Azithromycin can induce SOS response and horizontal gene transfer of SXT element in Vibrio cholerae.

BACKGROUND: The emergence and spread of drug resistance in Vibrio cholerae are mainly attributed to horizontal gene transfer of mobile genetic elements, especially the SXT (sulfamethoxazole and trimethoprim) element, an integrative conjugative element carrying multiple drug resistance genes. SOS (save our souls) bacterial stress response in Vibrio cholerae plays a pivotal role in inducing the SXT element transfer and induction of the CTX prophage, carrying the important virulence factor cholera toxin encoded by the ctxAB gene. METHODS: This study investigated whether the subinhibitory concentration of antibiotics like ciprofloxacin, tetracycline, and azithromycin induce SOS response by detecting the expression of recA and lexA, the key genes of SOS response by reverse transcriptase real time PCR (RT-qPCR). We also studied the transfer of SXT element in response to these three antibiotics by bacterial conjugation. Transfer of SXT elements was confirmed by detecting the SXT element-specific conserved genes. RESULTS: The results of the real-time PCR showed that all three antibiotics induced SOS response with more robust induction by tetracycline and azithromycin relative to ciprofloxacin. We observed a higher frequency of transfer of SXT elements in cultures exposed to these antibiotics and the control mitomycin C compared to unexposed cultures. CONCLUSION: Our study indicates that antibiotics including azithromycin, ciprofloxacin, and tetracycline activate SOS response in Vibrio cholerae and demonstrates a robust mechanism for wide dissemination of drug resistance.

Impact of Therapeutic Drug Monitoring on Once-Daily Regimen of Amikacin in Patients With Urinary Tract Infection: A Prospective Observational Study.

BACKGROUND: Amikacin is a semisynthetic antibiotic used in the treatment of gram-negative bacterial infections and has a narrow therapeutic index. Although therapeutic drug monitoring is recommended for amikacin, it is not routinely performed because of the use of a less toxic once-daily regimen. Only few studies have evaluated the role of therapeutic drug monitoring in patients treated with amikacin. The objective of our study was to find an association between the pharmacokinetic parameters of amikacin and the time required for a clinical cure, creatinine clearance, and frequency of ototoxicity in patients with urinary tract infection treated for 7 or more days. METHODS: A prospective study was conducted on patients with urinary tract infections who were administered amikacin for 7 or more days. Blood samples were obtained from the patients to measure the maximum drug concentration (Cmax) and trough concentration (Ctrough). Minimum inhibitory concentration (MIC) values were determined for patients with positive urine cultures. Serum creatinine levels were estimated every 3 days. The auditory assessment was performed using pure tone audiometry at baseline and weekly until the patients were discharged. Levels of amikacin were analyzed using a validated liquid chromatography-tandem mass spectrometry method. RESULTS: Of 125 patients analyzed, the median time required for a clinical cure was less in the group of patients who achieved a Cmax/MIC ratio ≥8 than it was in those who did not achieve this level [7 versus 8 days (P = 0.02)]. The Ctrough of amikacin was associated with the change in serum creatinine level (P = 0.01) and the incidence of nephrotoxicity (P = 0.004). CONCLUSIONS: In patients receiving short-term amikacin therapy, Cmax/MIC value can be used to predict the time required for a clinical cure. Ctrough can be used to predict the occurrence of nephrotoxicity in patients receiving amikacin therapy.

Antibodies to Zika virus, Campylobacter jejuni and gangliosides in Guillain-Barre syndrome: A prospective single-center study from southern India.

