Study of antibiogram pattern of organisms isolated from urinary tract infections from a tertiary care hospital from central India

Introduction: Urinary tract infections (UTI) are the most common infection encountered in the life of an individual irrespective of age and sex. Varying degree of antibiotic resistance shown by uropathogens against the routinely used antibiotics in the therapeutic regimen is a serious concern in treatment of UTI. This study was aimed at the isolation and demonstration of antibiogram pattern of organism responsible for acute UTI. Materials and methods: Clean catched mid stream urine (CCMSU) specimens collected from patients of different age group were cultured on blood agar and Mac Conkey’s agar by standard loop culture method. These organisms were further identified by standard methods and antibiotic sensitivity was evaluated using Kirby Bauers disc diffusion method in accordance with CLSI guidelines. Results: A total of 1230 samples were collected from both males and females of age group 5 to 80. Among the total, 443 (36%) samples were positive for culture and showed significant bacteruria. Gram negative isolates were responsible for majority of infection and 86.9% isolates and Escherichia coli was the predominant among them (37.2%). Conclusion: Majority of the isolates showed resistance to drugs commonly used to treat UTI. Variations in sensitivity may be due to the inappropriate exposure of different localities as to antibiotics which can drive the development of resistance. From the results of this study, it is certain that choosing drugs for empiric treatment will be challenging as no single common drug can conveniently be recommended for UTI.

Simultaneous quantification of pyrethroid metabolites in urine of non-toilet-trained children in Japan

BACKGROUND@#Pyrethroid (PYR) insecticides are widely used for controlling various pests. There are two types that differ in terms of usage: agricultural-purpose PYR (agriculture-PYR) and hygiene purpose PYR (hygiene-PYRs). Few studies exist on the exposure to these chemicals in small children. In this study, we conducted biomonitoring of urinary pyrethroid metabolites in 1.5-year-old children throughout the year.@*METHODS@#Study subjects were 1075 children participating in an Aichi regional sub-cohort of the Japan Environment and Children's Study as of 18-month health check-up. The concentrations of four specific hygiene-PYR metabolites including 2,3,5,6-tetrafluoro-1,4-benzenedimethanol (HOCH2-FB-Al), and five common metabolites of hygiene- and agriculture-PYRs including 3-phenoxybenzoic acid (3PBA) and cis- and trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carboxylic acid (DCCA), were measured in urine samples extracted from soiled diapers using a triple quadrupole gas chromatograph-mass spectrometer.@*RESULTS@#The highest detection frequencies were for 3PBA, followed by DCCA, 1R-trans-chrysanthemum dicarboxylic acid, and HOCH2-FB-Al. Among the six metabolites, urinary concentrations were seasonally varied. However, this variation was not observed in the most studied PYR metabolite, 3PBA. Spearman's correlation analysis demonstrated a significant positive correlation between FB-Al and DCCA (r = 0.56) and HOCH2-FB-Al and 4-methoxymethyl-2,3,5,6-tetrafluorobenzyl alcohol (r = 0.60).@*CONCLUSIONS@#This biomonitoring survey found widespread and seasonally specific exposure to multiple hygiene- and agriculture-PYRs in 1.5-year-old Japanese children.

Review and Proposal of Alternative Technologies for Comprehensive and Reliable Newborn Screening Using Paper Borne Urine Samples for Lysosomal Storage Disorders: Glycosphingolipid Disorders

Abstract Few current methods are efficient to detect a high number of lysosomal storage disorders (LSDs) in newborn screening. Therefore, we propose a stepwise procedure that starts with the use of paper borne urine samples (Berry-Woolf specimen) for the inexpensive detection of elevated lysosomal content and the identification of which of the three majors biochemical groups -mucopolysaccharides, oligosaccharides, and glycosphingolipids- is detected. Urine samples are preferable to blood samples because of their higher concentrations of the relevant analytes. Subsequent steps would precisely determine which enzyme deficiency is involved. As a summary, following our previous papers on the detection of elevated oligosaccharides and mucopolysaccharides, here we describe how elevated urinary glycosphingolipids (GSLs) could be fluorometrically detected using the reagent 5-hydroxy-1-tetralone (HOT) and subsequently identified with precision by continuous thin layer chromatography or other techniques. We also outline the steps required for the validation of this procedure for its introduction in newborn screening programs.

Performance of the cobas u 701 Analyzer in Urinary Tract Infection Screening

BACKGROUND: Negative urine cultures to rule out urinary tract infections (UTI) generate a considerable laboratory workload; thus, a rapid screening test is desirable. We evaluated the performance of a new automated microscopy analyzer, cobas u 701 (Roche Diagnostics International, Rotkreuz, Switzerland) for the screening of UTI, and developed a rule-out strategy to reduce the number of samples requiring culture. We also assessed squamous epithelial cell (SEC) count as a predictor of culture contamination. METHODS: In total, 1,604 urine samples from outpatients were analyzed with cobas u 701 and culture. Bacterial (BAC) and white blood cell (WBC) counts were used for sample interpretation. To determine a useful cut-off point to predict negative cultures, we selected the highest sensitivity and specificity values obtained from ROC curves. Diagnostic accuracy by age and gender was evaluated. RESULTS: Urine culture showed growth of ≥104 colony forming units (CFU)/mL in 256 samples (16.0%). The highest sensitivity (91.8%) and specificity (68.4%) were obtained for cut-off points of 119 BAC/µL and 22 WBC/µL. The combination of BAC and WBC improved the performance of the rule-out strategy with a low rate of false-negative results (1.5%) and a high negative predictive value (NPV, 97.3%). Fifty-seven percent of the samples would not have required culture. SEC count was a poor predictor of culture contamination. CONCLUSIONS: cobas u 701 can substantially reduce the number of urine samples requiring culture, with a low false-negative rate and a high NPV.

