Background: Genetic changeability of hepatitis B virus (HBV) signifies a challenge for the sensitivity of immunologic and molecular diagnostics. Therefore, knowing the spread of HBV genotypes (GENs) and mutation has considerable impacts on treatment strategies, vaccination program, diagnosis, and prevention. The present study aimed to detect HBV GENs and mutants in HBsAg-positive patients. Methods: The study conducted on 4927 patients in Meerut, India, between March 2013 and April 2017. The blood specimens were analyzed for HBsAg using an ELISA kit, then the blood samples from HBsAg-positive patients were subjected to HBeAg assay and DNA isolation. Amplification of the HBV DNA of pre-S gene and pre-core or basal core promoter region were performed by RT-PCR and sequenced to analyze both GEN and mutation. Results: According to the results, 245 cases were positive for HBsAg, and 55 were HBeAg-positive. With regard to HBV DNA levels, 16 samples were found positive in PCR assay with 7 (43.8%) less than 2000 IU/mL, 4 (25%) between >2000 and 20,000 IU/mL, and 5 (3.25%) >20,000 IU/mL. No mutations were detected in GENs B and A. The prevalence of HBV GENs B and A were 68.8% (n = 11) and 31.25% (n = 6), respectively. Conclusion: GEN-B was more prevalent in comparison to GEN-A. The genetic diversity of HBV and distribution of its GENs and mutation improve the current knowledge of epidemiological, clinical and virological patterns of hepatitis B in this region, which help physicians to prescribe proper antiviral/interferon therapy according to current genotyping pattern.

Objective:-Hepatitis B is noteworthy medical issues that may include the late continuation of liver cirrhosis and hepatocellular carcinoma. The present study aimed to know the seroprevalence of hepatitis B Virus in Meerut with different clinical conditions. Methods: The studyconducted on 4927 patients in Meerut, India, which was performed in central research center labortory of Microbiology at netaji subhash Chandra Bose Medical College and Hospital Between January 2013 to April 2017 The sera were separated and screened for HBsAg byimmune-chromatographic card test then HBsAg positive serum sampleswere tested again for HBsAg using ELISA kit. Positive samples for HBsAgwere tested for HBeAg ELISA kit. Results: In 245 positive cases 118 (48.16 %) were male and 127 (51.84%) were female. The seroprevalence rate of HBV was 4.97% in Meerut, India. Age group21–30 was the highest seroprevalence (7.46%) when compaired to other age group. Of the 245 HBsAg Positive case 55 (1.12%) were HBeAg positive. 20 were male, Highest HBeAg positive male were found in 21- 30age group. Out of 55 HBeAg cases, 35 were female and 92 female were negative for HBeAg, this was stastically significant (P< 0.039) by using Z test. Conclusions: The study uncovered that the seroprevalence of HBV was alarmingly higher in such a populace, which presumably mirrors a high foundation predominance of HBV contaminations ought to be mulled over and Implementation of group based preventive measures and enhanced procedures for safe blood supply may demonstrate valuable to diminish the seroprevalence Bangladesh Journal of Medical Science Vol.18(2) 2019 p.329-333

Objective:- Hepatitis B is noteworthy medical issues that may include the late continuation of liver cirrhosis and hepatocellular carcinoma. The present study aimed for the detection and diffrentiation of Hepatitis B virus HBsAg inactive non-replicative carriers, HBeAg-positive inactive replicative carriers, active carriers & HBeAg-negative chronic hepatitis B by Real Time PCR and their genotyping Methods: This research conducted on 245 positive for HBsAg, 118 (48.16 %) were male and 127 (51.84%) were female patients, which was performed in central research station labortory of Microbiology at netaji subhash Chandra Bose subharti Medical College and Hospital, Meerut Between march 2016 to November 2017 The sera were separated and screened for HBsAg by ELISA kit. Positive samples for HBsAg were tested for HBeAg ELISA kit and DNA Viral load then sequenced for genotying Results:. Of the 245 HBsAg Positive case 55 (1.12%) were HBeAg positive. In 16 PCR positive and HBV genotyping, In HBsAg inactive Non-Replicative 37.5% (n=6) genotype-B and 6.25% (n=1) genotype-A, In HBeAg inactive Replicative 12.5% (n=2) genotype-B and 12.5% (n=2) genotype-A and In HBeAg Active Chronic Hepatitis B 18.75% (n=3) genotype-B and 12.5% (n=2) genotype-A were detected Conclusions: Management strategy, using HBsAg, HBeAg and HBV DNA viral load, seems adequate for the confirmation and diffrentiation of Hepatitis B virus inactive, active carriers & HBeAg-negative chronic hepatitis B patients and genotype B was more prevalent in comparission to genotype A. Distribution of carriers & genotypes, help physicians to prescribe proper antiviral/interferon therapy according to current genotyping pattern in this region Keywords: Hepatitis B virus, Carrier State, HBsAg, HBeAg, RT-PCR