BACKGROUND: The objective of this article was to study the presence of antibodies against Zika virus (ZIKV), Campylobacter jejuni, and gangliosides in patients with Guillain-Barre syndrome (GBS). MATERIALS AND METHODS: Ninety consecutive patients (age more than 12 years) with GBS admitted to a tertiary care center in southern India were included in this study. Data on clinical manifestations, nerve conduction studies, and response to therapy were collected. The following tests were done in stored serum samples - anti-ZIKV (IgM) antibodies, anti-C. jejuni (IgG) antibodies, and anti-ganglioside antibodies (IgG). Those samples which were positive to anti-Zika antibodies were tested for conventional polymerase chain reaction for ZIKV and IgM antibodies against dengue, and Japanese encephalitis virus. RESULTS: Of the 90 patients, 3 died and 8 had persistent weakness. Acute inflammatory demyelinating polyradiculoneuropathy was the most common type of GBS (56.7%). Anti-ganglioside antibodies were present in 62.2% patients with GT1b being the most common. Anti-C. jejuni antibodies were present in 46.6%. Anti-Zika antibodies (IgM) were present in 14 patients (15.5%). Four of these patients also had anti-dengue antibody (IgM) positivity. CONCLUSION: This is one of the largest studies on GBS from India and the first one to report on the presence of Zika virus antibodies from this geographical area. Our study had a high prevalence of anti-C. jejuni and anti-ganglioside antibodies. Evidence of recent ZIKV infection, as evidenced by anti-IgM antibodies, was present in 14 patients, with 4 of them being tested positive for anti-dengue IgM antibody. Whether this represents cross-reaction with dengue or prior/co-infection with dengue virus could not be addressed in this study.

Comparison of nested-multiplex, Taqman & SYBR Green real-time PCR in diagnosis of amoebic liver abscess in a tertiary health care institute in India.

BACKGROUND & OBJECTIVES: Amoebiasis is a common parasitic infection caused by Entamoeba histolytica and amoebic liver abscess (ALA) is the most common extraintestinal manifestation of amoebiasis. The aim of this study was to standardise real-time PCR assays (Taqman and SYBR Green) to detect E. histolytica from liver abscess pus and stool samples and compare its results with nested-multiplex PCR. METHODS: Liver abscess pus specimens were subjected to DNA extraction. The extracted DNA samples were subjected to amplification by nested-multiplex PCR, Taqman (18S rRNA) and SYBR Green real-time PCR (16S-like rRNA assays to detect E. histolytica/E. dispar/E. moshkovskii). The amplification products were further confirmed by DNA sequence analysis. Receiver operator characteristic (ROC) curve analysis was done for nested-multiplex and SYBR Green real-time PCR and the area under the curve was calculated for evaluating the accuracy of the tests to dignose ALA. RESULTS: In all, 17, 19 and 25 liver abscess samples were positive for E. histolytica by nested-multiplex PCR, SYBR Green and Taqman real-time PCR assays, respectively. Significant differences in detection of E. histolytica were noted in the real-time PCR assays evaluated ( P<0.0001). The nested-multiplex PCR, SYBR Green real-time PCR and Taqman real-time PCR evaluated showed a positivity rate of 34, 38 and 50 per cent, respectively. Based on ROC curve analysis (considering Taqman real-time PCR as the gold standard), it was observed that SYBR Green real-time PCR was better than conventional nested-multiplex PCR for the diagnosis of ALA. INTERPRETATION & CONCLUSIONS: Taqman real-time PCR targeting the 18S rRNA had the highest positivity rate evaluated in this study. Both nested multiplex and SYBR Green real-time PCR assays utilized were evaluated to give accurate results. Real-time PCR assays can be used as the gold standard in rapid and reliable diagnosis, and appropriate management of amoebiasis, replacing the conventional molecular methods.

Profile of urinary tract infections in paediatric patients.

