Indian Journal of Microbiology Research

126-130

Authors: Norah Saad J. Bin Saad.

Antimicrobial agents are compounds which inhibit the growth, or kill microorganisms. Antimicrobial agents can be used for either treatment or prevention of infectious disease but must be used properly and accurately. The use of antimicrobials in the prevention of disease is called prophylaxis. There are four occasions when prophylactic antimicrobials are indicated and these are: · When the risk of post-operative infection is high. · When wounds are contaminated with soil or dirt (e.g. after road traffic accidents) and there is a risk of infection (e.g. Clostridium tetani). · Where the consequences of infection are serious or life threatening. · When a person’s defences against infection are compromised.

134-137

Authors: Hitesh Assudani, Jigar K. Gusani, Sanjay J. Mehta.

Introduction: Infections remain one of the major problems in pediatric intensive care unit and are leading cause not only of admission but also mortality in developing countries. Mortality from pediatric sepsis ranges from 9% to 35%. Objective: To study the prevalence & bacteriological profile of septicemia in post neonatal (29 days to 1 year) age group and older children (> 1- 12 year). Material and Methods: Venous blood was collected (2 ml in post neonatal and 5 ml in older children) and inoculation in blood culture bottle. Further processing was done by standard microbiological guidelines. Results: In present study out of 84 clinically suspected 52 (61.90%) were males and 32 (38.10%) were females. Among 84 cases, 31 (36.90%) were post neonatal cases and 53 (63.10%) were older children. Out of 31 post neonatal cases, in 6 (19.35%) cases pathogen were isolated and among them 3 were S.aureus & 1(33.33%) was MRSA. Out of 53 older children, in 14 (26.42%) cases pathogen were isolated and among them 5 S.aureus, 3E.coli, 3Klebsiella spp. were isolated out of which 3(60%), 6 (50%) were MRSA & ESBL respectively. Conclusion: Prevalence of septicemia is 9.83% in post neonatal age group and 22.95% in older children. Septicemia is more common in male age group. Most common clinical presentation in post neonatal age group and older children was fever of unknown origin.

144-149

Authors: RV Shinde, RV Mohite, AR Shinde, ST Mohite, GS Karande.

Aims & Objectives: To assess and compare the feasibility, availability, affordability, stability of In-house and commercial third party internal control of HIV antibody ELISA test. Methods: A comparative study was conducted in NABL Accredited Diagnostic Laboratory at Krishna Institute of Medical Sciences Karad during June 2014 to July 2016. Sample size includes total of 457 ELISA tests which were carried out on 11425 patients along with internal, external commercial and in-house control as per NACO guidelines. Descriptive and inferential statistical tests were applied to find out significant difference between in-house and commercial control. Results: There is significant difference in mean values of positive kit control and commercial positive control (p<0.05). There is also significant difference observed in mean values of commercial positive control and in-house positive control (p<0.05). The coefficient variance of positive in -house control is better (3.32) than commercial positive control (4.60) which indicates, in-house control is more precise than commercial control. The kit cut off values remains constant throughout the month indicates best technical competence, less environmental influences and reagent stability. The average cost of screening for HIV with ELISA and in-house control was Rs. 57/ per person. Conclusion: There is no significant difference in mean values of positive kit control and in-house positive control, the diagnostic cost, feasibility and percentage CV% is better observed in in-house control as compared to commercial control indicating in-house control could be considered as third party internal control for HIV antibody ELISA test.

154-158

Authors: Shreevidya Kinila, Mehnaz Zephyr, Vinay Padumane Gopalakrishna.

Background: To study the bacteriological profile of infections in patients admitted to ICU and to determine the antibiotic susceptibility patterns of the bacterial isolates. This study was done as Prospective study of 2 months period with a study population of patients admitted to Intensive care unit of a tertiary care hospital. Materials and Methods: Selection criteria for this study was with an inclusion criteria of patients admitted to ICU for various reasons and developing infection within 48 hours of admission and the exclusion criteria was patients admitted to ICU and not developing infection and patients admitted to ICU with an already existing infection. All the samples were processed as per standard microbiology guidelines. Results: Gram negative bacilli were predominant with 73.35% as compared to Gram positive cocci of 9.97% of the total aerobic bacteria grown from various samples of patients admitted in Intensive care unit. Escherichia coli were more common with 26.67% of the total bacteria isolated. This was followed by Acinetobacter species 16.67%, Pseudomonas aeruginosa 16.67%, Klebsiella pneumoniae 6.67% and Enterobacter species6.67%. Among the Gram positive cocci, Staphylococcus aureus was more commonly isolated with 16.67% followed by Enterococcus species 6.67% and Streptococcus species 3.30%. Antibiotic resistance was observed by most bacteria to Penicillins, third generation Cephalosporins, Fluoroquinolones like Ciprofloxacin, Cotrimoxazole. Conclusion: Multi-drug resistance is a major hurdle in treating patients admitted to ICU setting in a hospital. Regular surveillance of antibiotic susceptibility patterns is very important for setting orders to guide the clinician in choosing empirical or directed therapy of infected patients.

