Isolation of ESBL-producing gram-negative bacteria and in silico inhibition of ESBLs by flavonoids

AbstractObjectiveTo evaluate nosocomial accounts of 426 extended spectrum β-lactamase (ESBL)-producing strains from 705 isolates of 9 pathogenic gram-negative bacteria in vitro. We analysed the genetic divergence of ESBLs by constructing a phylogenetic tree and modelled flavonoid inhibition of ESBLs with in silico molecular docking to determine effective control options.MethodsNine ESBL-producing bacteria were isolated from urine samples and their antibiograms were determined by the disc-diffusion method. Comparative models of the 9 ESBL enzymes were generated computationally using reference sequences, and validated by Ramachandran plots. Molecular docking with 11 flavonoids was conducted against the ESBL models.ResultsIsolated strains were floridly multidrug-resistant. From the docking study, the predicted minimum energy value of amikacin was −8.108 kcal/mol against the wild type TEM-1 ESBL of Acinetobacter baumannii, while the docking value against the mutant type Escherichia coli was −7.388 kcal/mol. The docking scores obtained corroborated the in vitro results showing that the antibiotic was incapable of controlling the ESBL of the mutant strain. Among 11 flavonoids tested against the mutant ESBL of E. coli, epigallocatechin 3-gallate and eriodictyol, with docking scores of −9.448 and −8.161 kcal/mol, respectively, were the most effective, with drug-likeness scores of 0.39 and 1.37, respectively, compared to 1.03 for amikacin.ConclusionDocking scores and drug-likeness scores indicated that flavonoids are compelling alternative antimicrobial agents that could serve as complementary therapy for newly arising ESBL-producing bacteria. Phylogenetic tree analysis elucidated the genetic relationship of the 9 ESBL serotypes

In vitro antibacterial efficacy of plants used by an Indian aboriginal tribe against pathogenic bacteria isolated from clinical samples

AbstractObjectivesTo evaluate antibacterial efficacies of 21 medicinal plants used by an Indian aboriginal tribe against infectious diseases caused by bacteria isolated from clinical samples.MethodsStandard biochemical procedures were followed for identifying bacteria that were isolated from several clinical samples. All of the bacterial strains were subjected to antibiotic sensitivity tests by Kirby–Bauer's disc diffusion method. From antibiograms of isolated Gram-positive and Gram-negative bacteria, it was discernible that samples were multidrug resistant (MDR). The methanol leaf-extract of Solanum xanthocarpum was subjected to thin layer chromatography (TLC) for phytochemical analysis. Molecular docking of β-lactamase enzyme of Escherichia coli with phytochemicals of S. xanthocarpum was performed to locate effective compounds.ResultsThe most effective 5 plants, which caused the size of the zone of inhibition to range from 21 to 27 mm, were Buchanania latifolia, Careya arborea, Ocimum tenuiflorum, Senna alata and S. xanthocarpum, for MDR bacteria. S. xanthocarpum had the lowest MIC value of 0.67 mg/ml and the lowest MBC value of 1.51 mg/ml against E. coli. In the TLC study, 9 spots of methanol leaf-extract of S. xanthocarpum were recorded with two solvent systems. The phytochemicals of S. xanthocarpum, solasodine and stigmasterol glucoside had the highest docking score values, −10.868 kcal/mol and −10.439 kcal/mol, respectively, against β-lactamase.ConclusionThis study could prove in vitro antimicrobial efficacy of 5 uncommon plants against MDR pathogenic bacteria. Solasodine and stigmasterol glucoside were computationally recorded as the best controlling chemicals from the plant S. xanthocarpum

A study on clinical and laboratory profiles of hemorrhagic and ischemic strokes in an Eastern Indian teaching Hospital

Background: Stroke is a frequent cause of death and disability. This study was carried out to establish the pattern of various types of cerebrovascular accident (CVA) and comparison between hemorrhagic and ischemic stroke in Eastern India.Methods: This is a retrospective study of sample size 367 CVA patients; among them, 218 were ischemic and 149 were hemorrhagic admitted to Department of General medicine and Neurology of the hospital.Results: Of the total 367 patients, there were 149 as hemorrhagic stroke patients and 218 patients as ischemic stroke patients. Ratio of hemorrhagic to ischemic stroke was 40.6: 59.4. Among patients in hemorrhagic patients male to female ratio was 1.6:1 and in ischemic stroke patients male to female ratio was 1.27:1. It was recorded that prevalence of ischemic stroke was higher than hemorrhagic type.Conclusions: The CVA was associated with blood pressure, cholesterol and use of tobacco as the common risk factor for ischemic stroke. It was recorded that ischemic stroke patients had reported frequently than hemorrhagic patients

Prevalence of multidrug resistant uropathogenic bacteria in pediatric patients of a tertiary care hospital in eastern India