Salmonella  urinary tract infections (UTIs) are rare. Salmonella Typhi bacteriuria,  an unusual  finding, can be seen following a recent episode of typhoid fever, or in chronic carrier state involving the urinary system, especially with local abnormalities of urinary tract. We report a rare case of Salmonella urinary tract infection in a young male patient having bladder dysfunction with vesico-ureteric reflux with history of fulguration for posterior urethral valves in his childhood, not reported so far to the best of our knowledge.

Urinary tract Infections caused by extended-spectrum β-lactamase (ESBL), producing Klebsiella pneumoniae are on a rise all over the world with high morbidity and mortality. This study was carried out to determine the presence of TEM, SHV and CTX-M genes in extended-spectrum β-lactamase (ESBL) producing Klebsiella pneumoniae. A total of 300 Klebsiella pneumoniae isolates were collected and identified using traditional culturing and biochemical tests. Antibiotic susceptibility testing was performed by disc-diffusion method according to the CLSI guideline. Isolates were screened for ESBL and confirmed by phenotypic confirmatory disc diffusion test (PCDDT). 100 randomly selected isolates tested for the presence of ESBL encoding genes using PCR with specific primers for the detection of CTX-M, SHV and TEM genes using a standard protocol. Imipenem showed the highest antibacterial activity against ESBL producing K. pneumoniae. Based on the results of PCR, the prevalence of TEM, SHV and CTX-M genes among ESBLs-positive isolates was 74%, 27%, and 44% respectively. In conclusion, the rate of ESBL-producing K. pneumoniae was high in the present study. The bacterial resistance to many classes of antibiotic leads to limited treatment options. Since the management of infections caused by these organisms is difficult, it is important to control such strains in order to prevent and reduce their spread.

BACKGROUNDThe term otomycosis simply refers to superficial fungal infection of the external auditory canal. Most patients suffering from early otomycosis complain of severe itching which often progress to pain, hearing loss and tympanic membrane perforation. Aims-To determine the predisposing factors, symptoms and to identify the fungus and secondary bacterial infection associated with it.MATERIALS AND METHODSA total of 100 consecutive patients with clinical diagnosis of otomycosis were considered as the study group and 100 consecutive patients with no diagnosis of otomycosis were considered as the control group from a total of 200 individuals attending the Outpatient Department of ENT at Chhatrapati Shivaji Subharti Hospital (CSSH), a tertiary care teaching hospital in Meerut City, were chosen for the study.RESULTSHistory of wooden stick or any other object used to remove wax (68%), use of oil (44%), swimming (28%) and history of trauma (23%) were the common predisposing factors in otomycosis. Pruritus and otalgia (89% each) were the most common complaints. Statistical association (p value<0.001) was found between isolation of fungi and otomycosis whereas no statistical association was found between otomycosis and bacterial association. The fungi isolated were mainly Aspergillus niger (77.0%) followed by A. flavus (12.6%), A. terreus (4.6%), Candida guilliermondii and C. krusei (1.1% each). Bacterial isolates were Staphylococcus epidermidis (21%), S. aureus (7%), Klebsiella pneumoniae (2%) and Enterobacter aerogenes (1%). Fungal pathogens were isolated from external auditory canal of 8% of controls.CONCLUSIONPruritis and otalgia (89% each) were the common complaints followed by discharge, tinnitus and hearing loss. Aspergillus (94.2%) was the most common isolate from otomycosis in our study, followed by Candida (2.3%) as in studies across the world. Isolation of Aspergillus spp. such as A. terreus isolate is important, as they are largely resistant to antifungal drug Amphotericin B. It usually resolves with aural toilet and installation of antifungal drugs. Recurrence of the disease is common after treatment. We had a recurrence rate of 20% as compared to 5% quoted in literature. This may be due to patients having poor hygiene, diabetes mellitus, community bathing and poor compliance to treatment.

