Using the polymerase chain reaction coupled with denaturing gradient gel electrophoresis to investigate the association between bacterial translocation and systemic inflammatory response syndrome in predicted acute severe pancreatitis

AIM: To investigate the use of PCR and DGGE to investigate the association between bacterial translocation and systemic inflammatory response syndrome in predicted severe AP.METHODS: Patients with biochemical and clinical evidence of acute pancreatitis and an APACHE Ⅱ score ≥8 were enrolled. PCR and DGGE were employed to detect bacterial translocation in blood samples collected on d1,3, and 8 after the admission. Standard microbial blood cultures were taken when there was clinical evidence of sepsis or when felt to be clinically indicated by the supervising team.RESULTS: Six patients were included. Of all the patients investigated, only one developed septic complications;the others had uneventful illness. Bacteria were detected using PCR in 4 of the 17 collected blood samples. The patient with sepsis was PCR-positive in two samples (taken on d 1 and 3), despite three negative blood cultures. Using DGGE and specific primers, the bacteria in all blood specimens which tested positive for the presence of bacterial DNA were identified as E coli.CONCLUSION: Our study confirmed thatunlike traditional microbiological techniques, PCR can detect the presence of bacteria in the blood of patients with severe AP. Therefore, this latter method in conjunction with DGGE is potentially an extremely useful tool in predicting septic morbidity and evaluating patients with the disease. Further research using increased numbers of patients, in particular those patients with necrosis and sepsis, is required to assess the reliability of PCR and DGGE in the rapid diagnosis of infection in AP.

In-vitro antimicrobial activity of marine actinobacteria against multidrug resistance Staphylococcus aureus

Objective:To investigate the antibacterial aclivily of marine actinobacteria against multidrug resistance Staphylococcus aureus(MDRSA).Methods:Fifty one actinobacterial strains were isolated from salt pans soil,costal area in Kothapattanam,Ongole,Andhra Pradesh.Primary screening was done using cross-streak method against MDRSA.The bioaclive compounds are extracted from efficient actinobacteria using solvent extraction.The antimicrobial activity of crude and solvent extracts was perfomied using Kirby-Bauer method.MIC for ethyl acetate extract was determined by modified agar well diffusion method.The potent actinobacteria are identified using Nonomura key,Shirling and Gottlieb 1966 with Bergey's manual of determinative bacteriology.Results:Among the fifty one isolates screened for antibacterial activity,SRB25were found efficient against MDRSA.The ethyl acetate extracts showed high inhibition against test organism.MIC test was performed with the ethyl acetate extract against MDRSA and found to be 1 000μg/mL.The isolaled actinobacteria are identified as Streptomyces sp with the help of Nonomura key.Conclusions:The current investigation reveals that the marine actinobacteria from salt pan environment can be able to produce new drug molecules against drug resistant microorganisms.

Association of Streptococcus bovis presence in colonic content with advanced colonic lesion

