Seema Mittal

Biofilm and multidrug resistance in uropathogenic Escherichia coli

Abstract Context: Escherichia coli is known as causative agent of urinary tract infections (UTIs) tends to form microcolonies in mucosa lining of urinary bladder known as biofilm. These biofilms make the organism to resist the host immune response, more virulent and lead to the evolution of antibacterial drug resistance by enclosing them in an extracellular biochemical matrix. Aims: This study was done to know the association of various virulence factors and biofilm production in uropathogenic E. coli (UPEC) and antibiotic susceptibility pattern. Settings and design: This study was conducted in Pt. B.D. Sharma PGIMS, Rohtak, Haryana during a period of 1 year from January 2011 to December 2011. Methods and material: Biofilm was detected by microtiter plate (MTP) method, and various virulence factors like hemolysin, hemagglutination, gelatinase, siderophore production, serum resistance, and hydrophobicity were detected. The antibiotic susceptibility testing was done by modified Kirby-Bauer disk diffusion and the disk diffusion method was used to confirm the ESBL, AmpC, MBL production by the UPEC statistical analysis used: The data were analyzed by using SPSS version 17.0. A two-sided P-value of less than or equal to 0·05 was considered to be significant. Results: Biofilm production was found in 18 (13·5%) isolates, more commonly in females (two times). These isolates were found to be resistant to antibiotics common in use and were 100% MDR. Conclusions: Biofilm production makes the organism to be more resistant to antibiotics and virulent as compared to non-biofilm producers.

Vancomycin and High Level Aminoglycoside Resistance in Enterococcus spp. in a Tertiary Health Care Centre: A Therapeutic Concern

Aims. This study was aimed at knowing the prevalence of vancomycin and high level aminoglycoside resistance in enterococcal strains among clinical samples. Study Design. It was an investigational study. Place and Duration of Study. It was conducted on 100 Enterococcus isolates, in the Department of Microbiology, Pt. BDS PGIMS, Rohtak, over a period of six months from July to December 2014. Methodology. Clinical specimens including urine, pus, blood, semen, vaginal swab, and throat swab were processed and Enterococcus isolates were identified by standard protocols. Antibiotic sensitivity testing of enterococci was performed using Kirby-Bauer disc diffusion method. Results. High level gentamicin resistance (HLGR) was more common in urine samples (41.5%) followed by blood (36%) samples. High level streptomycin resistance (HLSR) was more common in pus samples (52.6%) followed by blood samples (36%). Resistance to vancomycin was maximum in blood isolates. Conclusion. Enterococci resistant to multiple antimicrobial agents have been recognized. Thus, it is crucial for laboratories to provide accurate antimicrobial resistance patterns for enterococci so that effective therapy and infection control measures can be initiated.

Acinetobacter lwoffii An Emerging Pathogen in Neonatal ICU

BACKGROUND Acinetobacter species are ubiquitous in the environment and are important causative agent for nososcomial infection especially in immunocompromised patients. Multi drug resistant Acinetobacter lwoffii are emerging as a pathogen in neoanatal sepsis. AIMS AND OBJECTIVE This study was aimed to evaluate the clinical and antibiotic profile of Acinetobacter lwoffii. MATERIAL AND METHODS This study was done on blood samples from neonates admitted to neonatal intensive care unit during a period of one year from January to December 2012, who developed Acinetobacter infection. The diagnosis of isolates and antibiotic susceptibility testing was done by both conventional as well as by automated system. RESULTS Out of total 13,133 blood samples received for culture, 1418(10.8%) were from NICU. Ninety (6.3%) isolates were found to be positive for the growth of Acinetobacter species. Of these isolates 31.11% were found to be Acinetobacter lwoffii, 68.9% were Acinetobacter baumannii calcaetius complex. Acinetobacter lwoffii isolates were most commonly sensitive to imepenem 16(57%), cotrimoxazole 9(32%), ciprofloxacin 6(21%) followed by amoxyclavulanic acid 2(7%) and cefuroxime 1(3.5%). CONCLUSION Multi drug resistant Acinetobacter lwoffii infection is increasing particularly in premature and very low-birth weight neonates. Judicious and timely antibiotic use in NICUs are one of the important key in controlling multi-drug resistant Acinetobacter infection and improving clinical outcome.

Comparison of ELISA and Microscopy for detection of Cryptosporidium in stool.

BACKGROUND Cryptosporidiosis, a diarrheal disease caused by the protozoan parasite Cryptosporidium spp. has become recognized as one of the most common causes of water borne diseases in humans. AIMS AND OBJECTIVES To compare the sensitivity of ELISA and Microscopy for detection of Cryptosporidium in stool samples Materials and Methods: The study was conducted in the Department of Microbiology of PT. B.D. Sharma PGIMS Rohtak, between January 2011 to june 2011 on 50 stool samples, which were processed for detection of cryptosporidial antigen by ELISA and detection of cysts by microscopy (Modified Ziehl and Nelsen staining). STUDY AND DESIGN This was a prospective study conducted in the Department of Microbiology in PT. BD Sharma, PGIMS, Rohtak, India. RESULT Out of total, 50 stool samples eighteen (36%) samples were found positive for Cryptosporidium cysts by microscopy in comparison to 3(6%) stool samples which were found positive for cryptosporidial antigen by ELISA. Samples found positive with ELISA were also positive with microscopy. Sensitivity, specificity, positive predictive value and negative predictive value for ELISA was 16.7%, 100%, 100% and 68% respectively. CONCLUSION The study concludes that stool microscopic Modified acid fast staining is more sensitive method than ELISA for detection of Cryptosporidium in stool samples but the specificity of ELISA was more than microscopy.

