Background: The diagnosis of chronic hepatitis B (CHB) infection has progressed from serological to molecular diagnostic methods. The newer sensitive technique of quantitation of hepatitis B virus (HBV) DNA by real-time polymerase chain reaction (RT-PCR) has helped in understanding the clinical stages of CHB, deciding on treatment and monitoring treatment response. Aim: This study aimed to determine the HBV DNA load by quantitative RT-PCR in the various clinical stages of CHB. Materials and Methods: Blood samples of CHB patients from the Gastroenterology Department received from June 2014 to December 2014 in the Microbiology Department were subjected to quantitative PCR analysis for HBV DNA. However, to facilitate analysis, only those patients' samples where hepatitis B e antigen (HBeAg) and alanine aminotransferase (ALT) status were known were selected for the study. Statistical analysis was done using SPSS (PASW statistics 18) for windows software. Chi-square test was used to analyse the differences in DNA level between the study groups. Results: A total of 71 CHB patients were included in the study. Of these, 29 (40.8%) were inactive carriers (HBeAg-negative ALT normal) and 42 (59.2%) were chronic active hepatitis B patients (ALT elevated with HBeAg-positive and HBeAg-negative cases). HBeAg was positive in 26 (36.6%) and negative in 45 (63.3%) patients. Among the 45 HBeAg-negative patients, 16 (22.5%) had CHB. Of the 71 CHB patients, 61 (85.9%) had detectable viral load. Serum HBV DNA load of 16 patients who were HBeAg-negative was significantly lower (median 5.5 × 10⁵) than that of 26 patients who were HBeAg-positive (median 2.4 × 10⁸) and higher than the 29 inactive carriers (median 1.6 × 10³). Based on HBV load, 14 CHB patients who were HBeAg-positive and seven who were HBeAg-negative were started on antiviral therapy. Conclusion: Quantitation of HBV DNA based on HBeAg and ALT status helps to determine the stages of CHB. It could play an important role in assessing the status of those patients who are HBeAg-negative and inactive carriers with respect to viral load, as the former require treatment. The major role of HBV DNA determination is to evaluate patients with CHB (HBeAg-positive or HBeAg-negative) and to decide on antiviral therapy.

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Background: The pathogenic bacterium on the hands of healthcare workers (HCWs) is considered as the main route of spread of nosocomial infections. The study was aimed to determine the bacterial contamination of hands of HCWs during routine patient care in a large teaching hospital. Material and Methods: This study includes a total of 200 consecutive, non-repetitive participants. Doctors, medical students, nurses and attendants from the Department of Anaesthesia and Department of Medicine of the hospital were included, who were actively providing health care. The informed consent was taken from all the participants of the study. The hand impressions of the participants were taken on 5% blood agar and processed as per guidelines. After taking the hand impressions, hand cleaning of each participant was done by standard preparation of 70% isopropyl alcohol-based hand rub. The hand impressions from the participants were again taken on 5% blood agar plates. Results: Out of 200 samples collected from HCWs, 95 (47.5%) samples showed growth of microorganisms. Most commonly isolated microorganism was Staphylococcus aureus present in 70% of nurses, 60% of students and 40% of attendants. Prevalence of Acinetobacter spp. was also found to be high among nurses (20%), followed by attendants, sweepers (16%) and doctors (4%). Conclusions: Hands of hospital staff are colonised with pathogenic bacteria, especially Gram-positive such as S. aureus. Hand hygiene is an effective method of reducing the bacterial flora on the hands of the HCWs.

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Background: Onychomycosis is most commonly caused by dermatophytes belonging to the genus Trichophyton. However, Candida spp. and non-dermatophyte moulds may also be responsible. Up to 10% of cases of onychomycosis are caused by non-dermatophyte moulds and these are becoming more common worldwide. Materials and Methods: A retrospective analysis for a period of one year was undertaken to know the prevalence of different fung causing onychomycosis in a tertiary care hospital. Nail samples of fingers and toes collected from patients presenting with clinically suspected onychomycosis were processed by standard methods. Results: Of the 87 clinically suspected cases of onychomycosis, non-dermatophyte moulds were the most common pathogens isolated in 24 (53.33%) patients, followed by dermatophytes in 12 (26.66%) patients and Candida spp. in 9 (20%). Among the non-dermatophyte moulds, Aspergillus flavus was the most common isolate found in 15 (62.5%) cases. Conclusion: The predominant isolate obtained in this study was non-dermatophyte moulds. Among moulds, Aspergillus spp. was the most common isolate.

