Journal of Microbiology and Related Research

Background: The glycopeptide vancomycin was considered to be the best alternative for the treatment of multi drug resistant MRSA. However, there are increasing numbers of reports indicating the emergence of vancomycinresistant Staphylococcus aureus (VRSA) strains exhibiting two different resistance mechanisms. The emergence and spread of resistance to vancomycin is a threat to the already challenging therapy of MRSA. Materials and Methods: The present study was carried out to find out the presence of VISA and VRSA in the tertiary hospital. A total 570 staphylococcus aureus isolates consisting of 340 MRSA and 230 MSSA were isolated from different clinical specimens from various outpatient departments and wards using the standard techniques. All MRSA isolates were subjected to disc diffusion testing and MIC testing against vancomycin. Result and Discussion: Out of the 340 MRSA isolates, 270 isolates were Vancomycin sensitive Staphylococcus aureus (VSSA) (MIC: 0.5-2µg/ml), 70 isolates were Vancomycin intermediate Staphylococcus aureus (VISA) (MIC: 4-8µg/ml) and none were Vancomycin resistant Staphylococcus aureus (VRSA) (MIC: >16µg/l). The present study reveals for the first-time emergence of VISA/VRSA from this part of Karnataka, India. Keywords: Staphylococcus Aureus; MRSA; Vancomycin; VRSA; VISA.

Introducion: Acinetobacter is a heterogeneous group of gram negative, oxidase negative, non-motile, nonfermenters emerged as an important nosocomial pathogen causing outbreaks of hospital infections. The high prevalence of multidrug resistant isolates makes initiation of effective empiric treatment challenging. Objective: This study was undertaken to isolate and characterise Acinetobacter species in various clinical specimens and to analyse the antibiotic susceptibility pattern. Materials and Methods: A total of 5395 clinical specimens were processed in the department of microbiology of a tertiary care hospital over the period of 2 years. Out of which 147 isolates were Acinetobacter species. Speciation and antibiotic susceptibility was determined by the standard conventional method. ESBL and MBL production was detected by disc potentiation test method and imipenem EDTA combined disc test, and MBL E-test respectively. Results: Prevalence was 2.72%. Most predominant species was Acinetobacter baumannii 128 (87.07%). Maximum isolataion was seen among ICU patients (31.97%). Most of strains were resistant to ciprofloxacin (87.5%), ceftazidime (85.94%). All strains were resistant to piperacillin (100%) and sensitive to colistin and polymyxin B. ESBL, MBL production and MDR was detected in 34.01%, 21.77% and 53.06% of the isolates respectively. Discussion and Conclusion: A high level of antibiotic resistance was observed in our study and maximum isolation rate of Acinetobacter was in the ICUs associated with respiratory tract infection. Acineobacter baumani was the most predominant species. Other species of Acinetobacter are also isolated and encountered in hospital acquried infection, though they are sensitive to presently used antimicrobials but in future have potential to acquire resistance. The analysis of susceptibility pattern will be useful in understanding the epidemiology of this organism in our hospital setup, which will help in treating individual cases and controlling the spread of resistant isolates to other individuals. Keywords: Acinetobacter Species; Antimicrobial Susceptibility Pattern.

Background:Enterococci are important causes of both communities acquired and nosocomial infections. They show intrinsic resistance to a number of commonly used antibiotics, particularly the cephalosporins. During the last few years, Enterococci have acquired resistance to a number of important antibiotics including glycopeptides. Enterococci resistant to all three antimicrobial agents (penicillin, aminoglycosides and Vancomycin) pose a serious challenge not only for clinicians but also for health care institutions. It results in treatment failure, selection and spreading of resistant strains in the health care institution. The increasing occurrence of Enterococcusspecies, worldwide, since late 1980s, is of particular concern due to the emergence of Vancomycin Resistant Enterococci (VRE). VRE has also been reported from some parts of India. The appearance of VRE has limited the therapeutic options available for clinicians. Materials and Methods: Study was carried out in the Department of Microbiology, Shimogga Institute of Medical Sciences, Shimoga, between November 2015 and October 2018. Enterococci were isolated from various clinical samples at a tertiary care hospital using the standard techniques. The isolated Enterococci are then tested for routine antibiotics sensitivity by disc diffusion method including Vancomycin sensitivity. Results and Discussion:A total 330 Enterococcus isolates were obtained from various clinical specimens such as C. Among 330 Enterococcus species, 235 species were Enterococcus faecalis and 95 species are Enterococcus faecium. The Enterococcal species showed 100% sensitivity to Vancomycin and Linezolid. The ability of the laboratory to identify enterococci and to detect Vancomycin resistance promptly and accurately is essential in recognizing VRE colonization and infection and avoiding complex, costly containment efforts that are required when recognition of the problem is delayed. Further, acquisition of Vancomycin resistance leaves few options for therapeutic management. Keywords: Enterococcus Faecalis; Enterococcus Faecium; Vancomycin; Vre.

