92 10 3494 https://omeka.ibu.edu.ba/files/original/bef59450528ddea5431fc3faed9f5273.pdf 04a04cef19a87d039daa95d539487772 PDF Text Text Effect of Gibberellic Acid Application Times on Earliness and Production of A-106 Artichoke Cultivar in Antalya Ecologic Conditions Meliha Temirkaynak, Sadettin Küçük Recep Çoşkun West Mediterrenean Agricultural Research Institute, Antalya-Turkey Abstract: Artichoke is one of the important vegetable crops in Mediterranean countries including Turkey. In our country, while artichoke production was undertaken in İzmir, Bursa and Yalova until 10 years ago, its area of production has broadened to include southern cities such as Antalya and Muğla. As early artichoke production, especially in Antalya region, is profitable for farmers, demand for growing this crop has increased. The present study investigated the effect of gibberellic acid application times (0, 1, 2, 3 and 4) on earliness and production of A-106 artichoke cultivar. The plants were grown in West Mediterrrenean Agricultural Research Institute in 2007-2008. The results show that GA3 applications are very effective to promote earliness in A-106 cultivar. Keywords: GA3 application, time, artichoke, A-106 and earliness. Introduction Artichoke is a vegetable whose forebear is thistle and is included in Cynara cenus in botanical classifying, which belongs to the Asteraceae family, and it is perannual, in rozetta shape. It is declared in literature that the home of this vegetable, which can be seen in Mediterranean basin, is middle and west Mediterranean and has firstly been consumed by the Romans in 1979 B.C. (Anonymous a 2008; Graifeirberg ve ark. 1995; Lanteri ve ark. 2001; Stamigna 2001; Hill 2001). In our country artichoke is grown by vegetative propagation and especially in Aegean and Mediterranean regions with our standard types for a long time. However, taking more and early products from unit area has been the main objective, as the other branches of agriculture where the production cost increases day by day. In this context, our region, especially Antalya and neighbours, has many advantages in earliness as in many other kinds of fruit and vegetables. Until a few years ago, it wasn’t possible to see other plantations that were established by other types than Sakız and Bayrampaşa. In the last years, the number of plantations that are established with hybrid artichoke cultivars that are produced by seed have increased. Production by seed has an important disadvantage, lateness, besides many important advantages like productivity, resistance to diseases(especially Verticillium spp. ve Fusarium spp.) and profitability (Macua ve ark. 2000; Gil ve ark. 2003; Calabrese ve ark. 2004; Bianco 2000). The elimination of lateness has an importance in production by seed if artichoke is going to be grown in Mediterranean region of our country, especially in Antalya where earliness is of importance. Artichoke needs vernelization 250 hours under 7 C to form a head (although changing from cultivar to cultivar). Foury ve Pecaut 1988; Welbaum 1994; Basnizki ve Goldschmidt 1994; Hill 2001). The effects of GA3 applications in artichoke on earliness and productivity has been known and applicated for a long time. It has been declared in many scientific works that GA3 applications has taken the place of colding need for head forming in many cultivars (Gerakis ve ark.,1969; Snyder ve ark.1971; Kocer ve Eser 1999; Miguel ve ark.,1997; Miguel ve ark.,2004; Ercan ve ark.2004). The effects of hormone applications on seed propagated artichoke cultivars depend on date of plantation, time of application, number of application, dosage of application, and cultivar. (Welbaum 1994; Schrader, 1992; Elia ve ark., 1994; Mauromicale ve Ierna 1995; Miguel ve ark., 1997; Calabrese ve Bianco 2000; Mauromicale ve Ierna 2000; Goreta ve ark.,2004; Eldin ve ark., 2007). Mauromicale ve Ierna (2000) has decided that the head characteristics of Orlando and Sicilia artichoke cultivars change by hormon applications. Calabrese (2000) examined the early productivity of 8 hybrid artichoke types which are produced by seed in Southern Italy. It has been decided that the period between seeding and harvest is 97-120 days. It is declared in the studies of these investigators that productivity changes according to cultivars between 1.75/2.20 ton/ha. These results show that artichoke types that are produced by seed can be grown in October-May like other types which are grown in early classical ways. 34 �Material In this study A-106 artichoke cultivar, which is included in Cynara cenus which belongs to Asteraceae family, has been used as plant material. A-106 artichoke cultivar is a type which is early, has a low vernelization need, with much productivity, which form qualified heads, and has many puple brakte leaves. This study was conducted between the years 2007-2008 in a artichoke plantation situated in West Mediterrenean Agricultural Research Institute in Antalya-Turkey (360 56 N', 300 51' E). Method In this research it is aimed to determine the most suitable GA3 application time for A-106 artichoke for earliness. In this research 4th, 6th and 8th weeks after transferring to soil was selected as the first application time for vegetable development regulatory. 25 ppm GA3 applications were made to the plants by spraying. Only water was sprayed to the plants that have been selected as the control group. In this study following criteria have been examined: 1. Number of days from transfer to soil until harvest (earliness) Number of days from transferring the vegetables to soil until harvest is an important criteria in determining the effectiveness of the applications on earliness. 2. Head weight (g) Differences among the applications are determined by weighing the heads at time of harvest. 3. Head length (mm) Differences among the applications are determined by measuring the length of heads at time of harvest. 4. Head diameter (mm) Differences among the application are determined by measuring the diameter of heads at time of harvest. 5. Productivity (kg/da) Productivity in da is calcuted in kg by the productivity per vegetable. All application groups were laid out with four replications and 10 plants in each replicants in the randomised block experimental design. Data were subject to analysis of variance by SAS statistical program (SAS Institute, Version 7) and means were compared by LSD’s (Least Significant Differences) test at 0.05 significance levels. Results and Discussion In this research, where the affects of GA3 application times on A-106 artichoke cultivar on earliness, yield and head quality is examined, it was determined that hormone applications have important affects on all criteria that have been examined, compared to the control vegetables with no applications. Although depending on the application time of hormone, average 1,5 months of earliness was provided at the vegetables with applications. The earliest harvest at the GA3 application made at the 4th week happened at the 98. day after transferring to soil. The latest harvest happened at 140. day at the control group with the plants with no hormone application. At other application times these periods were very close, between 100-110 days. The effects of hormon application times on productivity was shown at Figure 1. As it can be seen in Figure 1, the effects of the applications were found statistically significant. The effects of application times on yield changed between 1.12-1.88 t/da; the highest yield was determined at the GA3 application on the 4. week as 1.88 t/da, and it was followed by the application at 6. week and the control apllication at the 8. week. The effects of applications on yield was found to be more succesful in all applications than control. Our trial results are in accordance with many other investigators. In many studies where the effects of hormone application times on artichoke was examined, it is reported that hormone application times increase productivity significantly. (Esteva ve Ayala, 2004; Ercan ve ark. 2004;Gil ve ark. 1998; Goreta ve ark. 2004) Besides, the effects of hormone application times on head quality (head weight, head width and length) are examined and the results are presented in Figure 2. The effects of the applications on the head quality were 35 �different. As a result of the statistical evaluations it is determined that the all hormone apllication times have important effects on all quality criteria that are examined, compared to the control group. Furthermore, the effects of hormone application times on head quality (head weight, head width and length) were examined in this study and the results are presented at Figure 2. The effects of the applications made on head quality were different. For example, the highest head weight was found to be 423 g at the 4th week, the lowest head weight was 410 g at the heads at the control group. Head weight values determined at other hormone application times took place in the same group statistically with the 4th week applications. The highest value of head width was found in the heads with hormone applications on 4th week. The highest head height was found at the vegetables with hormon applications on 8th week. The effect of GA3 applications on head length can be explained by the fostering of the GA’s to the cells for growing. In this study, where the effects of GA3 application times on earliness, yield and head quality of A-106 artichoke cultivar were examined, considering all the criteria investigated it can be said that the optimum GA3 application time is the 4th week after transferring the vegetables to soil. Table 1. Effect of GA3 Application Times on Yield (tonnes/da) of A-106 Artichoke Cultivar Application Time Yield(tonnes/da) Control 1.12 C 4.th 1.88 A 6. th 1.64 B 8. th 1.58 B LSD%5 0.18 Table 2. Effect of GA3 Application Times on Head Quality of A-106 Artichoke Cultivar Application Time Head Weight (g) Head Diameter (mm) Head Lenght mm) Control 382 B 98.98 C 89.48 C 4.th 423 A 130.28 A 98.74 B 6. th 418 A 122.48 B 112.46 B 8. th 420 A 120.22 B 118.34 A LSD%5 22 8.42 6.32 36 �References: ANONYMOUS, 2008. Artichoke-Spring Season. http://www.mmdtkw.org BASNIZKI Y. and E.E. GOLDSCHMIDT, 1994. Further Examination of Gibberelline A3 Effects on Flowering of Globe Artichokes (Cynara scolymus L.) Under Controlled Environment and Field Conditions. Isr. J. Plant Sci. (42):159166. BIANCO V.V., 2000. Present situation and future potentials at artichoke in the Mediterranean basin. http://www. area.ba.cnr.it CALABRESE N. And V.V. BIANCA, 2000. Effect of Gibberellic Acid on Yeld and Quality of Seed Grown Artichoke (Cynara cardunculus var. Scolymus (L.) Fiori). Acta Hort. 514:25-32. CALABRESE N., E. DE PALMA and V.V. BIANCA, 2000. Gibberellic acid and earliness of new seed propagated artichoke cultivars grown for four years (Abstract). IV. International Congress on Artichoke, October 17-21. Valenzanı-Bari, Italy. CALABRESE N., E. DE PALMA and V.V. BIANCA, 2004. Yields and quality of new commercial seed grown artichoke hybrids (Abstract). V. International Congress on Artichoke 5-8 may, 2003. Tudela-Navarra, Spain. ELDİN A.S., W.H. SCHNİTZLER, G. NİTZ, A.M. RAZİN and I.I. El-OKSH, 2007. The Effect of Gibberellic Acid (GA3) on Some Phenolic Substances in Globe Artichoke (Cynara cardunculus var. scolymus (L.) Fiori). Scientia Horticulturae Volume 111, Issue 4, 16 February 2007, Pages 326-329. ELIA A., N. CALABRESE and V.V. BIANCO, 1994. Sowing Time, Gibberellic Acid Treatments and Cultivars of “Seed” Propagated Artichoke. Acta.Hort. 371:347-354. ERCAN N., A.N. ONUS, F. AYAR, M. TEMİRKAYNAK, A.S. ŞENSOY and E. POLAT, 2004. Determination of optimum GA3 concentrations and awakining irrigation time for globe artichoke (Cynara scolymus L. Cv. Sakız) grown in mediterranean region of Turkey. V. International Congress on Artichoke 5-8 may, 2003. Tudela-Navarra, Spain.197-203. FOURY C., 2003. Common vegetables for seed and fruit. Chapter: Artichoke or globe artichoke and cardoon. http://gears.tucson.ars.ag.gov. (03.05.2008) GERAKIS P.A., D. MARKAIN and S. HONMA, 1969. Vernalization of Globe Artichoke, Cynara scolymus. J. Amer. Hort. Sci. 94:254-258. GIL O.R., J.J. MACUA and P. ARCE, 1993. Differentes ciclos de produccion en variedades de alcacchofa de multiplicacion por semilla. Agricola Vergel 139 :379-380. GORETA S., L. BUCAN and G. DUMİNİC, 2004. Effect of Environment and Gibberellic Acid (GA3) on Earliness and Yield of Globe Artichoke. V. International Congress on Artichoke 5-8 may, 2003. Tudela-Navarra, Spain.155-159. GRAIFEIRBERG A., L. GIUSTINIANI, O. TEMPERINI and M. LIPUCCI DI PAOLA, 1995. Allocation of Na, Cl, K and Ca within plant tissues in globe artichoke (Cynara scolymus L.)under saline-sodic conditions. Scientia Horticulturae 63 (1995) 1-10. HILL, D.E, 2001. Globe artichoke trials 1998, 2000 Management of yield using ınduced or natural vernalization. The Connecticut Agricultural Experiment Station, New Haven Bulletin 975, may 2001. 7 pages. KOÇER, G. ve B. ESER, 1999. Köklü Dip Sürgünü Özellikleri ve GA3 Uygulamalarının Enginarda Verim Dağılımına Etkileri. Turkish Journal of Agriculture&Forestry. Türk Tarım ve Ormancılık Dergisi. 23(2): 325-332. LANTERI S., I. D. LEO, L. LEDDA, M.G. MAMELI and E. PORTIS, 2001. RAPD variation within and among populations of globe artichoke cultivar ‘Spinosa Sardo’. Plant Breeding 120, 243-246. MACUA J. I, L. LAHOZ, A. MALUMBRES, J. GARNİCA, I. URMENETA and A. ARRONDO, 2000. Agricultural Behavior of Two Cardoon Varieties at Different Planting Dates in Navarra. V International Congress on Artichoke (abst.) MAUROMİCALE G. and A. IERNA, 1995. Effects of Gibberellic acid and Sowing Date on Harvest Time and Yields of Seed-Grown Globe Artichoke (Cynara scolymus L.). Agronomie. 15:527-538. MAUROMİCALE G. and A. IERNA, 2000. Characteristics of Head of Seed-Grown Globe Artichoke (Cynara cardunculus L. var. scolymus (L.) Fiori) as effected by Harvest Period, Sowing Date and Gibberellic Acid MIGUEL A., J.V. MAROTO, B. IRANZO and S. LOPEZ, 1997. Acido Giberelico en alcachofa. Horticultura, 120:111113.Agronomie 20: 197-204. MIGUEL A., C. BAIXAULI, J.M. AGUILAR, A. GINER, J.V. MAROTO, S. LOPEZ, A SAN BAUTİSTA and B. PASCUAL, 2004. Gibberellic Acid Concentrations in Seed Propagated Artichoke. V. International Congress on Artichoke 5-8 may, 2003. Tudela-Navarra, Spain.167-172. SCHRADER W.L., 1992. Growth Regulator Effects on Earliness and Yield in Artichokes Grown as Annuals from Seed.HortScience. 27:643. SNYDER M.J, N.C. WELCH and V.E. RUBATZKY, 1971. Influence of Gibberellin on Time of Bud Development in Globe Artichoke. HortScience, 6 (5):484-485. STAMIGNA C., G. PANDOZY, P. CRINO and F. SACCARDO, 2001. Male-sterility in Artichoke (Cynara scolymus L.) (Abstract). Proceedings of the XLV Italian society of Agricultural Genetics SIGA Annual Congress. Salsomaggione Terme, Italy-26/29 September 2001. WELBAUM, G.E., 1994. Annual Culture of Globe Artichoke from Seed in Virginia. HortTechnology. 4 (2(:147-150. 37 � Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Extent The size or duration of the resource. 