Incidence of Significant Bacteriuria among Pregnant Women in a Teaching Hospital of Rural Setup

Aim This study was carried out to determine the prevalence of asymptomatic bacteriuria in pregnant women attending a teaching hospital in rural setup. Materials and Methods A total of 600 women were recruited for this study. A total of 200 antenatal women were clinically identified to have no signs and symptoms of UTI, 200 antenatal women were having signs and symptoms of UTI, and 200 were non-pregnant women that were studied as controls. Clean catch midstream urine samples were collected from each patient into a sterile universal container from Gynecology and obstetrics department and general medicine department. The urine samples were examined microscopically and by the cultural method. Identification of isolates was carried out by a standard microbiological technique. Result A total of 272 (45.3%) were positive for significant bacteriuria. There was a significant difference in the prevalence of asymptomatic bacteriuria with respect to age (P< 0.0001). Trimester did not show any significant difference (P = 0.2006) in the prevalence of asymptomatic bacteriuria. Escherichia coli was the most predominant organism followed closely by Klebsiella species. Nitrofurantoin, ampicillin, and levofloxacin were mostly used antibiotics. Conclusion Asymptomatic bacteriuria is not uncommon among antenatal patients in the population studied. Routine urine cultural test should be carried out on all antenatal patients in order to identify any unsuspecting infection. This measure will go a long way in reducing maternal and obstetric complications associated with pregnancy.

Correlation between Muscle Tone and Motor Delay and Biochemical Levels of Carbohydrates in Urine.

Aims: To establish a possible correlation between children’s clinical status and carbohydrates in their urine samples as they could be a useful additional tool for objective assessment of muscle tone and motor development. Place and Duration of Study: The Department of Child, Adolescent and Developmental Neurology, University Children’s Hospital in Ljubljana between December 1, 2011 and August 31, 2013. Methodology: The retrospective part of the study comprised children aged 0-5 years (average age, 29.3 months) with motor developmental delay and/or abnormal muscle tone. A group of healthy individuals comprised the prospective part. Besides clinical assessment, electrochemical detection of ten sugars in the urine samples was performed using ion chromatography combined with electrochemical detection and tandem mass spectrometry. Results: 72.0% of 82 hospitalised children had a disorder of muscle tone and/or DQ < 70, 28.0% had a pathological urine sample and an abnormal clinical examination, while 44.0% of children had a clinical abnormality and a normal urine sample. In the control group, 22.7% had pathological urine samples. There was a statistically significant difference between patients and control subjects in the value of fucose (p = 0.020), sucrose (p = 0.051) and ribose (p = 0.000). Conclusion: A significant correlation was found between abnormalities in clinical status and pathological urine samples (p = 0.002). Absolute and relative values of specific carbohydrates and the number of elevated sugars showed a strong correlation with disease severity.

Determinación cualitativa de algunos metabolitos de seis glucocorticoides en orina a través de cromatografía líquida de alta eficiencia acoplada a masas-masas. / Qualitative determination of metabolites in urine of six glucocorticoids by high performance liquid chromatography coupled to mass-sprectrometry

El uso de glucocorticoides está prohibido en el campo deportivo durante las competencias si son administrados por vías sistémicas, para lo cual, WADA definió un nivel de reporte de 30ng/mL para los compuestos padre y sus metabolitos. En este trabajo ha sido desarrollada una metodología analítica para la determinación cualitativa de los principales metabolitos de algunos glucocorticoides sintéticos: Betametasona, Budesonida, Dexametasona, Metilprednisolona, Prednisolona, y Triamcinolona Acetónido, presentes en muestras de orina conservadas por el Laboratorio de Control al Dopaje de COLDEPORTES. La metodología incluyó una comparación entre la extracción de los analitos libres y conjugados con glucurónido (para determinar cuál de las dos fracciones era más útil en el análisis de estas sustancias), su separación por cromatografía liquida de alta eficiencia con ionización positiva y la detección por medio de trampa iónica (HPLC-MS-MS). Se seleccionaron los metabolitos mayoritarios a través de sus tiempos de retención, posteriormente se fragmentaron para obtener sus espectros y a través de la abundancia se determinó la cantidad presente en cada fracción (libre y conjugada). Se identificaron los seis glucocorticoides y un total de doce metabolitos, que permitirán hacer un rastreo más prolongado del consumo de las sustancias. Además se confirmó que las cantidades para la mayoría de los compuestos de interés, fueron más altas en la fracción libre de las muestras de orina, pero que no es posible eliminar el proceso de hidrolisis porque algunos metabolitos se excretan conjugados.

Glucocorticoids use is prohibited on the sport during competitions when they are administered by systemic routes for which WADA defined a reporting level of 30ng/ml for parent compounds and their metabolites. In this paper has been developed an analytical methodology for the qualitative determination of the main metabolites of some synthetic glucocorticoids: Betamethasone, Budesonide, Dexamethasone, Methylprednisolone, Prednisolone and Triamcinolone Acetonide, present in urine samples preserved by the Laboratory of Doping Control of COLDEPORTES. The methodology included a comparison between the extraction of free and conjugated with glucuronide analytes (to determine which of the two fractions was more useful in the analysis of these substances), their separation by high performance liquid chromatography with positive ionization and detection using ion trap (HPLC -MS- MS). Major metabolites were selected by their retention times, later fragmented to obtain its spectra and through their abundance determine the amount present in each fraction (free and conjugated). There were identified the six glucocorticoids and twelve metabolites, that will allow to make a more extended screening of the consumption of these substances. Also it was confirmed that the amounts for most of the compounds of interest, were higher in the free fraction of the urine samples, but it is not possible to eliminate the hydrolysis process because some metabolites are excreted as conjugates.