SummaryToday, because systemic infections such as urinary tract infection (UTI) affect even pediatric patients, antibiotic resistant bacteria have become a constant clinical challenge. In the present study, a total of 1054 urine samples were collected from pediatric patients over 18 months. From these samples, 510 isolates of pathogenic bacteria were collected using HiCrome UTI agar. Antibiotic sensitivity tests of isolates were performed using the Kirby–Bauer method. Two Gram-positive bacteria (Enterococcus faecalis and Staphylococcus aureus) and 7 Gram-negative bacteria (Citrobacter freundii, Enterobacter aerogenes, Escherichia coli, Klebsiella oxytoca, K. pneumoniae, Proteus vulgaris and Pseudomonas aeruginosa) were isolated. Antibiograms of isolated bacteria were ascertained using antibiotics of 4 classes: aminoglycosides, β-lactams, fluoroquinolones and 2 stand-alones (co-trimoxazole and nitrofurantoin). Based on percent values of antibiotic resistance, isolated bacteria were (in decreasing order of number of isolated isolates): E. coli (109)>S. aureus (65)>E. faecalis (82)>E. aerogenes (64)>C. freundii (41)>P. aeruginosa (32)>K. pneumoniae (45)>K. oxytoca (50)>P. vulgaris (22). Surveillance results show that MDR isolates of 9 pathogenic bacteria were prevalent in the environment around the hospital. Thus, revisions to the antimicrobial stewardship program in this area of the country are required to increase clinician confidence in empiric therapy, which is often used for UTI cases

Surveillance of bacteria Pseudomonas aeruginosa and MRSA associated with acute suppurative otitis media (ASOM)

Background: Otitis media particularly with suppuration is a critical disease-causing perforation of the tympanic membrane associated with changes of the mucoperiosteum of the middle ear cleft. This surveillance includes isolation and antibiotic profiles of causative bacteria from ear discharges of patients in 3years attending outpatients of a hospital.Methods: Bacterial strains were grown in suitable media and were subjected to antibiotic profiling by the Kirby-Bauer’s method with most antibiotics of the day.Results: In total there were 1164 colonies with 1043 bacterial and 121 fungal isolates from 1230 ear discharge samples. Among 371 Pseudomonas aeruginosa isolates, tobramycin 30 µg/disk had the highest susceptibility rate as 93.2%, followed by ceftazidime 30µg/disk 91.5% and amikacin 10µg/disk 64.4%. From 359 Staphylococcus isolates, there were 236 coagulase negative Staphylococcus (CONS) + methicillin sensitive S. aureus (MSSA) and 123 methicillin resistant S. aureus (MRSA). Staphylococcus including MRSA isolates were most susceptibility to cloxacillin 15µg/disk 95.2%, followed by erythromycin 15µg/disk 83.3% and gentamicin 30µg/disk 78.5%. Of 1164, 49 patients presented post aural abscess, 12 patients had intracranial complications, 9 patients had facial palsy and 3 patients had labyrinthitis.Conclusions: Isolated bacteria, P. aeruginosa and MRSA were multidrug resistant. P. aeruginosa was most common followed by S. aureus. More than 90% P. aeruginosa and 90% S. aureus isolates were sensitive to tobramycin 30 µg/disk and cloxacillin 30 µg/disk, respectively. Therefore, these two antibiotics may be included in the formulary regimen to overcome bacterial infections involved in ASOM

Radiological significance of isolated ethmoid sinus infections in asymptomatic patients of recurrent acute rhinosinusitis

Background: Recurrent acute rhinosinusitis (RARS) being a low form chronic rhinosinusitis (CRS), is relatively under diagnosed many times because of lack of awareness regarding pattern of the disease presentation. Aim of the study is to evaluate the radiological significance of isolated ethmoid sinusitis in patients of RARS during the asymptomatic interval in between acute episodes of rhinosinusitis.Methods: Total 124 patients with history suggestive of RARS who had undergone computed tomography study of brain during April 2015 to October 2016 for indications other than sinonasal diseases, were evaluated for isolated ethmoid sinusitis in the department of Radiology. Patients having any inflammatory or expansile sinonasal masses were excluded.Results: Of total 124 patients of RARS aged from 18 to 70 years (mean age of 38.6 years), the study population included only seventy-two (58%) patients having isolated ethmoid sinus infections, were classified in to mild unilateral group I (46%) cases with grade I or II sinus infections and relatively severe bilateral group II (54%) cases having grade III or IV infections. Infective process involving two to three ethmoid cells each on either side is the most common variety encountered in 39% cases. Prevalence of infective process was fairly equal among all age groups. Statistical correlation between the severity of RARS in patients having group II variety of isolated ethmoid sinus infections was found highly significant.Conclusions: Isolated ethmoid sinus infections are commonly encountered CT findings in adult RARS patients while they are asymptomatic from acute episodes of rhinosinusitis

