive in Ag-RDT test, while 118 tested positive for SARS-CoV-2 RNA genome via Real time PCR assay. The overall diagnostic performance of Ag-RDT showed 63.6% sensitivity and 97.9% specificity. The diagnostic accuracy of Ag- RDT was 78.9% with κ value 0.590, showing moderate agreement with RT-PCR. Significant difference (p value <0.001) was observed between Ag- RDT+ and Ag- RDT- results when compared to Cq values obtained from RT- PCR. Conclusion: The promising performance of Ag-RDT renders it useful as screening tool alongside RT-PCR to reduce transmission via improving contact tracing, implementation of local mitigation strategies, and refining existing testing protocol for diagnosis of COVID-19.

Anti-SARS-CoV-2 Antibody Screening in Healthcare Workers and Its Correlation with Clinical Presentation in Tertiary Care Hospital, Kathmandu, Nepal, from November 2020 to January 2021.

BACKGROUND: Antibody titer and the life span of antibodies against SARS-CoV-2 have been found to be associated with the clinical presentation in individuals. The extent of exposure of healthcare workers and the general public to SARS-CoV-2 needs to be assessed to monitor the COVID-19 pandemic. Thus, this study is an attempt in assessing the anti-SARS-CoV-2 antibody in health care workers. METHODS: This laboratory-based cross-sectional study was performed in Manmohan Memorial Medical College and Teaching Hospital, Kathmandu from November 2020 to January 2021. A total of 185 HCWs were enrolled in this study. Their serum samples were screened for anti-SARS-CoV-2 antibodies, and a structured questionnaire was administered to collect further information. Anti-SARS-CoV-2 antibody screening was performed using lateral flow immunoassay. The data were analyzed using SPSS version 20. RESULTS: Among 185 HCWs that participated in the study, 41 (22.2%) tested positive for the anti-SARS-CoV-2 antibody. Of these 41 HCWs, 37 tested positive for IgG only and 4 of them tested positive for both IgM and IgG antibodies. The presence of the previous history of SARS-CoV-2 infection (p < 0.001), the presence of flu-like symptoms within the last 6 months (p < 0.001), and the presence of positive contact history (p=0.002) were statistically significant with the presence of the antibody among HCWs. CONCLUSION: Healthcare workers carry a high burden of SARS-CoV-2 infection and are at risk of acquiring infection from their workplace. Anti-SARS-CoV-2 antibody screening among healthcare workers is highly recommended in multiple healthcare settings as it can help in monitoring transmission dynamics and evaluation of infection control policies.

Nasal Carriage of Methicillin-Resistant Staphylococcus aureus among Healthcare Workers in a Tertiary Care Hospital, Kathmandu, Nepal.

INTRODUCTION: Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most common causes of nosocomial infections. One of the potential risk factors for nosocomial staphylococcal infections is colonization of the anterior nares of healthcare workers (HCWs). Our study aimed to determine the rate of nasal carriage MRSA among HCWs at Manmohan Memorial Medical College and Teaching Hospital, Kathmandu. METHODS: Two hundred and thirty-two nasal swabs were collected from HCWs of Manmohan Memorial Medical College and Teaching Hospital, Kathmandu, Nepal, within six months (February 2018-July 2018). Nasal swabs were cultured, and S. aureus isolates were subjected to the antimicrobial susceptibility test by the modified Kirby-Bauer disc diffusion method. MRSA and iMLSB (inducible macrolide lincosamide streptogramin B) resistance was screened using the cefoxitin disc (30 µg) and D-test (clindamycin and erythromycin sensitivity pattern), respectively, following CLSI (Clinical and Laboratory Standard Institute) guidelines. Risk factors for MRSA colonization were determined using the chi-square test considering the p value ˂0.05 as significant. RESULTS: A total of 34/232 (14.7%) S. aureus were isolated, out of which 12 (35.3%) were MRSA. The overall rate of nasal carriage MRSA among HCWs was 5.2% (12/232). Colonization of MRSA was higher in males (8.7%) than in females (4.3%). MRSA colonization was found to be at peak among the doctors (11.4%). HCWs of the postoperative ward were colonized highest (18.2%). All MRSA isolates were sensitive to linezolid and tetracycline. iMLSB resistance was shown by 7(20.6%) of the isolates. MRSA strains showed higher iMLSB resistance accounting for 33.3% (4/12) in comparison to methicillin-susceptible strains with 13.6% (3/22). Smoking was found to be significantly associated with MRSA colonization (p=0.004). CONCLUSION: Rate of nasal carriage MRSA is high among HCWs and hence needs special attention to prevent HCW-associated infections that may result due to nasal colonization.

Vitamin D deficiency and cardiovascular risk in type 2 diabetes population.

