Keywords: Bryophytes, Antibacterial, Antiviral, Cucumber, Pathogenic bacteria, bryophytes for antimicrobial activity. Journal of. Medicinal Plants, 7, ). Antibacterial Activity of Extracts from Some Bryophytes. Vizma Nikolajeva1*, Ligita Liepina1, Zaiga Petrina1, Guntra Krumina1. The antimicrobial activity of aqueous and ethanolic extracts of 11 Bryophyta and Naturally Grown Bryophytes,” Journal of Medicinal Plant Research, Vol. 5, No.
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Fourteen crude methanolic and ethanolic extracts of bryophytes from South Western British Columbia were screened for antibiotic activity against three bacterial strains with the disk diffusion method. None of the 14 species of bryophytes extracts, from either of the alcoholic extractions, showed any visible activity againstGram-negative Escherichia coli or Klebsiella pneumoniae.
The occurrence of antibiotic substances appears to be more frequent in hepatics. Lunularia cruciata also demonstrated the most significant antibiotic activity of all the bryophytes tested. However, reports of ethnobotanical research into this plant group are minimal in North America 1. Recently, the public demand for herbal medicine and the rise of antibiotic-resistant bacteria have motivated scientists to look for new natural sources with potential pharmaceutical capabilities 3.
Historically, a large number of medicinal plants were discovered and used by the aboriginal people to treat illnesses 4. The application ranged from topical applications, to prevent infection, to internal consumption, facilitating relief from nausea 5. However, there have been few studies to screen and confirm the medicinal potential of bryophytes in North Americans for antibiotic activity.
Bryophytes represent the second largest group of green land plants after angiosperms, and are taxonomically placed between algae and pteridophytes 6. The bryophytes group consists of three subgroups: Bryophyta mossesMarchantiophyta liverworts or [hepatics] and Anthocerotophyta hornworts 7. These groups exhibit an abundant distribution in China, Europe and North America and current estimates suggest that approximately species of mosses, species of liverworts and species of hornworts exist worldwide 9.
Bryophytes are an exotic and captivating species with a unique combination of distinguishing characteristics.
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Consequently, bryophytes remain small and flourish in places where there is an abundant water supply The climate of South Western British Columbia is very wet and humid, with an average of mm of rain per year Therefore, a plethora of different bryophyte species have evolved in this regions forests and wetlands Bryophytes in this region are herbaceous plants that grow closely packed together in mats or cushions on rocks, soil, or as epiphytes on the trunks and leaves of forest trees They lack roots and attach to their environment with rhizoids However, unlike vascular plants, water and minerals are absorbed chiefly by the thin leaves of the plant as the rain washes through the moss, not from roots Mosses and liverworts usually have leaves with a thickness of one cell, except along the costa and the alar cells, which facilitate diffusion Lastly, the characteristic that distinguishes bryophytes from tracheophytes, vascular plants, is the persistent photosynthetic phase of the life cycle is haploid; gametophyte generation occurs rather than the diploid sporophyte generation The sporophyte typically consists of an unbranched seta, and a single terminal sporangium It is short lived and attaches to the gametophytes for all its nutritional needs.
Furthermore, bryophytes utilize the sporangium for reproduction via sporulation but they neither flower nor produce seeds Hundreds of medicinal bryophytes have been identified and classified in ethnobotanical literature as potential antimicrobial agents 6,8,9, Many of these species are prevalent in British Columbia. However, few studies have been concerned with an in-depth analysis of the antimicrobial activity and efficacy of phytochemicals in different species of bryophytes from South Western British Columbia To explore the bryophytic potential in pharmaceutical applications, a quantitative and qualitative screening of S.
British Columbian bryophytes against Gram-positive and Gram-negative bacteria will be used as evidence for antimicrobial action and efficacy. Lastly, this study aims to determine whether ethanolic and methanolic extracts from similar bryophytes show significant differences in the relative antimicrobial activity.
Samples of all tested plants were collected by M. Ellis from the field in Vancouver, British Columbia. The collections took place from October to November Bryophytes typically grow in close association with each other. Therefore, homogenous patches were desired with fresh tissue void of dead or dying material.
Reproductive organs were bbryophytes in identification. Other tested plant species, were collected from homogenous plant populations.
Any part of the plant may contain active components. Therefore, the whole organism was collected wherever possible.
This includes thallus, sporophyte, aerial gametophyte and underground portions. Since different populations of the same species may exhibit different antimicrobial activity, the time and place of collection, and habitat were recorded Table 1.