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Background and Objectives: Effectiveness of prophylactic antibiotics is based on the susceptibility of the common potential pathogens to those antibiotics. Irrational use of drugs has an adverse impact on treatment outcome and financial burden to patients. Aims: Objective of this study was to evaluate the effect of antibiotic prophylaxis on surgical site infections and wound colonisation. Materials and Methods: A cross-sectional study was conducted among the patients of general surgery wards of a tertiary care hospital of central Kerala. Swabs collected from surgical wound sites after 48 h of surgery were analysed for bacteriological profile and antibiotic prophylaxis given. Statistical analysis was done by Epi Info. Results: Three hundred and ten cases were studied including 75.5% elective and 24.5% emergency cases. Cefotaxime (55.2%) and third generation cephalosporin combination with β-lactamase inhibitors (35.5%) were commonly used for antibiotic prophylaxis. Infection rates were 2.1% in clean surgeries, 8.1% in clean-contaminated and 22.8% in contaminated types of surgeries with an overall infection rate of 8.1%. Escherichia coli (45%) and Staphylococcus aureus (34.5%) were the predominant wound pathogens. Colonisation in 28.1% wounds with S. aureus as the major coloniser (72.8%) was noticed. Susceptibility of E. coli to Cefotaxime was 15.4% in infections and 20% in colonisation. Infection rates were higher in the group with multiple antibiotic usages for 5 days (40%) or above (52%). Colonisation increased on prolonged administration of multiple antibiotics (40%) and without using antibiotics (39%). Clean wounds without antibiotic prophylaxis were not infected. Conclusions: This study revealed that antibiotic prophylaxis is not required for clean wounds. Short duration of antibiotics usage based on the local susceptibility pattern is to be adopted. Antibiotic usage has to be restricted, and prophylactic regime with third-generation cephalosporin has to be curtailed.

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Chromobacterium violaceum is a rare pathogen that causes fatal infections in humans. Prompt diagnosis and specific treatment is important because infections caused by C. violaceum have a propensity to develop into fatal septicaemia unless appropriately treated. Here, we report a case of catheter-related blood stream infection caused by C. violaceum in a 55-year-old diabetic female patient.

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Melioidosis is a suppurative chronic infection caused by a Gram-negative bacterium, Burkholderia pseudomallei. A 41-year-old male diabetic patient presented with fever and abdominal pain. On computed tomography scan, he was found to have splenic abscess. Culture of splenic tissue yielded B. pseudomallei. He was treated with Ceftazidime and Cotrimoxazole.

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Background: An increase in the prevalence of infections caused by non-albicans Candida (NAC) has been reported from many parts of the world. The increased isolation rates of NAC and a gradual shift in the antifungal susceptibility profile underlines the need for early and accurate diagnosis of Candida infections along with antifungal susceptibility testing. Aim: The aim of this study is to characterise Candida spp. isolated from various clinical samples and determine the antifungal susceptibility pattern. Settings and Design: This study was conducted in the Department of Microbiology, at a tertiary care referral centre, over a period of one year and included 200 Candida spp. Materials and Methods: Candida isolates were characterised by conventional techniques and CHROMagar. Antifungal susceptibility test was performed using disc diffusion method. Clinical details and risk factors were recorded and analysed. Results and Conclusions: Candida albicans was the most common species isolated, followed by Candida tropicalis. The Candida isolates were more susceptible to Amphotericin B than other antifungal agents tested. Diabetes mellitus appeared to be the most common predisposing factor for the Candida infections, followed by indiscriminate drug usage.

