between atoms and molecules - Students Science Conference

1 downloads 214 Views 13MB Size Report
The method involves measurements of the solid waste streams gen- ...... PEDOT (Poly(3,4-ethylenedioxythiophene)-poly(sty
Key words: toxicity, cell culture, adenocarcinomic human alveolar basal epithelial cells

Maciej K. BEŁCIK*

THE INFLUENCE OF PARTICULATE MATTER FRACTION 2.5 µM ON THE CELL LINE A549

In the past few years in Poland is observer increasing air pollution. One of many pollution observed in the air are solid particles called particulate matter. Particulate matter is responsible for formation of smog over large metropolis. Among the cities for witch this problem is particularly observer are among others Wrocław. Inhalation of particulate matter adverse negative effects on human population causing asthma, chronic obstructive pulmonary disease, arteriosclerosis and cardiac ischemia. The main sources of particulate matter in atmosphere are exhaust from road transport and gases emitted from individual coal furnace. Other big issue is low awareness of the society, which use lowgrade coal and waste for heating in houses. The following article presents the initial studies of particulate matter air pollutions on adenocarcinomas human alveolar basal epithelial cells A549. Conducted research allow to check the suitability cell lines to research particulate matter fraction 2.5 µm, determine dose limits and impact of solvents concentration on A549 cell line.

1. INTRODUCTION In the atmospheric air there is more than 2,000 chemical substances. Some of them are absorbed can be adsorbed on the surface of matriculate matter. Substances in the air may form a complicated mixtures with unknown properties and activity. Among the substances adsorbed in the surface of particulate matter can be found inter alia metals, polycyclic aromatic hydrocarbons (PAH), aromatic hydrocarbons, phenols, organic compound containing chlorine and more [1].

__________ * Faculty of Environmental Engineering, Wrocław University of Technology, Wybrzeże Wyspiańskiego 27.

1

M.K. BEŁCIK

Particulate matter pollutions cause many health problems in human body including premature death causing cardiovascular and respiratory problems [2-4]. Research conducted by Burns and Lim in 2010 showed that air pollutions are responsible for more than 3.2 million premature deaths, which classifies them in the second position among environmental and ninth among global risk factors in the world [2, 5]. In addition particulate matter air pollutions are also responsible for airways disease such as asthma attacks, pneumonia, decreased lung function and cardiovascular system like hearth attack. This disease, caused by particulate matter, are observed not only in groups exposed on the environmental risk but also in potential healthy part of the population [2-3, 6]. In October 2013 group of 24 experts from 11 different countries affiliated to the International Agency for Research on Cancer (IARC) based on all available knowledge and evidences shown in studies, classifies air pollution, including particulate matter, to group 1 of genotoxic compounds. Group 1 of International Agency for Research on Cancer contains substances which studies proved carcinogenicity to humans. Except air pollutants there are substances adsorbed on the particulate matter like benzene, benzo[a]pyrene, formaldehyde, diesel exhaust. In groups 2A and 2B which contains substances probably and potentially carcinogenic are for example benzo[a]anthracene, benzo[k]fluoranthren, benzo[c]phenanthrene, dibenzo[a,h]pyrene [7-9]. Table 1 shows the classification of carcinogenic substances using by IARC. Table 1. IARC classification of carcinogenic substances August 24, 2015 [8] Group 1 2A 2B 3 4

Description Substances carcionogenic for humans Substances probably carcionogenic for humans Substances potentialy carcionogenic for humans Substances which cannot be classified as carcinogenic for humans Substances probably not carcinogenic for humans

Number of substances 117 74 287 503 1

The most frequently used methods of assessing genotoxicity of particulate matter air pollutions are tests based on the bacteria Salmonella typhimurium. The modified test strains under the influence of substances of genotoxic activities returns to the form which have ability to synthesize histidine. On this basis, there were developed wide range of strains possible to determine type of mutation and with different sensitivity to specific substances. Currently Salmonella test can be also carried out as a microplate test, which significantly simplifies and speeds up the examination of samples [7, 9]. Recently more often for test genotoxicity and cytotoxicity samples used cell lines. This article presents the initial studies of particulate matter air pollutions on adeno-

2

The influence of particulate matter fraction 2.5 µm on the cell line A549

carcinomas human alveolar basal epithelial cells A549. The aim of this studies was to determine the doses of particulate matter and check the influence of solvents in the cell line.

2. MATERIALS AND METHODS 2.1. COLLECTING SAMPLES

Material for the study was the particulate matter of ambient air fraction 2.5 μm collected in Wroclaw agglomeration (about 630 thousand residents, 2.1 thousand people/km2). Samples were collected on the filters using a high-flow air aspirator during different seasons. The filter was replaced every 24 hours. In that way particulate matter from one filter obtained one day of collection. 2.2. A549 CELL LINE

For tests was used A549 cell line, which is adenocarcinomas human alveolar basal epithelial cells (American Type Culture Collection, Cell Culture Line 185). The choice of adherent cells A549 allows to stimulate conditions close to real. The finest fractions of particulate matter penetrate deep parts of lungs going into alveoli. A549 cell line is used as an in vitro model of various studies including toxicity and genotoxicity particulate matter air pollutants. A549 cell line is conducted in culture medium Dulbeco containing 10% calf serum, 2 mM L-glutamine, 100 units/cm3 penicillin and 100 μg/cm3 streptomycin. Cell lines were incubated at 37OC and atmosphere 5% of carbon dioxide. 2.3. BIOLOGICAL ANALYSIS