All samples were preliminarily washed to remove contaminants. Once rinsed, plants were kept in sealed paper bags, and placed in the refrigerator to prevent the subsequent drying or decay.
Screening antimicrrobial potential antimicrobial plants requires bryoophytes aqueous or alcohol extracts and fine organic extraction methods. Since nearly all of the identified biologically active or antibiotic substances from plants have been identified as aromatic or saturated organic compounds, it was most efficient to obtain the compounds through ethanolic or methanolic extractions.
Before extraction all plant material collected was washed with distilled water to remove any adhering soil or extraneous material. One and half grams of the ground material were extracted in 25ml of methanol over 24 hours.
Antibacterial Activity of Extracts from Some Bryophytes
The homogenous surface liquid of the crude methanolic extracts was used to impregnate the diffusion discs with antibiotic activity. The homogenous surface liquid of the crude ethanolic extracts was used to impregnate the diffusion discs with antibiotic activity. Six mm sterilized filter paper discs were then impregnated with the approximately nine drops of the homogenous extract.
They were then allowed to dry at room temperature. All extracts were bryophhytes at room temperature. The disc diffusion assay technique was used to evaluate antimicrobial activity. The bacteria used were Bacillus subtilis, Escherichia coli, and Klebsiella pneumoniae. Six mm sterile filter paper discs were impregnated with extract and placed on seeded agar.
Antibiotics were used as positive controls against the bacteria. All assays were carried out in triplicates. Fourteen crude methanolic and ethanolic extracts of bryophytes from South Western British Columbia were screened for antibiotic activity against three bacterial strains. The botanical names of the plants tested, habitat and collection date are listed in Table 1.
The screening results are listed in Table 2 and 3 for ethanolic and methanolic extractions respectively. All extract activity was standardized journql comparable amounts of dried plant material and quantities of alcoholic extraction medium. Table 2 Antibiotic activity in disc diffusion assays of bryophytes from South Western British Columbia and reference antibiotics.
All extractions were performed with ethanol. Gm -ve Escherichia coli. Gm —ve Klebsiella pneumoniae. Table 3 Antibiotic activity in disc diffusion assays of bryophytes from South Western British Columbia.
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All extractions were performed with methanol. None of the mosses showed any antibiotic activity when extracted with methanol. Bartramia pomiformis, Leucolepsis acanthoneuron, Frullania nisquallensis showed antibiotic activity when ethanolic extractions where used but brryophytes when methanolic extracts were used. All other species showing antibiotic activity were common among ethanolic and methanolic extracts. The average inhibition zone in ethanolic extracts ranged from 1.
The average inhibition zone in methanolic extracts ranged from 1. The onlyexception antimicronial Polytrichastrum alpinum, which showedindeterminate antibiotic response. Alcoholic extracts from Conocephalum conicum, Sphagnum palustre, Hylocomium splendens and Atricum selwynii showed no visible activity against any Gram-positive or Gram-negative bacteria. All the reference antibiotics showed dramatic activity against all the Gram-positive and Gram-negative bacteria. Streptomycin jpurnal most effective against E.
The reference antibiotics showed a range of 5. Methanolic extractions The preliminary results of screening the alcoholic extracts of 14 species of bryophyte tested against three species of bacteria showed that there is a distinct difference between the antibiotic activities of the tested species in relation to the extraction method used. In addition, none of the mosses showed any antibiotic activity when methanolic extractions were used. For example, Bartramia pomiformis, Leucolepsis acanthoneuron, Frullania nisquallensis showed antibiotic activity only using ethanolic extraction.
This suggests that specific antibacterial compound seffective against the selected bacterial species, tend to be isolated more effectively from liverworts and mosses using ethanol. Table 4 lists some of the known compounds which have been extracted from methanolic and ethanolic extracts. The bolded compounds are partitioned exclusively by the particular solvent. Further investigation would be required to determine if a polyacetylene, sterol or propolis found in the mosses, and liverworts are responsible to the antibiotic nature of the species.
Furthermore, this also suggests that one antibiotic compound may be responsible for the antibacterial qualities of a genus or species. This clearly wntimicrobial the importance of using a multitude of extraction compounds and methods to isolate antibiotic compounds.
A failure to do so could result in an inability to retrieve valuable information, and incorrectly dismissing potentially beneficial species. This suggests that detectable antibacterial compounds are present in most taxa of liverworts Liverworts also demonstrated the greatest inhibition zone. For example, Lunularia cruciata, Plagiochila porelloides and Marchantia polymorphia showed inhibition zones greater than 2. These values are approximately one third of the inhibition of reference antibiotics.