Antimicrobial activity of the substances received from raw materials of Birch family plants

Abstract

Introduction. In accordance with the last events in Ukraine (considering military operations in anti-terrorist operation in the Luhansk and Donetsk regions) the domestic medicine is in great need in preparations with antimicrobial activity. Our attention as the sources of receiving biologically active substances with antimicrobial activity was drawn with birch Betulaceae family plants – hazel ordinary Corylus avellana L. and black alder Alnus glutinosa (L.) Gaertn. It is known that in medicine the leaves of hazel ordinary are used as antiseptic, anti-inflammatory, vesselrestorative drug, and the leaves of black alder reveal the antiinflammatory, astringent, wound healing, spasmolytic and choleretic action. However, the drugs with antimicrobial action received from the leaves of these plants are absent on the market of Ukraine. Therefore the studying of antimicrobial activity of this type of raw materials received from hazel ordinary and black alder, for creation of new medicines, is now one of the main directions in pharmacy. For this purpose we have revealed tinctures, spirit, lipophilic and polysacharid fractions received from the leaves of hazel ordinary and black alder. The purpose of our research is studying of antimicrobial activity of revealed substance received from the leaves of black alder and hazel ordinary. Materials and methods. There were being examined tinctures, lipophilic, spirit and polysacharid fractions received from the leaves of hazel ordinary and black alder. The test of antimicrobial effect of substances was carried out by means of serial dilution concerning the following six reference cultures: Staphylococcus aureus ATCC 6538-P, Candida albicans ATCC 885-653, Escherichia coli ATCC 25922, Bacillus subtilis ATCC 6833, Bacillus cereus ATCC 10702, Pseudomonas aeruginosa ATCC 9027, according to the State Pharmacopoeia of Ukraine, in the Department of Microbiology and Immunology of KMAPE. For the experiment there was prepared tenfold dilution of extracts on meat-peptone broth 1:10 and 1:100 to which there were added the referential cultures which are grown up on appropriate differential diagnostic medium depending on a type of cultures. The crops were incubated in the thermostat at t - 35-37 °C within 24-48 hours. For identification of test-strain growth of E. coli there was used Endo medium, for S. aureus – Chistovich medium, for B. subtilis and B. cereus - meat-peptone agar, for P. аeruginosa - 5% blood agar and for C. albicans – Saburo medium. As the solutions remained muddy after incubation, for the assessment of antimicrobial action of the received fractions, there were being made the cloning on differential and diagnostic mediums. Results and discussion. Data of the conducted researches of antimicrobial activity of various substances of hazel ordinary and black alder leaves – tinctures, lipophilic, spirit and polysacharide fractions are given in the table. The obtained data demonstrate that lipophilic fractions of hazel ordinary and black alder leaves possessed the bactericidal activity when it was diluted in the ratio of 1:10 and 1:100 concerning P. аeruginosa and E. coli. These fractions also have antimycotic activity concerning C. albicans. Concerning E. сoli it is revealed that the spirit fractions from black alder leaves had the bactericidal activity, and concerning P. аeruginosa and C. albicans – all studied spirit fractions possessed. Among polysacharide fractions the antimicrobial activity is revealed only for this substance of hazel ordinary leaves in both dilutions concerning E. coli, S. aureus, P. aeruginosa. As concerning B. subtilis and B. cereus, bacteria of these strains were resistant to all studied fractions. Conclusions. Antimicrobial activity of a number of substances of hazel ordinary and black alder leaves is studied. As a result of the conducted researches it is established that lipophilic and spirit fractions of hazel ordinary and black alder leaves, tinctures from these types of raw materials revealed antimicrobial activity concerning P. аeruginosa, C. аlbicans, lipophilic fraction of hazel ordinary leaves, spirit fraction and tincture of black alder leaves– also concerning E. coli. The polysacharide fraction of black alder leaves doesn't possess antimicrobial action, and the polysacharide fraction of hazel ordinary leaves has shown the activity only concerning E. coli, S. аureus and P. аeruginosa. 