BACKGROUND & OBJECTIVES: This cross-sectional study was conducted at a tertiary care centre in Puducherry, south India, with the aim of finding the profile of the paediatric urinary tract infection (UTI), bacterial pathogens involved, and also to observe vesicoureteric reflux (VUR) and renal scarring in these patients. METHODS: A total of 524 paediatric patients ≤13 yr, suspected to have UTI, were included in the study. Urine samples were collected, processed for uropathogen isolation and antibiotic susceptibility test was performed as per the Clinical and Laboratory Standards Institute (CLSI) guidelines. Thirty two culture proven children with UTI underwent micturating cysto-urethrography (MCU) and dimercaptosuccinic acid (DMSA) scanning was done for 69 children. RESULTS: o0 f the 524 children, 186 (35.4%) had culture proven UTI with 105 (56.4%) being infants, 50 (27.4%) between 1-5 yr, 30 (16.12%) between 5-13 yr and 129 (69.35%) males. Posterior urethral valve (PUV) was noted in three, hydronephrosis in one, VUR in 18 and renal scarring in 33. VUR as well as renal scarring were more in males >1 yr of age. A significant association (P=0.0054) was noted with a combined sensitivity and specificity of these investigations being 83 and 90 per cent, respectively of the MCU and DMSA scans for detecting VUR. Escherichia coli was the most common pathogen isolated, sensitive to nitrofurantoin, followed by cefoperazone-sulbactam, aminoglycosides and meropenem. INTERPRETATION & CONCLUSIONS: Our results indicate that UTI varies with age and gender and extensive evaluation is required in boys over one year of age with UTI. This study also highlights the better efficacy of aminoglycosides, cefoperazone-sulbactam and nitrofurantoin in vitro compared with meropenem in Gram-negative uropathogens.

Fungal profile of funguria cases at a tertiary care hospital in southern India.

BACKGROUND & OBJECTIVES: Candida albicans is the most common aetiological agent in funguria cases commonly observed in hospitalized patients. But a few reports are available from India where non-albicans Candida species have accounted for >50 per cent of urinary Candida isolates. We undertook this study to know the fungal profile amongst funguria cases. METHODS: A total of 123 consecutive fungal isolates obtained from clinically suspected cases of urinary tract infection from April to September, 2013, were included. Yeast species was identified by standard phenotypic methods. Antifungal susceptibility testing of yeast was performed for fluconazole (25 µg) by disc diffusion method as per Clinical and Laboratory Standards Institute (CLSI) guidelines. RESULTS: The male to female ratio was 0.92. The mean age of patients was 42.7 ± 18.9 yr. C. tropicalis (58.5%) was the most common fungal agent followed by C. albicans (30.1%). Only one isolate of C. tropicalis was resistant to fluconazole. INTERPRETATION & CONCLUSIONS: C. tropicalis and C. albicans were the predominant fungal pathogens responsible for urinary tract infection. Less resistance to fluconazole observed in the study may be due to restricted use of fluconazole in this area.

Non O1 non O139 Vibrio cholerae septicemia in a patient with carcinoma pancreas-a case report and literature review, 2020-2023.

Non-O1/non-O139 Vibrio cholerae, a comparably poorly studied pathogen is culpable of sporadic but serious infections. We report a case of non O1 non O139 Vibrio cholerae septicemia in a middle aged male recently diagnosed with carcinoma pancreas. He underwent biliary tract interventional procedure for hematemesis three weeks before the presentation. Now, he presented with fever, abdominal pain, hematemesis and melena. Endoscopy revealed severe portal hypertensive gastropathy and mild hemobilia. Blood culture grew Vibrio cholerae, identified as non O1 non O139 by serogrouping. He recovered successfully with timely diagnosis, appropriate antibiotics and supportive measures.

Exploring the use of Congo red agar in improving the serotyping of non-serotypeable Shigella with In-silico evidence.