162-166

Authors: Rajesh Bareja, Iqbal Singh, Subhash Chand Jaryal, Savita Mahajan, Hiba Sami.

Background: Importance of non-fermenting gram-negative bacilli (NFGNB) in the aetiopathogenesis of human infections has increased considerably in recent years. Their higher rate of isolation from a wide variety of clinical specimens necessitates their identification and characterization. Material and Method: A total of 5,369 bacterial isolates were obtained from 12,591 various clinical samples. All the samples were processed on Blood agar and MacConkey agar medium and identified according to standard protocol. Result: Out of 5,369 isolates, 478 were identified as non-fermentative bacteria. Out of NFGNB strains isolated, the predominant numbers were that of P. aeruginosa 337 (70.5%) followed by other Pseudomonad’s 63 (13.1%) and other Non-fermenters 78 (16.4%). Out of strains of other non-fermenters (78) predominant number was that of Acinetobacter anitratus (34), followed by Acinetobacter lwoffii (17), Alcaligenes xylosoxidans (10), Alcaligenes denitrificans (10) and Alcaligenes faecalis (7). The most sensitive drugs against non-fermenter isolates were found imipenem followed by amikacin. Conclusion: There is enough reason to believe that they are associated with human disease process. Therefore, careful attempts must be made for their isolation and identification from various clinical specimens.

171-176

Authors: Preethi Venkatesan, Priyadarshini Shanmugam, Shameem Banu Abdul Satter.

Introduction: Bacteremia is a serious infection with high morbidity and mortality. Knowledge of the etiological agents of blood-stream infections and their antibiotic resistance pattern could help in the choice of empirical antibiotics. Aim: To determine the causative agents of blood stream infection along with their antibiotic resistance pattern in our hospital between 2013 and 2016 and also to identify the uncommon blood culture isolates. Materials and Method: A retrospective study that included isolates obtained from blood culture between January 2013 and December 2016. The identification and drug resistance pattern of the organisms were performed using standard microbiological techniques. For those patients whose blood yielded uncommon isolates, the medical records were looked into to identify the risk factors and outcome. Results: Of a total of 9,926 blood samples received for culture, 534 (5.38%) yielded positive results, of which 277 were Gram-positive organisms and 257 were Gram-negative organisms. The most common etiological agents of bacteremia in the present study were Coagulase Negative Staphylococcus species (37.27%) followed by Salmonella typhi (10.11%), Escherichia coli (9.36%) and Staphylococcus aureus (8.05%), of which 53.49% were MRSA. Conclusion: The most effective antibiotics against Gram positive organisms were vancomycin, teicoplanin and linezolid. The Gram negative organisms showed least resistance to antibiotics like amikacin, tobramycin and carbapenems. The uncommon isolates were all non-fermenting Gram negative bacilli and among these Burkholderia species and Stenotrophomonas maltophila were the most frequently isolated.

182-186

Authors: Marcia Jaqueline Mendonca Maciel, Karol de Souza Barbosa, Luciana Araujo Cassa-Barbosa, Spartaco Ast.

New strains of the yeasts Meyerozyma guilliermondi CBA-524 and Pichia kudriavzevii CBA-519, capable of growing on sugarcane bagasse hydrolysate, have a great potential for second-generation bioethanol processes. Meyerozyma guilliermondi CBA-524, achieved a maximum ethanol concentration of 3.01g/L corresponding to a yield of 0.26 g ethanol per g of sugar and a productivity of 0.125 g/l/h, after 24 h of cultivation. Pichia kudriavzevii CBA-519 exhibited a prolonged fermentation time on bagasse hydrolysate resulting in an ethanol yield of 0.26 g/g and a productivity of 0.015 g/l/h after 192 h of fermentation. The ethanol yield and the volumetric productivity using the adapted P. stipitis CBS6054 standard were 0.34 g/g and 0.083 g/l/h, respectively, after 48 h of cultivation. Furthermore, the newly adapted M. guilliermondi CBA-524 demonstrated significantly enhanced tolerance to sugarcane bagasse hydrolysate, exhibiting a maximum in ethanol concentration after 24 h of fermentation. More adapted yeasts and improved fermentation conditions for this yeast can be of great importance for the increased production of ethanol using sugarcane bagasse hydrolysate.