Introduction. In accordance with the last events in Ukraine (considering military operations in anti-terrorist operation in the Luhansk and Donetsk regions) the domestic medicine is in great need in preparations with antimicrobial activity. Our attention as the sources of receiving biologically active substances with antimicrobial activity was drawn with birch Betulaceae family plants – hazel ordinary Corylus avellana L. and black alder Alnus glutinosa (L.) Gaertn.

It is known that in medicine the leaves of hazel ordinary are used as antiseptic, anti-inflammatory, vesselrestorative drug, and the leaves of black alder reveal the antiinflammatory, astringent, wound healing, spasmolytic and choleretic action. However, the drugs with antimicrobial action received from the leaves of these plants are absent on the market of Ukraine. Therefore the studying of antimicrobial activity of this type of raw materials received from hazel ordinary and black alder, for creation of new medicines, is now one of the main directions in pharmacy. For this purpose we have revealed tinctures, spirit, lipophilic and polysacharid fractions received from the leaves of hazel ordinary and black alder. The purpose of our research is studying of antimicrobial activity of revealed substance received from the leaves of black alder and hazel ordinary. Materials and methods. There were being examined tinctures, lipophilic, spirit and polysacharid fractions received from the leaves of hazel ordinary and black alder. The test of antimicrobial effect of substances was carried out by means of serial dilution concerning the following six reference cultures: Staphylococcus aureus ATCC 6538-P, Candida albicans ATCC 885-653, Escherichia coli ATCC 25922, Bacillus subtilis ATCC 6833, Bacillus cereus ATCC 10702, Pseudomonas aeruginosa ATCC 9027, according to the State Pharmacopoeia of Ukraine, in the Department of Microbiology and Immunology of KMAPE. For the experiment there was prepared tenfold dilution of extracts on meat-peptone broth 1:10 and 1:100 to which there were added the referential cultures which are grown up on appropriate differential diagnostic medium depending on a type of cultures. The crops were incubated in the thermostat at t - 35-37 °C within 24-48 hours. For identification of test-strain growth of E. coli there was used Endo medium, for S. aureus – Chistovich medium, for B. subtilis and B. cereus - meat-peptone agar, for P. аeruginosa - 5% blood agar and for C. albicans – Saburo medium. As the solutions remained muddy after incubation, for the assessment of antimicrobial action of the received fractions, there were being made the cloning on differential and diagnostic mediums. Results and discussion. Data of the conducted researches of antimicrobial activity of various substances of hazel ordinary and black alder leaves – tinctures, lipophilic, spirit and polysacharide fractions are given in the table. The obtained data demonstrate that lipophilic fractions of hazel ordinary and black alder leaves possessed the bactericidal activity when it was diluted in the ratio of 1:10 and 1:100 concerning P. аeruginosa and E. coli. These fractions also have antimycotic activity concerning C. albicans. Concerning E. сoli it is revealed that the spirit fractions from black alder leaves had the bactericidal activity, and concerning P. аeruginosa and C. albicans – all studied spirit fractions possessed. Among polysacharide fractions the antimicrobial activity is revealed only for this substance of hazel ordinary leaves in both dilutions concerning E. coli, S. aureus, P. aeruginosa. As concerning B. subtilis and B. cereus, bacteria of these strains were resistant to all studied fractions.

Conclusions. Antimicrobial activity of a number of substances of hazel ordinary and black alder leaves is studied. As a result of the conducted researches it is established that lipophilic and spirit fractions of hazel ordinary and black alder leaves, tinctures from these types of raw materials revealedantimicrobial activity concerning P. аeruginosa, C. аlbicans, lipophilic fraction of hazel ordinary leaves, spirit fraction and tincture of black alder leaves– also concerning E. coli. The polysacharide fraction of black alder leaves doesn't possess antimicrobial action, and the polysacharide fraction of hazel ordinary leaves has shown the activity only concerning E. coli, S. аureus and P. аeruginosa.