Background: Diabetes mellitus is a metabolic disorder in which there is increase in the levels of blood glucose because of insulin deficiency. Diabetic foot ulceration and infections are major medical, social, economical problem and it is the leading cause of morbidity and mortality in developing countries like India. The present study is an attempt made to know the aerobic bacteriological profile of diabetic foot ulcers. Material and methods: A total of 100 patients with diabetic ulcers admitted in surgical wards were studied. Pus was collected using two swabs from each patient one for staining and the other for aerobic culture. The organisms isolated were identified using standard techniques. Antimicrobial susceptibility of the bacterial isolates was done by Kirby-Bauer disc diffusion method. Result: Polymicrobial etiology was observed in 59% and monomicrobial etiology in 41%. A total of 165 organisms were isolated. Most common isolates were staphylococcus aureus 38 (23.03%), followed by Klebsiella spp 34 (20.6%), Pseudomonas aeruginosa 28 (16.96%), Escherichia.coli 26 (15.75%), Proteus spp 23 (13.93%), Enterococcus faecalis 8 (4.84%), Citrobacter spp 4 (2.42%) and Staphylococcus epidermidis 4(2.42%). Most sensitive antibiotics were Imipenem, amikacin, ciprofloxacin and Gentamicin. Conclusion: Culture and sensitivity from the wound plays an important role in prescribing the appropriate antibiotic at the time of admission itself rather than starting empirical treatment. Thus proper antibiotics policy and measures to restrict the indiscriminate use of antibiotics should be taken to minimize the emergence of drug resistant pathogens. Keywords: Diabetic Foot Ulcers; Staphylococcus aureus; Klebsiellaspp; Citrobacter Spp; Polymicrobial etiology.

Introduction: Group B streptococcus (GBS) is an important cause of illness in newborns, pregnant women as well as in non-pregnant elderly patients particularly those with significant underlying diseases. The study was carried out to determine the clinical spectrum, antibiogram and outcome of Group B Streptococcus infection as it can cause invasive infections in non-pregnant adults that can lead to substantial morbidity and mortality in them. Materials and Methods: A retrospective observational chart based study was conducted in a tertiary care centre for a period of one year (June 2016 to May 2017). Multisite samples like pus, wound swabs, blood, body fluids and urine samples were collected from male and female adult patients (adults were defined as >18 years old) with Group B streptococcus infections. The samples were processed for culture and sensitivity aerobically according to the standard operating guidelines. Statistical analysis of the data was performed using SPSS v.21 by frequency, percentage, Fisher’s exact test and a p value less than 0.5 was considered as significant. Results: In 156 cases of GBS infections among non-pregnant adults, the prevalence was higher in the age group of 50-64 years (37.8%) with female to male ratio of 1.1:1. The most frequent clinical presentations were urinary tract infections (65.4%) with a significant female gender predominance (p value <0.0001). All tested isolates were susceptible to Ampicillin, Cephalosporins, Penicillin and Vancomycin. Conclusion: Elderly people with underlying diseases are more prone to GBS infections. Therefore, evaluation of risk factors and development of an antibiotic policy is useful for the successful treatment of GBS infections. Keywords: Noninvasive; Non-Pregnant; Streptococcus Agalactiae; Group B Streptococcus.

