Evaluation of role of Tigecycline among clinically significant multidrug resistant pathogens from a tertiary care hospital.

Background: Tigecycline, a glycylcycline antibiotic is a promising option for the treatment of single or multidrug resistant pathogens. The aim of the study was to evaluate the in-vitro Tigecycline susceptibility of various pathogens from clinical samples received at the tertiary care hospitals in South India. Methods: The analysis of specimens from patients admitted were carried out in this prospective cross sectional study. The identification and antimicrobial susceptibility testing was performed by semi-automated Vitek 2 systems and Kirby Bauer method. Pattern of data analysis was done by descriptive statistics. Results: Among 2574 isolates, 812 isolates were Gram positive pathogens and 1762 isolates were Gram negative pathogens. Resistance to Tigecycline was more common among Gram negative pathogens (18.62%) in comparison to the Gram positive pathogens (0.49%). Among 740 Extended Spectrum Beta Lactamases (ESBL) producers such as Klebsiella species & E coli, 629 isolates were susceptible, and 93 isolates were resistant to the tigecycline. All the methicillin resistant Staphylococcus aureus (MRSA) isolates were susceptible to tigecycline. Conclusion: Multidrug resistant (MDR) pathogens like Acinetobacter species, and Klebsiella species were found to be highly effective in vitro to tigecycline for elimination of infections caused by both Gram positive and Gram negative pathogens. The use of combination therapy becomes crucial to prevent the development of Pan Drug resistance.

Heterogeneous Vancomycin Intermediate Staphylococcus aureus Infections in Diabetic and Non-Diabetic Patients - A Hospital-Based Comparative Study.

Purpose: To study the infections caused by methicillin resistant Staphylococcus aureus (MRSA) with emphasis on heterogeneous vancomycin intermediate S. aureus (hVISA) in diabetic and non-diabetic patients and their comparison. Patients and Methods: S. aureus strains isolated from diabetic and non-diabetic patients admitted in four tertiary care hospitals in Coastal Karnataka, South India, were tested for methicillin resistance and included in the present study. Demographic and clinical data of the patients were collected using structured proforma. Antimicrobial susceptibility testing was done using the Kirby-Bauer disc diffusion method, and MLSB phenotypes were identified using the D-test. The minimum inhibitory concentration (MIC) of vancomycin was determined using agar dilution. MRSA isolates were tested for hVISA using vancomycin screen agar and population analysis profile - area under the curve (PAP-AUC) test. Statistical analysis of the results was done using the chi-square test. SPSS version 29.0 was used for this purpose. Results: Out of 665 strains of S. aureus isolated, 220 (33.1%) were MRSA. Of these 220 MRSA strains, 122 (55.5%) and 98 (44.5%) were isolated from diabetic and non-diabetic patients, respectively. There was no significant difference in the antimicrobial resistance patterns of MRSA strains isolated from diabetic and non-diabetic patients. Foot infections and osteomyelitis caused by MRSA were significantly more among diabetic patients. Out of 220 strains of MRSA, 14 (6.4%) were hVISA. The rates of hVISA among MRSA isolated from diabetic and non-diabetic were 9.0% and 3.1%, respectively. This difference was statistically not significant. Conclusion: The rate of hVISA among all MRSA isolates was 6.4%. The risk of hVISA infection was three times more in diabetic patients. The results emphasize the importance of the detection of hVISA among MRSA isolates especially from diabetic patients.

India-discovered levonadifloxacin & alalevonadifloxacin: A review on susceptibility testing methods, CLSI quality control and breakpoints along with a brief account of their emerging therapeutic profile as a novel standard-of-care.

