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At present there are used two methods of standardization of the propolis preparations: the chemical methods and the microbiological ones. The first methods are based on the determination of a sum of either phenolic compounds or flavonoids, whereas the second ones comprise the determination of MIC (Minimal Inhibitory Concentration) or antibacterial activity of the propolis preparations in units. Both liquid and agar media as well as the standard strains of Staphylococcus aureus, Bacillus cereus and Bacillus subtilis are applied for these purposes. It seems to be worth to introduce pharmacological standardization into practice, for the internal preparations in particular.
Studies on the antibacterial action of 592 samples of propolis from various regions of Poland showed that ethanolic extracts of propolis differ by their antibacterial activity. The minimal inhibitory concentration (MIC) of the extracts ranged from 60 to 490 µg/ml and the minimal bactericidal concentration (MBC) ranged from 110 to 1680 µg/ml for Staphylococcus aureus 209P (Oxford). Samples of propolis of a determined bacteriostatic activity differ clearly by the MBC values. Therefore, standardization of propolis should be based both on the MIC and MBC values. Authors suggest that the most active propolis should characterize the MIC below 300 µg/ml and the MBC not more than 450 µg/ml. The methods of extraction and antibacterial activity determination have to be strictly defined because they affect both the MIC and MBC of propolis.
Bacteriostatic activity against Staphylococcus aureus of eleven single flavonoids, pinostrobin chalcone, cinnamic acid and four of its derivatives, hydrocinnamic acid, benzoic acid and six of its derivatives and vanillin occurring in propolis has been determined quantitatively by the method of serial dilutions. Moreover, the determination of bacteriostatic activity has been performed using ethanolic extract of propolis as well as that of three fractions obtained separately from propolis and identified earlier by GC-MS method. These were the following fractions: 1) the water fraction, 2) the fraction volatile with water vapour, 3) the flavonoids fraction. Bacteriostatic activities were also established for four flavonoids and eight organic acids using BIO method and the results of the used methods are compared and discussed. The highest bacteriostatic activity against Staphylococcus aureus displayed the following flavonoids: kaempferid, galangin, apigenin, quercetin, rhamnetin, pinocembrin and kaempherol.
The experimental materials comprised 400 Ross 308 chickens (200 ♂ and 200 ♀), divided randomly into four groups kept in 16 pens, as follows: a negative control group (I) – a diet without additives, a positive control group (II) – a diet supplemented with a combination of the antibiotic growth promoter flavomycin (10 mg kg⁻¹ feed) and the coccidiostat robenidine (500 mg kg⁻¹ starter and grower feed), and two experimental groups – diets supplemented with chemically standardized propolis – 10 mg kg⁻¹ feed in group 3 and 50 mg kg⁻¹ feed in group IV. The body weights of chickens were determined once a week. Feed intake, mortality and culling rates were monitored regularly. At the end of the experiment, on day 42, 12 chickens (6 ♂ and 6 ♀) were selected from each group for slaughter and carcass quality analysis. The final body weights of chickens, feed intake (kg) per kg body weight and carcass weight during the rearing period did not differ significantly between groups. The lowest feed intake per kg lean meat was noted in chickens fed a diet with 50 mg propolis/kg feed (3.457 kg), and the highest – in broilers fed a diet with 10 mg propolis/kg feed (3.611 kg). The weight of carcass and selected carcass parts (neck, breast, legs) tended to increase in chickens that received propolis, although the noted differences were statistically non-significant. The percentage content of legs in the carcass was significantly lower in chickens fed a diet with an antibiotic growth promoter (AGP). The legs of chickens fed 50 mg propolis had a significantly higher muscle content and a lower fat content, compared with the legs of birds fed a diet with AGP.
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