OBJECTIVES: To assess if congo red could make non-serotypeable Shigella strains serotypeable and to employ molecular docking to determine the basis of the same phenomenon. METHODS: We used 42 bacterial strains of non-serotypeable Shigella collected from 2012 to 2019 for this study. Each bacterial strain was freshly inoculated onto congo red agar and incubated at 37° C for 18-24 h. Bacterial colonies obtained were re-subjected to biochemical tests followed by serotyping and serogrouping. In-silico studies to investigate the interaction between MxiC protein of T3SS and O-antigen LPS with congo red were performed. RESULTS: Of the total 42 non-serotypeable Shigella studied, (26/42)62% were capable of being serotyped following the use of congo red agar, 65% were Shigella flexneri, 19% were Shigella dysenteriae, while 2 strains (7%) each of Shigella boydii and Shigella sonnei were detected. We observed no change in their biochemical properties. The in-silico molecular docking studies revealed high binding affinity between congo red and the B-Chain of Mxi C. Out of the 5 chains of the O-Antigen, congo red showed robust binding with the B-chain with the involvement of a cluster of hydrophobic interactions between them. This may have a crucial role in the conversion of non-serotypeable strains to serotypeable strains on exposure to congo red as observed in our study. CONCLUSION: Congo red agar as a medium converts a sizeable percentage of non-serotypeable Shigella strains to serotypeable Shigella strains.

Burden of biocide resistance among multidrug-resistant bacteria isolated from various clinical specimens in a tertiary care hospital.

BACKGROUND: Most studies on biocide resistance and its genetic determinants arise from environmental or food-borne microbial isolates and only a few from clinically relevant isolates. OBJECTIVES: This study determines the proportion of biocide resistance against five commonly used biocides and detects biocide resistance genes among MDR bacterial isolates using PCR. METHODS: Consecutive MDR isolates (n â€‹= â€‹180) were included (30 each of Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, Staphylococcus aureus, and Enterococcus species) from clinical specimens of various inpatient units at JIPMER. The isolates were challenged at 0.5,1 and 2 Macfarland (McF) inoculum with discrete dilutions of disinfectants. The minimum bactericidal concentrations (MBCs) for 70% Ethanol, 1.5% Cresol, 2% Glutaraldehyde, 1% Cetrimide, and 1% Chlorhexidine were determined for the isolates using ATCC reference strains as controls. PCR was performed targeting qac A/B, G; smr; and nfx B genes. RESULTS: For all biocides, MDR isolates had MBCs less than the maximum MBCs of ATCC strains. For MDR K. pneumoniae, A. baumannii, and P. aeruginosa, the highest MBCs of chlorhexidine and cetrimide were ≥75 and â€‹≥ â€‹150 â€‹µg/ml respectively at 0.5 McF inoculum; whereas these organisms grew at higher inoculum (2McF) even at commercially recommended biocidal concentration (1%) corresponding to 750 and 1500 â€‹µg/ml of chlorhexidine and cetrimide respectively. Meanwhile, the highest MBCs of MDR E. coli were 75 and 150 â€‹µg/ml for chlorhexidine and cetrimide respectively. Interestingly, the Gram-positive cocci survived the action of up to 35% ethanol. The nfxB and qacG genes were detected in 87% and 6.67% of MDR P. aeruginosa isolates respectively with no biocide resistance genes detected among the other organisms. CONCLUSIONS: Biocide dilutions challenged with higher inoculum indicated a narrow margin of effectiveness for certain biocides. Although a significant proportion of clinical MDR isolates of P. aeruginosa harbored biocide resistance genes, this finding had no phenotypic correlation.

Bacteriuria due to Typhoidal and Nontyphoidal Salmonella: A Report of Three Cases from South India.

Urinary tract infection (UTI) with Salmonella is uncommon, accounting for merely 0.01% to 0.1% of cases of UTIs. It is reported more frequently in the presence of predisposing factors such as structural abnormalities of the urinary tract or weakened immune system. We present a case series of three patients with Salmonella bacteriuria and their susceptibility patterns. All three patients had underlying urologic features such as neurogenic bladder, chronic kidney disease, and urethral stricture, and two presented with urinary tract involvement symptoms.

Third-generation cephalosporin-resistant Vibrio cholerae, India.

Vibrio cholerae resistance to third-generation cephalosporins is rarely reported. We detected a strain that was negative for extended-spectrum ß-lactamase and positive for the AmpC disk test, modified Hodge test, and EDTA disk synergy test and harbored the blaDHA-1 and blaNDM-1 genes. The antimicrobial drug susceptibility profile of V. cholerae should be monitored.