AIM:To prospectively examine the association between presence of Streptococcus bovis(S.bovis)in colonic suction fluid and the endoscopic findings on colonoscopy.METHODS:From May 2012 to March 2013,203 consecutivepatients who underwent colonoscopy for any reason were enrolled in the study.Exclusion criteria included:antibiotic use in the previous month,age younger than18 years,and inadequate preparation for colonoscopy.The colonoscopy was performed for the total length of the colon or to the occluding tumor.The endoscopic findings were registered.Samples were obtained proximal to the colonoscopic part of the suction tube from each patient and sent to the clinical microbiology laboratory for isolation and identification of S.bovis.Samples were incubated in enrichment media with addition of antibiotic disks for inhibition of growth of Gram-negative rods.The samples were seeded on differential growth media;suspected positive colonies were isolated and identified with Gram staining,catalase,and pyrrolidonyl arylamidase tests,and further identified using a VITEK2 system.Statistical analyses were performed using the Student’s t andχ2 tests.RESULTS:Of the 203 patients recruited,49(24%)patients were found to be S.bovis carriers;of them,the endoscopic findings included:17(34.7%)cases with malignant tumors,11(22.4%)with large polyps,5(10.2%)with medium-sized polyps,6(12.2%)with small polyps,4(8.1%)with colitis,and 6(12.2%)normal colonoscopies.Of 154 patients found negative for S.bovis,the endoscopic findings included:none with malignant tumors,9(5.8%)cases with large polyps,11(7.1%)with medium-sized polyps,26(16.9%)with small polyps,7(4.5%)with colitis,and101(65.6%)normal colonoscopies.S.bovis(Grampositive coccus)is considered part of the normal intestinal flora.There is an association between S.bovis bacteremia and colonic neoplasia.It is not well understood whether the bacterium has a pathogenetic role in the development of neoplasia or constitutes an epiphenomenon of colorectal neoplasms.There was a clear relationship between positivity for S.bovis in colonic suction fluid and findings of malignant tumors and large polyps in the colon.CONCLUSION:There is an association between S.bovis bacteremia and malignant colonic lesions;this should prompt for development of a reliable screening method for advanced colonic lesions.

Phytochemical composition and in vitro antioxidant activity of aqueous extract of Aerva lanata(L.) Juss.ex Schult.Stem(Amaranthaceae)

Objective:To analyze the phytochemical composition and in vitro antioxidant properties of aqueous extract of Aerva Lanata（A.lanata） stem.Methods:During the preliminary phytochemical analysis,the aqueous extract of A.Ianata was screened for the presence of carbohydrates,proteins,phenolic compounds,oil and fats,saponins,flavonoids,alkaloids. tannins and phytosterols.Antioxidant activity of the extract was determined by 2.2-dipbenyl- 1-picrylhydrazyl radical scavenging activity,metal chelating activity,reducing power activity and DNA damage inhibition activity.Analysis of phenolic compounds was performed by FolinCiocaiteau reagent method and gradient high performance liquid chromatography technique. Results:Preliminary phytochemical analysis exhibited the presence of phenolic compounds, saponins.flavonoids.tannins and phytosterols as major phytochemical groups.The extract exhibited high 2.2—diphenyl-1-picrylhydrazyl radical scavenging activity(IC50= 110.74μg/ mL).metal chelating activity(IC50= 758.17μg/mL).reducing power activity and DIA damage inhibition efficiency.The extract was reported to possess a high amount of total phenolic content and some of them were identified as gallic acid（3,4,3-OH）,apigenin-7—O-glucoside tapigetrin), quercetin-3—O-rutinoside（rutin） and myricetin（3,5,7,3,4,5-OH）by high performance liquid chromatography analysis.The extract was found non toxic towards human erythrocytes in the hemolytic assay(IC50= 24.89 mg/mL).Conclusions:These results conclud that A.lanata stem possesses high antioxidant activity and can he used for the development of natural and sale antioxidant compounds.

Ethnomedicinal plants used in the treatment of skin diseases in Hyderabad Karnataka region,Karnataka,India

Objective:To document traditional medicinal plants knowledge used in treating skin diseases at Hyderabad Karnataka Region.Methods:The information on the use of medicinal plants in the treatment of skin diseases was gathered from traditional herbal healers and other villagers through interviews.Results:A total of 60 plants species belonging to 57 genera and 34 families were found useful and herewith described them along with the method of drug preparation,mode of administration,probable dosage and duration of treatment.Several new findings on the traditional rural practices were reported.Conclusions:The present study revealed that the Hyderabad Kamataka rural people is primarily dependent on medicinal plants for treating skin diseases.

Vermicompost and Cow Dung Admixture Increases Rhizosphere Bacterial Population and Promotes Rapid Physiological Maturity in Maize (Zea mays L.)

