Plant-derived secondary metabolites as the main source of efflux pump inhibitors and methods for identification

The upsurge of multiple drug resistance(MDR)bacteria substantially diminishes the effectiveness of antibiotic arsenal and therefore intensifies the rate of therapeutic failure.The major factor in MDR is efflux pump-mediated resistance.A unique pump can make bacteria withstand a wide range of structurally diverse compounds.Therefore,their inhibition is a promising route to eliminate resistance phenomenon in bacteria.Phytochemicals are excellent alternatives as resistance-modifying agents.They can directly kill bacteria or interact with the crucial events of pathogenicity,thereby decreasing the ability of bacteria to develop resistance.Numerous botanicals display noteworthy efflux pumps inhibitory activities.Edible plants are of growing interest.Likewise,some plant families would be excellent sources of efflux pump inhibitors(EPIs)including Apocynaceae,Berberidaceae,Convolvulaceae,Cucurbitaceae,Fabaceae,Lamiaceae,and Zingiberaceae.Easily applicable methods for screening plant-derived EPIs include checkerboard synergy test,berberine uptake assay and ethidium bromide test.In silico highthroughput virtual detection can be evaluated as a criterion of excluding compounds with efflux substrate-like characteristics,thereby improving the selection process and extending the identification of EPIs.To ascertain the efflux activity inhibition,real-time PCR and quantitative mass spectrometry can be applied.This review emphasizes on efflux pumps and their roles in transmitting bacterial resistance and an update plant-derived EPIs and strategies for identification.

Characterization of the Novel Insecticidal Crystal Protein Cyt3Aa1 from <i>Bacillus thuringiensis</i>TD516

Bacillus thuringiensis (Bt) produces two families of insecticidal crystal proteins, i.e., crystalline (Cry) and cytolytic (Cyt) toxins. Cyt3Aa1, the newest Cyt family member, is produced by Bt TD516. Bioassay results have shown that Cyt3Aa1 has weak hemolytic activity against human red blood cells and is not toxic to A. aegypti larvae, but causing a teratogenic effect. The three-dimensional structure of Cyt3Aa1 has a typical cytolysin fold containing a β-sheet held by two surrounding α-helical layers, resembling the previously reported Cyt1Aa and Cyt2Aa structures, which indicated that Cyt3Aa1 might be a membrane-perforation toxin and could induce synergism with Cry protein. This study provides a new source of insecticidal crystal proteins, and presents a foundation for understanding the biological characterization of it, which will aid in the development of strategies to cope with the potential problem of insect resistance.

Antibacterial and antioxidant effects from seaweed,Sargassum wightii(Greville,1848)against marine ornamental fish pathogens

Objective:To screen seaweed Sargassum wightii(S.wightii)for bioactive natural substance against marine ornamental fish bacterial pathogens,and also study the antioxidant properties,brine shrimp toxicity effect.Methods:Crude extract was made using three solvents(acetone,ethanol and methanol)and screened for antibacterial activity and purified by column chromatography,purified fractions obtained were tested for the activity.The 1st fraction of acetone extract showed maximum activity,this was again subjected for purification and obtained three sub-fractions also tested for the activity.Total phenols and flavonoid contents,reducing power,free radical scavenging activities(DPPH and H_(2)O_(2))and brine shrimp toxicity were also studied using purified acetone extract followed by standard methods.Results:The purified acetone extract showed maximum activity against eight pathogens among ten.GC-MS results revealed two major compounds such as 24-methylene cholesterol(79.9%)and methyl oleate(30.3%)which presented in higher percentage in purified extract and had highest phenols and flavonoid contents,reducing power,free radical scavenging activities,and also showed less toxicity effect.In he present study,the purified extract of S.wightii had potential antibacterial activity against Aeromonas hydrophila[(22.25±0.35)mm]and minimum activity against Streptococcus sp.[(10.00±0.00)mm].The purified extract of S.wightii also had potential total antioxidant activity of(3.87±0.04)μg at 100μg/mL concentration and the lowest activity was exhibited(1.52±0.01)μg at 25μg/mL.Conclusions:The present study concluded that the brown seaweed,S.wightii has potential antimicrobial and antioxidant activities,which can be used in aquaculture industry for treated bacterial diseases in infected fishes.