646 Title A name given to the resource Effect of Gibberellic Acid Application Times on Earliness and Production of A-106 Artichoke Cultivar in Antalya Ecologic Conditions Author Author Temirkaynak, Meliha Küçük, Sadettin Çoskun, Recep Abstract A summary of the resource. Artichoke is one of the important vegetable crops in Mediterranean countries including Turkey. In our country, while artichoke production was undertaken in İzmir, Bursa and Yalova until 10 years ago, its area of production has broadened to include southern cities such as Antalya and Muğla. As early artichoke production, especially in Antalya region, is profitable for farmers, demand for growing this crop has increased. The present study investigated the effect of gibberellic acid application times (0, 1, 2, 3 and 4) on earliness and production of A-106 artichoke cultivar. The plants were grown in West Mediterrrenean Agricultural Research Institute in 2007-2008. The results show that GA3 applications are very effective to promote earliness in A-106 cultivar. Date A point or period of time associated with an event in the lifecycle of the resource 2009-06 Keywords Keywords. Conference or Workshop Item PeerReviewed Q Science (General) https://omeka.ibu.edu.ba/files/original/68a146321b51ccd878cd57463e1cc495.pdf 8ab71f08257f7082d8774e467dc33f02 PDF Text Text 1st International Syposium on Sustainable Development, June 9-10 2009, Sarajevo Effect of IBA and Bacteria (Agrobacterium rubi ve Bacillus OSU 142) on the Rooting of M9 Apple Rootstock Cuttings Lütfi Pirlak Selcuk University, Faculty ofAgriculture Department of Horticulture, Konya, Turkey lpirlak@hotmail.com Yüksel Baykal Celal AKIN Primary School,Yakutiye Erzurum, Turkey yukselbaykal@hotmail.com Abstract: This study was carried out to investigate the effects of IBA and bacteria (Agrobacterium rubi ve Bacillus OSU 142) applications on the rooting of M9 rootstock cuttings. The cuttings treated with IBA alone at different concentrations (2000, 4000 and 6000 ppm) and in combinations of three Agrobacterium rubi (A1, A16, A18) and Basillus OSU 142 strains. The treated cuttings were placed mist propagation system including perlite medium. After three months the cuttings were uprooted and evaluated for rooting percentage (%), callusing (%) and viability rate (%). There were determined inadequate results about rooting of cuttings. Rooting in cuttings was observed in 2000 ppm IBA (6.66 %), 4000 ppm IBA (13.33 %), and 2000 ppm IBA + Basillus OSU 142 combination (6.66 %), respectively. Callus were observed in all treated. Callus rate was 66.66 % in control while the rate was 84.61 % in A16 + A18 applications. Callus rate decreased depending on increasing of IBA doses. There were determined significantly differences in bacteria alone applications. According to application of bacteria combinations, the highests callus rate was A16 + A18 while the lowest was A1 + OSU 142. The best result of hormones + bacteria combinations were taken from 4000 ppm IBA + A18 application. Cutting living rate was similar to callus rating results. Many of the cuttings were live after application. Living rate was low at alone IBA application compared to that of the control. Introduction Apple is one ofthe most commonly cultivated produces around the world. Today, commercial apple (Malus domestica Borkh.) production is carried outthrough outtemperate regions of both north and southern hemispheres. To date, world’s apple production is 64 255 520 t(Anon. 2009). Such a widespread cultivation potential would be attributed to rich germplasm diversity ofmalus species in both cultivarand rootstock genotypes. Besides,recentimprovementsin utilitation of dwarf rootstocks in intensive cultivation contributed to enhance apple production. In orderto match present reguirements of modern apple orchards,rootstocks with high potentialin clonal propagation apptitude are recommended to use. Dwarf and semidwarf rootstocks, particularly, provide easyness in cultivation practices,reduce production cost and increase the proportionalincome. M9 is the most frequently used apple rootstock in the world to obtain dwarf plants. Malling 9 has for a long time been used as apple rootstock in Europe. Trees grafted on M9 do not exceed 2.7 m and are the 20-40% size of those grafted on seedling. Cultural processes are made without using a ladder. Thanks to excellent light exposure it provides, fruits are often well-colored and of high quality. However, it needs a backing for life because itsroot system is weak. Under available conditions,itsroots grow up well on beds, butitis quite hard to provide root formation via woody cuttings. Production of M9 apple rootstock is usually conducted with the “Stool Bed Layering” layering method (Howard 1985; Ferree & Carlson 1987; Hartmannn et al. 1990). M9 rootstock can be produced withlayering method, but can hardly be increased with cutting (Ferree & Carlson 1987). Increasing with cutting is an increasing method of great importance for all fruits as it is economical and fast. Despiteits all positive features,the biggest problem thatrestrictsitsuse isthat cuttings can not take root because regenerating abilities of some species are very deficient (Rugini & Fedelli 1990; Webster 1997). The cuttings belonging to the fruit species found in temperate climate group do not easily rooting. Thereby, cuttings are subject to various applications (plant growth regulators, different chemicals, bacteria 129 �1st International Syposium on Sustainable Development, June 9-10 2009, Sarajevo applications, carbohydrates etc.), one of which isthe use of plant growth regulator,in orderto remove problems about rooting in the kinds of these species (Dubeiovsky et al. 1993; Wiesman & Lavee, 1995; Grange et al. 1998; Eşitken et al. 2003). The most common application to increase the rooting ratio in cuttings is the use of auxins and especially of IBA (Hartmann et al. 1990; Howard 1985). In a study on determining the effects of IBA application with doses of 3000 and 8000 ppm on rooting properties of M9 apple root-stocks,it was reported thatthe maximum rooting (29.63%) was obtained from apple cuttings treated with 8000 ppm IBA (Ülger & Baktır 1995). In recent years, bacteria applications to cutting rootings have gained greatimportance. A. tumafaciens, E. milletiae, P. syringae pv. savastanoi, and P. syringae pv. myricae bacteria encourage cell divisions or gal formations in plants. These bacteria except A. tumafaciens are produced on intercell surfaces as indole-3acetamide and indol-3-pyruvate from oxine group by IAA (Goto 1990). These bacteria produce hormones in colonized areas on planttissues or encourage hormone production in plants. Therefore,these types of organisms cause cell divisions growth as depending on hormone production. Recently, especially some strains of A. rhizogenus (Bassilet al. 1991; Hatta et al. 1996) and A. rubi strains have been successfully been used for cutting rooting (Eşitken et al. 2003). In a study, 2000, 4000 and 6000 ppm IBA and three different types (A1, A16, A18) of Agrobacterium rubi bacteria were applied for determining cutting rooting of Kütahya sour cherry. W hile no rooting was determined in the control group, the highest rooting rate (70%) was obtained with application of 2000 ppm IBA+A16 (Ercisli et al. 2000).In a similar study, Ercisli et al.(2000a) applied 2000, 4000 and 6000 ppm IBA and three differenttypes (A1, A16, A18) of Agrobacterium rubi bacteriato rose hip and the highestrooting rate (95%) was obtained with 2000 ppm IBA + A18 application. Eşitken et al. (2003) applied IBA (250, 500, 750 ppm), Agrobacterium rubi and Bacillus OSU-142 to wild cherry, and reported that the best rooting rate (80%) was obtained with 250 ppm IBA + OSU 142. The objective ofthis study wasto determine the individual and combination effects of IBA, Agrobacterium rubi and Bacillus OSU 142 applications on rooting rate of M9 apple rootstock cuttings. Material and Methods Material This study was carried out at Atatürk University Faculty of Agriculture Department of Horticulture to investigate the effects of different applications on the rooting of hardwood M9 cutting. By this aim, twenty six different applications were performed using various doses ofIBA alone and/or distinct strains of Agrobacterium rubi and Bacillus OSU 142 bacteria. Applications are listed in Table 1. Bacteria strains were obtained from Atatürk University Faculty ofAgriculture Department of Plant Protection. 1. Kontrol 8. OSU 142 15. 2000 ppm IBA + A1 2. 1000 ppm IBA 3. 2000 ppm IBA 4. 6000 ppm IBA 5. A1 9. A1 + A16 16. 2000 ppm IBA + A16 17. 2000 ppm IBA + A18 18. 2000 ppm IBA + OSU 142 19. 4000 ppm IBA + A1 6. A16 7. A18 130 10. A1 + A18 11. A16 + A18 12. A1 + OSU 142 13. A16 + OSU 142 14. A18 + OSU 142 20. 4000 ppm IBA + A16 21. 4000 ppm IBA + A18 22. 4000 ppm IBA + OSU 142 23. 6000 ppm IBA + A1 24. 6000 ppm IBA + A16 25. 6000 ppm IBA + A18 26. 6000 ppm IBA + OSU 142 �1st International Syposium on Sustainable Development, June 9-10 2009, Sarajevo Methods Cuttings were collected from M9 clone root stocks and one-year shoots at dormancy stage. The bottom of cuttings was submitted into IBA solutions for 3-5 seconds, following evaporation then submitted into Agrobacterium rubi strains and Bacillus OSU 142 bacteria suspension with a concentration of 1x108 bacteria per milliliter. The bottom of control cuttings was submitted into distilled water only (Eşitken et al. 2003).In orderto smear bacteria on cutting bottoms,they were mixed for 30 minutes at 75 rounds per minute. Then cuttings were planted in fogging area in greenhouse (Figure 2.1). The growing media is automatically controlled for heating (26 o C) and relative humidity (90-95%). Perlite was used as growing media. Rooting rates (%), callus formation rates (%) and survival rates (%) of cuttings were determined at the end of 3 month growing period. The experimental design was completely randomized design with 3 replications (10 cuttings at each replication). Duncan’s multiple comparison tests was used for mean comparisons after arcsine transformation of raw data (Düzgüneş et al. 1987). Results The findings on the effects of IBA and bacteria applications on rooting, callus formation and survival cuttings percentages are presented in Table 1. Root formation did not ocur in cuttings of most treatments including control. However, 2000 ppm IBA, 4000 ppm IBA and 2000 ppm IBA + OSU 142 treatments resulted in root formations, 6.66, 13.33 and 6.66 %, respectively. Differences among the treatments were statistically significant. 131 �1st International Syposium on Sustainable Development, June 9-10 2009, Sarajevo Applications Rooting (%) Control 2000 ppm IBA 4000 ppm IBA 6000 ppm IBA A1 A16 A18 OSU 142 A1 + A16 A1 +A18 A16 +A18 A1 + OSU142 A16 + OSU142 A18 + OSU142 2000 ppm IBA + A1 2000 ppm A1 + A16 2000 ppm IBA + A18 2000 ppm IBA + OSU142 4000 ppm IBA + A1 4000 ppm A1 + A16 4000 ppm IBA + A18 4000 ppm IBA + OSU142 6000 ppm IBA + A1 6000 ppm A1 + A16 6000 ppm IBA + A18 6000 ppm IBA + OSU142 0b* 6.66 ab 13.33 a 0b 0b 0b 0b 0b 0b 0b 0b 0b 0b 0b 0b 0b 0b 6.66 ab 0b 0b 0b 0b 0b 0b 0b 0b 12.85 LSD .01 LSD .05 Callus Formation (%) 66.66 b 46.66 bcd 20.00 ef 13.33 f 21.42 ef 26.66 def 53.33 bc 53.33 bc 53.33 bc 60.00 bc 84.61 a 40.00 cde 46.66 bcd 46.66 bcd 20.00 ef 13.33 f 13.33 f 21.42 ef 26.66 def 46.66 bcd 60.00 bc 40.00 cde 0g 0g 0g 0g Survival Ratio (%) 60.00 c 13.33 ef 13.33 ef 13.33 ef 21.42 de 40.00 cd 60.00 c 66.66 bc 66.66 bc 86.66 ab 92.30 a 66.66 bc 53.33 cd 53.33 cd 13.33 ef 13.33 ef 21.42 de 40.00 cd 40.00 cd 73.33 bc 93.33 a 73.33 bc 0f 0f 0f 0f 18.29 4.78 • Statistical analysis have been carried out using arc sin values. Table 3.1. The effects of IBA and bacteria applications on the rooting, callus formation and survival ratio on M9 apple rootstock cuttings. Callus formation occured in cuttings of most applications, except forthe combinations of6000 ppm IBA with bacteria. There were statistically significant differences between the applications for callusing. The highest callusing rate was obtained from A16 + A18 (84.61%), followed by control(66.66%). The findings relevant to survival percentages of cuttings are generally similar to those of callus formation rates.In most cases the cuttings maintained their survives up to the end of study. However,there was no surviving cutting in combination of 6000 ppm IBA with bacteria as was seen in callusing rates. Differences in survival percentages were statistically significant. The highest percentage on this criterion was observed in 4000 ppm IBA + A18 treatment (93.33 %), while control group demonstrated considerably higherrate (60.00 %). Discussion Considering the overallrooting percentages,the results appear dissatisfactory. This results conform the previous statement of Parmar & Aier (1989) who clarified the general difficulties in rooting of temperate fruits as a results of their hard tissues. Besides this, rooting ağabeylity of apple cuttings as well as other temperate fruits, are solely affected by external treatments along with genotypic apptitude and environmental conditions. Auxin treatments, out of externalfactors, have a specialinterest(Goto 1990). Literatureinvestications revealthat 132 �1st International Syposium on Sustainable Development, June 9-10 2009, Sarajevo auxin application do not have significantinfluence on rooting of M9 cuttings (Ülger & Baktır 1995). Callus development was observed in mosttreatments. Cutting ratio with callus was found as 66.66% in control and increased up to the 84.61% after A16 + A18 applications. When the cuttings are placed to the suitable media for rooting, callus layer occurs at the lower part of the cuttings. Protective layer resulted from callus tissue delays the rot formation at the lower part of the cuttings. In some cases, callus layer helps water uptakes ofthe cuttings (Hartmann et al. 1990). On the other hand,there has been different information related to the effect of callus layer on root formation. Hartmann et al. (1990) reported that callus formation and rooting formation are independent from each other. Kantarcı & Ayfer (1989) reported similar result in one study conducted in hazelnut plant. Similarly, Tayfon (1995) obtained low rooting in hard wood cuttings of kiwi, but callus formation was high in same conditions. In general,the results obtained from being alive ratio of cutting look like almost the callus formation ratios. Most cuttings were observed alive aftertreatments. For example, after pull offthe cuttings, alive cutting ratios for control and 4000 ppm IBA+A18 application were 60% and 93.33%, respectively. By evaluation of the results of alive cutting ratios, it can be observed that alive cutting ratio and callus formation ratio were close to each other. The result was also supported by Hartmann et al.