Incessant application of chemical fertilizers to the agricultural fields may alter the composition and activities of soil microbiota.Thus,the shift of cultivation practices from chemical to organic is considered to be the need of the hour in order to maintain soil health.A study was conducted in the agricultural fields of the University of Burdwan,India to observe the impact of organic manure on the rhizosphere bacterial community.The experiments were conducted on maize plants,supplemented with the recommended dose of chemical fertilizer and organic manure(vermicompost and cow dung mixture).Corresponding changes in the plant phenological events and soil health in terms of soil physico-chemical factors and rhizosphere bacterial groups up to the level of CFU g-1×105 dry soil was noted.The results showed a significant increase in population of phosphate solubilizing bacteria during 30DAS.However,at 90 DAS,significant increase in the population of phosphate solubilizing bacteria,nitrifying bacteria,asymbiotic nitrogen-fixing bacteria and protein hydrolyzing bacteria was observed in the organically treated plots.The growth of rhizosphere bacteria was attributed to the type of organic manure supplied to the agricultural fields.In addition,a strong correlation was observed between Zn and protein hydrolyzing bacteria.The soil organic carbon and available nitrogen were strongly correlated with nitrifying,fat solubilizing and phosphate solubilizing groups of bacteria.

Evaluating the effectiveness of marine actinobacterial extract and its mediated titanium dioxide nanoparticles in the degradation of azo dyes

Aim of the present study was to synthesize titanium dioxide nanoparticles （YiO2 NPs） from marine actinobacteria and to develop an eco-friendly azo-dye degradation method. A total of five actinobacterial isolates were isolated from Chennai marine sediments, Tamilnadu, India and analyzed for the synthesis of TiO2 NPs using titanium hydroxide. Among these, the isolate PSV 3 showed positive results for the synthesis of TiO2 NPs, which was confirmed by UV analysis. Further characterization of the synthesized TiO2 NPs was done using XRD, AFM and FI＇-IR analysis. Actinobacterial crude extract and synthesized TiO2 NPs was found efficient in degrading azo dye such as Acid Red 79 （AR-79） and Acid Red 80 （AR-80）. Degradation percentage was found to be 81% for AR-79, 83% for AR-80 using actinobacterial crude extract and 84% for AR-79, 85% for AR-80 using TiO2 NPs. Immobilized actinobacterial ceils showed 88% for AR-79 and 81% for AR- 80, dye degrading capacity. Degraded components were characterized by FT-IR and GC-MS analysis. The phytotoxicity test with 500 μg/mL of untreated dye showed remarkable phenotypic as well as cellular damage to Tagetes erecta plant. Comparatively no such damage was observed on plants by degraded dye components. In biotoxicity assay, treated dyes showed less toxic effect as compared to the untreated dyes.

Isolation and characterization of tyrosinase produced by marine actinobacteria and its application in the removal of phenol from aqueous environment

The present study was focused on screening and characterization of tyrosinase enzyme produced by marine actinobacteria and its application in phenolic compounds removal from aqueous solution. A total of 20 strains were isolated from marine sediment sample and screened for tyrosinase production by using skimmed milk agar medium. Among 20 isolates, two isolates LK-4 and LK-20 showed zone of hydrolysis and these were taken for secondary screening by using tyrosiue agar medium. Based on the result of secondary screening LK-4 was selected for further analysis, such as tyrosinase assay, protein content and specific activity of the enzyme. The tyrosinase enzyme was produced in a SS medium and was partially purified by ammonium sulfate precipitation, dialysis and SDS PAGE. The isolate （LK-4） was identified as Streptomyces espinosus using 16S rRNA gene sequencing and named as ＂Streptomyces espinosus strain LK4 （KF806735）＂. The tyrosinase enzyme was immobilized in sodium alginate which was applied to remove phenolic compounds from water. The enzyme efficiently removed the phenolic compounds from aqueous solution within few hours which indicated that tyrosinasc enzyme produced by Streptomyces espinosus strain LK-4 can be potently used for the removal of phenol and phenolic compounds from wastewater in industries.