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Extended spectrum beta lactamase (ESBL) production is a major resistance mechanism of Gram-negative bacteria. Phenotypic tests for ESBL detection only confirm whether an ESBL is produced or not, but it cannot detect the ESBL subtype. There are various genotypes of ESBLs. Of these, the most common are the TEM, SHV and CTX-M types. Fifty phenotypically ESBL-producing Escherichia coli and Klebsiella pneumoniae urinary isolates were subjected to polymerase chain reaction to look for the presence of bla_TEM, bla_SHVand bla_CTX-Mgenes. Forty-one isolates were positive for TEM gene, eleven isolates were positive for SHV gene and 10 isolates were positive for CTX-M gene. Both TEM and SHV genes were present in six isolates. Two isolates showed the simultaneous presence of CTX-M and TEM genes. All the three genes were present in two isolates.

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Background and Objectives: Exogenous osteomyelitis frequently follows traumatic or surgical inoculation of bacteria into bone and surrounding tissue. It is usually associated with open fractures, surgical implants, orthopaedic fixation devices and vascular insufficiencies such as diabetes mellitus, peripheral vascular diseases and presence of foreign bodies. The aim of this study was to identify the pathogenic bacteria causing osteomyelitis and to evaluate the prognosis and outcome of patients with exogenous osteomyelitis. Materials and Methods: Patients admitted with clinical and radiological features of exogenous osteomyelitis were selected for the study from June 2005 to October 2006. Bone curettings or aspirated materials from the sinus tract were collected. The specimens were cultured on appropriate media for bacterial culture. The patients who consented were followed up for 6 months to record their progress and outcome. Those followed up included both culture positives and negatives. Results: Out of 125 cases of osteomyelitis, 75 (60%) were positive by culture, 59 (78.7%) were monomicrobial infections and 16 (21.3%) were polymicrobial infections. Staphylococcus aureu s, 56 (60.8%) in number, was the predominant organism isolated followed by Escherichia coli, eight (8.7%); Klebsiella pneumoniae, seven (7%); Pseudomonas aeruginosa, five (5.4%); β-haemolytic streptococci, four (4.4%); Proteus mirabilis, three (3.2%); Enterococcus faecalis, two (2.2%); Acinetobacter baumannii, two (2.2%); Corynebacterium jeikeium, one (1.1%); Staphylococcus epidermidis, one (1.1%); Proteus vulgaris, one (1.1%); Kingella species one (1.1%) and Arcanobacterium species, one (1.1%). Out of the coagulase-positive staphylococci, 29 (51.8%) were Methicillin-resistant S. aureus. Of the 61 cases that were followed up, 44 cases were culture positive. The total cure rate was 60.2%. Conclusion: Appropriate antibiotic therapy after culture and sensitivity has a major role in treating exogenous osteomyelitis in the presence of fracture and instability of bone. In spite of appropriate surgical correction, the major risk factors that led to recurrence of infections and amputations were overcrowding in wards, extensive tissue injury, poor economic status of patients, non-availability of drugs in the hospital pharmacy, poor compliance of patients and drug toxicity.

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Pneumocystis jirovecii is widely known as an important cause of pneumonia in immunocompromised hosts. It remains a leading cause of opportunistic infection, morbidity and mortality in these patients. Here, a case of severe P. jirovecii pneumonia in a post-renal transplant patient is described. Giemsa and Gomor methenamine silver stain of sputum showed cysts of P. jirovecii. Despite treatment with Trimethoprim- Sulfamethoxazole, the patient developed severe respiratory failure and expired.

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Propionibacterium species are a part of the normal microbial flora of the body and are usually found as contaminants of blood culture. Here, we report a case of Propionibacterium spp. causing lung abscess in a 69-year-old male smoker, alcoholic and diabetic. After taking treatment for about 1 year, he was referred to the Government Medical College, Thiruvananthapuram where Propionibacterium species, sensitive to Penicillin was isolated from his sputum repeatedly. He recovered completely after 6 weeks of oral Penicillin therapy.

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An 8-month-old infant presented with history of recurrent urinary tract infection and was found to have a perinephric abscess on investigation. Enterococcus faecium resistant to Vancomycin and sensitive to Linezolid was obtained from the baby's urine and perinephric collection. The patient was treated successfully with Linezolid. This case report describes the emergence of Vancomycin-resistant enterococc in this hospital.

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