(1990)thatthe protective layerresulted from callus tissue delays the rotformation atthe lower part ofthe cutting and in some cases, calluslayer helps water uptake of cutting. In result,the study performed to research the effects of IBA and bacterium applications on rooting ratio of cuttings in M9 apple rootstock showed that sufficient rooting ratio was not obtained. The present study was mainly focused on recentthe effect of bacterium on cutting rooting thatis very popularin recently. The increase or decrease in rooting may be determined properly with use ofthe different bacterium races or cuttings taken in different period by future studies. References Anon., (2009). www.fao.org Bassil, N.V., Proebsting, W.M., Moore, L.W., & Lightfoot, D.A. (1991). Propagation of hazelnut stem cutings using Agrobacterium rhizogenes. HortScience, 26(8), 1058-1060. Dubeikovsky, A.N., Mordukhova, E.A., Kochetkov, V., Polikarpova, F.Y., & Boronin, A.M. (1993). Growth promotion of blackcurrant softwood cuttings by recombinant strain Pseudomonas fluorescens BSP53a synthesizing an increased amount of indole-3-acetic acid number. Soil Biology-Biochemistry, 25(9), 1277-1281. Düzgüneş, O., Kesici, T., Kavuncu, O., & Gürbüz, F. (1987). Araştırma ve Deneme Metodları (Đstatistik Metodları-II). Ankara Üniv. Ziraat Fak. Yay: 1021, Ders Kitabı: 295, Ankara, 381 s. Ercişli, S., Eşitken, A., & Şahin, F. (2000). IBA ve bakteri (Agrobacterium rubi) uygulamalarının Kütahya vişne çeşidi çeliklerinin köklenmesi üzerine etkisi. Bahçe, 29(1-2), 75-80. Ercişli, S., Eşitken, A., & Şahin, F. (2000a), IBA ve bakteri (Agrobacterium rubi) uygulamalarının kuşburnu çeliklerinin köklenmesi üzerine etkisi. II. Ulusal Fidancılık Kongresi, 25-29 Eylül 2000, Bademli /Ödemiş-Đzmir, www.agr.ege.edu.tr/fitekno Eşitken, A., Ercişli, S., Şevik, Đ., & Şahin, F. (2003). Effect of indole-3-butyric acid and different strains of Agrobacterium rubi on adventive root formation from softwood and semi-hardwood wild sour cherry cuttings. Turkish Journal of Agriculture and Forestry, 27, 37-42. Ferree, D.C., & Carlson, R.F. (1987). Apple Rootstocks. Rootstocks for Fruit Crops, Rom, C.R. and Carlson, R.F. (eds). John Wiley and Sons Inc., New York, U.S.A., pp 107-144. Goto, M. (1990). Fundamentals of Bacterial Plant Pathology. Academic Press. Inc, 339 pp. Grange, O., Bartschi, H., & Gay, G. (1998). Effect of the ecto-mycorrhizal fungus Hebeloma cylindrosporum on in vitro rooting of micropropagated cuttings of arbuscular mycorrhiza forming Prunus avium and Prunus cerasus. Trees: Structure and Function, 12(1), 49-56. Hartmann, H.T., Kester, D.E., & Davies, F.T. Jr. (1990). Plant Propagation, Principles and Practices. Prentice-Hall, Inc. New Jersey, 647 p. Hatta, M., Beyl, C.A., Garton, S., & Diner, A.M. (1996). Induction of roots on jujube softwood cuttings using Agrobacterium rhizogenus. Journal of Horticultural Science, 71(6), 881-886. Howard, B.H. (1985). The contribution to rooting in leafless winter plum cuttings of IBA applied to the epidermis. Journal of 133 �1st International Syposium on Sustainable Development, June 9-10 2009, Sarajevo Horticultural Science, 60(2), 153-159. Kantarcı, M., & Ayfer, M. (1989). Bazı önemli fındık çeşitlerimizin çelikle çoğaltılmaları üzerine araştırmalar. Turkish Journal of Agriculture and Forestry, 13(3b), 1096-1109. Parmar, S.D., & Aier, N.B. (1989). Seasonal rooting behaviour of cuttings of plum cultivars as influenced by IBA treatments. Scientia Horticulturae, 40, 297-303. Rugini, E., & Fedelli, E. (1990). Olive (Olea europaea L.) as an oilseed crop. Ed. J.P.S. Bajaj, Biotechnology in Agriculture and Forestry, Vol. 10: Legumes and oilseed crops. I. Springer-Verlag Berlin, Heiderberg. Tayfon, A. (1995). Kivi’nin Çelikle Üretilmesi Üzerine Araştırmalar. Ege Üniv. Fen Bil. Ens. (Y.Lisans Tezi). Ülger, S., & Baktır, Đ. (1995). Bodur M9, J9 ve Colt anaçlarının fog serasında köklenme özelliklerinin saptanması. Türkiye II. Ulusal Bahçe Bitkileri Kongresi, Cilt I (Meyve):21-24. Webster, A.D. (1997). A review of fruit tree rootstock research and development. Acta Horticulturae, 451(1), 53-74. Wiesman, Z., & Lavee, S. (1995). Relationship of carbohydrate sources and Indole-3-Butyric Acid in olive cuttings. Australian Journal of Plant Physiology, 22(5), 811-816. 134 � Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Extent The size or duration of the resource. 492 Title A name given to the resource Effect of IBA and Bacteria (Agrobacterium rubi ve Bacillus OSU 142) on the Rooting of M9 Apple Rootstock Cuttings Author Author Pirlak, Lütfi Baykal, Yüksel Abstract A summary of the resource. This study was carried out to investigate the effects of IBA and bacteria (Agrobacterium rubi ve Bacillus OSU 142) applications on the rooting of M9 rootstock cuttings. The cuttings treated with IBA alone at different concentrations (2000, 4000 and 6000 ppm) and in combinations of three Agrobacterium rubi (A1, A16, A18) and Basillus OSU 142 strains. The treated cuttings were placed mist propagation system including perlite medium. After three months the cuttings were uprooted and evaluated for rooting percentage (%), callusing (%) and viability rate (%). There were determined inadequate results about rooting of cuttings. Rooting in cuttings was observed in 2000 ppm IBA (6.66 %), 4000 ppm IBA (13.33 %), and 2000 ppm IBA + Basillus OSU 142 combination (6.66 %), respectively. Callus were observed in all treated. Callus rate was 66.66 % in control while the rate was 84.61 % in A16 + A18 applications. Callus rate decreased depending on increasing of IBA doses. There were determined significantly differences in bacteria alone applications. According to application of bacteria combinations, the highests callus rate was A16 + A18 while the lowest was A1 + OSU 142. The best result of hormones + bacteria combinations were taken from 4000 ppm IBA + A18 application. Cutting living rate was similar to callus rating results. Many of the cuttings were live after application. Living rate was low at alone IBA application compared to that of the control. Date A point or period of time associated with an event in the lifecycle of the resource 2009-06 Keywords Keywords. Conference or Workshop Item PeerReviewed Q Science (General) https://omeka.ibu.edu.ba/files/original/a9c60f561a4c20da19f9c16a5a311733.docx f620604759bfc76f080585af4886ce43 Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Journal of Natural Sciences and Engineering Identifier An unambiguous reference to the resource within a given context 2637-2835 DOI Digital object identifier 10.14706 Publisher An entity responsible for making the resource available International Burch University Description An account of the resource Journal of Natural Sciences and Engineering (JONSAE) is a peer-reviewed, biannually published international journal focusing on empirical and theoretical research in all branches of Engineering and Natural Sciences. It is published on the behalf of Faculty of Engineering and Natural Sciences of International Burch University and aims to provide the best content regarding by publishing original research papers, review articles, special issues, feature articles, and book reviews. All manuscript submissions are subject to initial appraisal by the Editor, and, if found suitable for further consideration, to peer review by independent, anonymous referees. All peer review is double-blind and submission is online. The journal welcomes theoretical, applied, interdisciplinary and methodological work, with preference on empirical research, critical approach and problem-solving methods in manuscripts. Language A language of the resource English Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Effect of initial bacterial innoculum density on zones of inhibition in disk diffusion antibiotic susceptibility testing Author Author Aldijana Hadžić Nudžejma Kudić Monia Avdić Lejla Smajlović-Skenderagić Alisa Smajović Samra Međedović Abstract A summary of the resource. Abstract - The primary objective of this study was to show how the density of the initial bacterial inoculum effects the zone inhibition in Disk diffusion Bauer Kirby (BK) antibiotic susceptibility testing. In this study, three strains of Staphylococcus aureus were tested: a methicillin sensitive clinical strain, a methicillin resistant clinical strain and ATCC 25923. A series of decreasing initial inoculum densities of the three tested strains were prepared and poured onto Mueller Hinton agar plates. After overnight incubation the zones of inhibition around tested antibiotics from different inoculum densities were measured in mm. The results showed that inoculum density does have an effect on the zones of inhibition in BK antibiotic susceptibility testing of S.aureus where in the case of gentamycin sensitivity category change occurred. Correlation analysis showed that there is significant negative correlation between tested inoculum densities and zones of inhibition clinical methicillin sensitive strain of S.aureus after using oxacillin and gentamycin (Pearson coefficient were -0.917 and -0.892, respectively), and between tested inoculum densities and zones of inhibition clinical methicillin resistant strain of S.aureus after using ampicillin (Pearson coefficient was -0.960). Hence, initial bacterial inoculum density can be of high relevance in Bauer-Kirby disk diffusion testing and ought to be precisely determined in purpose of adequate therapy ordination. Keywords Keywords. Keywords: Staphylococcus aureus, susceptibility testing, McFarland standards, antibiotics, zones of inhibition. Publisher An entity responsible for making the resource available International Burch University Source A related resource from which the described resource is derived Journal of Natural Sciences and Engineering https://omeka.ibu.edu.ba/files/original/f4323d165e548514650b5b329a74df0b.pdf 8c90e5535ebc053d642fcd3d071d3bbe PDF Text Text Effect of Marble Dust on Consolidation Characteristics of Clay Soils Ismail ZORLUER Department of Construction Education, AKU, Afyonkarahisar, Turkey izorluer@aku.edu.tr Ismail MURATOGLU Department of Construction Education, AKU, Afyonkarahisar, Turkey Abstract : The usage of waste materials as an additive material has became widespread, in soil stabilization. This case was positive effects on environment by means of recycling, regains to economy and reducing environmental pollutions. In this study, marble dust had been used as an additive material in clay soil. Marble dust is a waste of the marble industry and despite its recycling in various industries, there is still a significant amount of marble dust left as waste. In this study, soil specimens were sampled from different locations in the ANS campus of Afyon Kocatepe University. These specimens were mixed with waste marble dust at ratios of 5%, 10% and 15%. Geotechnical experiments were carried out on specimens. Test results shows that marble dust have affected consolidation characteristics of clay soils. Especially, swelling index and consolidation index of specimens were decreased. This decrease is important in point of swelling potential of clay soils. Introduction Marble dust is a waste of the marble industry and despite its recycling in various industries, there is still a significant amount of marble dust left as waste. Marble dust has been used as an additive for soil stabilization. Okagbue and Onyeobi (1999) showed that the geotechnical parameters of red tropical soils are substantially improved by adding marble dust: plasticity is reduced by 20 to 33% and strength and CBR increased by 30 to 46% and 27 to 55%, respectively. The effect of marble dust on the swelling potential of Na-bentonite and Meşelik clays and unconfined compressive strength was investigated by Zorluer (2003, 2006). Specimens were mixed with marble dust at different percentages (3,5,8,10%) of dry soil weight, and compacted at standard proctor compaction energy. For swelling tests, specimens were obtained using oedometer floating ring from compacted mixtures and then tests were carried out with oedometer. Swelling potential reduced from 25.6% to 21% at 5-8% marble dust additive. For compression test, specimens were sampled with coring tube from compacted mixtures. Unconfined compressive tests were performed to these specimens and were cured for 1, 7 and 28 days. At the end of 28 days of curing time, strength increased from 20.1 to 57.3 N/cm2. Waste marble dust was used as an additive material by Zorluer and Taspolat (2009) in landfill liner. Mixtures of kaolinite-bentonite were mixed with waste marble dust for design of landfill liner. This process was performed at marble dust ratio of 5%, 10% and 15%. Freezing-thawing tests were carried out in these mixtures. At the end of the tests, it was observed that waste marble dust increased strength of liner in conditions of freezing and thawing. The objective of this study was to investigate use of marble dust as an additive in clay soils. For this purpose, soil specimens were sampled from 3 locations at ANS campus of Afyon Kocatepe University. These specimens were mixed with waste marble dust (proportions of 5, 10 and 15% dust to dry soil by weight). Index properties of the specimens were determined by liquid limit, plastic limit, sieve, hydrometer and buoyancy analysis tests. Standard proctor and odeometer tests were carried out in these specimens. Materials Afyonkarahisar region is known as one of the most important marble production and processing centre in Turkey. Yearly production of marble is about 80,000 m3 in this region. About 24,000 m3 marble dust occurs 514 �from this production. Marble dust is minimum sized marble waste that occurs with sawing of marble blocks and plates. This dust is carried by water to sedimentation ponds. Sediment dust is removed from the pond to wasteland, but this forms serious problems for the environment. Waste marble dust is used in very small quantities despite being used in widely variable industries, such as construction, ceramics and cement, paint, agriculture and fertilizer; as a result, a lot of marble dust ends up as waste (2003). The marble dust used in this study, was obtained from a marble processing factory in Afyonkarahisar-Turkey. It was dried and sieved, resulting in marble dust grains smaller than 300 microns. Table 1 are shown chemical compound percentage (%) of marble dust. SiO2 0.01 Al2O3 Fe2O3 0.85 0.04 CaO 55.30 MgO 0.24 P2 O 3 ----- K2 O 0.20 Na2O 0.03 SO3 ----- Mn2O3 LOIa ----- 43.51 Table 1. Chemical Compound Percentage of Marble Dust (%) Afyon Kocatepe University Campus area was formed clay. Clay specimens were sampled from three points at campus area. These points were named as BH1, BH2 and BH3. Properties of specimens are in the table 2. Soil classification and definition tests were applied according to the TS 1900 standard. wp Ip class Specimen Gs wl BH1 2,65 59,5 25,6 33,9 CH BH2 2,72 37,2 20,4 16,8 CL BH3 2,76 29,4 21,6 7,8 CL Gs:Specific Gravity, wl: Liquid limit, wp: Plastic Limit, Ip: Plasticity index Table 2. Geotechnical Properties of Clay Specimens 100 BH1 90 BH2 Percent finer (%) 80 BH3 70 60 50 40 30 20 10 10 1 0,1 0,01 0,001 Grain size (mm) Figure 1. Grain size distribution curves of specimens. Experimental Study and Results Standard proctor test was performed on clay specimens. Compaction characteristics of clay soils were determined from this Proctor test. Maximum dry density and optimum water contents were obtained from figure 2. The specimens were mixed with waste marble dust at ratio of 5%, 10% and 15%. These ratios were obtained 515 �from other studies (Okagbue&Onyeobi, 1999; Zorluer, 2003, 2006, 2009). Then, these mixtures were compacted with optimum water content at the standard compaction mold. 17,5 BH1 BH2 BH3 Dry Unit weight (kN/m2) 17 16,5 16 15,5 15 14,5 14 10 12 14 16 18 20 22 24 26 28 Water content (%) Figure 2. Compaction curves of soil specimens For consolidation tests, specimens were sampled from compacted mixtures using odeometer ring. Consolidation tests were carried out on these specimens. e-log p graphs were plotted from consolidation tests results (fig. 3 a, b, c). In addition, compression index (cc) and expansion index (ce) were obtained from figure 3. 