Background: Pseudomonas species are the commonest pathogens causing nosocomial infections. Pseudomonas is basically resistant to many antibiotics and they are known to produce Extended Spectrum Beta Lactamase and Metalo beta lactamase. AIM: To detect Metalo beta lactamase producing Pseudomonas spp. from clinical samples in tertiary care hospital. Material and Methods: The study was agreed over a period of 6 months from January 2015 to June 2015. A total non repetitive of 329 Pseudomonas spp were isolated from unusual clinical samples like blood, pus and wound swabs, urine, body fluids, sputum, endo tracheal tube and secretions from the patients attending the hospital. Antimicrobial susceptibility test of all the isolates was performed by the disc-diffusion (Kirby Bauer disc diffusion method) according to CLSIs guidelines. All imipenem resistant isolates were tested for MBL production by Imipenem - EDTA double- disc synergy test (DDST) and Imipenem- EDTA combined disc test (CDT). Result: Of 329 samples, majority of the Pseudomonas spp were isolated from Pus/Wound 173 (52.58%) followed by Blood 103 (31.31%). The isolation rate was highest from pediatrics wards 112 (34.04%) and surgical wards 112 (34.04%). Total 329 samples, 24 (7.29%) isolates were MBL producer by CDST and DDST. Majority of the MBL producing Pseudomonas spp. were isolated from Pus/Wound 11 (45.83%) followed by Urine 10 (41.66%). The isolation rate was highest from Surgical wards 9 (37.5%) followed by the Pediatric wards 7 (29.16%). Conclusion: Detetion of Metallo beta lactamase isolates of Pseudomonas spp. will help to implement rational use of antibiotics and strictly adhere to the concept of “reserve drugs” are important to identify because it poses therapeutic problems and serious concern for infection control management. Keywords: Metalo Beta Lactamase (MBL); Pseudomonas Species; Nosocomial Infections.

Fungal infection is alarming phenomenon now a day because of increasing number of patients. Lactophenol Cotton Blue is widely used for staining, but there is a necessity to develop an alternate equipotent stain which can replace it due to its high level of tumorigenic and hazardous nature. iodine glycerol is better alternative for it. Iodine glycerol is safe, eco-friendly, good visual clarity, with better staining properties. Hence, we got here various clinical sampleswhichwere analysed simultaneously using Lacto-phenol Cotton Blue and Iodine Glycerol by using different techniques like teasing technique, slide culture technique and adhesive tape technique. Parameters like degree of transparency, visual clarity, resolution, contrast, staining characteristics like uniformity, formation of artefacts were analysed for better demonstration of fungal morphology. Iodine-Glycerol is a better alternative to Lacto-phenol Cotton Blue for the demonstration of fungal morphology in the clinical microbiological laboratories. It is eco-friendly, noncarcinogenic and much potent staining reagent. It is necessary to carry further research as there are no specific guidelines regarding the preparation of the Iodine-Glycerol staining reagent. Keywords: LPCB, Iodine Glycerol; Lugol’s Iodine; Fungal Morphology; Transparency; Staining Characters.

Introduction: Burn patients are at a risk of infection because of their destroyed skin barrier, suppressed immune system compounded by prolonged hospital stay and invasive therapeutic & diagnostic procedure. Despite various advances in infection control measures like early detection of causative agent and use of newer and broad spectrum antibiotics, management of burn septicemia still remains a big challenge. Aim and Objective: To study prevalence of various aerobic bacterial isolates among burn wound infection and to study their antimicrobial susceptibility pattern. Material and Methods: During study period of Jan- June 2017, a total of 369 pus samples were received and inoculated on different media as per standard protocol. Isolate were identified and antimicrobial susceptibility testing was done as per CLSI guidelines (2016). Results and observation: Out of 369 samples, 241 (65.31%) samples were from female and 128 (34.69%) from male. Among these 209 (56.64%) samples were having single isolate and in 125 (33.88%) samples, two types of bacteria were isolated. Out of 459 isolates, 133 Gram positive cocci and 326 Gram negative bacilli were isolated. Among various bacterial isolates Pseudomonas aeruginosa was most commonly isolated bacteria followed by Staphylococcus spp. and Klebsiella pnemoniae. Most of the isolates were resistant to routinely used antimicrobial agents. Conclusion: P. aeruginosa and S. aureus are the leading cause of infection in burn patients and isolation of multidrug resistant organism should be considered as a serious risk in burn unit. Early identification of infection caused by multidrug resistant organisms might help to modify treatment and outcome in burn patient. Keywords: Burn Wound Infection; P. Aeruginosa; S. Aureus and Multidrug Resistance.