Presentation Title

Assessment of Antimutagenic Action of Trametes versicolor (Turkey Tail), Inonotus obliquus (Chaga Mushroom), and Ophiocordycetps sinensis Extracts by Ames Test

Format of Presentation

Poster to be presented the Friday of the conference

Presenter Information

Iris Romero AndradaFollow

Abstract

Inonotus obliquus, also known as Chaga mushroom is a fungus that belongs to the Hymenochaetaceae family of Basidiomycota. This species has been used in folk medicine for treating cancer in North Europe, Western Siberia, Asia, and North America. For the past years, it has been studied since it produces a large number of compounds which show an array of biological activities, including antibacterial, hepato-protective and antitumor properties.

Ophiocordyceps sinensis (or Cordyceps sinensis) belongs to the Ophiocordycipitaceae family of Ascomycota. This fungus is an entomopathogen which has been used in traditional Chinese medicine for over 300 years. Some studies have shown that O. sinensis has functional activities such as immunomodulating, antiinflammation, antioxidant, antitumor, antimetastatic, among others.

Trametes versicolor, also known as Coriolus versicolor or “turkey tail”, is a fungus which belongs to the Polyporaceae family of Basidiomycota. It is a medicinal mushroom widely prescribed for the treatment of cancer and infections in China; and its polysaccharopeptides have shown biological properties, including stimulatory effects of the immune system and inhibition of cancer growth.

In accordance with current research into the different biological activities of these three fungi, and considering their antitumor and antimetastatic properties, there may be value in elucidating if these fungi present antimutagenic activity.

Objectives

The purpose of this project is to assess the antimutagenic activity of Trametes versicolor, Inonotus obliquus and Ophiocordyceps sinensis extracts. Furthermore, applications of various concentrations of each extract will be used to determine if the effect is quantity dependent.

Materials and Methods

Fungi extracts will be acquired from those collected in a parallel study where the antimicrobial activity of these fungi is going to be assessed.

For the Ames test assay, Salmonella typhimurium strains TA98 and TA100 will be used, which have a frameshift and base-pair substitution mutations in the histidine operon, respectively. In addition, both strains carry rfa, uvrB and pKM101 mutations, which will make the bacteria more permeable to chemicals and DNA lesions will be repaired by error-prone DNA repair mechanisms. The mutagenic agents that will be used in this project are 2-Nitrofluorene (2-NF) and 4-Nitroquinoline-N-oxide (4-NQO) in order to revert frameshift and base-pair substitution mutations, respectively.

Bacteria will be exposed to a constant concentration of the mutagenic agents and to 6 different concentrations of each of the extracts for 90 minutes in medium containing histidine. After exposure, the cultures will be diluted in pH indicating medium (lacking histidine) and aliquoted into 48 wells of a 384-well plate. Within two days, cells that have undergone mutation reversal will grow, and bacterial metabolism will reduce the pH of the medium, changing the color of the wells; those wells which does not change color will indicate antimutagenic activity of the fungi extracts. Each dose is done in triplicate for posterior statistical analysis of the data

Positive control will consist on bacteria with the mutagen agents (mutagenic activity will be shown) and two different negative controls are going to be used. The first one will be bacteria without mutagen and without extract, therefore, no mutagenic nor antimutagenic activity will be shown; the second one will consist on bacteria with the extracts, which will allow to assess if any of the three studied fungi have mutagenic activity.

Finally, data will be collected and statically analyzed.

Expected results

Through completing this project, I expect to gain information on the antimutagenic activity of these three species of fungi. Specifically, how incubation with varying concentrations of the extracts influences the antimutagenic effect.

Department

Biological Sciences

Faculty Advisor

Joanna Urban

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Assessment of Antimutagenic Action of Trametes versicolor (Turkey Tail), Inonotus obliquus (Chaga Mushroom), and Ophiocordycetps sinensis Extracts by Ames Test

Inonotus obliquus, also known as Chaga mushroom is a fungus that belongs to the Hymenochaetaceae family of Basidiomycota. This species has been used in folk medicine for treating cancer in North Europe, Western Siberia, Asia, and North America. For the past years, it has been studied since it produces a large number of compounds which show an array of biological activities, including antibacterial, hepato-protective and antitumor properties.

Ophiocordyceps sinensis (or Cordyceps sinensis) belongs to the Ophiocordycipitaceae family of Ascomycota. This fungus is an entomopathogen which has been used in traditional Chinese medicine for over 300 years. Some studies have shown that O. sinensis has functional activities such as immunomodulating, antiinflammation, antioxidant, antitumor, antimetastatic, among others.

Trametes versicolor, also known as Coriolus versicolor or “turkey tail”, is a fungus which belongs to the Polyporaceae family of Basidiomycota. It is a medicinal mushroom widely prescribed for the treatment of cancer and infections in China; and its polysaccharopeptides have shown biological properties, including stimulatory effects of the immune system and inhibition of cancer growth.

In accordance with current research into the different biological activities of these three fungi, and considering their antitumor and antimetastatic properties, there may be value in elucidating if these fungi present antimutagenic activity.

Objectives

The purpose of this project is to assess the antimutagenic activity of Trametes versicolor, Inonotus obliquus and Ophiocordyceps sinensis extracts. Furthermore, applications of various concentrations of each extract will be used to determine if the effect is quantity dependent.

Materials and Methods

Fungi extracts will be acquired from those collected in a parallel study where the antimicrobial activity of these fungi is going to be assessed.

For the Ames test assay, Salmonella typhimurium strains TA98 and TA100 will be used, which have a frameshift and base-pair substitution mutations in the histidine operon, respectively. In addition, both strains carry rfa, uvrB and pKM101 mutations, which will make the bacteria more permeable to chemicals and DNA lesions will be repaired by error-prone DNA repair mechanisms. The mutagenic agents that will be used in this project are 2-Nitrofluorene (2-NF) and 4-Nitroquinoline-N-oxide (4-NQO) in order to revert frameshift and base-pair substitution mutations, respectively.

Bacteria will be exposed to a constant concentration of the mutagenic agents and to 6 different concentrations of each of the extracts for 90 minutes in medium containing histidine. After exposure, the cultures will be diluted in pH indicating medium (lacking histidine) and aliquoted into 48 wells of a 384-well plate. Within two days, cells that have undergone mutation reversal will grow, and bacterial metabolism will reduce the pH of the medium, changing the color of the wells; those wells which does not change color will indicate antimutagenic activity of the fungi extracts. Each dose is done in triplicate for posterior statistical analysis of the data

Positive control will consist on bacteria with the mutagen agents (mutagenic activity will be shown) and two different negative controls are going to be used. The first one will be bacteria without mutagen and without extract, therefore, no mutagenic nor antimutagenic activity will be shown; the second one will consist on bacteria with the extracts, which will allow to assess if any of the three studied fungi have mutagenic activity.

Finally, data will be collected and statically analyzed.

Expected results

Through completing this project, I expect to gain information on the antimutagenic activity of these three species of fungi. Specifically, how incubation with varying concentrations of the extracts influences the antimutagenic effect.