Samples collected during winter season were fractionated to give 4 fractions containing the whole of pollutants (A), PAHs (B), nitro-PAHs (C) and dinitro-PAHs (D). Each of the samples were diluted, based on the previous research, 4-times (1-4) form an initial concentration 50 m3/cm3 using geometric progression with a common ratio q=1/2 to 6.25 m3/cm3. To each dilution was added a predetermined amount of DMSO solvent ranging from 0.5 to 2%. In addition it will be tested pure solvent. Concentrations DMSO taken for test are 2%, 1.5%, 1% and 0.5%. Notation of samples are presented in Table 2.

3

M.K. BEŁCIK

All the samples were applied on a 24-hour A549 cell culture with a density of cells/cm3 and incubated for 48 hours. After this time results was performed using microscope and spectrophotometer at a wavelength of 550-630 nm. Table 2. The samples taken for research Name of sample A B C D E F G H

Fraction

whole pollutants

PAHs

Solvent concentration 2% 1.5% 1% 0.5% 2% 1.5% 1% 0.5%

Name of sample K L M N P R S T

Fraction

NitroPAHs

DinitroPAHs

Solvent concentration 2% 1.5% 1% 0.5% 2% 1.5% 1% 0.5%

3. RESULTS Results of test measuring by spectrophotometer at a wavelength of 550-630 nm was presented in the pictures 1 to 4. It was also presented absorbance of cell lines A549 without addition any substances and with addition DMSO in mentioned earlier doses.

Picture 1: The absorbance of the samples A-D

4

The influence of particulate matter fraction 2.5 µm on the cell line A549

Picture 2: The absorbance of the samples E-H

Picture 3: The absorbance of the samples K-N

Picture 4: The absorbance of the samples P-T

5

M.K. BEŁCIK

4. DISCUSSION The results showed toxic effects of the samples on cell line A549. The toxic effects of particulate matter extracts is diferent depending in the fraction and solvent dose. The highest toxicity showed samples with all collected pollutants regardless of the amount of solvent addition. High toxicity was also shown for the fraction of PAHs, little lower for the nitro-PAHs and dinitro-PAHs. As expected, the results show a reduction in toxic effect with the decrease in the concentration of the samples. Moreover, almost all samples showed a higher toxicity of the samples with a higher content of DMSO in relation to the corresponding samples with less solvent dose. Toxic effects have been also reported for DMSO solvent. It has been shown that the addition of the extracts to the solvent in any case reduces the absorbance of the test samples. The lowest toxic effect was achieved at a concentration lower than 1.5% DMSO. Based on the obtained results it was conclude that during the planed research, the initial concentration of extracts should not exceed 50 m3/cm3 and the maximum solvent should not be higher than 1%.

5. CONCLUSIONS 1. 2. 3. 4. 5.

All tested extracts have shown toxicity in relation to cell line A549. It was observed lower toxicity of DMSO then the test samples. The highest toxicity shows a fraction containing all pollutants and the lowest dinitro-WWA fraction. DMSO at a dose less than 1% showed relatively low toxicity in tests. It has been shown, that the initial concentration for cell line A549 test should not exceed 50 m3/cm3 and not more than 1% solvent control. REFERENCES

[1] Alves D.K.M., KUMMROW F., CARDOSO A.A., MORALES, D.A., UMBUZEIRO, G.A., Mutagenicity profile of atmospheric particulate matter in a small urban center subjected to airborne emission from vehicle traffic and sugar cane burning, Environmental and molecular mutagenesis, 2015, Vol. 57, No. 1, 41-50. [2] BURNS J., BOOGAARD H., TURLEY R., PFADENHAUER L.M., VAN ERP A.M., ROHWER A.C., REHFUESS E. et al., Interventions to reduce ambient particulate matter air pollution and their effect on health (Protocol), Cochrane Database of Systematic Reviews 2014, No. 1. [3] POPE III C.A., DOCKERY D.W., Health effects of fine particulate air pollution: lines that connect, Journal of the Air & Waste Management Association, 2006, Vol. 56 No. 6, 709-742. [4] STIEB D.M., JUDEK S., BURNETT R.T., Meta-analysis of time-series studies of air pollution and

6

The influence of particulate matter fraction 2.5 µm on the cell line A549 mortality: effects of gases and particles and the influence of cause of death, age, and season, Journal of the Air & Waste Management Association, 2002, Vol. 52 No. 4, 470-484. [5] LIM S. S., VOS T., FLAXMAN A.D., DANAEI G., SHIBUYA K., ADAIR-ROHANI H., ARYEE M., et al. A comparative risk assessment of burden of disease and injury attributable to 67 risk factors and risk factor clusters in 21 regions, 1990–2010: a systematic analysis for the Global Burden of Disease Study 2010, The lancet, 2014, Vol. 380 No. 9859, 2224-2260. [6] RÜCKERL R., SCHNEIDER A., BREITNER S., CYRYS J., PETERS A, Health effects of particulate air pollution: a review of epidemiological evidence, Inhalation toxicology, 2011, Vol. 23 No. 10, 555-592. [7] BEŁCIK M., TRUSZ-ZDYBEK A., GALAS E., PIEKARSKA K., Mutagenicity of organic pollutants adsorbed on suspended particulate matter in the center of Wrocław (Poland), Atmospheric Environment, 2014, Vol. 95, 620-628. [8] http://monographs.iarc.fr/ENG/Classification/ [access: 8.10.2015]. [9] PIEKARSKA K., Modyfikacje testu Salmonella do oceny mutagenności pyłowych zanieczyszczeń powietrza atmosferycznego, [w:] Prace Naukowe Instytutu Inżynierii Ochrony Środowiska Politechniki Wrocławskiej, 2008, Nr 87, Seria: Monografie Nr 52.