BACKGROUND: Levonadifloxacin (intravenous) and alalevonadifloxacin (oral prodrug) are novel antibiotics based on benzoquinolizine subclass of fluoroquinolone, licensed for clinical use in India in 2019. The active moiety, levonadifloxacin, is a broad-spectrum antibiotic with a high potency against methicillin-resistant Staphylococcus. aureus, multi-drug resistant pneumococci and anaerobes. OBJECTIVE: This review, for the first time, critically analyses the antimicrobial susceptibility testing methods, Clinical Laboratory & Standards Institute (CLSI)-quality control of susceptibility testing and breakpoints of levonadifloxacin. Further, the genesis, discovery and developmental aspects as well as therapeutic profile of levonadifloxacin and alalevonadifloxacin are briefly described. CONTENTS: In order to aid the scientific and clinician communities with a single comprehensive overview on all the key aspects of levonadifloxacin and alalevonadifloxacin, the present article covers the reference MIC and disk diffusion methods for levonadifloxacin susceptibility testing that were approved by CLSI and the reference ranges for quality control strains published in the CLSI M100 document. The breakpoints of levonadifloxacin were derived in concordance to US FDA, European Committee on Antibiotic Susceptibility Testing (EUCAST) and CLSI approaches. Further, the article provides a brief account of challenges encountered during the discovery stages of levonadifloxacin and alalevonadifloxacin, activity spectrum and safety benefits accruing from structural novelty-linked mechanism of action. Further, the review also covers in vitro and in vivo activities, registrational clinical studies and patient-friendly features of levonadifloxacin/alalevonadifloxacin. Cumulatively, levonadifloxacin has a potential to offer a long awaited new standard-of-care treatment for the resistant Gram-positive bacterial infections.

Treatment of Hospital-Acquired Infections in Patients with Cirrhosis - New Challenges.

Background: Hospital acquired infections (HAI) in the cirrhotic patients contribute to hepatic decompensation. With emergence of bacterial drug resistance, designing the treatment protocol of HA infection has become the foremost challenge. Purpose: To analyze the resistance pattern of organisms isolated from hospital-acquired (HA) infections and determine appropriate antibiotics treatment protocols for these infections. Study Design: A prospective hospital based observational study was undertaken. Patients and Methods: The present study was conducted over 18 months at Kasturba Medical College, Mangalore, Karnataka, India. Patients with suspected HA infections were subjected to clinical, hematological and microbiological evaluation. Antibiotic sensitivity evaluation was undertaken for the bacteria isolated from these patients. Results: During the study period, 398 patients with cirrhosis were 472 times admitted to the hospital for treatment. Out of these patients, 40 patients were diagnosed with 50 HA infections. Fifty five different organisms were isolated from these infections. It was found that these 55 bacteria isolates comprised 30 (54.54%) gram-negative (GN) and 25 (45.45%) gram-positive (GP) bacteria. Quite seriously, extended-spectrum beta-lactamase (ESBL) producers and methicillin-resistant Staphylococcus aureus (MRSA) were detected in 40% and 58% of GN and GP infections respectively. A total of 36 (65.4%) and (14.5%) 8 out of 55 isolated organisms exhibited multi-drug resistance (MDR) and extensive drug resistance (XDR) behavior, respectively. Conclusion: Cirrhosis patients with HA infection possess higher prevalence of MDR and XDR infections. In such sick patients, cephalosporin and quinolones are not the appropriate empirical antibiotics. Herein, we propose a tigecycline with carbapenem like meropenem and vancomycin based empirical antibiotics protocol to be prescribed for such patients. De-escalation is advised after the culture sensitivity report is obtained.

Discriminant value of automated leucocyte VCS parameters in the detection of tropical infections.