Antibiotic resistance pattern among common bacterial uropathogens with a special reference to ciprofloxacin resistant Escherichia coli.

BACKGROUND & OBJECTIVES: The resistance of bacteria causing urinary tract infection (UTI) to commonly prescribed antibiotics is increasing both in developing as well as in developed countries. Resistance has emerged even to more potent antimicrobial agents. The present study was undertaken to report the current antibiotic resistance pattern among common bacterial uropathogens isolated in a tertiary care hospital in south India, with a special reference to ciprofloxacin. METHODS: A total of 19,050 consecutive urine samples were cultured and pathogens isolated were identified by standard methods. Antibiotic susceptibility was done by Kirby Bauer disk diffusion method. The clinical and demographic profile of the patients was noted. RESULTS: Of the 19,050 samples, 62 per cent were sterile, 26.01 per cent showed significant growth, 2.3 per cent showed insignificant growth and 9.6 per cent were found contaminated. Significant association (P<0.001) of prior use of antibiotics in males, UTI in adults, gynaecological surgery in females, obstructive uropathy in males and complicated UTI in females with the occurrence of UTI with ciprofloxacin resistant Escherichia coli was noted. Significant association was noted in females with prior antibiotics, with prior urological surgery and in males with prior complicated UTI. There was no significant association with diabetes mellitus with the occurrence of UTI with ciprofloxacin resistant E. coli. Fluoroquinolone resistance was found to increase with age. INTERPRETATIONS & CONCLUSIONS: Ciprofloxacin resistance has emerged due to its frequent use. This resistance was seen more in the in-patients, elderly males and females. Also the resistance to other antibiotics was also high. Increasing antibiotic resistance trends indicate that it is imperative to rationalize the use of antimicrobials in the community and also use these conservatively.

Increasing antimicrobial resistance of Vibrio cholerae OI biotype E1 tor strains isolated in a tertiary-care centre in India.

The antimicrobial susceptibility patterns are on constant change with the recent emergence of multidrug-resistant strains of most bacteria. Results of recent studies in India showed that most isolates of Vibrio cholerae O1 were resistant to the commonly-used antibiotics. The study was conducted to determine the antibiotic susceptibility patterns of V. cholerae O1 isolated during 2008-2010 at the hospital of the Jawaharlal Nehru Institute of Post Graduate Medical Education and Research, Puducherry, India. In total, 154 strains of V. cholerae O1 from 2,658 stool specimens were reported during January 2008-December 2010--34 in 2008, 2 in 2009, and 118 in 2010. The isolates of V. cholerae O1 were subjected to antimicrobial susceptibility testing using the Kirby-Bauer method. The antibiotic disks tested were tetracycline (30 microg), furazolidone (100 microg), ampicillin (10 microg), ceftriaxone (30 microg), and ciprofloxacin (5 microg). Escherichia coli ATCC 25922 was used as the control organism. The minimum inhibitory concentrations (MICs) of ceftriaxone, ciprofloxacin, and tetracycline were determined using the agar dilution method for all the strains. The E-test method was used for the strains which had either intermediate resistance or were resistant to the antibiotics by the agar dilution method. The results of the agar dilution corroborated the results of the E-test. The MIC of ceftriaxone in 151 strains was <2 microg/mL while it was 16 microg/mL in three strains; the latter three strains were resistant to ceftriaxone by the disc-diffusion test. The MIC of ciprofloxacin in 150 strains was <0.5 microg/mL while the MIC of tetracycline was <1 microg/mL. In the remaining four strains, the MIC of tetracycline was >32 microg/mL, and the MIC of ciprofloxacin was >8 microg/mL. These four strains were resistant to both tetracycline and ciprofloxacin by the disc-diffusion test and were exclusive of the three ceftriaxone-resistant strains. The majority of the isolates were obtained from children aged 0-5 year(s)-70.3% (83 of 118) and 41.2% (14 of 34) were reported in 2010 and 2008 respectively. Since treating severe cases of cholera with antibiotics is important, the continuing spread of resistance in V. cholerae to the most important agents of therapy is a matter of concern. Also, chemoprophylaxis with antimicrobial agents is likely to become even more difficult.