7

Keywords: cambium, dormancy, limiting factors, rings formation,

Katarzyna GÓRKA*

SEASONAL CHANGES OF CAMBIAL ACTIVITY IN SEED PLANTS IN RELATION TO CLIMATIC ZONES

Wood is a natural raw material that is commonly used is human societies. It is composed of the cells of xylem – a plant tissue that is formed during the divisions of cambial cells. Because of this, it is important to understand the processes in the cambium. Problems related to the functioning of the cambium from different climatic zones (its activity, dormancy and periods of transition) are still an open issue. Cambial cells are influenced by external factors (temperature, photoperiods, water availability, type of soil, pollution) and internal ones (the age of a plant, gene expression, plant hormones and the arrangement of mechanical tension). Some trees shed their leaves during seasons and the cambium becomes dormant. The limiting factors in cold climates are photoperiods and temperature. It is commonly observed in hot and warm climates that there is also one other important factor – a decrease in water availability. Evergreens have an active cambium all year round because they grow in the tropics, where the conditions are close to optimum during the entire year. In areas where weather conditions are variable, changes in cambial activity can be observed. Woody plants are divided into three main groups: (1) plants with the formation of one annual growth ring, (2) plants forming false rings and (3) plants that do not form clearly visible growth rings.

1. INTRODUCTION Issues connected with cambial activity have widely been described, but they are rarely described in relation to different climatic zones. This tissue is strongly connected with climatic and weather conditions [1]. Without wide knowledge about its structure and the processes that occur inside the vascular cambium, it is hard to use one of the most important natural raw materials – wood – wisely. Wood is an extensively used material that is crucial for the functioning of modern society. It is not without significance that its availability may be severely limited due to the __________ * Laboratory of Applied Mycology and Plant Systematics, Department of Biosystematics, Opole University, Oleska 22, 40-052 Opole, Poland

8

Katarzyna Górka

progressive deforestation around the world [2]. The aim of this paper is to review information about cambial activity in plants from different areas of the world. The term “cambium” is used in the meaning of the meristem in plants that have a typical secondary growth, which is formed after the primary growth, which is reflected by the deposition of layers of the secondary phloem and the secondary xylem.

2. CAMBIAL ACTIVITY IN DIFFERENT CLIMATIC ZONES Plants are found in most of the climatic zones around the world. When they were spreading around the world, they had to adapt to habitats that had different conditions. Their environments differ in the amount of precipitation, daily and annual average temperature, sunlight and type of soil substrate. These factors affect the length of the growing season, which in turn affects the growth of woody plants. There are five basic climatic zones: the equatorial, tropical, subtropical, temperate and polar zone [3]. The effects of cambial activity in woody plants can be observed by the naked eye as growth rings. These visible rings result from changes in the speed of growth through the seasons of the year. Depending on how the secondary growth occurs, all woody plants are classified into one of three groups, as follows: 1. Plants with only one ring formation each year (equal to “annual ring formation”), 2. Plants with more than one ring formation each year (with a false ring formation), 3. Plants without a clear ring formation [4]. 2.1. EQUATORIAL ZONE

The cambium in the equatorial climatic zone is active all year round because most plants in these places have the ability to produce leaves continuously [5]. There is a lack of ring formation in cross sections of their trunks [6]. However, there are exceptional plants for which the production of leaves is not continuous during a year. Their cambium occurs as a seasonal activity because this tissue is dormant when woody plants do not have leaves. Here, the main factors that affect changes in the cambial divisions are dry and rain seasons. Droughts and water shortages can deactivate these divisions, which can cause in the formation of false rings [7]. It is interesting that cambial activity is not the same in different parts of a plant. Only one ring is usually formed inside trunks, while many rings are created inside branches [4].

9

Cambial activity in different climatic zones

Brachystegia spiciformis is a common tree species in the miombo forests in Zambia, where both dry and rainy seasons occur. Cambium is only active in the middle of the rainy season and it lasts two or three months. Even if rains fall during the dry season, the cambium remains dormant, so there is no false ring formation [8]. In the case of Araucaria angustifolia, a common species in the southwest of South America, the tendency to form an annual growth ring was observed [9]. However, if the changes in weather were strong enough, false rings or even no rings were created during a year. The seasonal activity of the cambium in Araucaria angustifolia was longer than in B. spiciformis. The xylogenesis took place from October to March (early wood formation) and from April to May (late wood formation). Cedrela fissilis in Brazil has a similar cambial activity. 2.2. TROPICAL ZONE