Introduction: Chronic Suppurative Otitis Media (CSOM) still is a common health care problem in developing countries like India with its potential for serious local damage and complications. Knowledge of causative microorganisms and their antimicrobial sensitivity pattern is essential so that early and effective therapeutic measures can be initiated for better patient outcome. Aim: The study was carried out to determine the clinicomicrobiological spectrum of CSOM from this geographical area and to evaluate the susceptibility pattern of aerobic bacterial isolates. Materials and Methods: The prospective study was carried out for a period of one year from December 2013 to November 2014 in Chhatrapati Shivaji Subharti Hospital, a Tertiary Care Hospital in Meerut city. A total of 113 ear discharge was collected from clinically suspected cases of CSOM. The discharge was cultured on blood agar and Mac Conkey’s agar plates for isolation of bacterial pathogen and Sabourauds Dextrose Agar (SDA) slants for isolation of fungal pathogens. Identification of the bacterial and fungal isolates was done using standard bacteriological and mycological methods respectively. Antimicrobial susceptibility testing of the bacterial isolates was performed by Kirby-Bauer’s disc diffusion method as per the Clinical Laboratory Standards Institute (CLSI) guidelines. Further, detection of Metallo Beta Lactamase (MBL) production, Methicillin Resistance in Staphylococcus aureus (MRSA) and Extended Spectrum Beta –Lactamase (ESBL) production was carried out by phenotypic methods. Result: A total of 73.45% cases were culture positive. Pure bacterial pathogen was isolated from 67.46 % followed by pure fungal pathogen 22.89% and mixed pathogen (bacteria and fungus) from 9.63% of cases. Pseudomonas aeruginosa and Staphylococcus aureus were the predominant bacterial pathogen and Aspergillus spp. and Candida species were the predominant fungal pathogens isolated. Overall the rate of MBL producers, MRSA and ESBL producers were 18.18%, 52.94% and 62.5% respectively. Conclusion: High level of resistance was observed in cases of CSOM from our hospital. Therefore, knowledge of the causative microorganisms and its susceptibility pattern is required and this data may contribute to an effective management of cases of CSOM.

INTRODUCTIONEnterococci are recognized as opportunistic pathogens, as well as commensals in both humans and animals. They are an important cause of nosocomial infections, difficult to treat, as the organism have intrinsic and acquired resistance to many antibiotics.AIMTo isolate and identify clinically relevant Enterococcus up to species level from all the clinical samples processed in the microbiology laboratory and also to study their resistance pattern.MATERIALS AND METHODSA prospective study was carried out for a period of one year from May 2014 to April 2015 at the Department of Microbiology, Subharti Medical College, Meerut, Uttar Pradesh, India. A total of 200 isolates of Enterococcus species from 15342 clinical samples obtained from IPD/OPD patients irrespective of age, having suspicion of bacterial infection were processed in the microbiology laboratory. Identification was done with standard biochemical methods. Antibiotic susceptibility testing was done on Muller Hilton agar plate by Kirby Bauer disc diffusion method. High-Level Gentamycin Resistance (HLGR) and High-Level Streptomycin Resistance (HLSR) were further confirmed by Agar dilution method and Broth microdilution method. Vancomycin and linezolid resistance was further confirmed by Agar dilution method and MIC was calculated by using VITEK 2, Biomerieux. All methodology was followed as per Clinical and Laboratory Standards Institute (CLSI) M100-S 24, 2014 guidelines.RESULTSE. faecalis (n=169, 84.5%) was the predominant species isolated, followed by E. faecium (n=27, 13.5%) and E. casseliflavus (n=4, 2%). A total of 25 (12.5%) isolates were HLGR, 13 (6.5%) isolates were HLSR and 62 (31%) isolates were HLGR+HLSR. Vancomycin resistance was found in 14 (7%) isolates of which 11 (78.5%) were Van A and 3 (21.4%) were Van B, detected phenotypically as per relative MIC of vancomycin and teicoplanin. Linezolid resistance was seen in 4 (2%) of isolates which were vancomycin as well as high-level gentamycin and high-level streptomycin resistant.CONCLUSIONIsolation of Enterococcus species resistant to most of the higher antibiotics like vancomycin and linezolid, with high prevalence of High-Level Aminoglycoside Resistance (HLAR), from hospitalized patients is a major concern as such isolates have limited or no therapeutic option.