190-193

Authors: Sathya Bhama MC, Saritha N, Jyothi R.

Introduction: Melioidosis is caused by Burkholderia pseudomallei which is endemic in South East Asia. Lack of proper microbiological services in many countries results in under-reporting of cases and the global burden of infection remains underestimated. This is a study to analyse the clinical presentation and risk factors for melioidosis in India. Materials and Method: A retrospective study was carried out on ten culture proven cases of melioidosis in a tertiary care hospital in South India between January 2009 and December 2014. Result: Ten cases were studied in five years of which nine were males. They had varying clinical presentations like loss of weight and loss of appetite(10%), fever(60%), headache(10%), abdominal pain(20%), joint pain(10%), weakness of limbs(10%), swelling of thigh(20%) and faecal and urinary incontinence((10%). The risk factors included diabetes mellitus (100%), alcohol consumption(90%)and tuberculosis(30%). All the isolates were sensitive to Ceftazidime, Piperacillin-tazobactam, Imipenem, Cotrimoxazole, Amoxycillin-clavulanate and Meropenem while only 80% were sensitive to Ciprofloxacin. Of the 10 cases, two expired and the rest responded to treatment. Conclusion: Melioidosis is an emerging infection in India especially in males and diabetes and alcoholism are identified as the commonest risk factors. Control of the disease requires close monitoring, improved clinical laboratory standards and aggressive therapy.

199-202

Authors: Prabha Poonusamy, Radhika Katragadda, Thyagarajan Ravinder.

Introduction: Asymptomatic bacteriuria is more common in pregnant females. Early screening and treatment avoids complications like acute pyelonephritis and chronic kidney failure. Escherichia coli is the most commonly encountered organism among them. Certain virulence factors associated with E.coli have a direct link in pathogenic mechanism of organism like adhesins, toxins and hemolysin. Aim: We aimed at identifying E.coli and demonstrating phenotypic and genotypic presence of mannose resistance by hemagglutination in urinary isolates. Materials and Method: A cross sectional study during 2014 – 2015 among pregnant females was conducted in a tertiary care hospital. About 1000 urine samples were included in study. Urine culture with antimicrobial susceptibility testing was performed as per standard microbiological conditions. Virulence factor were identified using phenotypic method i.e., haemagglutination test and subsequently confirmed by polymerase chain reaction by looking for Pap A gene expression. Results: Out of 1000 urine samples, 118 culture positives were seen. Out 118 culture positivity, 54 (45.76%) Escherichia coli isolates causing asymptomatic bacteriuria was identified. Among the 54 isolates, phenotypic method for mannose resistance haemagglutination showed 9 positivity (16.66%), which was confirmed with PCR for Pap A expression. Conclusion: As asymptomatic bacteriuria is more common in pregnancy, it is a must to screen pregnant females in every trimester even without any specific symptoms. Virulence factors need to be explored in order to understand the pathogenic mechanism and importance of screening the asymptomatic women.

207-209

Authors: Anjana Gopi, Swati Jain, Faiza Samreen.

Introduction: Onychomycosis is a fungal infection of nails caused by dermatophytes, yeasts or non-dermatophyte moulds and represents about 30% of mycotic cutaneous infections. Onychomycosis manifests itself in various forms, notably onychodystrophy, onycholysis, subungual hyperkeratosis, or nail-plate discoloration. The objective of this study was isolation and speciation of various fungi by culture and to determine its prevalence. Materials and Method: The study was conducted from Jan 2014 - June 2016 at a tertiary care hospital, Bangalore. Nail-clipping and subungual debris of clinically suspected patients were subjected to Potassium hydroxide (KOH) preparation. Culture was done on Sabouraud’s dextrose agar (SDA) media. Species identification was done by colony character, pigment production, Lactophenol cotton blue (LPCB) staining, some special tests like germ tube test, inoculation into chrome agar in case of Candida and slide culture wherever necessary. Result: Out of 400 cases, the KOH preparation was positive for fungal elements in 288 (72%) cases and culture grew fungi in 268 (67%) cases. Among those 268 cases, the infective fungal agents were predominantly dermatophytes (59.7%) and the rest were due to moulds (22.38%) and yeasts (17.91%). Among the different species, Trichophyton rubrum (T. rubrum-37.5%) accounted for the majority of dermatophytes, Fusarium (60%) was the commonest mould and Candida albicans (37.5%) the predominant yeast. Conclusion: Direct microscopy and culture are extremely important in the diagnosis of onychomycosis and dermatophytes are the most common etiological agent in our region.