There is a wide annual diversity of weather in the tropical climatic zone. This causes multiple ring formations in many plant species during a year. One ring formation is only observed in areas with dry and rainy seasons, for example in the southeastern part of Brazil (in Schizolobium parahyba) and Mexican forests (in Tabebuia donnell-smithii and Ceiba pentandra), where these trees shed their leaves once a year [1011]. This is different in woody plants growing on river banks. The dry season has to be longer than normal and rivers have to dry up in order to slow down cambial divisions or to make the cambium dormant [11]. There are plantations of Tectona grandis in the southern parts of the Indian Peninsula where cambium activation is connected with the summer monsoons. During three years without a dry season, the cambium of T. grandis did not create any late wood structures and the cambial divisions were maintained throughout this time. Their wood looks like a diffuse porous wood, while the xylem of this species is characterized as a ring porous wood [12]. The cambium of Ficus religiosa from the northern area of the Indian Peninsula lasts one month longer during the year than T. grandis [13]. Thus, plants from different genera that grow in the same areas within the same climatic variation zone may have different cambial activities. 2.3. SUBTROPICAL ZONE

The cambium of Pinus radiata from New Zealand never goes dormant, which is normal for the species that grow in the European temperate zone [14]. Plants in Israel, where the climate has some features of a subterranean climate, have two growing strategies. The first one is to stop cambial divisions during droughts, which can last up to eight months. The second one concerns plants with strongly expanded root systems that can reach water storage in deep layers of soil. In this way, they have access

10

Katarzyna Górka

to water that is not available for plants from the first group and therefore, their cambium is active all year round [15]. The cambial divisions of Cryptomeria japonica were investigated in Kyoto, Japan where climatic seasons are connected with monsoons. The number of cambial cell layers was the highest during periods of monsoon activity, while there were only two layers during the dormant period. The most intensive cambial divisions were initialized by two main factors – rainfall and an increase in temperature. The dormancy period was in May and June, which was a consequence of the small amount of precipitation [16]. 2.4. TEMPERATE ZONE

Cambium is active through the entire growing season in the temperate climatic zone [5]. It only goes dormant in the second part of August [6]. The dormancy of Robinia pseudoacacia, which grows in Wisconsin, occurs from October to the first days of April, after which a one-month-long period of cambial reactivation occurs. Active cambium is observed from the end of April to August, after which the cambial divisions become slower [14]. The cambial activity period of Fagus sylvatica from Slovenia starts one month sooner and it usually takes 100 days. Cambium divides the most efficiently from May to July, which is in accordance with the optimal temperature and precipitation [17]. There are many modifications of cambial activity in the subterranean variety of the temperate zone. For example, Pinus halapensis grows in eastern Spain, where the climate is dry. Thus, cambial activity is strongly connected with the weather conditions. It is common that cambial activity is strongly variable over a period of years. Spanish summers are hot and dry, while the winters are mild and wet. The cambium may be reactive in spring, stay active in summer and become dormant after winter, as is the case with most plants in the temperate zone. If the winter is warm and the summer is not hot and wet, secondary growth may occur all year round. If weather conditions are extremely hard, the cambium may remain dormant and no new growth rings will appear [18]. Weather conditions (especially temperature and water availability) are also important factors for the Pinus sylvestris that grows in Poland. Water availability is an important factor, but heavy rains in wetlands inhibit cambial activity [1920]. 2.5. POLAR ZONE

Cambial activity in polar zone seems to be related to temperature, although there is a correlation between the width of growth rings and the number of sunspots [15]. The growing season is very short. It lasts from May to July in the northern parts of the

11

Cambial activity in different climatic zones

forests in Quebec in Canada. New growth rings are formed after rainfall and a rise in temperature [21]. Similar conditions are observed in the Alps in Italy where the climate is described as subpolar. What is interesting is that temperature is not the most important factor that causes cambial divisions. It is photoperiods. Cambial activity was the most intensive during the longest (not the warmest) days in the year [22]. The cambial activity of Larix decidua, Pinus cembra and Picea abies starts in April or May and stops in August [23]. There is a strong tendency to form only one growth ring during a year [24].

3. SUMMARY The seasonal activity of cambium is associated with the formation of annual growth rings. The type of climate, as well as weather anomalies, can change cambial activity. Climatic zones have many subtypes of climates and therefore the cambium of the plants that grow within these areas has a level of different activity. It is commonly believed that evergreens have active cambium all year round. These occur in warm climates and in the tropics, where the conditions are close to optimum during the whole year. Trees that shed their leaves during some seasons have a dormant period in annual cambial activity. This is observed in hot climates (with a dry and rainy season) and in warm ones (with a warm summer and cold winter). Precipitation, which is connected with an increase of water availability, is an important factor that causes cambial divisions in warm climates. The limiting factors in cold climates are photoperiods and temperature. REFERENCES