INTRODUCTION: In India, infectious diseases are a leading treatable cause of morbidity and mortality. Mangalore being endemic to many vector-borne diseases, their incidence is known to show seasonal variations with sharp increase during monsoon. Leucocytes have substantial role in the immunological pathogenesis of infections. METHODS: The present series was a hospital-based cross-sectional study performed in a tertiary care hospital for a period of three months from June-August wherein the cell population data of cases of malaria, dengue, leptospirosis, typhoid and rickettsial infections along with equal number of healthy controls were collected and analysed. Effectiveness of leucocyte-related volume (V), conductivity (C) and scatter (S) parameters by Coulter®DXH800 haematology analyser in predicting these infections was appraised. RESULTS: A total of 324 cases comprising of malaria (50%), dengue (30.9%), leptospirosis (13.9%), typhoid (4.0%) and rickettsial infections (1.2%) were included. There was statistically significant differences (P < 0.05) in the mean values of complete blood count parameters-haemoglobin, total leucocyte count, red blood cell count, haematocrit, red cell distribution width, differential leucocyte count, platelet count and plateletcrit between cases and controls and also between specific infections. The mean volumes of neutrophil, monocyte and lymphocyte were considerably increased in malaria and dengue fever compared to leptospirosis, typhoid and rickettsial infections. VCS parameters were the least altered in typhoid fever, except for a strikingly high conductivity and scatter of eosinophils. CONCLUSIONS: Haematological analysis is a part of routine evaluation of any case of febrile illness. This study showed that there are specific alterations in VCS parameters in different types of infections such as malaria, dengue, leptospira, typhoid and rickettsia, the information and analysis of which comes without any additional cost.

Profile of Secondary Bacterial Respiratory Infections in H1N1 Patients Admitted in a Tertiary Care Centre - A Four-year Retrospective Study.

BACKGROUND: H1N1 is known to cause periodic seasonal flu in the Indian subcontinent since 2009. The clinical course and the underlying immunity of the host contribute to the development of secondary bacterial infections in the infected patients. OBJECTIVES: This study aims to analyze the secondary bacterial infections in confirmed H1N1 cases admitted in our hospital (from 2015 to 2018) with respect to the comorbidities, complications, associated bacteria with its antibiotic susceptibility pattern, and the outcome of such episodes. MATERIAL AND METHODS: Data of 164 patients admitted in a tertiary care hospital with H1N1 was extracted from medical records using a semi-structured case report form. Data were entered and analyzed with SPSS version 17. A p-value of <0.05 was considered statistically significant. RESULTS: Most patients were aged above 40 years with female preponderance. In our study, 42% of patients had comorbidities. Only 14 (8.53%) had secondary bacterial infection confirmed by culture. Klebsiella pneumoniae and Acinetobacter baumannii were the most common bacteria that were isolated. They were treated based on the culture reports. There was no mortality in patients with a secondary bacterial infection. CONCLUSION: The early start of the antiviral agents and adherence to the antibiotic policy of the hospital contributed to lower secondary bacterial infections and zero mortality.

Reprocessing of N95 masks: Experience from a resource-limited setting in India.

OBJECTIVES: Due to the surge in demand for N95 masks during the Covid-19 pandemic, and considering the situation in countries grappling with acute shortages of N95 masks, this study investigated the possibilities of decontamination and reuse of masks. METHODS: Three N95 masks of different makes (A, B and C) were subjected to six decontamination methods: ultraviolet (UV) irradiation, isopropyl alcohol (IPA) dip, plasma sterilization (Sterrad®), ethylene oxide (ETO, 3M®), dry heat sterilization, and moist heat sterilization (autoclaving). The integrity of the N95 masks was assessed by measuring their particle filtering efficiency at particle sizes ranging 0.3-0.5 microns. RESULTS: All the masks decontaminated with ETO and plasma sterilization retained over 95% particle filtering efficiency. Masks decontaminated using IPA dip and autoclaving showed a drop, and UV irradiation showed variations in particle size efficiency degradation after decontamination. CONCLUSIONS: Plasma sterilization is recommended for decontamination of N95 masks in low-resource settings. ETO is not recommended due to hazards associated with handling of ethylene oxide, although the filtering efficiency was retained. Since the UV irradiation method showed variations in results, evaluation of UV decontamination for N95 masks needs to be performed on a case-by-case basis.

Comparative Analysis of Change in pH, Oral Health Status, and the Count of Streptococcus mutans and Lactobacillus Species in the Oral Cavity in Patients with Gastroenteral Diseases Using Saliva: A Pilot Study.