1,3 BH2 5% 10% 15% 1,8 1,6 Void ratio (e) 1,2 1,1 1,4 1,2 1 1 0,9 0,8 0,8 0,6 1 10 100 Pressure (kN/m 2) 1000 1 10 100 Pressure (kN/m 2) a b 2,2 2,1 Void ratio (e) Void ratio (e) 2 BH1 5% 10% 15% BH3 5% 10% 15% 2 1,9 1,8 1,7 1,6 1,5 1,4 1,3 1,2 1,1 1 1 10 100 Pressure (kN/m 2) 1000 c Figure 3. (a, b, c) e-log p curves of specimens 516 1000 �Discussion Compression index (cc) is the slope of the linear portion of the e-log p plot and dimensionless. It was seen that cc has decreased with marble dust increasing for all specimens (fig 4 a). For example, this decreasing is from 0,304 to 0,220 for BH3. Similarly, void ratios of specimens have decreased with marble dust increasing (fig 3). Consolidation settlement of soils is fewer when soil voids decreased. 0,320 0,060 BH1 BH2 0,055 BH3 BH3 Expansion index Compression index BH1 BH2 0,300 0,280 0,260 0,240 0,050 0,045 0,040 0,035 0,220 0,200 0,030 0% Additive 5% Additive %10 Additive %15 Additive 0% Additive 5% Additive %10 Additive %15 Additive a b Figure 4. Change of compression and expansion indices with increasing marble dust. Decreasing of expansion index (ce) is same with other studies (Zorluer, 2003). The findings show that the expansion index of specimens decreases when the amount of the added marble dust increases (fig 4.b). Therefore, swelling potential reduces when the amount of the added marble dust increases. This case shows that marble dust can be used at stabilization of swelling soils. Also, at the other study of Zorluer (2003), swelling potential was reduced by adding marble dust. Besides, swelling potential values was measured from swell pressure test. Conclusion Marble dust affects the properties of clay like strength, swelling potential, freeze-thaw strength. This case was expressed at previous studies. In this study, consolidation characteristic of clay were affected from waste marble dust. Compression index (cc) and expansion index (ce) of specimens decreases when the amount of the added marble dust increases. Furthermore, void ratio decreases with increasing of marble dust. This result shows that consolidation settlement reduced when marble dust mixed to clay soil. Use of marble dust in soil stabilization, provide the protection of the environment. In addition, it is gained an economical material for soil stabilization. References Okagbue, C.O. Onyeobi, T.U.S. (1999). Potential of marble dust to stabilize red tropical soils for road construction. Engineering Geology, 53. 371-380. Zorluer, I. (2003). Effect of waste marble dust to swelling potential of clay soils. XI. National Clay Symposium. Izmir, Turkey. 475-482. TS 1900-1 (Turkish Standard) (2006) Methods of testing soils for civil engineering purposes in the laboratory - Part 1: Determination of physical properties Ankara, Turkey. TS 1900-2 (Turkish Standard) (2006) Methods of testing soils for civil engineering purposes in the laboratory – Part 2: Determination of Mechanical Properties Ankara,Turkey. Zorluer, I. (2006). The Effect of waste marble dust on unconfined compression strength of clay soils. GAP V, Engineering Congress. Sanlıurfa, Turkey, 1042-1046. Zorluer, I. & Taspolat, L.T. (2009). Reuse of waste marble dust in the landfill layer. First International Symposium on Sustainable Development. Sarajevo, Bosnia and Herzegovina. 301-305. 517 � Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Extent The size or duration of the resource. 581 Title A name given to the resource Effect of Marble Dust on Consolidation Characteristics of Clay Soils Author Author ZORLUER, Ismail MURATOGLU, Ismail Abstract A summary of the resource. The usage of waste materials as an additive material has became widespread, in soil stabilization. This case was positive effects on environment by means of recycling, regains to economy and reducing environmental pollutions. In this study, marble dust had been used as an additive material in clay soil. Marble dust is a waste of the marble industry and despite its recycling in various industries, there is still a significant amount of marble dust left as waste. In this study, soil specimens were sampled from different locations in the ANS campus of Afyon Kocatepe University. These specimens were mixed with waste marble dust at ratios of 5%, 10% and 15%. Geotechnical experiments were carried out on specimens. Test results shows that marble dust have affected consolidation characteristics of clay soils. Especially, swelling index and consolidation index of specimens were decreased. This decrease is important in point of swelling potential of clay soils. Date A point or period of time associated with an event in the lifecycle of the resource 2010-06 Keywords Keywords. Conference or Workshop Item PeerReviewed Q Science (General) https://omeka.ibu.edu.ba/files/original/75d3d0af9ea8e10805ad5c3282bba357.pdf 7331ebb69ef6ccfd5bd7b7b6a4f1b3ca PDF Text Text Journal of Natural Sciences and Engineering, Vol. 1, (2019) DOI number: 10.14706/JONSAE2019114 Effect of metals on antibiotic sensitivity, growth, and biofilm-forming capacity of B. subtilis subsp. spizizenii Selma Cifric1 1 International Burch University, Sarajevo, Bosnia and Herzegovina selma.cifric@stu.ibu.edu.ba Abstract – B. subtilis is normally considered a soil organism, it can be also found in the animal and human gastrointestinal tract. Bacillus subtilis subsp. spizizenii is a type of Bacillus subtilis complex. It shares up to 99% of homology with B. subtilis CU1, which can be represented as a probiotic strain. Metal compounds found in soil or used in agriculture can easily enter the food chain and end up in our gut. Gram-positive bacteria (e.g. Bacillus spp.) have good adsorptive capacity for metals due to high peptidoglycan and teichoic acid content in cell walls. There is some evidence that certain metals inside the intestine play an important role in influencing growth and functionality of specific probiotic strains. Some of them have inhibitory, while others have an activating effect on bacteria. This study revealed that metal compounds increased antibiotic susceptibility of B. subtilis subsp. spizizenii. Higher concentrations of metal solutions inhibited growth of tested bacteria. Culture did not show affinity to form biofilms before or after addition of metal solutions. Keywords – antibiotic susceptibility, biofilms, MIC, metals. 1. Introduction Various bacteria reside in the gut or arrive there by food consumption. A microbiome is the overall collection of the genetic material of all microorganisms that live on or inside our body or collection of the genetic material of microorganisms in a particular environment (e.g., in your gut). Bacteria within our gut have an important role in digesting food, modulating the immune system, providing protection against harmful microbes, and more. Multiple factors including genotype, antibiotics, mode of delivery, dietary habits, lifestyle, social interactions and environmental factors shape the gut microbiota to make everyone’s microbiome unique [1, 2, 3]. Metal compounds can cause alterations in the composition of the gut microbiota. Usually, decrease in richness as well as the diversity of gut microbiota, is observed after exposure to metals [4, 5]. Gram-positive bacteria (e.g. Bacillus spp.) have good adsorptive capacity for metals due to high peptidoglycan and teichoic acid content in cell walls, in contrast to Gram-negative bacteria [6]. The phylum Firmicutes found in colon is mostly composed of gram-positive species, such as Clostridium and Bacillus. There is some evidence that certain metals inside the intestine play an important role in influencing growth and functionality of specific probiotic strains. Some of them have inhibitory, while others have an activating effect on bacteria. It has been concluded that many effects of metals are strain-specific [7]. �Journal of Natural Sciences and Engineering, Vol. 1, (2019) DOI number: 10.14706/JONSAE2019114 Bacillus subtilis is a gram positive and catalase positive rods. It is spore-forming bacteria. Although normally considered a soil organism, it is also found in the animal and human gastrointestinal tract [8]. Bacillus subtilis subsp. spizizenii is a type of Bacillus subtilis complex. It shares up to 99% of homology with Bacillus subtilis CU1, which can be represented as a probiotic strain that can have specific outcomes on the immune system of the elderly [9, 10]. Probiotics are commensal bacteria in the gut that have a health beneficial effect on the host organism. However, there are still a few unresolved questions regarding the safety of certain Bacillus strains, which is the main reason for their still limited application as probiotics [11, 12]. Biofilms are communities of bacteria joined together by a sticky extracellular matrix. This extracellular matrix is also responsible for adherent biofilms to various surfaces. Probiotic bacteria in the gut also use biofilm attachment to bind to the mucosa layer of the intestine. Biofilm attachment improves their survival rate. Specifically, biofilms provide protection against antibiotics and enzymes [13, 14, 15]. Antibiotics are antimicrobial agents active against bacteria. Their mode of action can be bactericidal or bacteriostatic. Application of antibiotics influences intestinal microbiota. It affects growth, diversity and antibiotic resistance of bacteria. Since Bacillus subtilis are partially considered as probiotic bacteria, normally found in the human gastrointestinal tract, this study will show their antibiotic susceptibility in the presence of metal compounds that can end up in our gut via food intake [16, 17]. In this paper, the effect of metal compounds on biofilm forming capacity, bacterial growth, and changes in antibiotic sensitivity is examined. It is assumed that metal compounds would increase antibiotic sensitivity and suppress growth. 2. Methods 1. Cultivation of B. subtilis subsp. spizizenii strain Bacillus subtilis subsp. spizizenii (ATCC 6633) was cultivated on solid and liquid media (trypticase soy broth (TSB) broth, TSB agar). After overnight incubation at 37 C, the turbidity of bacterial density is adjusted to 0.5 McFarland standard, as such was used for further tests. 2. Determination of antibiotic susceptibility before the addition of metal supplements Bacteria is previously cultivated on TSB agar. Susceptibility to fifteen types of antibiotics will be performed using the standard Kirby-Bauer disk diffusion method [18]. Antibiotics (Liofilchem) are listed in Table 1 below. 3. Microbroth dilution method Microbroth dilution method will be used to determine the minimal dose of metal supplement necessary to inhibit the growth of bacteria (minimum inhibitory concentration - MIC). It is accomplished through the standardized broth microdilution assay procedure [19, 20]. 96-well microtiter plates were used. The metal salts were aseptically diluted in TSB broth in the following w/V solutions: 1%, 0.5%, 0.25%, 0.12%, 0.06%, 0.03%, 0.015%, 0.007%, 0.003%, 0.0018%, 0.0009%. The 96-well plate contained 100 ul of different concentrations of metal solutions (CuSO4, ZnSO4 x 7H2O, Fe(NO3)3, and Mg), 100 ul TSB broth, and 20 ul of B. subtilis subsp. spizizenii (0.5 McFarland standard). This test was done in triplets. The �Journal of Natural Sciences and Engineering, Vol. 1, (2019) DOI number: 10.14706/JONSAE2019114 purpose was to determine the exact concentration of each metal that inhibits bacterial growth. After overnight incubation at 37 C visible growth of bacteria is recorded and MICs have been determined. 4. Determination of biofilm forming capacity This test determines how different concentrations of CuSO4 - copper (II) sulfate pentahydrate (Sigma-Aldrich), ZnSO4 x 7H2O - zinc sulfate heptahydrate (Sigma-Aldrich), Fe(NO3)3 - iron (III) nitrate (Fisher Scientific), and magnesium complex (Twinlab - dietary supplement from local pharmacy) will facilitate the biofilm formation. This test will be performed using TCP method. The 96-well plate contained different concentrations of metals, TSB medium, and 20 ul of B. subtilis subsp. spizizenii (0.5 McFarland standard). The inoculated plate should be covered with a lid and incubated for 24 h at 37 C. After incubation the content of the plates is discarded and washed. Crystal violet assay is used as a method of indirect biofilm quantification. Each microtiter-plate well is stained with 120 ul of 0.1% crystal violet and set aside for 10 minutes. Microliter-plate is decanted again and washed with distilled water. The test is done in triplets [21, 22]. 5. Determination of antibiotic susceptibility after addition of metals Susceptibility to fifteen types of antibiotics (Table 1) after addition of metal solutions will be performed using the Kirby-Bauer disk diffusion method [18]. Table 1. List of fifteen antibiotic discs used for antibiotic susceptibility testing. Name of antibiotic Micrograms Abbreviation Cefoxitin 30 FOX30 Gentamicin 10 CN10 Oxacillin 1 OX1 Amoxicillin 10 AML10 Ceftazidime + clavulanic acid 40 CAL40 Ciprofloxacin 5 CIP5 Streptomycin 10 S10 Vancomycin 30 VA30 Erythromycin 15 E15 Ceftazidime 10 CAZ10 Amoxicillin-clavulanic acid 30 AUG30 Azithromycin 15 AZM15 Kanamycin 30 K30 Tetracycline 30 TE30 Ampicillin 2 AMP2 �Journal of Natural Sciences and Engineering, Vol. 1, (2019) DOI number: 10.14706/JONSAE2019114 3. Results After testing the effect of metal compounds on growth, antibiotic susceptibility, and biofilm forming capacity, the following results were obtained. Table 2 shows results obtained after performing antibiotic susceptibility test for B. subtilis subsp. spizizenii. It compares diameters of the inhibition zone (in millimeters), before and after addition of four different metal compounds. Table 2. Antibiotic susceptibility test for B. subtilis subsp. spizizenii. Diameter of the zone of inhibition is in millimeters. (* - partially bactericidal) B. spizizenii B. spizizenii + Mg B. spizizenii + ZnSO4 x 7H2O B. spizizenii + Fe(NO3)3 B. spizizenii + CuSO4 FOX30 25 28 27 28 24 CN10 20 22 21 20 21 OX1 15 18 18 17 15 AML1 0 9 12* 15* 14* 11 CAL40 0 0 0 0 0 CIP5 29 32 32 32 35 S10 19 20 18 19 19 VA30 18 21 20 20 20 E15 21 25 28 24 25 CAZ10 0 0 6 8 0 AGU30 21 23 25 24 22 AZM1 5 20 22 21 23 22 K30 22 24 24 23 24 TE30 26 31 29 30 27 AMP2 0 0 0 0 0 Since diameters of inhibition zones for fifteen antibiotics were measured manually, Figure 1 visualizes sizes of diameters and possible manual errors during the measurement process. �Journal of Natural Sciences and Engineering, Vol. 1, (2019) DOI number: 10.14706/JONSAE2019114 Figure 1. Antibiotic susceptibility to fifteen antibiotics measured by zone of inhibition (in millimeters). Results of microbroth dilution tests are presented in Table 3 and Figure 2. Table 3 shows how different concentrations of metal (w/V) solutions affect growth of B. subtilis subsp. spizizenii, while minimum inhibitory concentrations of metals are summarized in Figure 2. Table 3. Growth of B. subtilis subsp. spizizenii under different concentrations of metal solutions w/V solution 1% 0.5% 0.25% 0.12% 0.06% 0.03% 0.015% 0.007% 0.003% 0.0018% 0.0009% Mg Fe(NO3)3 CuSO4 ZnSO4 x 7H20 No growth No growth No growth No growth No growth No growth No growth No growth No growth Growth No growth No growth No growth Growth No growth No growth Growth Growth No growth No growth Growth Growth Growth No growth Growth Growth Growth Growth Growth Growth Growth Growth Growth Growth Growth Growth Growth Growth Growth Growth Growth Growth Growth Growth �Journal of Natural Sciences and Engineering, Vol. 1, (2019) DOI number: 10.14706/JONSAE2019114 Figure 2. Minimum inhibitory concentration (MIC) of metal compound for growth of B. subtilis subsp. spizizenii B. subtilis subsp. spizizenii did not show affinity to form biofilms before (visible to the naked eye) or after addition of metal solutions at any concentration (w/V). The limitation of this study might be that the optical density of each microplate was not measured using ELISA reader. 