[1] JEZIK, M., BLAZENEC M., STRELCOVA K., DITMAROVA, Ľ., The impact of the 2003–2008 weather variability on intra-annual stem diameter changes of beech trees at a submontane site in central Slovakia, 2011, Dendrochronologia 29(4): 227-235. [2] HARGRAVE J., KIS-KATOS K., Economic causes of deforestation in the Brazilian Amazon: a panel data analysis for the 2000s. 2013, Environmental and Resource Economics 54(4): 471-494. [3] OKOŁOWICZ W., Klimatologia ogólna, PWN, Warszawa 1969, 1-395. [4] ZIMMERMANN M.H., BROWN C.L., Trees. Structure and Function, Springer-Verlag, Heidelberg, New York, Berlin 1974, 1-336. [5] SZWEYKOWSKA A., SZWEYKOWSKI J., Botanika t. 1 Morfologia, PWN, Warszawa 2007, 1342. [6] TOMANEK J., Botanika leśna, Państwowe Wydawnictwo Rolnicze i Leśne, Warszawa 1997, 1-506. [7] HEJNOWICZ Z., Anatomia i histogeneza roślin naczyniowych t. 1 i 2, PWN, Warszawa1985, 1-859. [8] TROUET V., MUKELABAI M., VERHEYDEN A., BEECKMAN H., Cambial Growth Season of

12

Katarzyna Górka

Brevi-Deciduous Brachystegia spiciformis Trees from South Central Africa Restricted to Less than Four Months, 2012, PLOS ONE 7(10): 1-9. [9] OLIVEIRA J.M., SANTAROSA E., PILLAR V.D., ROIG F.A., Seasonal cambium activity in the subtropical rain forest tree Araucaria angustifolia, 2009, Trees 23: 107–155. [10] MARCATI C.R., MILANEZ C.R.D., MACHADO S.R., Seasonal development of secondary xylem and phloem in Schizolobium parahyba (Vell.) Blake (Leguminosae: Caesalpinioideae), 2008, Trees 22: 3–12. [11] HAYDEN B., Annual growth rings in dry tropical forest trees. Master thesis, Concordia University, Montreal, Quebec, Canada 2008, 1-124. [12] PRIYA P.B., BHAT K.M., Influence of rainfall, irrigation and age on the growth periodicity and wood structure in teak (Tectona grandis), 1999, IAWA Journal 20(2): 181–192. [13] SIDDIQI T.O., Impact of seasonal variation on the structure and activity of vascular cambium in Ficus religiosa, 1991, IAWA Bulletin 12(2): 177 – 185. [14] FARRAR J.J., EVERT R.F., Ultrastructure of cell division in the fusiform cells of the vascular cambium of Robinia pseudoacacia, 1997, Trees 11: 203–215. [15] FAHN A., Plants Anatomy, Pergamon Press, Oxford, London, Edinburgh, New York, Toronto, Sydney, Paris, Braunschweig 1967, 1-534. [16] ITOH T., HAYASHI S., KISHIMA T., Cambial Activity and Radial Growth in SUGI Trees (Japanese Cryptomeria), 1968, Wood Research 45: 23–35. [17] CUFAR K., PRISLAN P., de LUIS M., GRINCAR J., Three-ring variation, wood formation and phenology of beech (Fagus sylvatica) from a representative site in Slovenia, SE Central Europe, 2008, Trees 22: 749–758. [18] de LUIS M., GRICAR J., CUFAR K., RAVENTOS J., Seasonal dynamics of wood in Pinus halapensis from dry and semi-arid ecosystems in Spain, 2007, IAWA Journal 28(4): 389–404. [19] COPENHEAVER C., GARTNER H., SCHAFER I., VACCARI F., CHERUBINI P., Droughttriggered false ring formation in a Mediterranean shrub, 2010, Botany, 88(6): 545-555. [20] CEDRO A., Dependence of radial growth of Pinus sylvestris L. from Western Pomerania on the rainfall and temperature conditions, 2001, Geochronometria 20: 69–74. [21] DESLAURIES A., MORIN H., URBINATI C., CARRER M., Daily weather response of balsam fir (Abies balsamea (L.) Mill.) stem radius increment from dendrometer analysis in the boreal forests of Quebec (Canada), 2003, Trees 17: 477–484. [22] ROSSI S., DESLAURIES A., ANFODILLO T., Assessment of cambial activity and xylogenesis by microsampling tree species: an example at the alpine timberline, 2006, IAWA Journal 27(4): 383– 394. [23] ROSSI S., DESLAURIES A., ANFODILLO T., MORIN H., SARACINO A., MOTTA R., BORGHETTI M., Conifers in cold environments synchronize maximum growth rate of tree-ring formation with day length, 2006, New Phytologist 170: 301–310. [24] RIXEN C., SCHWOERER C., WIPF S., Winter climate change at different temporal scales in Vaccinium myrtillus, an Arctic and alpine dwarf shrub, 2010, Polar Research 29(1): 85-94.

13

Keywords: fungi, fruit bodies, medicinal properties, traditional use

Katarzyna GÓRKA*

ETHNOMYCOLOGY: EXAMPLES OF THE FOLK USE OF SOME MEDICINAL FUNGI People have instinctively used fungi since ancient times. Many of them were chosen because of a belief in their health-promoting effects which could treat simple diseases. However, their basic role was as a food that was rather easy to get. Thanks to this, they were used as items of everyday use. Some of them were used in religious ceremonies and helped shamans to enter into a mystical trance. Generally, mushrooms were an important part of life and they were valued because of their comprehensive applications. The lack of scientific knowledge about their properties, chemical composition, and even about many illnesses, were not obstacles for using fruit bodies. Information about their folk use is still being collected. Modern medicine and chemistry research has often confirmed the correctness of the traditional use of many mushrooms, and it also allows folk myths to be dispelled. All of the information about the folk use of fungi is gathered and described by ethnomycology. This term confirms data about the use of mushrooms in all of the historic aspects of human life. However, ethnomycology is easily forgotten in modern societies that are dominated by technology.