CRISPR-cas heterogeneity and plasmid incompatibility types in relation to virulence determinants of Shigella.

Introduction. Virulence factors (VFs) are the most potent weapon in the molecular armoury of Shigella. In bacteria, the mobile genetic elements (MGEs) are contributors to the evolution of different types of clustered regularly interspaced short palindromic repeats-CRISPR associated genes (CRISPR-cas) variants and plasmid incompatibility types. The present study explored the virulence potential of Shigella in relation to the CRISPR-cas pattern and incompatibility types among the isolates.Hypothesis/Gap Statement. The profile of the CRISPR-cas systems among clinical isolates of Shigella in India has not been reported earlier. Limited knowledge is available on the pattern of plasmid incompatibility groups among clinical isolates Shigella. The bias is always towards studying the genetic elements associated with AMR, but the present study highlights CRISPR-cas and incompatibility types among Shigella in association with virulence.Aim. We aimed to investigate the distribution of virulence factors, CRISPR-cas pattern followed by plasmid incompatibility types among Shigella isolates.Methodology. Between 2012-2017, a total of 187 isolates of Shigella were included in the study. The virulence genes' distribution was carried out. CRISPR-cas profiling followed by analysis of the repeats and spacers was carried out. PCR-based replicon typing was used to determine the incompatibility types. The interplay was statistically determined using STATA.Results. The distribution of virulence genes showed varied pattern with ipaH present in all the isolates followed by ompA (93.6 %), virF (66.8 %), ial and sen (60.4 %), set1A (39.6 %) and set1B (39 %). CRISPR 1, CRISPR 3 and Cas6-Cas5 region were dominantly conserved. Twenty-two types of spacers were identified. The CRISPR3 repeat appeared to have a highly conserved sequence. CRISPR2 being the least common CRISPR type showed a strong association with an array of virulence genes (ial-set1A-set1B-virF) while CRISPR1 being the most dominant showed the least association with virulence genes (sen-virF). The dominant plasmids were found to be belonging to the inc FII group. The incompatibility groups FII, IncIγ, U, FIIS, FIIK, K, A/C, I1alpha was found to be associated with a greater number of virulence genes.Conclusion. The isolates showed increasing diversity in their gene content that contributes to increasing heterogeneity among the isolates, which is a known virulence strategy among pathogens.

Association Between Helicobacter pylori and Periampullary and Pancreatic Cancer: a Case-Control Study.

INTRODUCTION: Helicobacter pylori has been implicated in the etiopathogenesis of various malignant conditions; however, there is a dearth of studies on the correlation between H. pylori infection and pancreatic cancers. Hence, this study was carried out to evaluate the association between H. pylori infection and periampullary and pancreatic cancer. METHODS: This was a single-centre, retrospective, case-control study in which all consecutive patients of periampullary or pancreatic cancer were included. The demographic details with tumour characteristics were recorded. Age and gender-matched controls were patients with other extra-abdominal benign conditions. H. pylori and the Cag A status were determined using IgG antibodies and Cag A antibodies respectively. The association between H. pylori infection and periampullary and pancreatic cancer was the primary outcome. RESULTS: A total of 155 patients, 61 in the study and 94 in the control group were included. The overall prevalence of H. pylori in the study group (78.6%) was similar to that of the control group (76.5%) (p = 0.76). Although a higher trend of IgG and Cag A seropositivity was seen in the study group, the difference was not significant. The correlation of H. pylori and Cag A seropositivity showed a higher trend with site-specificity, differentiation, and nodal status. However, the difference was not significant. CONCLUSION: There was no association between H. pylori infection and Cag A seropositivity with periampullary and pancreatic cancers. The various tumour characteristics were also not associated with H. pylori infection. Thus, routine eradication of H. pylori infection may not be recommended in periampullary and pancreatic cancers.

