Validation Of Reversed-Phase High-Performance Liquid Chromatography Method For Simultaneous Determination Of 6-, 8-, 10-Gingerols And 6-Shogaol From Ginger Extracts

Objective: Development and validation of a High-Performance Liquid Chromatography (HPLC) method for the simultaneous estimation of 6-, 8-, 10-Gingerols and 6-Shogaol in ginger extract using authentic standards. Methods: The chromatographic separation was achieved by using a C18 column and a mobile phase composed of acetonitrile, ortho-phospohoric acid in water and methanol. The proposed method was validated in terms of the analytical parameters such as specificity, accuracy, precision, linearity, range, the limit of detection (LOD) and limit of quantification (LOQ) according to ICH guidelines. Results: Linear calibration curves were obtained over concentration ranges of 10-250 µg/ml for 6-, 8-, 10-gingerols and 6-shogaol with determination coefficients more than 0.99 for each analyte. Intra and inter-day precisions of the method were found to be below 2% for each analyte, with relative standard deviation (% RSD) values in the range of 0.47 to 1.55% for 6-gingerol, 0.44 to 1.51% for 8-gingerol, 0.24 to 1.90% for 10-gingerol and 0.25 to 1.67% for 6-shogaol. The percentage recovery of gingerols and shogaol was obtained with an average of 99.53%, 99.97%, 100.13% and 100.53% respectively, which was well within acceptance range. Conclusion: Simple, accurate, precise and rapid HPLC method was developed for the simultaneous analysis of 6-, 8-, 10-gingerols and 6-shogaol and validated in accordance with ICH guidelines. The developed method was found to be suitable for the standardization of herbal extracts and polyherbal formulations for the content of 6-, 8-, 10-gingerols and 6-shogaol.

Clinico-microbiological analysis of toxigenic clostridium difficile from hospitalised patients in a tertiary care hospital, Mangalore, Karnataka, India

Purpose: Prevalence of Clostridium difficile, an anaerobic, Gram-positive, spore-forming bacillus, is very much underestimated in India. The present study was intended to assess the burden of toxigenic C. difficile in hospitalised patients with clinically significant diarrhoea and analysis of their clinical picture. Materials and Methods: This cross-sectional study was conducted in a tertiary care teaching hospital, South India, from January 2012 to December 2014. Stool samples were collected consecutively from 563 inpatients from various wards. The prevalence of toxigenic C. difficile was determined by toxigenic culture and a two-step algorithm. The clinical spectrum of these patients was also analysed. Associated pathogens were identified using standard procedures. Statistical analysis was done by frequency, percentage, Chi-square test and z-test. Results: Out of the 563 stool samples analysed, the prevalence of toxigenic C. difficile was 12.79% and that of non-toxigenic C. difficile was 10.83%. The prevalence of toxigenic C. difficile among oncology patients was highly significant (HS). Antibiotic treatment, prolonged hospital stay and underlying diseases/conditions were the risk factors which were HS, and fever was the significant clinical feature among the patients. Escherichia coli was the predominant associated pathogen isolated (18.47%). Conclusion: The presence of toxigenic C. difficile in our locality is a matter of concern. Constant supervision, appropriate treatment and preventive measures are crucial in controlling C. difficile infection.

Comparative analysis of virulence factors & biotypes of Gardnerella vaginalis isolated from the genital tract of women with & without bacterial vaginosis

Background & objectives: Bacterial vaginosis (BV) involves the presence of a thick vaginal multispecies biofilm, where Gardnerella vaginalis is the predominant species. The reason for an increase in the number of G. vaginalis which are usually present as normal flora of the female genital tract in cases of BV, is not known. Hence, the objective of the present study was to compare the biotypes and virulence factors of G. vaginalis isolated from the genital tract of women with and without BV. Methods: High vaginal swabs collected from 811 women of reproductive age were cultured. G. vaginalis isolates were biotyped and tested for adherence to vaginal epithelial cells, biofilm formation, agglutination of human red blood cells (RBCs), protease production, phospholipase production and surface hydrophobicity. Results: Of the isolates from women with BV, 83.3 per cent (60/72) showed good adherence, 78.4 per cent (58/74) produced biofilm, 82.9 per cent (63/76) produced phospholipase, 67.1 per cent (51/76) produced protease, 77.3 per cent (58/75) were positive for surface hydrophobicity and 61.6 per cent (45/73) were positive for haemagglutination of human RBC. In case of G. vaginalis from non-BV women, 25 per cent (15/60) isolates showed good adherence, 18.4 per cent (9/49) biofilm production, 35 per cent (21/60) phospholipase, 36.6 per cent (22/60) protease, 41.7 per cent (25/60) surface hydrophobicity and 10.1 per cent (6/59) agglutination of human RBCs. Maximum number of isolates belonged to biotypes 6, 2 and 3. Biotype 3 was more associated with non-BV rather than BV; biotype 6, 2 and 1 were more associated with cases of BV. Maximum virulence factors were expressed by biotypes 6, 2 and 1. Interpretation & conclusions: Virulence factors were more expressed by G. vaginalis isolates obtained from women with BV rather than from non-BV. Biotypes 6, 2 and 1 were more associated with cases of BV and expressed maximum virulence factors.