BACKGROUND: Seventy million people are affected by gastroenteral (GI) disturbances throughout the world. Oral cavity possesses various bacteria that remain as healthy commensals or turn pathogenic due to shift of balance with disturbances in health, which is reflected in the oral cavity too. Studies have shown a possible oro-systemic link. This study aimed at assessing the effect of GI disease on oral health comparing levels of pH, microbiological counts, and oral health status between test and control groups. MATERIALS AND METHODS: This pilot study consisted of two groups: test group containing 14 participants (GI disease) and control group (healthy) containing 3 participants. Two saliva samples were collected per patient. One sample was inoculated onto Mitis Salivarius and Rugose agar plates at 37oC in the CO2 incubator for 2 days. The second sample was used for recording pH. Parameters such as decayed, missing, and filled teeth, plaque index, gingival index, probing pocket depth, and clinical loss of attachment were also recorded. The results were analyzed using Statistical Package for Social Sciences (SPSS) version 11.5. Regression analysis was applied to predict the three-microbe culture based on the pH and GI disease. RESULTS: The oral health parameters showed a higher number of missing teeth, higher bleeding on probing, higher values of plaque and gingival index, a higher amount of clinical loss of attachment, and acidic pH of saliva in the test group. Microbiological analysis showed more Streptococcus mutans in the control group (7,500-10,000 cfu/mL), with a mean of 8,833.33±1,258.31 cfu/mL; S. salivarius was more in the test group (2,000-25,000 cfu/mL) with a mean of 15,866.67±6,697.76 cfu/mL. Candida was seen only in the test group (2,166.67±2,549.51 cfu/mL) and absent in the control group. Lactobacillus was absent in both the groups. CONCLUSION: The present study suggests the relation between oral health and GI diseases. Hence, saliva could be used as an easy, non-invasive biomarker to analyze the gastroenteric status of the patient.



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Assessment of antibacterial activity of levonadifloxacin against contemporary gram-positive clinical isolates collected from various Indian hospitals using disk-diffusion assay.

Objectives: Levonadifloxacin is a novel benzoquinolizine subclass of quinolone with broad-spectrum activities against problematic pathogens such as methicillin-resistant Staphylococcus aureus, quinolone-resistant S. aureus, vancomycin intermediate S. aureus, and vancomycin-resistant S. aureus. Levonadifloxacin and its oral prodrug, alalevonadifloxacin, have been recently approved in India for the treatment of acute bacterial skin and skin structure infections, including concurrent bacteraemia and diabetic foot infections. The aim of the study is to assess the activity of levonadifloxacin against Gram-positive clinical isolates collected from various Indian hospitals using the disc-diffusion method. Materials and Methods: Nonduplicate isolates of S. aureus and other Gram-positive isolates collected from June 2019 to March 2020 were subjected to levonadifloxacin susceptibility testing (disk diffusion method) as per the Clinical and Laboratory Standards Institute guidelines (Year 2019). Levonadifloxacin 10 µg impregnated disks were used during the testing. Results: A total of 664 diverse Gram-positive clinical isolates collected from six different hospitals in India were analyzed. Majority (65.5%) of the isolates were S. aureus. All the S. aureus and other Gram-positive isolates were found to be susceptible to levonadifloxacin as per the prespecified interpretive criteria identified based on population pharmacokinetic model and Monte Carlo simulation enabled probability of pharmacodynamic target attainment analysis. Conclusions: The present study showed that levonadifloxacin was highly active against contemporary Gram-positive pathogens and furthermore demonstrated that levonadifloxacin susceptibilities can be reliably determined using the disc-diffusion method.

Occurrence of blaTEM and blaROB in Haemophilus species Causing Respiratory Tract Infections.