In-vitro study of the effect of Centella asiatica on cholera toxin production and the gene expression level of ctxA gene in Vibrio cholerae isolates.

ETHNOPHARMACOLOGICAL RELEVANCE: Centella asiatica (L.) Urb or Indian pennywort is a plant of ethnopharmacological relevance, commonly called as Brahmi in South India known for its antimicrobial property in gut and for the treatment of other gut ailments. Natural anti-virulence drugs that disarm pathogens by directly targeting virulence factors or the cell viability and are thus preferred over antibiotics as these drugs impose limited selection pressure for resistance development. In this regard, an in-vitro experimental study was conducted to know the effect of extract of Centella asiatica(L.) Urb. on cholera toxin, gene expression and its vibriocidal effect on five standard strains of Vibrio cholerae; IDH03097 (El Tor variant), N16961 (El Tor), O395 (Classical) as well as five clinical strains (Haitian variant). AIM OF THE STUDY: To study the effect of extract of Centella asiatica on Vibrio cholerae. MATERIALS AND METHODS: Crude extract was prepared from the leaves and stem part of the plant. The vibriocidal concentration was tested at different concentrations of the extract. The amount of cholera toxin released from the strains before and after exposure to the extract of Centella asiatica to Vibrio cholerae was measured using Bead ELISA. ctxA gene expression in the strains before and after exposure to extract of Centella asiatica was measured using quantitative real time PCR. All the above assays were performed with commercially obtained asiaticoside as well. RESULTS: The vibriocidal activity was tested at the different concentration of the extract, where 1g/mL of crude extract and 12.5mg/mL of asiaticoside was found to be vibriocidal. The amount of cholera toxin released before and after the exposure to extract of C. asiatica was measured using Bead ELISA, showing a reduction of 70%, 89% and 93% toxin produced by classical, El Tor and variant respectively. ctxA gene expression before and after exposure to extract of Centella asiatica as well as asiaticoside was measured using qRT-PCR. We found a decrease in expression of ctxA gene transcription by 6.19 fold in classical strain, 4.29 fold in El Tor, 1.133 fold in variant strains and about 10.13-10.20 fold for the clinical strains of V. cholerae using the extract of C.asiatica while, the reduction with the exposure to the asiaticoside were 2.762 fold in classical strain, 4.809 in El Tor, 24.1 in variant strain and 34.77 - 34.8 for the clinical strains. CONCLUSION: Centella asiatica extract inhibited the CT production in Vibrio cholerae as well as decreased the transcription of ctxA gene expression.

Diagnostic Accuracy of Point-of-care Nitrite and Leukocyte Esterase Dipstick Test for the Screening of Pediatric Urinary Tract Infections.

Our study aimed to evaluate the diagnostic performance of point-of-care nitrite and leukocyte esterase (LE) dipsticks in the diagnosis of suspected urinary tract infection (UTI) in infants <6 months (young infants) versus older children. The secondary objectives were to study the dipstick efficacy in children with congenital anomalies of the kidney and urinary tract (CAKUT) versus those without CAKUT; in children with simple UTI versus complicated UTI; and to evaluate the clinico-microbiological profile of children presenting with UTI. In this prospective observational study, cases with suspected UTI were enrolled from pediatric emergency or outpatient departments. Urine was collected for performing the urine dipstick and culture. Descriptive data regarding CAKUT, age, gender, etc., were recorded in a predesigned pro forma. We screened 506 children with suspected UTI, of whom 221 had urine culture positive. Approximately 38.4% of the children with UTI had underlying CAKUT, while 7.6% had renal scars. The most common CAKUT was vesicoureteric reflux (VUR). About 12 patients (2.3%) were known to have CAKUT at the time of enrollment in the study. In infants <6 months, LE dipstick had sensitivity 92%, specificity 89.7%, positive predictive value (PPV) 86.7%, negative predictive value (NPV) 93.8%, likelihood ratio (LR) + 8.9, LR- 0.09. In infants <6 months, nitrite dipstick had sensitivity 38%, specificity 97%, PPV 90.4%, NPV 68%, LR+ 12.6 and LR-0.63. In the age group 6 months to 12 years, the efficacy was better for both dipsticks. In age group more than 6 months to 12 years, LE dipstick had sensitivity 96.4%, specificity 95.8%, PPV 94.8 %, NPV 97.2%, LR+ 22.9, LR- 0.04. In age group more than six months to 12 years, nitrite dipstick had sensitivity 94.7%, specificity 99.5%, PPV 99.3%, NPV 96%, LR+ 189.4, and LR-0.05.