Prevalence and identification of Candida sp. in pregnant women using VITEK-2

Background: Candida sp. is seen in several areas of body such as, mouth, groin area including vagina and digestive tract as thrush or gastroenteritis. The slide-culture technique and the VITEK-2 automated system were used for species-identification of the fungus; nonetheless, a gold standard or any first identification method would have inherent errors in arriving at a correct identification of a microorganism at species level.Methods: Morphological fungal criteria were ascertained with germ tubes, glucose agar, sugar fermentation and sugar assimilation tests Candida from vaginal swabs and other clinical samples of 85 infected pregnant women with diabetes, by growing swab lots on Sabouraud’s Dextrose Agar (SDA) plates, the slide culture technique and the VITEK-2 automated system.Results: Of 85 patients, 122 isolates in SDA culture were determined as 7 Candida sp.  with number of isolates of each species, as follows: 47 C. albicans, 9 C. famata, 11 C. glabrata, 13 C. guilliermondii, 8 C. krusei, 3 C. parapsilosis and 37 C. tropicalis from vaginal swabs. From 60 vaginal swabs, 46 urine samples and 12 throat swabs it was seen that C. albicans was most prevalent. However, withVITEK-2, 201 fungal strains were identified; Candida sp. was isolated in all samples: 59 C. albicans, 19 C. famata, 21 C. glabrata, 23 C. guilliermondii,18 C. krusei, 13 C. parapsilosis and 48 C. tropicalis.Conclusions: The most prevalent species among the isolated fungi was C. albicans, causing VC in diabetic pregnant women

Radiological significance of the sinonasal anatomical variants in recurrent acute rhinosinusitis patients

Background: Recurrent acute rhinosinusitis (RARS), a low form chronic rhinosinusitis is frequently under evaluated. The significance of sinonasal anatomical variants of osteomeatal complex (OMC) and spheno-ethmoidal (SE) recess regions in patients of RARS is assessed.Methods: Retrospective analysis of coronal sinonasal computed tomography images of 120 RARS patients presented with sinonasal anatomical variants during November 2013 to October 2016 was carried out. Patients with acute and expansile sinonasal lesions are excluded.Results: Sinonasal anatomical variants in the regions of OMC and SE recess are responsible for obstruction of normal mucociliary drainage of corresponding paranasal sinuses in presence of inflammation predisposing to RARS. Anatomical variants of nasal septum were, deviated nasal septum (DNS) in 86 (72%) and septal spur in 58 (48%) patients. Anatomical variants in OMC region were, pneumatized middle turbinate (concha bullosa) in 50 (48%), paradoxical middle turbinate in 38 (28%), giant ethmoid bulla in 35 (32%), agger nasi cell in 29 (38%), Haller cell in 23 (15%), pneumatised uncinate process in 20 (18%), medialized uncinate process in 18 (22%) and septated maxillary sinus in 5 (4%) patients. Anatomical variants in SE recess were superior concha bullosa in 14 (12%) and septal bullosa in 11 (9%) patients.Conclusions: OMC pattern of recurrent rhinosinusitis is often prevalent in patients of RARS due to abundance of anatomical variants in the OMC region. DNS or ethmoid bulla when gets associated with concha bullosa increased the incidence of RARS. Present analysis would help surgeons to evaluate RARS patients for selective endoscopic sinus surgery

Surveillance of acute community acquired urinary tract bacterial infections

Objective: To record the antibiotic resistance of community acquired uropathogens over a period of 24 months (May 2011–April 2012). Methods: Urine samples from patients of outpatient department (OPD) were used for isolating urinary tract infection (UTI)-causing bacteria that were cultured on suitable selective media and identified by biochemical tests. Their antibiograms were ascertained by Kirby–Bauer's disc diffusion method, using 17 antibiotics of 5 different classes. Results: From 2137 urine samples 1332 strains of pathogenic bacteria belonging to 11 species were isolated. Two Gram-positives, Staphylococcus aureus and Enterococcus faecalis and nine Gram-negatives, Acinetobacter baumannii, Citrobacter sp., Escherichia coli, Enterobacter aerogenes, Klebsiella oxytoca, Klebsiella pneumoniae, Proteus mirabilis, Proteus vulgaris and Pseudomonas aeruginosa were isolated. Both Staphylococcus aureus and Enterococcus faecalis were vancomycin resistant, and resistant-strains of all pathogens increased in each 6-month period of study. Particularly, all Gram-negatives were resistant to nitrofurantoin and co-trimoxazole, the most preferred antibiotics of empiric therapy for UTI, but were moderately resistant to gentamicin, ampicillin, amoxyclav, ofloxacin and gatifloxacin. Most Gram-negatives produced extended spectrum β-lactamase. Conclusions: It was concluded that periodic surveillance of pathogens is an essential corollary in effective health management in any country, as empiric therapy is a common/essential practice in effective clinical management