BACKGROUND: Emergence of resistance to some antibiotics in Haemophilus influenzae, a respiratory pathogen is a cause of concern. The aim is to study the antibiotic susceptibility pattern of Haemophilus isolates from respiratory infections with reference to beta-lactam resistance. METHODS: This is a laboratory based prospective study done in the department of microbiology in a tertiary care center after institutional ethics committee clearance. Haemophilus influenzae isolates from respiratory tract specimens over a period of one year were subjected to antibiotic susceptibility tests. Beta-lactamase production was detected by nitrocefin disc. hpd gene, blaTEM and blaROB genes were detected by PCR. The data was analysed using SPSS 11.5 version. RESULTS: Of the 162 isolates, 89.5% were from sputum specimens. Ampicillin resistance was seen in 5 (3.09%) isolates. The ampicillin resistant strains were positive for beta-lactamase enzyme and blaTEM gene. BLNAR and isolates with blaROB gene were not found. CONCLUSION: In case of Haemophilus influenzae respiratory tract infection empirical treatment with amoxicillin clavulanate or third generation cephalosporin may be the drugs of choice in our geographic area.

Correlation of Clinical Severity and Laboratory Parameters with Various Serotypes in Dengue Virus: A Hospital-Based Study.

OBJECTIVES: Dengue fever, being hyperendemic with analogous presentations as in many other acute febrile illnesses, poses a challenge in diagnosis during the acute stage. Additionally, the coexistence of multiple serotypes further complicates the disease prognosis. The study was undertaken to determine the dengue virus serotypes, clinical, and laboratory markers as predictors in the severity of infection. METHODS: A prospective study was conducted among 106 patients admitted with acute febrile illness having positive NS1 antigen/IgM ELISA. Clinical data were extracted from medical records including demographics, presence of comorbid conditions, clinical presentation, laboratory investigations, and course including length of hospital stay and outcome. Detection of dengue serotypes was done by multiplex reverse transcriptase polymerase chain reaction (RT_PCR). RESULTS: Out of 106 RT-PCR-confirmed cases, DENV-3 was the most common serotype found in 56 (52.8%) patients, followed by DENV-3 and DENV-4 coinfection in 27 (25.4%) patients. Coinfection with more than one serotype was witnessed in our study. Raised liver enzymes and increased ferritin are good biomarkers in differentiating dengue from severe dengue with cutoff levels for AST (134 U/L), ALT (88 U/L), and ferritin (3670 ng/ml). Musculoskeletal, followed by gastrointestinal, manifestations were comparatively higher than respiratory and cutaneous manifestations. CONCLUSION: This study provides more information on the dengue serotypes. The clinical spectrum along with laboratory parameters such as ferritin, liver enzymes, platelet can be used as potential biomarkers in prediction of dengue severity. The data demonstrated will be useful in early detection and monitoring of the disease.

In vitro activity of a novel antibacterial agent, levonadifloxacin, against clinical isolates collected in a prospective, multicentre surveillance study in India during 2016-18.

BACKGROUND: Levonadifloxacin is a novel antibiotic belonging to the benzoquinolizine subclass of fluoroquinolones with potent activity against MRSA and quinolone-resistant Staphylococcus aureus. IV levonadifloxacin and its oral prodrug alalevonadifloxacin have recently been approved in India for the treatment of acute bacterial skin and skin structure infections (ABSSSIs) including diabetic foot infections. OBJECTIVES: To investigate the in vitro activity of levonadifloxacin against contemporary clinical isolates collected from multiple tertiary care hospitals across India in the Antimicrobial Susceptibility Profiling of Indian Resistotypes (ASPIRE) surveillance study. METHODS: A total of 1376 clinical isolates, consisting of staphylococci (n = 677), streptococci (n = 178), Enterobacterales (n = 320), Pseudomonas aeruginosa (n = 140) and Acinetobacter baumannii (n = 61), collected (2016-18) from 16 tertiary hospitals located across 12 states in India, were included in the study. The MICs of levonadifloxacin and comparator antibiotics were determined using the reference agar dilution method and broth microdilution method. RESULTS: Levonadifloxacin exhibited potent activity against MSSA (MIC50/90: 0.5/1 mg/L), MRSA (MIC50/90: 0.5/1 mg/L) and levofloxacin-resistant S. aureus (MIC50/90: 1/1 mg/L) isolates. Similarly, potent activity of levonadifloxacin was also observed against CoNS including MDR isolates (MIC50/90: 1/2 mg/L). Against Streptococcus pneumoniae, levonadifloxacin (MIC50/90: 0.5/0.5 mg/L) showed superior activity compared with levofloxacin (MIC50/90: 1/2 mg/L). Among levofloxacin-susceptible Enterobacterales, 80.6% of isolates were inhibited at ≤2 mg/L levonadifloxacin. CONCLUSIONS: Levonadifloxacin displayed potent activity against contemporary MRSA and fluoroquinolone-resistant staphylococcal isolates, thus offering a valuable IV as well as an oral therapeutic option for the treatment of ABSSSIs. Furthermore, levonadifloxacin exhibited a broad-spectrum activity profile as evident from its activity against streptococci and levofloxacin-susceptible Gram-negative isolates.