Asymptomatic bacteriuria among medical inpatients: Data from an Indian teaching hospital.

Studies indicate that asymptomatic bacteriuria in medical inpatients is often inappropriately treated with antibiotics. We prospectively studied the proportion of asymptomatic bacteriuria among 200 positive urine cultures which were ordered in hospitalised medical inpatients of a teaching hospital in southern India. We used pre-defined criteria to classify patients as urinary tract infection and asymptomatic bacteriuria. Median age of patients was 53.5 (42-65) years, and 51% were male. In all, 157 (78.5%) patients had urinary tract infection (131 [66.5%] definite and 26 [13%] probable) and 43 (21.5%) had asymptomatic bacteriuria. In patients with asymptomatic bacteriuria, 18 (41.8%) received urinary tract infection-directed antibiotics; broad spectrum antibiotics were used in 10 (23%). Patients with asymptomatic bacteriuria were younger, more likely to be on a urinary catheter, had higher prevalence of chronic kidney disease and congestive cardiac failure and had lower prevalence of pyuria and lower total leucocyte counts. Urine cultures should be ordered only in indicated patients. Inappropriate antibiotic treatment in patients with asymptomatic bacteriuria should be avoided.

Surveillance Stool Culture and its Association with Microbiologically Documented Infection During Febrile Neutropenia in Patients with Acute Leukemia (AL) Undergoing Induction Chemotherapy.

The study aimed at identifying the profile of gut colonization of patients with acute leukemia who underwent induction chemotherapy and its association with induction events and outcome. Baseline bacterial stool culture with resistance pattern of isolates were recorded. Multi-drug resistance was defined as resistance to at least two antibiotic classes including beta lactam and fluoroquinolones. During induction chemotherapy, blood and clinically indicated cultures were taken during febrile neutropenic episodes. Association studies were done between gut colonization and induction events/outcome. Among 109 patients enrolled, 71 (65.13%) patients undergoing induction chemotherapy were colonized with bacteria, with nearly 50% of colonizers harboring multi-drug resistant bacteria. Organisms isolated from stool pre-induction were mostly gram negative (98%), with Escherichia coli and Klebsiella pneumoniae being the commonest. 65.13% patients developed febrile neutropenia. Overall multi-drug resistant positivity during febrile neutropenia was 70.14%. Concordance of 8.45% was observed between isolates from stool and organisms isolated from cultures during febrile neutropenia. There were significant proportion of gut colonized gram-negative multi-drug resistance bacteria among patients with acute leukemia. There was a low concordance rate between baseline stool isolates and subsequent cultures during the induction. There was no significant association between gut colonization and induction events/outcomes studied.

Diarrhea and Acute Tubular Necrosis Mimicking Hemolytic Uremic Syndrome in a Man With Immunoglobulin A (IgA) Nephropathy.

Immunoglobulin A (IgA) nephropathy is a heterogeneous disease with variable clinical presentations ranging from asymptomatic hematuria to advanced renal failure. A young male diagnosed with IgA vasculitis (skin, joints, and gastrointestinal) one month ago and placed on oral steroids presented with acute diarrhea, hemolytic anemia, renal failure (non-dialysis requiring), altered sensorium, and thrombocytopenia. The stool was found to be positive for Shiga toxin. He improved with methylprednisolone pulse alone, and renal biopsy showed acute tubular injury.