1. INTRODUCTION There are two meanings of the term “ethnomycology”. It is used in order to describe the traditional knowledge and customs of a people concerning fungi or as the name of the study (or description of this) [1]. People have known how to use the fruit bodies of macromycetes since ancient times, despite the fact that the knowledge about their chemical compositions and the precise way that they influenced the human organism was very poor. The fruit bodies of fungi were a part of everyday life. Because of their relatively easy and cheap collection, they also became a part of social life. They were valued for their unique properties. There were many traditional ways to take advantage of edible as well as inedible fungi. While some species were primarily used as food, others were a cheap substitute for items of everyday use or even a part of religious ceremonies. Nowa__________ * Laboratory of Applied Mycology and Plant Systematics, Department of Biosystematics, Opole University, Oleska 22, 40-052 Opole, Poland

14

Katarzyna GÓRKA

days, they are mainly known as sources of food and/or for their bioactive compounds. Knowledge about their chemical composition often confirms the correctness of their application in folk medicine [2]. Thus, much of the mycological folk data should be appreciated. This knowledge is often transmitted orally from one generation to the next, and therefore ethnomycology, which has become dominated by science and technology in the modern world, is easily forgotten. European and American medicine have provided an opportunity to confirm the value of ethnomycology by creating written records about the experimental verification of the properties of fungi, which were (and sometimes still are) used.

2. FOLK USE OF SOME MEDICINAL FUNGI 2.1. TRADITIONAL USES OF EDIBLE FUNGI

Today, one of the most popular mushrooms in markets are champignons (Agaricus sp.). They are valued for their taste, but they also contain health-promoting compounds, about which the people who ate those centuries ago did not know. The fruit bodies of different Agaricus spp. (class – Agaricomycetes) are used as folk drugs or simply as a part of healthy nourishment. In Bangladesh, it is believed that A. albolutescens can increase energy, treat sexual and heart disorders, and even the bites of snakes and tigers [3]. In Japan and Brazil, the fresh fruit bodies of A. blazei are eaten, while dry ones are prepared as infusions [4]. Another edible species that supports health is Hericium erinaceum (class – Agaricomycetes), which has been used as an anti-inflammatory medicine as well as an immune system booster for centuries. H. erinaceum is administered as tablets from dried and powdered fruit bodies. Its fruit bodies and the preparations that are produced from them were treated as excellent medicinal material in Chinese medicine [5]. Nowadays, these products and dry fruit bodies are commonly available in “eco shops”. The first cultivations of Agrocybe cylindracea (class – Agaricomycetes) took place in ancient Greece and Rome, although there was no knowledge about their bioactive compounds. Today, A. cylindracea is used in modern medicine and in the food industry [6]. Traditionally, hypogeous macrofungi (desert truffles; class - Pezizomycetes) have been used as folk medicine in Africa and in the Middle East for over two millennia. In general, most of known desert truffles species belong to genera such as Terfezia, Delastreopsis, Delastria, Leucangium, Mattirolomyces, Phaeangium, Picoa, Tirmania and Tuber. Boiled truffle water extract has been recommended for the treatment of eye and skin diseases. Desert truffles are also known to be one of the most famous aphrodisiac agents [7].

15

Folk use of some medicinal fungi

2.2. TRADITIONAL USES OF SOME INEDIBLE FUNGI

Not only edible fungi were (or still are) used. There are tribes in Siberia that use Amanita muscaria (class – Agaricomycetes)as the centerpiece of their religious ceremonies to this day. This poison mushroom is used by shamans to induce religious trances [8]. Psilocybe aztecorum (class – Agaricomycetes), which is known for its hallucinogenic properties, is currently used in religious rites in San Pedro Nexapa [9]. Fungi from the Phellinus genus (class – Agaricomycetes) are valuable sources of bioactive compounds and they are used in traditional Asian medicine. P. rimosus was used by a local tribe in Kerala (India) as a folk drug for the treatment of mumps [10]. P. igniarius is one of the most popular medicinal mushrooms in the world. New health-promoting preparations are created that are based on its compounds, spores or parts of its fruit bodies. Traditionally, it is known as a health booster and an ancient herbal medicine. It has been described as being effective on a wide diversity of diseases, including improving blood circulation, enhancing the detoxification and hepatoprotection of the human body, combating allergies and diabetes, curing oral ulcers and alleviating gastroenteric disorders or lymphatic diseases in Asian herbal medicine literature. Its extraordinary capacity for suppressing cancer and enhancing a body’s immunity has been scientifically proven (for example it has antiproliferative and antimetastatic effects) [11]. The basidiocarps of Phellinus nigricans were burned and their ashes were mixed with water. In Siberia, people chew on these wet ashes to take care of their teeth [12]. Cordyceps sinensis (class - Sordariomycetes), which consists of the dried fungus that grows on the larva of caterpillars, is one of the most valued traditional Chinese medicines. Both dry fruit bodies and infected caterpillars are eaten as a versatile remedy [13]. The next fungus that has been well known for centuries and is used by non-specialists is Ganoderma lucidum (class – Agaricomycetes). It is widely consumed in the belief that it promotes health and longevity [14]. Some fungi were not only used as food and/or folk medicine, but also as items of everyday use. Ganoderma applanatum is often called “Artist’s Conk” because its pilei have a white pore surface that instantly turns brown where touched, and thus enables an artist to draw a permanent picture [15]. Arboreal fungi refer to all fungi that live in trees (for example Ganoderma spp. and Phellinus spp. mentioned above). They are still used in order to kindle and keep a fire burning. It was easier to move from place to place with a smoldering fruit body than to try to kindle a new fire. The smoke from burning basidiocarps is still used by beekeepers in apiaries to keep the bees calm when the beekeeper has to enter the beehives [16].