Salmonella enterica serotype dublin bacteraemia mimicking enteric fever.

Salmonella enterica serotype Dublin, a bovine adapted serotype, is the commonest cause of salmonellosis in cattle. Salmonellosis in animals always presents a potential zoonotic threat. Infected cattles serves as a source of infection to humans. We present here Salmonella Dublin Bacteraemia in an elderly patient, with all the clinical details, due to the rarity of its occurrence. He was treated successfully with ciprofloxacin and his follow up period was uneventful.

Food poisoning due to Salmonella enterica serotype Weltevreden in Mangalore.

An outbreak of food poisoning due to Salmonella enterica serotype Weltevreden ( S.weltevreden ) involving 34 students has been reported from a tertiary care hospital in Mangalore. The symptoms developed 8-10 hours, after consuming a non- vegetarian dish, probably fish, from an outside caterer. The identity of the organism was confirmed at Central Research Institute, Kasauli. This report emphasizes the geographical distribution of this organism in the Coastal Karnataka region. S.Weltevreden may be overlooked due to the biochemical similarity to S. Paratyphi B & S. Typhimurium.

Burkholderia pseudomallei septicaemia - A case report.

Burkholderia pseudomallei, a natural saprophyte widely distributed in soil, stagnant waters of endemic areas, is said to infect humans through breaks in the skin or through inhalation causing protean clinical manifestations including fatal septicaemia. A case of septicaemia in a elderly female diabetic due to B. pseudomallei following a history of fall is being reported with complete details.

Bacteriological spectrum of cholecystitis and its antibiogram.

Bile Cultures for aerobic and anaerobic bacteria were carried out on 125 samples from patients with chronic cholecystitis with cholelithiasis. Cultures grew 71(56.8%) aerobes and 17(13.6%) anaerobic microorganisms. Polymicrobial infection was seen in 7(16.2%) cases. E. coli (45.07%) and Klebsiella (25.35%) were predominant among the aerobes and Bacteroides fragilis (58.82%) was predominant among the anaerobes. Highest incidence of the disease was observed in the fourth decade of life and females predominated in this study.

Intraperitoneal abscess in a new born secondary to Klebsiella septicaemia--a case report.

A five day old female baby was admitted with distension of abdomen since birth and nonbilious vomiting, fever of one day duration. Blood culture grew Klebsiella pneumoniae. Abdominal exploration revealed thick walled cavity containing purulent fluid grew klebsiella pneumoniae which was sensitive to various antibiotics including gentamycin. The child was treated with injection gentamycin and ceftazidime. The child had uneventful recovery and is doing well 3 years post operatively.

Meningitis due to Xanthomonas maltophilia.

During 1st week of post-operative period, a 28 year old female patient operated for left cerebellopontine angle tumor, continued to get fever. Lumbar puncture did not reveal any organisms. She responded to ciprofloxacin. Two months later, she was readmitted with signs and symptoms of meningitis. The CSF tapped on lumbar puncture grew Xanthomonas maltophilia, Gram negative bacilli, sensitive to various antibiotics, ciprofloxacin being one of them. The patient was given ciprofloxacin for 3 weeks. On follow up, a year later she was found to be asymptomatic.

Bacteriology of peritoneal fluid in paediatric surgical emergencies.

This study deals with the incidence of aerobic and anaerobic organisms of the peritoneal fluid of 60 consecutive children subjected to emergency celiotomy. Two-thirds of the cases were below the age of year. Peritoneal fluid was collected immediately after entering the peritoneal cavity and inoculated both for aerobic an anaerobic cultures. Bacterial growth occurred in 34 (57%) of the specimens. Anaerobic bacteria alone grew in 3 (5%) of the specimens, aerobes alone in 22 (37%) of specimens. A mixed aerobic and anaerobic bacteria grew in 9 (15%) of the specimens. There were 44 aerobic and 19 anaerobic isolates. All anaerobes were sensitive to metronidazole while most of the aerobes were sensitive to gentamycin. Culture was positive in 20 out of 42 (47.6%) specimens, where the peritoneal fluid was clear and there was no evidence of peritoneal contamination at the time of surgery.