4. Discussion In order to test the antibiotic sensitivity and growth, for this particular experiment, different metals had been taken to test this effect. In this particular experiment, one of the metals that had been used was zinc sulfate, a specific solid that can have a colorless crystalline structure. In a historical approach, it is known that zinc could be found in soil where different plants are harvested, but in different areas there is something known as solid deficiency, where plants cannot develop properly and grow because of the lack of zinc. And in order for this to be corrected, people have experimented and found out that in order to correct this deficiency, zinc sulfate can be added to the soil in order to have the proper growth of different crops. Because these metals are used in order to grow crops, this may have a different effect when the crops are consumed as a food source. [23, 24, 25]. Copper (II) sulfate pentahydrate is most commonly described as an inorganic compound that could be found in copper in the form of salt. It is highly soluble in water. This type of salt has a usage as an additive in order to recover pentose sugars from the fronts of palm oils. It had been used as well to prove specific antimicrobial properties when working with specific types of bacteria, but most importantly here with Bacillus subtilis [26, 27]. Copper (II) sulfate is used as fungicide in agriculture, as an additive for fertilizers and food [28]. �Journal of Natural Sciences and Engineering, Vol. 1, (2019) DOI number: 10.14706/JONSAE2019114 Iron (III) nitrate, or in other words ferric nitrate, is a type of metal that can be used in many fields. This type of compound can be used to treat different sludges and wastewaters, it can be used to remove nitrogen from different plants and it can also be used in analytical chemistry [29, 30]. All three of metals aforementioned, zinc sulfate, copper (II) sulfate pentahydrate, and iron (III) nitrate, can be found in soil or are used in agriculture. In that way they can get into the food chain and enter the human gut. One of the most abundant minerals that are important for different metabolic processes in the human body is magnesium. It can be found in over 300 enzymes as a cofactor and it regulates different biochemical reactions that are processed in the human body. Usually, magnesium is provided as a type of dietary supplement, people consume it in order for their body to function properly, and different amounts of these minerals are given to people based on various factors [31]. For example, magnesium citrate helps with constipation, it acts as laxative, while magnesium aspartate is important for digestion of macronutrients [32]. B. subtilis subsp. spizizenii showed visible growth at 0.06% (w/V) magnesium solution (Table 3). Dietary supplement was used as a source of magnesium. No significant changes were recorded in antibiotic susceptibility tests in presence of Mg, except with amoxicillin. Addition of Mg solution slightly changed property of B. subtilis subsp. spizizenii. According to obtained results amoxicillin was partially bactericidal (a few colonies appeared within the inhibition zone) for tested bateria, in the presence of magnesium. Susceptibility to fifteen types of antibiotics (Table 1), before and after addition of metal solutions, will be performed using Kirby-Bauer disk diffusion method. This test showed that B. subtilis subsp. spizizenii is completely resistant to ampicillin (AMP2), as well as to ceftazidime+clavulanic acid (CAL40). Antibiotic sensitivity of B. subtilis subsp. spizizenii did not significantly change for the following antibiotics: gentamicin (CN10), streptomicin (S10), vancomycin (VA30), azithromycin (AZM15), kanamycin (K30), cefoxitin (FOX30), oxacillin (OX1), ciprofloxacin (CIP5). Change in diameter was less or equal to 3 mm. Note that diameters were measured manually, and manual errors (gross errors) should be taken into account. Difference in diameter of zone inhibition of erythromycin (E15) with addition of zinc sulfate heptahydrate and without metal solution is 7 mm. There was an increase in diameter size of the inhibition zone for tetracycline (TE30) and amoxicillin (AML10) in presence of magnesium, zinc sulfate heptahydrate, and iron (III) nitrate solutions, compared to diameters of inhibition zones before addition of metal compounds. Besides that, a few colonies of bacteria were observed within amoxicillin zones of inhibition. Amoxicillin was partially bactericidal for B. subtilis subsp. spizizenii, in presence of magnesium, zinc sulfate heptahydrate, and iron (III) nitrate solutions. �Journal of Natural Sciences and Engineering, Vol. 1, (2019) DOI number: 10.14706/JONSAE2019114 B. subtilis subsp. spizizenii without presence of metal solutions was resistant to ceftazidime (CAZ10). With addition of zinc sulfate heptahydrate, and iron (III) nitrate solutions, zones of inhibition were 6 and 8, respectively. Since B. subtilis subsp. spizizenii shares the biochemical similarities with Bacillus subtilis subsp. subtilis results for these two strains can be compared. There is up to 58 to 68% is the DNA relatedness between these two bacteria [10, 33, 34]. According to Silman et al. vancomycin showed great bactericidal effect for B. subtilis in general [35]. Our data shows that zones of inhibition obtained by vancomycin (VA30) are ~20 mm, while the largest zones of inhibition were recorded in presence of ciprofloxacin (CIP5) ranging from 32-35 mm in diameter (Figure 1). Sim et al. obtained similar results about CIP5 and TE30, where zones of inhibition were 32 and 31, respectively [36]. Bacterial growth was registered for all four metal compounds at different concentrations (Table 3). No bacterial growth was registered for Mg at the concentrations 0.1%, 0.5%, 0.25%, 0.12%, while bacterial growth occured at all other tested w/v solutions (Table 3). B. subtilis spizizenii growth occurred at all other w/v solutions of iron (III) nitrate except at the concentrations 0.1% and 0.5%. Growth of bacteria in the presence of copper (II) sulfate w/v solution occurred at concentrations 0.03-0.0009%. The lowest growth rate was observed in the presence of zinc sulfate heptahydrate solution, bacterial growth occurred only on concentrations 0.015-0.0009% (Table 3). The lowest concentration of chemical (drug, antimicrobial) that inhibits visible growth of microorganism (in this case bacteria) in overnight culture is known as minimum inhibitory concentration (MIC) [37]. After overnight incubation at 37 C MICs were recorded (Figure 2). Obtained MICs of metal solutions that inhibit growth of B. subtilis subsp. spizizenii are: magnesium 0.12%, iron (III) nitrate 0.50%, 0.06% copper (II) sulfate, 0.03% zinc sulphate heptahydrate. Considering that, growth of tested bacteria is slightly inhibited by iron (III) nitrate solution (bacteria is growing in presence of metal solution whose concentration is <0.50%), while it is tolerating much lower concentrations of zinc sulfate heptahydrate solution (<0.03%). For this experiment laboratory strain of B. subtilis subsp. spizizenii was used. This strain did not form biofilms at all. According to other studies, during domestication of laboratory strains of B. subtilis accumulation of mutation can occur which can lead to their inability to form well-structured biofilms. Compared to the laboratory strains, undomesticated strains of B. subtilis usually form rich and strong biofilms [38, 39]. 5. Conclusion B. subtilis complex is normally found in soil, however it is also found in the human gut as harmless bacteria. Further research is needed for its wider application on the probiotic market due to safety concerns. Metal traces can be found in soil, wastewaters, products used in agriculture, fungicides, etc. as �Journal of Natural Sciences and Engineering, Vol. 1, (2019) DOI number: 10.14706/JONSAE2019114 such they can easily enter our food chain and end up in the human gut. This study investigated how specific metal compounds influence growth, antibiotic susceptibility, and biofilm forming capacity of B. subtilis subsp. spizizenii. Based on the results that have been retrieved, we can conclude that higher concentrations of metal solutions inhibited growth of tested bacteria, while it showed good tolerance to majority of lower concentrations of metals. Generally, culture showed increased sensitivity against antibiotics after addition of metal solutions. B. subtilis subsp spizizenii used in this experiment was laboratory strain and was not able to form biofilms. No influence of metals was recorded there. 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Determination of minimum inhibitory concentrations. Journal of antimicrobial Chemotherapy, 48(suppl_1), 5-16. [38] Bate, A. R., Bonneau, R., & Eichenberger, P. (2016). Bacillus subtilis systems biology: applications of‐omics techniques to the study of endospore formation. The Bacterial Spore: from Molecules to Systems, 129-144. [39] McLoon, A. L., Guttenplan, S. B., Kearns, D. B., Kolter, R., & Losick, R. (2011). Tracing the domestication of a biofilm-forming bacterium. Journal of bacteriology, 193(8), 2027-2034. � Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Journal of Natural Sciences and Engineering Identifier An unambiguous reference to the resource within a given context 2637-2835 DOI Digital object identifier 10.14706 Publisher An entity responsible for making the resource available International Burch University Description An account of the resource Journal of Natural Sciences and Engineering (JONSAE) is a peer-reviewed, biannually published international journal focusing on empirical and theoretical research in all branches of Engineering and Natural Sciences. It is published on the behalf of Faculty of Engineering and Natural Sciences of International Burch University and aims to provide the best content regarding by publishing original research papers, review articles, special issues, feature articles, and book reviews. All manuscript submissions are subject to initial appraisal by the Editor, and, if found suitable for further consideration, to peer review by independent, anonymous referees. All peer review is double-blind and submission is online. The journal welcomes theoretical, applied, interdisciplinary and methodological work, with preference on empirical research, critical approach and problem-solving methods in manuscripts. Language A language of the resource English Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Effect of metals on antibiotic sensitivity, growth, and biofilm-forming capacity of B. subtilis subsp. spizizenii Author Author Selma Cifric Abstract A summary of the resource. B. subtilis is normally considered a soil organism, it can be also found in the animal and human gastrointestinal tract. Bacillus subtilis subsp. spizizenii is a type of Bacillus subtilis complex. It shares up to 99% of homology with B. subtilis CU1, which can be represented as a probiotic strain. Metal compounds found in soil or used in agriculture can easily enter the food chain and end up in our gut. Gram-positive bacteria (e.g. Bacillus spp.) have good adsorptive capacity for metals due to high peptidoglycan and teichoic acid content in cell walls. There is some evidence that certain metals inside the intestine play an important role in influencing growth and functionality of specific probiotic strains. Some of them have inhibitory, while others have an activating effect on bacteria. This study revealed that metal compounds increased antibiotic susceptibility of B. subtilis subsp. spizizenii. Higher concentrations of metal solutions inhibited growth of tested bacteria. Culture did not show affinity to form biofilms before or after addition of metal solutions Keywords Keywords. antibiotic susceptibility, biofilms, MIC, metals Identifier An unambiguous reference to the resource within a given context 2637-2835 DOI Digital object identifier 10.14706/JONSAE2021323 https://omeka.ibu.edu.ba/files/original/3779a878e77391b47785b5e50a894e24.pdf 28033a8277dae49558f6c4b5dd1aa64e PDF Text Text Effect of Organic and Inorganic Manganese Supplementation in Diets on Performance and Some Organ Weights of Japanese Quails (Coturnix coturnix japonica) Yusuf CUFADAR Selçuk University, Agricultural Faculty, Department of Animal Science, Konya-TURKEY ycufadar@selcuk.edu.tr Osman OLGUN Selçuk University, Agricultural Faculty, Department of Animal Science, Konya-TURKEY oolgun@selcuk.edu.tr Alp Önder YILDIZ Selçuk University, Agricultural Faculty, Department of Animal Science, Konya-TURKEY aoyildiz@selcuk.edu.tr Abstract: This study was carried out to determine the effects of diets containing different levels of inorganic and organic manganese sources on fattening performance and some organ weights of Japanese quails. In this study, 256 one day old quail chicks were fed four levels of inorganic and organic Mn in factorial arrangement design for 35 days. The dietary treatments consisted of the supplementation of the basal diet with 30, 60, 90 and 120 mg/kg Mn supplied from manganese sulphate and manganese bioplex. Dietary Mn sources as a main factor did not effect investigated parameters. But dietary Mn levels had significant effect on all parameters except for feed intake. Supplementation of 60 mg/kg Mn to diets resulted in an increase BW, BWG, liver and pancreas weights, also improved feed conversion ratio. Keywords: Manganese, performance, Quail Introduction Manganese (Mn) is essential for normal bone formation, enzyme function, and amino acid metabolism in poultry (Scott et al. 1976). The utilization of Mn has become an increasing concern because of extremely rapid growth rate of commercial broiler strains (Ji et al. 2006). National Research Council (NRC, 1994) recommended 60 ppm Mn in broiler and quail chicks diet. However, under practical conditions the diets are formulated to contain higher concentrations of Mn to overcome the possibility of its deficiency. This is because of relatively low absorption of dietary Mn in birds (Sunder et al. 2006). Organic Mn sources were more bioavailable than inorganic sources. One of the possible reasons is that there are less chelating or other unwanted reactions with dietary constituent in the gastrointestinal tract for organic mineral complexes compared with those for inorganic minerals (Yan and Waldroup 2006). Diet supplementation with trace elements of high bioavailability, also known as specific amino acid metal compounds of Mn and Zinc which bind themselves to a specific amino acid show a capacity to increase the immune response and diminish negative effects in turkey and laying hens in respect of these minerals inorganic forms (Menocal et al. 2004). However, according to some research results, the organic and inorganic Mn sources reported no difference in terms of bioavailability (Baker and Holpin 1987, Scheideler 1991). The aim of the this study to determine the effect of diets containing different levels of inorganic and organic manganese sources on fattening performance and some organ weights of Japanese quails. Materials and Methods A 5-wk experiment, 256 mixed sex day-old quail chicks (Coturnix coturnix japonica) were used. Four replicate groups of 8 chicks were assigned to each of 8 dietary treatments. The dietary treatments consisted of the supplementation of the basal diet with 30, 60, 90 and 120 mg/kg Mn supplied from manganese sulphate (MnSO4) and manganese bioplex. Dietary treatments were prepared from a corn-soybean common diet without additional Mn contained 21.52 mg/kg. All birds received feed and water ad-libitum. Lighting was treated as a 23 489 �h/day. Compositions of nutrients in the diets were adjusted according to the recommendation of NRC (1994; Table 1). In quails, body weights (BW) and feed intake (FI) were recorded on a pen basis as weekly intervals. Mortality was recorded daily. At the end of the experiment (at five weeks of age), four quails that randomly selected were slaughtered at a processing plant from each replicate and processed, and then the carcass yield were calculated to used warm carcass weight. A general linear model (GLM) was used for the analysis of variance of the data (Minitab 2000). Significant differences among means were tested by Duncan’s multiple range tests. Differences were considered as significant when P values were less than 0.05 (Duncan 1980). Nutrients Corn Soybean meal ( % 47.6 CP)* Vegetable oil (7800 kcal/kg ME)* Limestone Dicalcium phosphate Salt Vitamin Premix1 Mineral Premix2 Methionine TOTAL Calculated nutrients Energy, kcal/kg ME CP, % Calcium, % Available phosphorus, % Lysine, % Methionine, % Methionine + Cysteine, % Crude cellulose, % Manganese, mg/kg* % 53.1 41.3 2.8 1.26 0.8 0.3 0.15 0.10 0.19 100.00 2901 24.06 0.80 0.31 1.32 0.51 0.95 2.25 21.52 * Analyzed value. CP: Crude protein, ME: Metabolizable energy 1 Vitamin premix (supplied the following per kg of diet): Vitamin A, 12000 I.U; Vitamin D3, 2400 I.U; Vitamin E, 25.0mg; Vitamin K3, 4.0 mg; Vitamin B1, 3.0 mg; Vitamin B2, 5.0 mg; Vitamin B6, 8.0 mg; Vitamin B12, 0.015 mg; Niacin, 25.0 mg; Calcium-D-Pantothenate, 8.0 mg, D-Biotin, 0.05 mg; Folic acid, 0.5 mg; Choline Chloride, 125.0 mg. 2 Mineral premix (supplied the following per kg of diet): Fe, 60.0 mg; Zn, 60.0 mg; Cu, 5.0 mg; I, 1.0 mg; Co, 0.2 mg; Se, 0.15 mg. Table 1. Composition of basal diet used in experiment (%) Results and Discussion The effects of diets containing different sources and levels of Mn on performances are shown in Table 2 and 3. The treatments as the main sources of inorganic and organic Mn were not significantly effect on BW, body weight gain (BWG), FI, feed conversion ratio (FCR), carcass yield, liver and pancreas weight (P> 0.05). The diets containing different levels of Mn had significantly effect on all parameters of quails except for feed consumption (P< 0.05). The best results of performance parameters, liver and pancreas percentage of BW were obtaining in quails fed with diet containing 60 mg/ kg Mn, but the lowest results of carcass yield obtaining the same diet. The interactions groups in the experiment, the diets containing different sources and levels of Mn had significantly effect on BW and BWG of quails (P< 0.05). The highest results of BW and BWG were obtaining fed with diet containing MnSO4 x 60 mg/ kg Mn. Quail studies on this subject with a limited number of studies but the results are in broilers. Quails and broilers are similar in terms of requirements of Mn (NRC 1994). The results of the experiment, supplemental Mn sources (inorganic and organic) there were no differences. The similar result, Berta et al. (2004) reported that the same level of supplementation of the two Mn sources there were no differences between the Mn concentrations of organs and tissue in broiler chicks. Additionally, these researchers stated that a corn-soybean diet supplemental with levels of 0, 30, 60 and 240 mg/kg Mn from organic and inorganic sources did not significant effect on the BW, FCR in broiler chicks. Collins and Moran (1999) reported that body weight and feed 490 �efficiency were not influenced by supplementary Mn (180 ppm). Also, supplemental Mn did not alter processed carcass weights, yield, or percentage abdominal fat in broilers. Gajula et al. (2010) stated that Mn (60 ppm) as recommended by NRC (1994) was sufficient for broiler performance and bone parameters. The results of this study with the contradiction between the results of previously conducted studies may be due to different Mn levels and animal material. It is concluded that, 60 mg/kg supplementation Mn to diet is suitable in growing Japanese quails. The number of research interest in this subject is very limited. Therefore, many studies are needed. Diets Sources MnSO4 Mn Bioplex Mn levels, mg/kg 30 60 90 120 Sources x levels MnSO4 x 30 MnSO4 x 60 MnSO4 x 90 MnSO4 x 120 Mn Bioplex x 30 Mn Bioplex x 60 Mn Bioplex x 90 Mn Bioplex x 120 A-D BW, g/bird BWG, g/bird FI, g/bird FCR, Feed/ Gain 169.3±2.75 170.1±1.36 161.0±2.39 161.9±1.36 524.3±06.41 522.3±05.54 3.26±0.042 3.23±0.032 164.4±1.84B 179.5±3.20A 166.9±1.87B 167.9±1.98B 156.2±1.79B 169.3±2.33A 158.8±1.86B 161.4±2.55B 519.5±06.39 520.5±07.62 519.1±11.57 534.0±07.32 3.33±0.043A 3.08±0.040B 3.27±0.055A 3.31±0.045A 160.3±0.73C 185.9±3.71A 163.9±2.36BC 167.3±1.15BC 168.6±1.96BC 173.2±2.63B 170.0±2.17BC 168.5±4.10BC 152.2±0.68D 173.7±2.45A 155.9±2.37CD 162.4±3.58BC 160.3±1.84BCD 165.0±2.63B 161.8±2.14BC 160.4±4.12BCD 515.7±08.77 532.2±13.16 516.6±16.38 532.9±14.57 523.4±10.35 508.9±10.29 521.6±18.77 535.2±06.10 3.39±0.041 3.06±0.063 3.31±0.080 3.28±0.058 3.27±0.067 3.09±0.058 3.22±0.079 3.34±0.074 : Means within a column with unlike superscript differ significantly (P< 0.05). Table 2. Effect of the experimental diets on performance of Japanese quails Diets Sources MnSO4 Mn Bioplex Mn levels, mg/kg 30 60 90 120 Sources x levels MnSO4 x 30 MnSO4 x 60 MnSO4 x 90 MnSO4 x 120 Mn Bioplex x 30 Mn Bioplex x 60 Mn Bioplex x 90 Mn Bioplex x 120 A-B Carcass yield, % of BW Liver, % of BW Pancreas, % of BW 63.25±0.41 62.64±0.23 2.05±0.087 2.02±0.103 0.24±0.019 0.25±0.009 64.02±0.37A 61.79±0.37B 63.28±0.50A 62.70±0.33AB 1.98±0.085AB 2.32±0.104A 2.01±0.070AB 1.82±0.040B 0.23±0.009B 0.29±0.012A 0.24±0.009B 0.23±0.009B 64.86±0.20 61.26±0.56 63.58±0.77 63.30±0.44 63.18±0.34 62.31±0.37 62.97±0.70 62.10±0.27 2.00±0.175 2.35±0.218 2.07±0.111 1.78±0.062 1.97±0.055 2.30±0.048 1.95±0.091 1.86±0.048 0.22±0.005 0.29±0.010 0.22±0.006 0.24±0.019 0.25±0.016 0.28±0.023 0.26±0.016 0.23±0.009 : Means within a column with unlike superscript differ significantly (P< 0.05). Table 3. Effect of the experimental diets on some organ weights of Japanese quails Acknowledgments This research was funded in part by a grant from the University of Selcuk (BAP) 491 �References Baker. D. H. and Hoplin, K. M. (1987). Efficacy of a manganese-protein chelate compared with that of manganese sulfate for chicks. Poult. Sci. 66:1561- 1563. Berta, E., Andrasofszky, E., Bersenyi, A., Glavits, R., Gaspardy, A. And Fekete, Gy. (2004). Effect of inorganic and organic manganese supplementation on the performance and tissue manganese content of broiler chicks. Acta Veterinaria Hungarica, 52 (29), pp. 199-209. Collins, N.E. and Moran, JR, E.T. (1999). Influence of supplemental manganese and zinc on live performance and carcass quality of diverse broiler strains. J. Appl. Poultry Res., 8, pp. 228-235. Duncan D.B. (1980). Multiple Range and Multiple F-tests. Biometrics. Gajula, S.S., Chalasani, V.K., Panda, A.K., Mantena, V. L. N. R and Savaram, R. R. (2010). Effect of supplemental inorganic Zn and Mn and their interactions on the performance of broiler chicken, mineral bioavailability and immune response. Biological Trace Element Research, doi: 10.1007/s12011-010-8647-8. Ji, F., Luo, X.G., Lu, L., Liu, B. and Yu, S.X. (2006). Effect of manganese source on manganese absorption by the intestine of broilers. Poultry Science, 85, pp, 1947-1952. Menocal, J.A., Gonzales, E.A., Coello, C.L., Fakler, TM., Rapp, C.J., Ward, T.L. and Vela, G. (2004). Use of zincmethionine and manganese-methionine in broilers diets:productive parameters and ascites syndrome incidence. Tec.Pecu Mex., 42 (1), pp, 113-119. Minitab (2000). Minitab Reference Manuel (release 13.0). Minitab Inc. State Coll.. P.A.. USA. National Research Council (NRC). (1994). Nutrient Requirements of Poultry. 9th ed. National Academy Press. Washington. DC. Scheideler, S.E. (1991) Interaction of dietary calcium, manganese and manganese source (manganese oxide or manganese methionine chelate) on chick performance and manganese utilization. Biological Trace Element Research. 29, pp. 217-223. Scott, M.L., Nesheim, M.C. and Young, R.J. (1976). Nutrition of the chicken. M.L. Ithaca, NY. Sunder, G.S., Panda, A.K., Gopinalth, N. C.S., Mantena, V.L.N., Savaram, R. R. and Chalasani, V.K. (2006). Effect of supplemental manganese on mineral uptake by tissue and immune response in broiler chickens. The Journal of Poultry Science, 43, pp, 371-377. Yan, F. and Waldroup, P.W. (2006). Evaluation of Mintrex manganese as a source of manganese for young broilers. International journal of Poultry Science, 5 (8), pp.708-713. 492 � Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Extent The size or duration of the resource. 570 Title A name given to the resource Effect of Organic and Inorganic Manganese Supplementation in Diets on Performance and Some Organ Weights of Japanese Quails (Coturnix coturnix japonica) Author Author CUFADAR, Yusuf OLGUN, Osman YILDIZ, Alp Önder Abstract A summary of the resource. This study was carried out to determine the effects of diets containing different levels of inorganic and organic manganese sources on fattening performance and some organ weights of Japanese quails. In this study, 256 one day old quail chicks were fed four levels of inorganic and organic Mn in factorial arrangement design for 35 days. The dietary treatments consisted of the supplementation of the basal diet with 30, 60, 90 and 120 mg/kg Mn supplied from manganese sulphate and manganese bioplex. Dietary Mn sources as a main factor did not effect investigated parameters. But dietary Mn levels had significant effect on all parameters except for feed intake. Supplementation of 60 mg/kg Mn to diets resulted in an increase BW, BWG, liver and pancreas weights, also improved feed conversion ratio. Keywords: Manganese, performance, Quail Date A point or period of time associated with an event in the lifecycle of the resource 2010-06 Keywords Keywords. Conference or Workshop Item PeerReviewed Q Science (General) https://omeka.ibu.edu.ba/files/original/dfab893bf48dd5ccaf8b8a2b56571fdd.pdf f99e51ded30f232896b6d68ef36e5041 PDF Text Text EFFECT OF SALICYLIC ACID ON SALINITY STRESS IN COWPEA (Vigna unguiculata L. Walp) Hüsnü ÜNLÜ Süleyman Demirel University, Faculty of Agriculture, Department of Horticulture, Isparta,TURKEY Nüket ALTINDAL Süleyman Demirel University, Faculty of Agriculture, Department of Field Crops, Isparta, TURKEY Halime ÖZDAMAR ÜNLÜ Süleyman Demirel University, Faculty of Agriculture, Department of Horticulture, Isparta,TURKEY hozdamar79@yahoo.com Demet ALTINDAL Süleyman Demirel University, Faculty of Agriculture, Department of Field Crops, Isparta, TURKEY Hüseyin PADEM International Burch University, Sarajevo, BOSNIA-HERZEGOVINA hpadem@ibu.edu.ba Abstract:This study was conducted to determine the effects of seed soaking in salicylic acid (0.25 mM, 0.50 mM, 0.75 mM, 1.0 mM and control) on the growth and some seedling properties in cowpea (Vigna unguiculata L. Walp) under different NaCl doses (2.5 ds m-1, 5.0 ds m-1, 7.5 ds m-1, 10.0 ds m-1 and control). In this work; seedling length, cotyledon width, cotyledon length, fresh-dry weight, chlorophyll a, chlorophyll b and total chlorophyll were investigated. In result, according to NaCl x SA interaction, seedling length, dry weight, total chlorophyll values changed between 5.05-13.58 cm, 0.055-0.138 g, 1.705-3.690 mg/g fresh weight, respectively. Keywords: Cowpea (Vigna unguiculata L. Walp), salinity, salicylic acid, seedling properties Introduction Cowpea, Vigna unguiculata belongs to the group of crops known as legumes. It is a important source of human food. Its vegetative part can be buried into the soil to improve its fertility, and also, cowpea is a well known leguminous cover crop (Onyesom et al. 2005). Cowpea has a good tolerance to salinity, heat, drought, and it has a high yield potential under irrigation (Murillo-Amador & Troyo-Diéguez 2000). Soil salinity has become an environmental problem and it adversely affects the growth and productivity of many crops. Salinity affects the crop physiology, and causes changes within cellular processes and plant levels (Yusuf et al. 2008). High salt concentrations have toxic effects on plant growth. Soil salinity level decreases availability of nutrients to plants and create high osmotic pressure which leads to physiological drought (Endris & Mohammed 2007). Soil salinity also affects seed germination and limit the agricultural production (Dantas et al. 2005). Salinity creates abiotic stress conditions for plants such as ionic and osmotic stress and these stress conditions cause production of reactive oxygen species and oxidative damage, metabolic toxicity and membrane deterioration. As a result plants developed complex responses to overcome harmful effects of salinity (Szalai & Janda 2009). Salicylic acid (SA) is a plant phenolic compound and now considered as a hormone-like endogenous regulator. It play important roles to defend plants against both biotic and abiotic stress conditions. Earlier reports show that SA plays important regulatory roles in plants against to a wide range of oxidative stresses (Choudhury & Panda 2004; Deef 2007). 