Study of virulence factors of uropathogenic Escherichia coli and its antibiotic susceptibility pattern.

CONTEXT Urinary tract infection (UTI) is one of the most common nosocomial infections, caused by Escherichia coli. This study determined the presence of virulence factors in the organism and correlates it with the multi-drug resistance (MDR). AIMS The aim of the following study is to assess the virulence factors of uropathogenic E. coli and antibiotic susceptibility pattern. SETTINGS AND DESIGN This was a prospective study conducted in the Department of Microbiology in PT. B. D. Sharma, PGIMS, Rohtak. SUBJECTS AND METHODS The study was conducted over a period of 1 year. Urine samples received were processed as per standard microbiological procedures. Virulence factors such as hemolysin, hemagglutination, cell surface hydrophobicity, serum resistance, gelatinase and siderophore production were studied. The antimicrobial susceptibility was done as per Clinical and Laboratory Standard Institute Guidelines. STATISTICAL ANALYSIS USED The data was analyzed by using SPSS(Statistical Package for the social sciences) IBM Corporation version 17.0. A two sided P ≤ 0.05 was considered to be significant. RESULTS Hemolysin production was seen in 47.4%, hemagglutination in 74.8%, cell surface hydrophobicity in 61%, serum resistance in 59%, gelatinase in 67.5% and siderophore production in 88% isolates. Nitrofurantoin was found to be most effective followed by, gatifloxacin and gentamicin. Twenty nine percent (29.62%) isolates were MDR. CONCLUSIONS Therefore, the knowledge of virulence factors of E. coli and their antibiotic susceptibility pattern will help in better understanding of the organism and in the treatment of UTI.

Role of Non Albicans Candida Spp. and Biofilm in Neonatal ICU

BACKGROUND Candida spp. remains the fungal species most commonly associated with biofilm formation. Increase in Candida infections in last decades has almost paralleled the increase and wide spread use of a broad range of medical implant devices mainly in population with impaired host defences. One of the most important characteristics of biofilms is their high level of resistance to antimicrobial drugs. AIMS AND OBJECTIVES This study was conducted to know the prevalence of different Candida spp. causing blood stream infections and ability to form biofilm and to evaluate the co relation of biofilm with antifungal drug resistance. MATERIAL AND METHODS The present study was conducted on 12464 blood samples for the identification and speciation of various Candida spp. causing blood stream infection over a period of one year. Antifungal susceptibility was performed as per clinical laboratory standard institute guidelines and biofilm formation was detected by method described by Christensens et al. RESULTS Out of total 12464 blood culture received, 1378 (11.05%) were culture positive rest and among culture positive 100 (7.25%) Candida isolates were recovered. C. tropicalis was the commonest (43%) species followed by C. albicans (41%), C. krusei (9%) and C. parapsilosis (7%). A total of 41 Candida isolates were biofilm producers and rest 59 isolates were non-biofilm producers. CONCLUSION A changing trend of increased prevalence of non albicans Candida spp. was observed which were resistant to commonly used antifungal fluconazole. Multi drug resistance was more common in biofilm forming Candida isolates.

Mupirocin resistance among Methicillin resistant Staphylococcus isolates in a tertiary health care center

INTRODUCTION The evolution of antibiotics in the last century has revolutionized the field of medicine and led this field to higher level of success in treating mild to severe infections, but the inappropriate use of these life saving drugs has been accompanied with the appearance of resistant strains to these agents. AIMS & OBJECTIVES The aim of the study was to determine the prevalence rate of high and low-level mupirocin resistance in methicillin resistant staphylococcus,and to find out resistance to other antibiotics. MATERIALS & METHODS This study was conducted on 100 Staphylococcus isolates recovered from pus samples. Conventional disc diffusion tests were used for the detection of high and low level mupirocin resistance (mupirocin 5µg and 200µg discs) and for various other antimicrobials for example cephalexin, erythromycin, doxycycline, oxacillin, linezolid etc. Results: Outof 100 Staphylococcus isolates processed during the study period in the department of microbiology, 74 were Staphylococcus aureus (S.aureus) and 26 were Coagulase negative Staphylococcus (CoNS). Among S.aureus 43.4% were Methicillin resistant Staphylococcus aureus (MRSA) and 56.6% were Methicillin sensitive Staphylococcus aureus (MSSA), whereas among CoNS 42% were methicillin resistant and 58% were methicillin sensitive. We observed 6.75% of high level mupirocin resistance among Staphylococcus aureus and 19.23% among Coagulase negative staphylococcus. CONCLUSION It was concluded that an inappropriate excessive use of mupirocin leads to a rapid increase in high-level resistance to mupirocin and other antibiotics in CoNS, affecting the treatment lines and success rate of infection control in hospital settings.

Case Report: Escherichia fergusonnii - Pathogen in Urinary Tract Infection.

Urinary tract infections are one of the leading cause of morbidity in admitted patients. Most commonly caused by Escherichia coli, but there are some variants which are commonly reported in urinary tract infection. This study was about to speciate such isolate like E.fergusonnii and find out its antibiogram.