This study aims to assess vitamin D deficiency-induced dyslipidemia and cardiovascular disease (CVD) risk in poor glycemic control among type 2 diabetes mellitus (T2DM) patients. This study was carried out among 455 T2DM patients involving poor glycemic control (n = 247) and good glycemic control (n = 208). Fasting plasma glucose (FPG) and HbA1c were measured to assess glycemic control. Cardiac risk ratio, atherogenic index plasma, and atherogenic coefficient were calculated to assess and compare the CVD risk in different groups. Patients with poor control had a significantly higher level of total cholesterol (TC), triglyceride (TG), and non-high-density lipoprotein lipase cholesterol (non-HDL-C), atherogenic variables, and lower level of high-density lipoprotein lipase cholesterol (HDL-C) as compared to patients with good glycemic control. We also observed significant negative correlation of vitamin D with lipid markers and atherogenic variables in poor glycemic control diabetic population. The serum vitamin D levels were inversely associated with HbA1c, FPG, TG, TC, and non-HDL-C. Furthermore, hypercholesterolemia, hypertriglyceridemia, and elevated non-HDL-C were the independent risks in hypovitaminosis D population. Vitamin D deficiency in poor glycemic control is likely to develop dyslipidemia as compared to vitamin D insufficient and sufficient groups. Thus, vitamin D supplementation and an increase in exposure to sunlight may reduce the risk of cardiovascular complications in diabetes.

Metabolic Syndrome and Diabetes Risk Among Young Adult Students in the Health Sciences from Kathmandu, Nepal.

BACKGROUND: The widespread dissemination of unhealthy dietary habits, childhood-teenage obesity, and sedentary lifestyle in young adults has paved the way for public health burden metabolic syndrome and early onset of type 2 diabetes mellitus. The aim of this study was to assess the prevalence and risk factors for metabolic syndrome and diabetes among young adult students. METHODS: This cross-sectional study was conducted among students of age group (18 to 25 years) studying at Manmohan Memorial Institute of Health Sciences and Central Institute of Science and Technology. The diabetes risk score of each individual was calculated by the Finnish Diabetes Risk Score (FINDRISC tool). Independent risk factors for diabetes and metabolic syndrome were measured by multivariable logistic regression analysis. The p-value of <0.05 was considered statistically significant in this study. RESULTS: A total of 825 students were recruited and 739 (89.6%) students completed the study with all the fulfilled criteria. The metabolic syndrome (Harmonized Joint Scientific Statement (HJSS) criteria) was present in 7.1%, and the most prevalent defining component was low HDL-C (78%); 74.8% of students were under low risk, 22.18% were at slightly elevated risk, 2.02% were at moderate risk, and 1.01% were at high risk of diabetes. The cardiometabolic risk factors like BMI, TC, and LDL-C were higher at a significant level (p<0.001) with an increased diabetes risk score. Independent lifestyle risk factor for metabolic syndrome was current smoking (AOR, 4.49, 95% CI 1.38-14.62) whereas, an independent lifestyle risk factor for diabetes was low adherence to physical exercise (AOR, 4.81, 95% CI, 2.90-7.99). CONCLUSION: Metabolic syndrome is present, although in low numbers in young adults putting them at risk to develop diabetes in the near future. Early assessment of metabolic syndrome and diabetes risk in young may provide insights for preventive and control plans for risk population.

Classical Dermatomyositis: A Case Report.

Dermatomyositis is an idiopathic myopathy involving progressive muscle weakness with skin manifestation. Diagnosis is based upon the progressive muscle weakness, skin rashes, elevated serum muscle enzymes, muscle biopsy, and abnormal electromyogram. The incidence of dermatomyositis is rare. In this case study, we report a case of classical dermatomyositis without incidence of malignancy. Timely diagnosis and administration of steroid led to better prognosis of the patient.

Self-Care Adherence And Barriers To Good Glycaemic Control In Nepalese Type 2 Diabetes Mellitus Patients: A Hospital-Based Cross-Sectional Study.

PURPOSE: The patient believes in adherence to medication rather than to self-care adherence and lifestyle changes for the management of diabetes. This study was carried out to establish the association of self-care adherence and their barriers in poor glycemic control in our diabetic population. PATIENTS AND METHODS: This cross-sectional study was conducted among 480 already diagnosed diabetes outpatients attended in our two hospitals. Glycaemic control was defined by levels of HbA1c. Socio-demographic data, lifestyle variables and anthropometric measurements were recorded using a standard questionnaire. Fasting blood glucose, HbA1c and lipid profiles were estimated using the manufacturer's guideline. Student's t-test and one-way ANOVA were used for comparison between different groups and the correlation was established by Spearman correlation. Risk factors associated with poor glycaemic control were verified by logistic regression analysis. RESULTS: The mean HbA1c of the study population was 7.4±1.3% and 65.4% had poor glycaemic control with mean 8.0±1.1%. Higher HbA1c levels were significantly associated with duration of diabetes, a number of drugs used, patient-physician relationship and knowledge about diabetes. The poor glycaemic control was significantly associated with low adherence of following the meal plan, regular medication and regular exercising (p<0.001). Among all the barriers, a too busy schedule for following the meal plan, taking medications and exercising regularly was significantly correlated with HbA1c levels. Multivariable logistic regression analysis showed irregular meal plan (OR=5.27), irregular exercise (OR=2.25), number of medication used (OR= 0.19) and lesser extent patient-physician relationship (OR=2.68) were independent risk factors for poor glycaemic control. CONCLUSION: The poor glycaemic control was associated with poor adherence to self-care adherence and their barriers in our diabetic population. Integrated knowledge on diabetes management should be targeted to improve glycaemic control in our communities.