3. SUMMARY

16

Katarzyna GÓRKA

The times when folk medicine was treated as a superstition have passed. Conventional medicine has not discovered any panacea, and because of this, people have started to separate the useful information about fungi from the folk myths. Ethnomycology concerns the simple, traditional and intuitive use of fungi. What is more, it is strongly connected to the things that can be created from mushrooms whose fruit bodies used to be much more important in everyday use than they are today. Not so long ago, many inedible fungi were valued, and unfortunately, today, they have been forgotten. REFERENCES

[1] Oxford Dictionaries, http://www.oxforddictionaries.com/definition/english/ethnomycology [Access: 21.07.2016.]. [2] SULLIVAN R., SMITH J.E., ROWAN N.J., Medicinal mushrooms and cancer therapy: translating a traditional practice into Western medicine, 2006, Perspectives in biology and medicine 49(2): 159170. [3] MOLLIK A.H., HASSAN A.I., PAUL T.K., SINTAHA M., KHALEQUE H.N., NOOR F.A., NAHAR A., SERAJ S., JAHAN R., CHWDHURY M.H., RAHMATULLAH M., Survey of Medicinal Plant Usage by Folk Medicinal Practitioners in Two Villages by the Rupsha River in Bagerhat District, 2010, American-Eurasian Journal of Sustainable Agriculture 4(3): 349-356. [4] BARBISAN L.F., SPINARDI-BARBISAN A.L., MOREIRA E.L.T., SALVADORI D.M.F., RIBEIRO L.R., da EIRA A.F., de CAMARGO J.L.V., Agaricus blazei (Himematsutake) does not alter the development of rat diethylnitrosamine-initiated hepatic preneoplastic foci, 2003, Cancer Science 94(2): 188-192. [5] SOKÓŁ S., GOLAK-SIWULSKA I., SOBIERALSKI K., SIWULSKI M., GÓRKA K., Biology, cultivation, and medicinal functions of the mushroom Hericium erinaceum, 2015, Acta Mycologica 50(2): 1-18. [6] SOBIERALSKI K., SIWULSKI M. (eds.), Polówka południowa Agrocybe cylindracea. Biologia, uprawa i właściwości prozdrowotne, Wydawnictwo Uniwersytetu Przyrodniczego w Poznaniu, Poznań 2015, 1-86. [7] ENSHASY H., ELSAYED E.A., AZIZ R., WADAAN M.A., Mushrooms and truffles: historical biofactories for complementary medicine in Africa and in the middle East, 2013, Evidence-Based Complementary and Alternative Medicine 2013: 1-10. [8] PAHNKE W.N., Drugs and Mysticism, 1966, The International Journal of Parapsychology VIII(2): 295-313. [9] MONTOYA, A., HERNANDEZ-TOTOMOCH O., ESTRADA-TORRES A., KONG A., CABALLERO J., Traditional knowledge about mushrooms in a Nahua community in the state of Tlaxcala, México, 2003, Mycologia 95(5), 793-806. [10] AJITH T.A., JANARDHANAN K.K., Antioxidant and Anti-inflammatory Activities of Methanol Extract of Phellinus rimosus (Berk) Pilat, 2001, Indian Journal of Experimental Biology 39: 11661169. [11] ZHU T., KIM S.-H., CHEN C.-Y., A Medicinal Mushroom: Phellinus Linteus, 2008, Current Medicinal Chemistry 15: 1330-1335. [12] SAAR M., Fungi in Khanty folk medicine, 1991, Journal of Ethnopharmacology 31(2): 175-179. [13] LI S.P., ZHAO K.J., JI Z.N., SONG Z.H., DONG T.T.X., LO C.K., CHEUNG J.K.H., ZHU S.Q.,

17

Folk use of some medicinal fungi

TSIM K.W.K., A polysaccharide isolated from Cordyceps sinensis, a traditional Chinese medicine, protects PC12 cells against hydrogen peroxide-induced injury, 2003, Life sciences 73(19), 25032513. [14] WACHTEL-GALOR S., TOMLISON B., BENZIE I.F., Ganoderma lucidum (‘Lingzhi’), a Chinese medicinal mushroom: biomarker responses in a controlled human supplementation study, 2004, British Journal of Nutrition 91(02): 263-269. [15] GINNS J., Annotated Key to Pacific Northwest Polypores, Pacific Northwest Key Council, Vancouver 2011, 04-17. [16] ROLFE R.T, ROLFE F.W., The romance of the fungus world: an account of fungus life in its numerousguises, both real and legendary, Dover Publications, New York 1974, 137-138.