61 �The aim of present work was to reveal whether cowpea plants pre-treated with different concentrations of SA could tolerant salt stress. Materials and Methods Cowpea seeds (Vigna unguiculata L. Karnıkara cv.) were used in this study. The experimental design consisted of 25 treatments replicated 3 times in a split plot design. Seeds of cowpea were surface-sterilized for 5 minute in sodium hypochlorite solution. They were then washed 3 times with sterilized distilled water. After sterilization, seeds were soaked in 0, 0.25, 0.5, 0.75 and 1 mM SA for 12 h in dark. The seeds were sown in plastic pots containing vermiculite and torf (1:2), maintained the same environmental condition and watered with different NaCl doses (0, 2.5, 5, 7.5 and 10 ds/mol) in total 50 ml during 10 days. After 10 days, the following were recorded: seedling length (cm), cotyledon width and length (cm), fresh and dry weight (g), chlorophyll a (mg/g fresh weight), chlorophyll b (mg/g fresh weight) and total chlorophyll (mg/g fresh weight). Chlorophyll concentration was determined from cotyledons. A leaf sample of 0.2 g was ground and extracted with 15 ml of 80% (v/v) acetone in the dark (Haklı 2008). The mixture was filtered and absorbancies (T80 UV/VIS, Spectrophotometer) were determined at 645 and 663 nm. Concentration of chlorophyll a, chlorophyll b and total chlorophyll were estimated by the equations of (Witham et al. 1971). All of the data were analyzed statistically using CoStat computer program. Duncan’s test was used to compare averages at the 0.05 significance level. Results NaCl (P<0.01) and SA (P<0.05) applications were found statistically important on seedling length. According to the salt concentration, seedling length was changed between 6.21-10.75 cm. The highest seedling length was obtained from 0 ds/mol (untreated control) of salt concentration and the lowest was obtained from 10.0 ds/mol NaCl dose. Under SA applications, seedling length was changed between 6.88-9.43 cm and the highest seedling length was determined in 0.50 mM SA application. NaCl and SA interaction showed that seedling length changed between 5.05-13.58 cm (Table 1). When Table 2 was investigated, NaCl and SA applications on cotyledon width were statistically important at the level 1%. According to NaCl and SA applications, cotyledon width varied between 2.34-3.82 cm and 2.513.49 cm, respectively. While cotyledon length values which were found statistically significant (P<0.01) changed between 3.205.77 cm in NaCl treatments, these values changed between 3.79-5.18 cm in SA treatments (Table 3). When the values of fresh and dry weight of seedlings were investigated, NaCl, SA applications and their interactions were found statistically significant. The 10 ds/mol dose of NaCl gave the lowest value of dry and fresh weight of seedlings. The highest values of fresh and dry weight were determined in 0.50 mM SA treatment (Table 4, 5). The effects of SA, NaCl x SA applications interactions on chlorophyll a were statistically significant at level 1%. Chlorophyll b values were statistically significant at level 5% under NaCl x SA applications interactions. According to NaCl, SA applications and their interactions, chlorophyll a values were changed between 2.075-2.297 mg/g fresh weight, 1.930-2.271 mg/g fresh weight and 1.006-2.500 mg/g fresh weight, respectively. Chlorophyll b values changed; between 0.836-0.995 mg/g fresh weight under NaCl treatment, between 0.817-0.981 mg/g fresh weight in SA treatments and between 0.641-1.903 mg/g fresh weight in NaCl x SA treatments interactions (Table 6,7). SA applications (P<0.05) and NaCl x SA applications interactions (P<0.01) were found statistically significant on total chlorophyll. It was determined that total chlorophyll values changed between 2.990-3.213 mg/g fresh weight in NaCl treatments and between 2.747-3.240 in SA treatments. According to NaCl x SA interactions total chlorophyll values varied between 1.705-3.690 mg/g fresh weight (Table 8). 62 �Discussion As NaCl concentrations were increased, a decrease in cowpea seedling length, cotyledon width, cotyledon length, fresh weight and dry weight (except for 2.5 ds/mol) was obtained. This situation can be explained by that salt stress unfavorably affected plant growth and productivity during all developmental stages (Tavili & Biniaz 2009). For example Hussein et al. (2007) reported that in general salinity decreases plant development. The same results were obtained by Okçu et al. (2005) who used pea, Kaya & Day (2008) who used sunflower. The highest seedling length, cotyledon width, cotyledon length, fresh weight and dry weight were obtained from 0.5 mM SA concentration. A correlation was observed between increases in the concentration of these SA (0.75 and 1.0 mM) and inhibition of these parameters. Salicylic acid may influence a range of different processes in plants, including seed germination, ion uptake and transport, and membrane permeability (Dolatabadian et al. 2009). These results are consistent with those of Çanakçı & Munzuroğlu (2007) who showed that SA application induced fresh weight and chlorophyll content. Chlorophyll a, chlorophyll b and total chlorophyll contents increased in SA treatments according to untreated control. The greatest chlorophyll (a, b, a+b) values were found from 7.5 ds/mol dose of NaCl x 1 mM dose of SA interaction. This case shows that SA treatments decreases the negative effects of salt. Chlorophyll content of cotyledons increased slightly in general under higher NaCl concentration (7.5-10.0 ds/mol) although their increases were not statistically significant. According to Tohma (2007), Çanakçı & Munzuroğlu (2007), Kuşvuran et al. (2008) the effect of NaCl concentrations on chlorophyll content was found statistically significant. This situation can be explained by differences in species, varieties, medium and concentration of NaCl. NaCl treatments have a negative effect on seedling length, cotyledon width, cotyledon length, fresh weight and dry weight and data were found statistically significant (P<0.01). In terms of all parameters were examined, 0.5 mM SA treatment reduce saline medium negative effects. This case show the necessity of studies about the application of SA in saline soil. Table 1. Effect of SA on NaCl stress in cowpea seedling length (cm) NaCl Doses (ds/mol) 0 2.5 5.0 7.5 10.0 SA Means 0 13.58 9.02 8.82 9.02 5.97 9.28 a* Salicylic Acid Doses (mM) 0.25 0.50 0.75 9.97 11.03 10.93 9.95 10.32 9.93 8.49 10.68 8.80 9.05 8.82 7.37 6.75 6.32 6.83 8.84 a 9.43 a 8.77 a 1.0 8.25 9.70 6.25 5.05 5.17 6.88 b NaCl Means 10.75 a** 9.78 ab 8.61 bc 7.86 c 6.21 d Significance Levels : *P<0.05, **P<0.01 Table 2. Effect of SA on NaCl stress in cowpea cotyledon width (cm) NaCl Doses (ds/mol) 0 2.5 5.0 7.5 10.0 SA Means Salicylic Acid Doses (mM) 0 3.73 4.00 3.22 3.18 2.25 3.28 a** 0.25 3.70 4.02 2.65 3.37 2.62 3.27 a 0.50 3.83 4.00 3.93 3.20 2.47 3.49 a 0.75 4.00 3.62 3.47 2.75 2.20 3.21 a 1.0 3.33 3.48 1.90 1.63 2.18 2.51 b NaCl Means 3.72 a** 3.82 a 3.03 b 2.83 b 2.34 c Significance Levels : **P<0.01 Table 3. Effect of SA on NaCl stress in cowpea cotyledon length (cm) NaCl Doses (ds/mol) 0 2.5 5.0 7.5 10.0 SA Means Significance Levels : **P<0.01 63 0 5.37 5.85 4.83 4.95 3.08 4.82 a** Salicylic Acid Doses (mM) 0.25 0.50 0.75 5.63 5.73 6.25 6.12 5.98 5.57 4.47 5.98 5.23 4.93 4.82 4.23 3.47 3.37 3.08 4.92 a 5.18 a 4.87 a 1.0 5.05 5.32 3.05 2.55 3.00 3.79 b NaCl Means 5.61 a** 5.77 a 4.71 b 4.30 b 3.20 c �Table 4. Effect of SA on NaCl stress in cowpea fresh weight (g) NaCl Doses (ds/mol) 0 2.5 5.0 7.5 10.0 SA Means 0 1.614 abc 1.563 abc 1.390 bcd 1.437 bcd 0.800 fg 1.361 a** Salicylic Acid Doses (mM) 0.25 0.50 0.75 1.555 abc 1.789 ab 1.972 a 1.747 abc 1.781 ab 1.626 abc 1.384 bcd 1.841 ab 1.471 bcd 1.472 bcd 1.306 cde 1.102 def 0.906 efg 0.804 fg 0.770 fg 1.413 a 1.504 a 1.388 a 1.0 1.614 abc 1.821 ab 0.849 fg 0.613 g 0.619 g 1.103 b NaCl Means 1.709 a** 1.708 a 1.387 b 1.186 c 0.780 d Significance Levels : **P<0.01 Table 5. Effect of SA on NaCl stress in cowpea dry weight (g) NaCl Doses (ds/mol) 0 2.5 5.0 7.5 10.0 SA Means 0 0.109 a-d* 0.117 abc 0.107 a-e 0.112 abc 0.074 d-g 0.104 a** 0.25 0.109 a-d 0.122 abc 0.105 a-e 0.121 abc 0.088 c-g 0.109 a Salicylic Acid Doses (mM) 0.50 0.75 0.122 abc 0.137 a 0.129 ab 0.123 abc 0.138 a 0.113 abc 0.109 a-d 0.092 b-f 0.072 efg 0.067 fg 0.114 a 0.106 a 1.0 0.115 abc 0.133 a 0.065 fg 0.055 g 0.059 fg 0.085 b NaCl Means 0.118 ab** 0.125 a 0.106 bc 0.098 c 0.072 d Significance Levels : *P<0.05, **P<0.01 Table 6. Effect of SA on NaCl stress in cowpea chlorophyll a (mg/g fresh weight) NaCl Doses (ds/mol) 0 0.25 Salicylic Acid Doses (mM) 0.50 0.75 1.0 0 2.5 5.0 7.5 10.0 SA Means 2.063 a-d** 2.122 a-d 2.229 abc 1.006 e 2.232 abc 1.930 b** 2.289 abc 1.996 a-d 2.102 a-d 2.100 a-d 2.382 ab 2.174 a 2.449 a 2.395 ab 1.634 d 2.479 a 2.334 abc 2.258 a 2.345 abc 2.122 a-d 2.244 abc 2.291 abc 2.353 abc 2.271 a 1.854 cd 1.881 bcd 2.440 a 2.500 a 2.193 abc 2.173 a NaCl Means 2.200 2.103 2.130 2.075 2.297 Significance Levels : **P<0.01 Table 7. Effect of SA on NaCl stress in cowpea chlorophyll b (mg/g fresh weight) NaCl Doses (ds/mol) 0 2.5 5.0 7.5 10.0 SA Means 0 0.789 b-f* 0.893 b-f 0.922 b-f 0.699 def 0.783 b-f 0.817 0.25 0.898 b-f 0.729 def 0.769 c-f 0.907 b-f 0.997 b-f 0.860 Salicylic Acid Doses (mM) 0.50 0.75 1.035 b-e 0.963 b-f 1.101 bc 0.732 c-f 0.670 ef 0.895 b-f 1.152 b 1.028 b-e 0.948 b-f 0.991 b-f 0.981 0.922 1.0 0.641 f 0.724 def 1.045 bcd 1.903 a 0.851 b-f 0.890 NaCl Means 0.865 0.836 0.860 0.995 0.914 Significance Levels : *P<0.05 Table 8. Effect of SA on NaCl stress in cowpea total chlorophyll (mg/g fresh weight) NaCl Doses (ds/mol) 0 2.5 5.0 7.5 10.0 SA Means Salicylic Acid Doses (mM) 0 2.852 a-e** 3.014 a-e 3.150 a-d 1.705 f 3.016 a-e 2.747 b* 0.25 3.187 a-d 2.725 b-e 2.872 a-e 3.007 a-e 3.378 abc 3.034 ab 0.50 3.485 ab 3.495 ab 2.304 ef 3.632 a 3.283 a-d 3.240 a 0.75 3.308 a-d 2.854 a-e 3.138 a-d 3.319 a-d 3.343 abc 3.193 a 1.0 2.495 de 2.605 cde 3.485 ab 3.690 a 3.043 a-e 3.064 ab NaCl Means 3.065 2.939 2.990 3.070 3.213 Significance Levels : *P<0.05, **P<0.01 64 �References Choudhury, S., & Panda, S. K. (2004). Role of salicylic acid in regulating cadmium induced oxidative stress in Oryza sativa L. roots. Bulg. J. Plant Physiol., 30 (3-4), 95-110. Çanakçı, S., & Munzuroğlu, Ö. (2007). Asetilsalisilik asit ’in mısır (Zea mays L.) fidelerinin taze ağırlık değişimi, pigment ve protein miktarları üzerine etkileri. Fırat Üniv. Fen ve Müh. Bil. Dergisi, 19 (3), 259-264. Dantas, B. F., Sá Ribeiro, L., & Aragão, C. A. (2005). Physiological Response of Cowpea Seeds To Salinity Stress. Revista Brasileira de Sementes, 27 (1): 144-148. Deef, H. E. (2007). Influence of salicylic acid on stress tolerance during seed germination of Triticum aestivum and Hordeum vulgare. Advances in Biological Research, 1 (1-2): 40-48. Dolatabadian, A., Modarres Sanavy S. A. M., & Sharifi, M. (2009). Effect of salicylic acid and salt on wheat seed germination. Acta Agriculturae Scandinavica Section B- Soil and Plant Science, 1-9. Endris, S., & Mohammed, M. J. (2007). Nutrient acquisition and yield response of Barley exposed to salt stress under different levels of potassium nutrition. Int. J. Environ. Sci. Tech., 4 (3): 323-330. Haklı, E. (2008). Alternatif sıcaklığın su stresi altındaki mercimek (Lens Culinaris Medik.) çeşitlerinin çimlenme ve bazı fizyolojik parametreleri üzerine etkisi. Çukurova Üniversitesi, Fen Bilimleri Enstitüsü, Biyoloji Anabilim Dalı, Yüksek Lisans Tezi, 1-75, Adana. Hussein, M.M., Balbaa, L. K., & Gaballah, M. S. (2007). Developing a salt tolerant cowpea using alpha tocopherol. Journal of Applied Sciences Research, 3(10): 1234-1239. Kaya, M. D., & Day, S. (2008). Relationship between seed size and NaCl on germination, seed vigor and early seedling growth of sunflower (Helianthus annuus L.). African Journal of Agricultural Research, 3 (11), 787-791. Kuşvuran, Ş., Yaşar, F., Abak, K., & Ellialtıoğlu, Ş. (2008). Tuz stresi altında yetiştirilen tuza tolerant ve duyarlı Cucumis sp.’nin bazı genotiplerinde lipid peroksidasyonu, klorofil ve iyon miktarlarında meydana gelen değişimler. Tarım Bilimleri Dergisi (J. Agric. Sci.), 18(1): 13-20. Murillo-Amador, A, B., & Troyo-Diéguez, E. (2000). Effects of salinity on the germination and seedling characteristics of cowpea [Vigna unguiculata (L.) Walp.]. Australian Journal of Experimental Agriculture, 40, 433–438. Okçu, G., Kaya, M. D., & Atak, M. (2005). Effects of salt and drought stresses on germination and seedling growth of pea (Pisum sativum L.). Turk J Agric For., 29, 237-242. Onyesom, I., Enaholo, A. T., & Mordi, J. (2005). Effect of processing techniques on the contents of flatulence factors and emulsion properties of cowpea (Vigna unguiculata). J. Appl. Sci. Environ. Mgt., 9 (2) 65– 72. Szalai, G., & Janda, T. (2009). Effect of salt stress on the salicylic acid synthesis in young maize (Zea mays L.) plants. J. Agronomy & Crop Science ISSN 0931-2250. Tavili, A., & Biniaz, M. (2009). Different Salts effects on the germination of Hordeum vulgare and Hordeum bulbosum. Pakistan Journal of Nutrition, 8 (1): 63-68. Tohma, Ö. (2007). Çilekte salisilik asit uygulamasının tuz stresine dayanıklılık üzerine etkisi. Atatürk Üniversitesi Fen Bilimleri Enstitüsü, Bahçe Bitkileri Anabilim Dalı, Y. Lisans Tezi, 1-73, Erzurum. Witham, F. H., Blaydes, D. F., & Deulin, R. M. (1971). Experiments in Plant Physiology. Van Nostrand Reinhold Company, Newyork, 245. Yusuf, M., Hasan, S. A., Ali, B., Hayat, S., Fariduddin, Q., & Ahmad, A. (2008). Effect of salicylic acid on salinity-induced changes in Brassica juncea. Journal of Integrative Plant Biology, 50 (9): 1096– 1102. 65 � Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Extent The size or duration of the resource. 