The “LMG 3770 LMG 3279 and LMG 14520. the ethanol extract

The “LMG 3770 LMG 3279 and LMG 14520. the ethanol extract of propolis inhibited the development of all analyzed micro-organisms with the best antimicrobial activity against Gram-positive bacterias and (25). The recurring usage of antibiotics in various areas (veterinary and individual medicine) boosts the introduction and occurrence from the level of resistance sensation in pathogenic bacterias. Some seafood bacterial pathogens may also be associated to illnesses in human beings (zoonotic or meals borne illnesses) producing the aquaculture items LY404039 being a potential risk to the clients (6). About the issue of microbial level of resistance there can be an urgent have to establish the guidelines for the logical usage of antibiotics as well as the breakthrough of brand-new drugs and substitute therapies to regulate bacterial illnesses in aquaculture field. Owing the capability to synthesize many different chemicals the propolis is among the top richest resources of new drugs (1 11 It is showed that ethanol extract of propolis has a high potential as an alternative source of antibacterial compounds (2 3 4 5 13 19 20 22 33 Propolis (bee glue) is usually a resinous hive product collected by honeybees ((16 30 Bees change propolis by β-glucodiases enzymes from hypopharyngeal glands during collection and LY404039 processing. Results of this enzymatic modification are hydrolyzation of phenolic compounds like flavonoid heterosides to free flavonoid aglycones and sugars and enhancement of the pharmacological action of HILDA the producing products. Chemically flavonoid aglycones from propolis are flavones flavonols flavanones dihydroflavonols and chalcones. Other phenolic compounds are phenolic aldehydes and polyphenolic derivates of cinnamic and benzoic acid including caffeic acid esters terpenes β-steroids sesquiterpenes naphthalene and stilbene derivatives (16). Several investigations on propolis have been carried out in Eastern Europe and South America but there is no statement about antimicrobial effect of propolis in aquaculture previously. Therefore the aim of the present study was to investigate the antimicrobial activity of ethanol extract of propolis from Iran against three fish pathogenic bacteria that are often the cause of bacterial diseases in aquaculture. MATERIALS AND METHODS Propolis samples Crude propolis samples were collected from honey bee LMG 3770 YLMG LY404039 3279 and LMG 14520. All micro-organisms were provided by Belgian Co-ordinated Selections of Micro-organism Belgium. All bacteria were cultured for 18 h at 28 °C in brain heart infusion broth (Merck Darmstadt Germany) and used as inoculums. Susceptibility assessments The following methods were used to evaluate the activity of the EEIP. All assessments were repeated three times using an 80% ethanol answer without propolis as a control to test the inhibitory effect of the solvent. LY404039 Micro-broth dilution method Minimum inhibition concentrations (MIC) of EEIP against the tested pathological bacterial strains were decided using micro-broth dilution method (26). Briefly serial two-fold dilutions of EEIP (10% w/v) were prepared in 96-well micro-titer plate ((from 1: 2 to 1 1: 8192) made up of cation-adjusted Mueller-Hinton broth (Merck Darmstadt Germany). Control micro-titer plates made up of medium and 80% ethanol at the same dilutions were also made. Bacterial suspensions were adjusted to the 0.5 McFarland standards (approximately 1 to 2 2 × 108 CFU/ml). A constant amount of bacteria were added to all wells and the plate was incubated at 28°C for 18-24 hour (final inoculate were adjusted to the 105 CFU per each well). Each well was examined for growth comparing each well to the control. The MIC was defined as the lowest concentration of propolis at which there was no visible growth of the organisms. For each test enrofloxacin and gentamycin were used as the control antimicrobial brokers. The minimal bactericidal concentration (MBC; the lowest concentration of propolis that resulted in a 99.9% reduction in CFU of the initial inoculums) was determined by plating count the contents of wells that showed no visible growth of.