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most common hospital-acquired infections associated with a significant morbidity and mortality. Healthcare workers (HCWs) act as carriers for MRSA and transmit the organism across individuals. Hence, we undertook this study to assess the prevalence of MRSA among HCWs in a tertiary care hospital and their hand washing hygienic practices. Materials and Methods: A cross-sectional study between March and November 2014 was conducted. Dry swabs were taken from the nasal mucosal membrane, both the palms, web space between the fingers from each of the participants and were subsequently cultured on mannitol salt agar and antibiotic susceptibility to methicillin was assessed using Kirby-Bauer disc diffusion method using cefoxitin. In addition, information on demographics, occupation, and use of antiseptics were collected from the study participants. Results: A total of 17/150 (11.3%) were found to be MRSA positive of which 10 were staff nurses, three each were laboratory technicians and class IV employees and one was a doctor. Of the 17, 11 (64.7%) had shown growth from nasal swabs, 5 (29.4%) from digital and 1 (5.9%) in both the regions. Of them, 8/15 (53.3%) reported the use of only soap for hand washing whereas 7/15 (46.7%) were using soap and hand sanitizers and the remaining two not responded to this question. Only 9/15 (60%) frequently washed their hands, and the rest rarely did. Conclusion: More awareness and training sessions on the importance of MRSA and hand hygienic measures to contain the carrier status of MRSA among hospital care workers is warranted.

Malaria is a heavy burden on tropical communities, a threat to non-endemic countries, and a danger to travellers. Keeping in mind the seriousness of the condition and the current availability of various diagnostic facilities; this study is proposed to compare the peripheral blood smear examinations (thick and thin), Quantitative Buffy Coat (QBC) examination, Rapid Antigen Detection Test (RDT), and Modified Centrifuged Blood Smear (MCBS) in clinically suspected cases of malaria. The study included 1982 blood samples of patients, clinically suspected to be of Malaria. All the samples were subjected to Thin Blood Smear, Thick Blood Smear, QBC, RDT and MCBS. Results were statistically compared considering MCBS as the standard method. Out of total 1982 samples, maximum 81(4.09%) were found positive by RDT, followed by MCBS 66(3.33%), QBC 65(3.28%), thick blood smear 63(3.18%) and thin blood smear 58(2.92%). Addition of centrifugation step in MCBS increases the sensitivity of the smear technique and at the same time preserves the morphology of the parasite, giving it more specificity. MCBS is an easy, cheap and accurate technique and could be adopted for more reliable diagnosis of malaria in resource-limited settings where expensive techniques like RDT and QBC are not feasible.