18

Key words: municipal solid waste, SWA-Tool, separation, waste composition, waste analysis

Agnieszka GURGUL*

SIEVED ANALYSIS AND COMPOSITION OF MUNICIPAL SOLID WASTE

Waste management is the process of treating solid wastes and offers variety of solutions. Municipal solid waste data can be a source of information that can determine the planning and selection of a suitable method of waste management in the area. A review of results of the research involving the sieved and morphological analyses of municipal solid waste conducted in Waste Treatment Plant in Zielona Gora is presented in this paper. For these analyses European methodology for the Analysis of Solid Waste was used (SWA-Tool). The SWA-Tool aims to provide a waste analysis methodology for use at a local and regional level. The method involves measurements of the solid waste streams generated in the examined area. The presented data indicates that wastes composition of representative sample of 100 kilograms. Waste stream was divided into six sieve fractions and in every fraction there were separated material groups of wastes. The results of the research are compared to literature data of wastes composition in Polish cities. That comparison indicates that waste composition strongly depends on many factors e.g. the type of housing and size of urban agglomeration.

1. INTRODUCTION In order to select the method of waste management is required to know the composition and properties of the waste generated in the region. Various alternatives of waste management may include: reduction of waste, recycling, incineration with energy recovery, com__________ Wroclaw University of Science and Technology, Faculty of Environmental Engineering, Wybrzeże Wyspiańskiego 27, 50-370 Wrocław, e-mail:[email protected]

19

Agnieszka Gurgul

posting and landfilling. Reliable data on the characteristics of the waste can be a source of a decision on the planning and selection of a suitable method of waste management in the area which can help to reduce the negative impact of waste on the environment. Properties of municipal waste may vary depending on geographical and socioeconomic factors, for example the wealth of the residents, selective collection of recyclable materials and backyard composting. Effects on the characteristic of the waste may also have a type of building, equipment and season of the year. Composition of the waste affects its properties which is important when choosing a method of processing waste. Completed analysis of waste includes: sieved analysis (fractional), morphology (composition) and analysis of chemical properties. Based on research that allows determine the morphological and sieved analysis and the physical and chemical properties it is possible to choose the most suitable method for waste disposal and to design the most efficient system for the region. The aim of the study is to present research methodology that allows to analyze sieved composition and morphology of municipal solid waste. The studies were performed in Department of Public Utilities and Housing in Zielona Gora. 2. METHODS

Size fraction of municipal solid waste is often varied and may result from the characteristics of certain components. Using several mechanical sieves enables to divide waste into the samples of different grain size class. Mechanical separation allows for example the following size fractions to be obtained: the fine fraction 100 mm. During studies on waste composition mostly manual sieves are used which is the simplest tool that enable to separate every sample into bottom and upper fraction. Waste with a size smaller than the size of the holes fall through the sieve and become a bottom fraction. The remaining material forms a coarse fraction. We can separate analyzed waste into fractions using several sieves with different sizes of holes. Morphological analysis of municipal solid waste has objective to determine the material composition of a sample of waste. This analysis is an essential source of information about municipal solid waste. The procedure consists of sorting the total mass of waste into the components that differ in the type of material or characteristics. Separation and determination of the individual components provides information about the possibilities of processing and recycling. Typically, a complete morphological analysis is used only for middle and coarse fraction and fine fraction is weighted. Methods of morphological analysis of waste used in the various countries differ mainly in the mass of waste sample and the number of sorted categories [1]. SWA-

20

Sieved analysis and composition of municipal solid waste

3

Tool method had been developed within the project of the European Commission to provide a tool for the study on municipal solid waste at local and regional level. This method is mainly applied to households waste and waste from other sources, which are collected and transported together with household waste. Important advantage of the method SWA-tool is that it can be adapted to different local conditions, eg. there may be added additional criteria, so that the waste analysis can be adapted to local requirements. However, despite the additional criteria, in this method must be meet basic principles such as the sorting guidelines category and statistical accuracy [2]. It is not possible to examine all waste generated in a given area so it is important to choose the appropriate sample for research. The sample should be representative, it means that it should reflect the waste steam generated in the study area. The size of one sample should be approximately 100 kg. Samples should be selected in a random way, then every sample should be weighed and divided into following fractions: 100 mm. Then, fractions 40-80 mm, 80-100 mm, > 100 mm are divided into 34 categories and fractions 10-20 mm and 20-40 mm are divided into 11 categories, which is illustrated in the figure 1.

Fig.1 Waste analysis according to SWA-Toll methodology [2].

21

Agnieszka Gurgul

3. RESULTS AND DISCUSSION

Specification of waste categories in every sieved fraction determines which waste management methods should be implemented in examined area. The aim of this study was to determine the fractional composition and morphology of municipal waste brought to the Department of Public Utilities and Housing in Zielona Gora. Analysis were done with using SWA-Tool methodology, total weight of the sample of 83.93kg was randomly chosen from waste stream of about 1ton which was transported to place of research by special car. Sieve composition of the waste is shown in table 1. Tab.1 Sieve composition of the analysed waste. Fraction, mm

>100 80-100 40-80 20-40 10-20