Evaluation of the antimicrobial efficacy of 20% Punica granatum, 0.2% chlorhexidine gluconate, and 2.5% sodium hypochlorite used alone or in combinations against Enterococcus faecalis: An in-vitro study.

OBJECTIVES: The study was aimed at evaluating the antimicrobial efficacy of 20% Punica granatum, 0.2% chlorhexidine (CHX) gluconate, and 2.5% sodium hypochlorite used alone or in combinations against Enterococcus faecalis. MATERIALS AND METHODS: Aqueous extract of pomegranate peel was prepared in the Pharmacology Departmental Laboratory. A total of 240 wells were prepared (40 for each group) with 5 wells per with a diameter of 6 mm and depth of 4 mm at equidistant from each other. Using a pipette, each well was filled with 50 µl of the test irrigant solution. CHX (0.2%), 2.5% sodium hypochlorite, and aqueous extract of pomegranate peel and their combinations were tested as root canal irrigants against a standard strain of E. faecalis (ATCC 29212) on sheep blood agar plate by calculating the zones of inhibition. The mean diameter of zones was calculated and tabulated. Data were analyzed using one-way analysis of variance and Tukey's post hoc test. Descriptive statistics was obtained using SPSS software (version 11.5) with P established at < 0.05. RESULTS: Combination of Punica granatum with sodium hypochlorite and CHX showed maximum mean zones of inhibition with mean of 23.9 and 25.7 mm, respectively, and showed significantly better results than all other groups either irrigants used alone or in combinations. CONCLUSIONS: Punica granatum and CHX was proved to be a very good combination among experimental groups against E. faecalis, and sodium hypochlorite was least effective against E. Faecalis.

Rapid method for detecting and differentiating Mycobacterium tuberculosis complex and non-tuberculous mycobacteria in sputum by fluorescence in situ hybridization with DNA probes.

OBJECTIVE: In resource-limited tuberculosis-endemic countries, Mycobacterium tuberculosis in sputum is mainly detected by acid-fast bacillus (AFB) staining and the identification of sputum-derived cultures. PCR techniques are practical only in well-resourced laboratories. This study investigated the application of a rapid, simple, and inexpensive fluorescence in situ hybridization (FISH) assay to identify and differentiate M. tuberculosis complex (MTBC) from non-tuberculous mycobacteria (NTM) in sputum. METHODS: The Mycobacterium/Nocardia Genus (MN Genus)-MTBC FISH assay performed in this study utilizes two different DNA probes labeled with different fluorescent molecules that hybridize respectively with 16S rRNA of the genus Mycobacterium and 23S rRNA of MTBC. The assay was tested on 202 patient sputum samples in Mangaluru, Karnataka State, India. Sputa were first liquefied and bacteria concentrated before performing the FISH assay and parallel culturing and AFB staining. The identities of cultured bacteria from DNA sequencing were compared with FISH assay findings from corresponding sputa. RESULTS: Of the 202 sputum samples tested, 67 reacted with both MN Genus-specific and MTBC-specific probes, none reacted only with the MTBC-specific probe, and 22 reacted only with the MN Genus-specific probe. The FISH assay yielded results in 2h and had a limit of detection of 2.2×104CFU/ml in sputum spiked with cultured M. tuberculosis. The diagnostic sensitivity, specificity, and positive and negative predictive values of the FISH assay for MTBC in patient sputa were 89.7%, 95.5%, 88.0%, and 92.6%, respectively. NTM were a significant cause of tuberculosis-like infections in Mangaluru. CONCLUSIONS: The MN Genus-MTBC dual probe fluorescence FISH assay previously applied to cultures can also be utilized in resource-limited tuberculosis-endemic countries for rapidly identifying and differentiating MTBC and NTM in sputum samples.