Introduction: Bio-Medical Waste (BMW) management is of utmost importance as its improper management poses serious threat to health care workers, waste handlers, patients, care givers, community and finally the environment. Simultaneously, the health care providers should know the quantity of waste generated in their facility and try to reduce the waste generation in day-to-day work because lesser amount of BMW means a lesser burden on waste disposal work and cost saving.Aim: To have an overview of management of BMW in a tertiary care teaching hospital so that effective interventions and implementations can be carried out for better outcome.Materials and Methods: The observational study was carried out over a period of five months from January 2016 to May 2016 in Chhatrapati Shivaji Subharti Hospital, Meerut by the Infection Control Team (ICT). Assessment of knowledge was carried out by asking set of questions individually and practice regarding awareness of BMW Management among the Health Care Personnel (HCP) was carried out by direct observation in the workplace. Further, the total BMW generated from the present setup in kilogram per bed per day was calculated by dividing the mean waste generated per day by the number of occupied beds.Results: Segregation of BMW was being done at the site of generation in almost all the areas of the hospital in color coded polythene bags as per the hospital protocol. The different types of waste being collected were infectious solid waste in red bag, soiled infectious waste in yellow bag and sharp waste in puncture proof container and blue bag. Though awareness (knowledge) about segregation of BMW was seen in 90% of the HCP, 30%-35% did not practice. Out of the total waste generated (57912 kg.), 8686.8 kg. (15%) was infectious waste. Average infectious waste generated was 0.341 Kg per bed per day. The transport, treatment and disposal of each collected waste were outsourced and carried out by 'Synergy' waste management Pvt. Ltd.Conclusion: The practice of BMW Management was lacking in 30-35% HCP which may lead to mixing of the 15% infectious waste with the remaining non-infectious. Therefore, training courses and awareness programs about BMW management will be carried out every month targeting smaller groups.

HIV/AIDS has turned out into global pandemic. Integrated Counselling and Testing Centres (ICTC) are entry points for a wide range of intervention in HIV prevention and care. To assess socio-demographic profiles, sero-positivity and risk behaviour pattern of clients attending ICTC. ICTC attached to Department of Microbiology, Subharti Medical College, Meerut. A retrospective studywas conducted among clients who visited ICTC either by self-initiative or referred by a doctor or non- governmental organization (NGO) from January 2012 to December 2014. Chi square test and Z- proportion test. Overall prevalence of HIV positivity was 1.12% of the clients attending an ICTC. A total of 81.08% males and 83.69% females who were seropositive were in the age group of 15-49years. A total of 88.04% of females were housewives, 31.08% males were having business, 72.15% were married and 66.77% were from rural areas. Hetero- sexual route was the most common route of transmission in 78.16% clients. Best antidote for HIV/AIDS remains increased awareness and adaptation of safe behavioural practices. Therefore, epidemiological studies should be carried out to find out behavioural, social and demographic factors which help in interruption and control of disease.

Shigella flexneri is the most commonly isolated species in the developing world. Prompt and appropriate therapy is necessary in shortening the duration of clinical symptoms, as well as to prevent transmission of infection to close contacts. Emerging multidrug resistant strain of Shigella is a growing concern across the globe and in India. Herein we report a case of dysentery in a child caused by an extended spectrum beta lactamase (ESBL) producing and multidrug resistant (MDR) S. flexeneri so as to alert the clinicians, microbiologists and epidemiologists regarding the emergence of resistance in Shigella spp. An outbreak of shigellosis by such resistant strains in overcrowded communities will be a tough challenge in the containment of the disease.

BACKGROUND Vancomycin may be ineffective against an increasing proportion of methicillin resistant Staphylococcus aureus (MRSA) with minimum inhibitory concentrations (MICs) well within the susceptible range. On the other hand it is common knowledge that determination of vancomycin MICs is method dependent. Therefore, given the apparent variability in vancomycin MIC results obtained with the different methods, the use of the vancomycin MIC to predict the outcome of serious S. aureus infections needs to take into account the method used and the results of studies using that particular method. AIM Comparative study was carried out to evaluate the MICs obtained by BMD method, E-test, and Vitek 2 method and to detect inconsistencies in these vancomycin for 66 MRSA isolates obtained from various samples of patients attending the OPDs & IPDs within a period of one year. MATERIALS AND METHODS A comparative study was carried out to evaluate the MICs obtained by BMD method, E-test, and Vitek 2 method to detect vancomycin susceptibility in 66 clinical isolates of MRSA obtained from various samples of patients attending the OPDs & IPDs within a period of one year. The study was conducted in Department of Microbiology, Subharti Medical College, Meerut from January to December 2012. RESULTS On determination of MICs for vancomycin for the MRSA isolates, all were identified as VSSA by BMD, E-Test & Vitek 2 methods. However, the vancomycin MIC values obtained by E-test correlated better with BMD method (correlation factor= 0.6727) than Vitek 2 (correlation factor=0.5316), indicating E-Test to be a better method for determination of vancomycin MICs as compared to Vitek 2. CONCLUSION MRSA isolates with higher vancomycin MICs, even within the susceptibility range, are being observed more frequently which result in treatment failures with vancomycin. Because of the discrepancy that exists in vancomycin MIC results from different methods, the prediction of outcome of serious S.aureus infections should take into account the method used & results of studies using that particular method.