216-219

Authors: Sudhindra KS, Sumanta A, Shubha DS, Narayana Murthy C, Srinivasa Setty TK.

Introduction: Rickettsial diseases though very commonly found in India, are often misdiagnosed due to lack of awareness and non-availability of diagnostic facilities. Aim: The aim of this study was to determine the prevalence of rickettsial infections in patients attending our hospital with complaints of fever. Materials and Method: A total of 150 serum samples were tested for rickettsial infections by Weil Felix test and the results were analysed. 25 serum samples from afebrile patients were also tested as controls. Results: Of the 150 samples, 59 samples showed significant titres, out of which 29 showed significant titres to OX-K, 10 to OX-2, 11 to OX-19 and mixed titres seen in other 09 samples. Conclusions: Clinical suspicion and routine testing for rickettsial diseases will be useful in patients presenting with fever for prompt diagnosis and management of these cases.

224-226

Authors: Ashwini Kacharu Dedwal, Sae Satish Pol, Sujata Sudhir Ranshing, Shobha Dattatray Chitambar, Renu Sat.

Introduction: Acute gastroenteritis is a major cause of illness and death among infants and young children worldwide. According to the World Health Organization (WHO), an estimated 2.5 billion cases of diarrhea occur among children under 5 years of age each year. Nearly one in five pediatric deaths (about 1.5 billion each year) is due to diarrhea. Present study was thus drawn to identify microbial etiology of acute gastroenteritis in pediatric patients in Western India. Materials and Method: This was Prospective laboratory based study approved by Institutional Ethical committee. 100 stool samples were collected from children admitted with acute diarrhea of 72 hours or less duration in 1- 60 months of age over a period of one year. Bacteria were identified by standard Microbiological methods and serotyping of isolated Escherichia coli (E. coli) was done. Rotavirus antigen was detected by Enzyme Linked Immunosorbant Assay (ELISA) followed by genotyping by Reverse Transcriptase Polymerase chain Reaction (RT-PCR) and Multiplex PCR. Parasitic identification was done by microscopy. Results: 74% of children with diarrhea were in the age group of 7 to 12 months. Watery diarrhea (94%) was the commonest clinical presentation, followed by vomiting (78%), fever (78%), and dehydration (74%). Pathogenic bacteria were isolated in 51% of samples. Escherichia coli was most common (48%) followed by Shigella flexneri (2%) and Vibrio cholerae (1%). The most prevalent E. coli type was Enterotoxigenic Escherichia coli (ETEC) (20.8%) followed by Enteropathogenic Escherichia coli (EPEC)(16.7%), Enterohaemorrhagic Escherichia coli (EHEC) (4.1%), and STEC (2.1%). The most prevalent serotypes of ETEC were O27, O23, and O169. Among EPEC most prevalent serotypes were O90, O26. The most prevalent EHEC strain found in this study was O71. Rotavirus was detected in 35% of patients. Most prevalent Rotavirus genotype was G9P[4] (28.6%) followed by G2P[4] (21.4%), G1P[8] (21.4%), G12P[6] (14.3%), G9P[8] (7.1%). Parasitic etiology was detected in 5% of cases. Coinfection of E.coli and Rotavirus was detected in 23% of children. Rotavirus was most commonly associated with EPEC (25.7%) followed by ETEC (17.1%). Conclusion: In the present study, E. coli was the commonest microorganism followed by Rotavirus. Thus, the importance of safe water and food hygiene would be most important intervention to prevent acute gastroenteritis in children along with Rotavirus vaccine.

231-233

Authors: B. Divya.

Sporotrichosis is a chronic pyogranulomatous fungal infection of cutaneous and subcutaneous tissues which remain localised or may occasionally disseminate to other parts of the body. The causative agent is a thermally dimorphic environmental saprotrophic fungus, Sporothrix schenckii which enters the body through scratches or cuts in the skin. Diabetes, alcoholism, AIDS, leukemia, patients on corticosteroids, chemotherapeutic agents and patients with pre-existing lung disease are important predisposing factors. Here, we report a 65 year old female patient with history of diabetes mellitus for the past 12 years, presented with a leg ulcer and pus discharge for the past 2 years, which did not respond to antibiotic therapy. On examination, a single ulcer of 4x3 cm size was present over the plantar aspect of left foot with pus discharge. Pus was sent for bacteriological and fungal culture identification which revealed growth of Staphylococcus aureus and Sporothrix schenckii respectively. Slide culture and histopathological examination confirmed Sporothrix schenckii. Patient was treated with saturated solution of potassium iodide(SSKI) oral drops and itraconazole and followed up for the regression of lesions.