Detection of human papilloma virus in potentially malignant and malignant lesions of the oral cavity and a study of associated risk factors.

BACKGROUND: Squamous cell carcinoma of the head and neck is the 6th most frequently occurring cancer worldwide, with over 400,000 cases projected annually. Multiple factors such as tobacco, alcohol, irradiation, virus, and chronic irritants are involved in the development of oral squamous cell carcinomas (OSCCs). The most important risk factors are chronic exposure to tobacco and alcohol. Although the evidence that implicates virus is increasing, particularly (human papillomavirus [HPV]), in the carcinogenesis process, the role of virus is not well established. AIM AND OBJECTIVE: This study is designed to assess the presence of HPV in potentially malignant and malignant lesions of the oral cavity as well as to correlate the presence of HPV with addictive habits and histopathological grading of the disease. MATERIALS AND METHODS: Biopsy samples of OSCC and potentially malignant lesions were obtained and 3, 5 µm thickness sections were cut using a microtome. The sections were collected using a sterile brush and transferred to an Eppendorf tube. DNA extraction and polymerase chain reaction for the detection of HPV were done. RESULTS AND CONCLUSION: The association between histopathological grading and presence of HPV was assessed using Chi-square test and the values thus obtained were found to be statistically significant. HPV was more predominantly seen in well-differentiated carcinomas and moderately differentiated carcinomas as compared to poorly differentiated carcinomas.

Clinico-microbiological study of Pseudomonas aeruginosa in wound infections and the detection of metallo-ß-lactamase production.

Pseudomonas aeruginosa is a common opportunistic pathogen of humans among the Gram-negative bacilli. Clinically, it is associated with nosocomial infections like burns and surgical-site wound infections and remains a major health concern, especially among critically ill and immunocompromised patients. This is a prospective laboratory-based 2 year study conducted to isolate P. aeruginosa from wound specimens and the antimicrobial susceptibility pattern with reference to metallo-ß-lactamase (MBL) production. Two hundred and twenty-four samples of P. aeruginosa isolated from wound specimens were included in the study. Antimicrobial susceptibility was done as per Clinical Laboratory Standard Institute (CLSI) guidelines. MBL-producing P. aeruginosa was detected using the EDTA disk diffusion synergy test. Statistical analysis was done using the SPSS 11 package (SPSS Inc., Chicago, IL). Out of the 224 P. aeruginosa isolates, 100% were susceptible to polymyxin B and colistin, 92·8% were sensitive to imipenem, 38% showed resistance to gentamicin followed by ceftazidime (31·69%) and meropenem (33·03). Sixteen (7·14%) isolates showed MBL production. Infection caused by drug-resistant P. aeruginosa is important to identify as it poses a therapeutic problem and is also a serious concern for infection control management. The acquired resistance genes can be horizontally transferred to other pathogens or commensals if aseptic procedures are not followed.

An Outbreak of Burkholderia cepacia Bacteremia in a Neonatal Intensive Care Unit.

OBJECTIVES: To identify the source of infection, to study the clinical profile and outcomes of neonates with Burkholderia septicemia and to determine the antimicrobial susceptibility patterns of the isolates. METHODS: The authors describe a 3 mo outbreak of nosocomial Burkholderia cepacia bacteremia involving 12 neonates. During the outbreak, ventilator humidifier water, intravenous solutions and other possible sources were taken from the concerned neonatal intensive care units (NICUs); cultured and isolates identified by standard microbiological techniques and VITEK system. Clinical details of affected babies were also obtained to ascertain the clinical significance of the isolates. RESULTS: All neonates had clinical and biochemical evidence of sepsis and the source could be tracked to intravenous solutions of 5% dextrose, normal saline (opened bottles) and continuous positive airway pressure humidifier water. Strain relatedness of the environmental isolates with the clinical isolates is likely as antibiotic susceptibility patterns were similar. CONCLUSIONS: The investigations revealed the source of the nosocomial outbreak which is crucial for initiating appropriate control measures.