Chronic hepatitis B affects over 350 million people worldwide. India represents the second largest pool of chronic hepatitis B virus (HBV) infection in the world with an estimated 40 million infected people. Vertical transmission is known to be the leading cause of infection and perinatal infection is associated with a very high rate of chronicity (up to 90%). Hepatitis B e antigen (HBeAg) positivity indicates that replicative form of HBV may play a role in immunotolerance in utero by crossing the placenta. Up to 40% of chronically infected individuals will die prematurely from complications such as acute liver failure, cirrhosis and hepatocellular cancer. In case of HBeAg positivity and high viral load of mother, HBV immunoglobulin is preferably given along with HBV vaccination. Antiviral therapy is recommended for use in the third trimester of pregnancy to reduce the perinatal transmission of HBV, however, use of antiviral therapy should be individualized during pregnancy. Addressing perinatal transmission through the use of immunoprophylaxis can help contain the spread of HBV. Pregnant mothers with chronic hepatitis B have unique challenges and require specialized management during and after pregnancy. This review will look at the screening of pregnant women for hepatitis B, passive and active immunoprophylaxis, mechanisms of perinatal viral transmission and therapeutic considerations in pregnancy including possible teratogenicity and efficacy of medication. Other issues such as the mode of transmission and breastfeeding will be covered.

A 40-year-old male patient presented to us with complaints of pain in abdomen for the past 2 weeks and fever for 3 days. The patient gave history of being previously operated for hydatid cyst of liver 15 years back. His chest radiograph and computed tomography scan revealed a cystic lesion in the right lobe of liver and a cyst in left lung. The patient was managed surgically. Aspirate from the cyst fluid showed plenty of hooklets and scolices of Echinococcus granulosus. An intact brood capsule was also seen. Diagnosis of hydatidosis was further confirmed by histopathological examination. Post-operative the patient had a good recovery.

Introduction: Biofilm forming microorganisms are related to chronic and recurrent human infections as well as are highly resistant to antimicrobial agents. Various methods have been used in medical field for the detection of biofilm production which includes visual assessment by electron microscopy and polymerase chain reaction, quantitative method like Tissue Culture Plate (TCP) method and two qualitative methods such as Tube Method (TM) and Congo Red Agar (CRA) method. Objectives: To detect the prevalence of biofilm formation in Staphylococci, to evaluate two different methods i.e. TM and CRA for the detection of biofilms and to see its relation with antimicrobial resistance. Materials and Method: The study was conducted at Department of Microbiology, Dr. D.Y. Patil Hospital and Research Centre during the period June 2011 to June 2013. A total of 130 clinical isolates were subjected to biofilm detection methods. Standard microbiological procedure was done to identify the isolates. Biofilm detection was tested by TM and CRA. Kirby-Bauer disc diffusion technique was performed to do antibiotic susceptibility test of biofilm producing bacteria according to CLSI guidelines. Results: From the total of 130 clinical isolates, biofilm productions among Staphylococci were 59.23%. And the percentage of biofilm detection of Staphylococci isolates by Congo red method were 31.54% and by test tube method 50%. Hence, the TM method was considered to be superior to CRA. We have also observed higher antibiotic resistance in biofilm producing bacteria than non-biofilm producers. Conclusion: We can conclude from our study that the TM method is a more suitable and reliable method as compared to CRA and can be recommended as a general screening method in laboratories for detecting biofilm forming bacteria.