Association of Risk Factors, Antimicrobial Resistance Trends and Occurrence of blaTEM, bla SHV and blaCTX M in Escherichia coli Causing Bacteremia.

PURPOSE: Escherichia coli are the most frequent cause of gram negative bloodstream infection. This study was done to evaluate the association of risk factors, antimicrobial susceptibility pattern and detection of TEM, SHV and CTX M genes in the extended spectrum beta lactamase (ESBL) producing E.coli. MATERIALS AND METHODS: 11,133 blood samples were processed in BacT/Alert bottles. Bacteria grown were identified and antibiotic susceptibility patterns were studied using VITEK2 system. ESBL production was tested using phenotypic method, VITEK system and by PCR. Statistical package SPSS Version 11.5 was used to do the analysis. RESULTS: Blood culture positive isolates were 1530 (13.7%), among which 108 were identified as Escherichia coli. The most common risk factor associated with E.coli bacteremia was diabetes and common source was UTI. E.coli were resistant to Ampicillin (86%), Piperacillin (82.4%), Ceftazidime (80.6%), Ceftriaxone (80.6%), Cefipime (77%), Aztreonam (80.6%) and Fluoroquinolones (80%). Isolates were sensitive to Carbapenems and combination drugs. ESBL was produced by 76%. CTX M bla (betalactamase) gene was common in all the ESBL isolates. CONCLUSION: ESBL producing E.coli isolates were observed to be multi drug resistant. Owing to the high prevalence of ESBL.Carbapenems are clearly the drug of choice for empirical treatment in these cases.Ertapenem may be used to prevent development of carbepenem resistant pseudomonas or acinetobacter isolates in the hospital settings. To limit the earlier development of resistance to carbepenems, it is better to de-escalate to combination drugs like piperacillin tazobactum with aminoglycosides after the susceptibility report.

Phaeohyphomycotic cyst in the Foot by Exophiala.

A 52-year-old male, presented to us with a swelling over plantar aspect of right foot following trauma. Clinically it was a cystic swelling diagnosed as an abscess; ultrasound showed thick walled multilocular collection with thick echogenic debris, following which complete excision of the swelling was done. A part of the swelling was sent for histopathological examination and cut section showed thick purulent material. Other part sent for culture sensitivity grew, Exophiala, which belongs to Dematiaceous group of fungi. Surgical excision with antifungal treatment is the management in general for fungal cyst, whereas in our case complete excision was done without antifungal treatment.

Comparison of various culture methods for isolation of group B streptococcus from intrapartum vaginal colonization.

AIMS: Group B Streptococcus (GBS) is one of the most common causes of neonatal sepsis throughout the world. Reports of vaginal colonization of GBS in India are few and variable. A study was conducted on pregnant women in a tertiary care hospital to compare various methods for isolation of GBS, to study the prevalence of GBS in pregnant women in third trimester, and to determine risk factors for GBS colonization. SETTINGS AND DESIGN: Observational descriptive study. MATERIALS AND METHODS: High vaginal swabs from 150 pregnant women in their third trimester were used to compare three methods for isolation of GBS viz. direct culture on 5% Sheep Blood agar, direct culture on selective Columbia Blood Agar and culture in LIM enrichment broth with subsequent culture on 5% Sheep Blood agar. A history of associated risk factors was also taken. STATISTICAL ANALYSIS USED: Statistical analysis was performed by Chi-square test. RESULTS: Isolation was best from LIM enrichment broth with subsequent culture on 5% Sheep Blood Agar. Prevalence of GBS colonization by using culture method was 12.67%. Most frequently associated risk factor was intrapartum fever (42.11%). CONCLUSIONS: Standard Culture Method using LIM enrichment should be adopted as standard practice for isolation of GBS from vaginal swabs.