651 Title A name given to the resource EFFECT OF SALICYLIC ACID ON SALINITY STRESS IN COWPEA Author Author ÜNLÜ, Hüsnü ALTINDAL, Nüket ÖZDAMAR ÜNLÜ, Halime ALTINDAL, Demet PADEM, Hüseyin Abstract A summary of the resource. This study was conducted to determine the effects of seed soaking in salicylic acid (0.25 mM, 0.50 mM, 0.75 mM, 1.0 mM and control) on the growth and some seedling properties in cowpea (Vigna unguiculata L. Walp) under different NaCl doses (2.5 ds m-1, 5.0 ds m-1, 7.5 ds m-1, 10.0 ds m-1 and control). In this work; seedling length, cotyledon width, cotyledon length, fresh-dry weight, chlorophyll a, chlorophyll b and total chlorophyll were investigated. In result, according to NaCl x SA interaction, seedling length, dry weight, total chlorophyll values changed between 5.05-13.58 cm, 0.055-0.138 g, 1.705-3.690 mg/g fresh weight, respectively. Date A point or period of time associated with an event in the lifecycle of the resource 2009-06 Keywords Keywords. Conference or Workshop Item PeerReviewed Q Science (General) https://omeka.ibu.edu.ba/files/original/ba506bbd231217a22d4fac647aea7a6e.pdf 5ae125e49742a908a886fa6d4cec4da1 PDF Text Text 3rd International Symposium on Sustainable Development, May 31 - June 01 2012, Sarajevo Effect of steel plant pollution on photosynthetic apparatus of some spontaneous plants by chlorophyll fluorescence imaging EriolaZhuri Hida1, VeledinÇako2,FatbardhaBabani3,Theodhor Karaja4 1University ‘AleksanderMoisiu’, Durres, Albania, 2 Department of Physics, University ‘Ismail Qemali’,Vlora, Albania, 3 Biotechnology Department, University of Tirana, Tirana, Albania, 4Physics Department, University of Tirana, Tirana, Albania, E-mails: eri_zhuri@yahoo.com, v.cako69@hotmail.com, fbabani@yahoo.com, theodhor.karaja@yahoo.com Abstract Environmental pollution by the steel plant in Elbasan, the largest metallurgical complex in Albania causes serious problems. Although the blast furnaces and basic oxygen converters closed in 1991, small scale steel production from scrap metal continued. This steel plant releasing chemicals and particulates into the atmosphere is considered a source of air pollution. Industrial siderurgic air pollution as other stressors affects growth and physiological functions of plants. Pollution has effects on the photosynthetic performance of leaves and can modify their optical and fluorescence properties either directly or indirectly. The chlorophyll (Chl) fluorescence provides ample information on the performance of photosynthetic apparatus. Chl fluorescence signatures of leaves as an efficient tool and a nondestructive method for the in vivo analysis of plant stress is applying to describe and investigate the photosynthetic light processes and quantum conversion at physiological conditions as well as to detect stress on the photosynthetic apparatus. The chlorophyll fluorescence images were measured on leaves using the FluorCam 700MF imaging system (Photon Systems Instrument) as a techniques that offer the possibility to study the distribution and patchiness of fluorescence signatures over the whole leaf area. Chl fluorescence images were measured on the leaves of some spontaneous plants grown in sites with different level of steel plant air pollution assessed on base of different distances and different directions from the source of the pollution. Efficiency of photosynthetic apparatus of analyzed plants was evaluated via chlorophyll fluorescence images during induction kinetics and various fluorescence ratio images. The images of Chl fluorescence ratios, acquired by pixel to pixel arithmetic operations performed by FluorCam software, were aplied as indicators of the functional state or the damage of the photosynthetic apparatus. The potential and effective quantum yields of photosynthetic electron transport were estimated too. Observed differences on imaging of chlorophyll fluorescence signature and photosynthetic pigment content of some spontaneous plants allowed characterizing of the photosynthetic performance in order to evaluate the damage by plant steel air pollution. 87 �3rd International Symposium on Sustainable Development, May 31 - June 01 2012, Sarajevo Keywords: chlorophyll fluorescence imaging, induction kinetics, plant steel air pollution, photosynthetic apparatus, potential/effective quantum yield, spontaneous plant. 1.INTRODUCTION Environmental pollution by the steel plant in Elbasan, the largest siderurgical complex in Albania causes serious problems. In this area operate some plants as steel plant operator "Kurum“, cement factory "ECF", Establishment of Ferro-Chrome "ACR“. This steel plant releasing chemicals and particulates into the atmosphere is considered a source of air pollution. Monitored elements in the Elbasan area are: - solids suspended in the air (LNP), rigid particles with dimensions smaller than 10 microns (PM10) - lead (Pb), sulfur dioxide (SO2), nitrogen dioxide (NO2), ozone (O3) in the air. Siderurgic air pollution as other stressors affects growth and physiological functions of plants. Pollution has effects on the photosynthetic performance of leaves and can modify their optical and fluorescence properties either directly or indirectly. The chlorophyll (Chl) fluorescence signatures of plants have been applied as an efficient tool to describe and investigate the photosynthetic light processes and quantum conversion to detect pollution effects on the photosynthetic apparatus. Image analysis of the Chl fluorescence signals offers the possibility to study the function of photosynthetic apparatus and to detect early stress damage in plants by screening the fluorescence emission over the leaf area as well as the gradients and local irregularities in fluorescence emission and ratios (Lichtenthaler 1996, Buschmann and Lichtenthaler, Krause and Weis, 1991; Lichtenthaler and Miehé, 1997; Lichtentaler and Babani, 2004; Schreiber1986). Various parameters and ratios of the Chl fluorescence determined from the induction kinetics (Kautsky effect) can be used as indicators of the functional state or stress damage of the photosynthetic apparatus and photosynthetic electron transport (Babani and Lichtenthaler, 1996; Buschmann and Lichtenthaler, 1998; Govindjee 1995, 2004). Imaging of Chl fluorescence kinetics correctly screening the emission heterogeneity reflects localized biotic or abiotic stress or heterogeneous metabolism. Offering the possibility to study distribution and patchiness of fluorescence signatures over the whole leaf area these techniques were developed as invaluable tool for determining the photosynthetic performance of plants (Buschmann and Lichtenthaler 1998, Lichtenthaler and Babani 2000; Lichtenthaler et al., 2000,Lichtenthaler and Babani 2004, Lichtenthaleret al., 2007, Nebdal et al. 2000, Schreiber 2004). The objective of the presented research is the evaluation of damage by plant steel air pollution on spontaneous plants characterizing the photosynthetic performance by imaging of chlorophyll fluorescence signature. Imaging of Chl fluorescence kinetics correctly screening the emission heterogeneity reflects localized biotic or abiotic stress or heterogeneous metabolism. Offering the possibility to study distribution and patchiness of fluorescence signatures over the whole leaf area these techniques were developed as invaluable tool for determining the photosynthetic performance of plants 88 �3rd International Symposium on Sustainable Development, May 31 - June 01 2012, Sarajevo 2.MATERIALS AND METHODS 2.1. Plant material Endemic-spontaneous plant Cercius siliquastrum grown in different steel plant pollution conditions were analyzed. Study areas were chosen in three different locations: Dajti area (site 1) characterized by optimal physiological conditions, Krrabe area (site 2 2.6 km) and Elbasan area(site3- 6 km)characterized by plant steel air pollution. Chlorophyll fluorescence imaging of induction kinetics Chlorophyll (Chl) fluorescence induction kinetics were measured using the FluorCam 700MF kinetics imaging system constructed by Photon Systems Instrumentto capture kinetics and 2dimensional maps of key fluorescence parameters. The fluorescence emission is induced by two sets of 325 super-bright orange light emitting diodes (LED’s) (wavelength 605nm) that provide excitation flashes or a continuous actinic irradiance controlled by defined protocol. Fluorescence images are captured by CCD camera. The images are taken at 12-bit resolution in 512 x 512 pixels of CCD chip. The size of an analyzed object is smaller than 10  13 cm. The chlorophyll (Chl) fluorescence images and induction kinetics were measured on predarkened leaves (30 min) using the FluorCam quenching protocol. The images of the measured Chl fluorescence intensity were obtained on false colour, whereby black is the lowest (zero) and red the highest fluorescence intensity. Chlorophyll fluorescence images of parameters as F0 and F0’(minimum fluorescence in the dark and in the light-adapted states), Fm and Fm’ (maximum fluorescence in the dark and in the light-adapted states), Fp, (initial fluorescence increase caused by the actinic light exposure) and Fs, (steady-state fluorescence in actinic light exposure) were recorded. Images of various Chl fluorescence ratios obtained by pixel to pixel arithmetic operations performed by FluorCam software were: maximum quantum yields of Photosystem II Fv/Fm and Fm/Fo; effective quantum yields of Photosystem II Fv'/Fm‘andFm’/Fo’; fluorescence decline ratio in steady-state which assess plant vitality Rfd=(Fp-Fs)/Fs; non photochemical quenching coefficients NPQ=(Fm-Fm’)/Fm and qN=(Fv-Fv’)/Fv; where Fv=Fm-Fo and Fv’=Fm’-Fo’. Kinetics of the fluorescence transient over the leaf area was performed by FluorCam software where each data point represents one image. The represented induction kinetics are the mean curves of six different kinetics from six different leaves grown in every study area. 3.RESULTS AND DISCUSSION 3.1.Image Fluorescence parameters Fully green leaves of Cercius siliquastrum grown in optimal physiological conditions (Dajti area-Site 1) from six different branches were analyzed. Images of the maximum fluorescence in the dark Fm of all analyzed leaves exhibited almost the same distribution as well as the same level of fluorescence signal over the whole leaf area. The images represented in Figure 1 (A) that belong to one of these leaves showed no irregularities and nearly uniform distribution of fluorescence signatures. 89 �3rd International Symposium on Sustainable Development, May 31 - June 01 2012, Sarajevo A B C Figure 1. Images at the maximum fluorescence in the dark Fm of a leaf grown in optimal physiological conditions A-site 1, and plan steel air pollution conditions B-site 2, C- site 3 (pseudoscale 0-700). Images at the maximum fluorescence in the dark Fm of leaves grown in plan steel air pollution conditions showed a different distribution of the fluorescence emission over the leaf area as compare to the leaves grown in optimal conditions, especially leaves grown in site 3 (Figure 1, B, C). Fluorescence images at Fm displayed at the same pseudoscale clearly showed changes of the values of this parameter and their distributions related to the heterogeneity over leaf area between leaves grown in different conditions. The shape of the fluorescence induction kinetics of green leaves grown in optimal conditions demonstrated the healthy physiologically state of these leaves (Figure 2, A). While the shape of the induction kinetics exhibiting an increase of the fluorescence signal at the steady state of fluorescence demonstrated the effect of pollution to which were exposed leaves during their growth (Figure 2, B, C). A B Figure 2.Induced fluorescence kinetics of leaves of Cercius siliquastrumgrown in optimal physiological conditions A-site 1, and plan steel air pollution conditions B-site 2, C- site 3. Image fluorescence parameters of every leaf were calculated by FluoCam software as the mean of the fluorescence signals of all pixels over the leaf area. The represent values of these parameters (Table 1) correspond to a mean of six different leaves. The mean values of image fluorescence parameters demonstrated the differences between leaves grown in different conditions. The values of standard deviations of fluorescence parameters of leaves grown in optimal conditions show no significant differences between analyzed leaves. The observed increase of the values of standard deviations from optimal growth conditions to pollution conditions can be illustrated the increase of the variability through the leaves as the effect to stress exposure. 90 C �3rd International Symposium on Sustainable Development, May 31 - June 01 2012, Sarajevo Table 1. Induced fluorescence image parameters of leaves of Cercius siliquastrum grown in area of optimal and plan steel air pollution conditions (mean of six leaves). Cerciussiliquastrum Image Fluorescence Fo parameters Fm Fv Fo' Fm' Fv' mean 130.8 406.8 276.3 141.7 181.4 40.0 std (3.36) (2.62) (5.39) (3.92) (4.87) (1.88) mean 140.5 484.4 343.9 161.3 209.5 48.3 std (4.35) (39.60 ) (39.62 ) (16.5) (18.29 ) (5.54) mean 118.4 333.2 214.8 149.9 193.4 43.4 std (5.54) (64.12 ) (61.32 ) (26.76 ) (23.52 ) (13.49 ) Site 1 Site 2 Site 3 3.2.Image Fluorescence ratios Images of the fluorescence decline ratio Rfd of a green leaf grown in optimal conditions showed almost no irregularities and a uniform distribution of the values of this ratios over the leaf area (Figure 3 A). The values of Rfd ratios as plant vitality indicator demonstrated that these full green leaves can be characterized by the high photosynthetic activity, as reflect by the mean values the ratio 1.63 (Table 2). A B C Figure 3. The fluorescence decline ratio image Rfdof leaves of Cerciussiliquastrum grown in optimal physiological conditions A-site 1, and plan steel air pollution conditions B-site 2, Csite 3 (pseudoscale 0-3). The images of the fluorescence decline ratio Rfd of leaves grown in pollution conditions (Figure 3 B, C) showed a non uniform distribution and increase of irregularities of the values of this ratios over the leaf area. 91 �3rd International Symposium on Sustainable Development, May 31 - June 01 2012, Sarajevo Table 2. Image fluorescence ratios of leaves of Cercius siliquastrum grown in area of optimal and plan steel air pollution conditions (mean of six leaves). Cerciussiliquastrum Image ratios Fluorescence Fm/Fo Fv/Fm Fm'/Fo' qN NPQ Rfd mean 3.15 0.684 1.27 0.856 1.240 1.63 std (0.09) (0.009) (0.02) (0.005) (.053) (0.10) mean 3.09 0.66 1.12 0.826 1.732 1.26 std (0.29) (0.02) (0.04) (0.007) (0.142) (0.18) mean 2.81 0.64 1.29 0.798 0.723 1.14 std (0.65) (0.080) (0.036) (0.145) (0.308) (0.377) Site 1 Site 2 Site 3 The values of Rfd ratios represented a lower mean values of the ratio comparing to the optimal conditions, mean values of 1.26. The values of Rfd ratios in pollution growth conditions represented the lowest mean values of the ratio comparing to two other growth conditions, mean values of 1.14 (Table 2, Figure 3). The distribution of the Rfd values over the leaf area as well as the values of the ratio demonstrated a lower activity of photosynthetic apparatus of the leaves grown in plan steel air pollution growth conditions in comparison of leaves grown in optimal conditions (Dajti area). The observed decrease of activity of photosynthetic apparatus in pollution conditions seem to be related to the distance of the source of pollution, (Figure 3, Table 2). Figure 4.Fluorescence decline ratio Rfd of leaves in different growth conditions (mean of six leaves). The distribution of the Rfd values showed a considerable reduction of the activity of the photosynthetic apparatus of the leaves grown in area of site 3 than of leaves grown in two other areas as plants were grown in severe stress-pollution conditions 92 �3rd International Symposium on Sustainable Development, May 31 - June 01 2012, Sarajevo The values of Rfd ratios as plant vitality indicator demonstrated that full green leaves can be characterized by the high photosynthetic activity, as reflect by the mean The fluorescence ratios that allow to estimate maximum and effective quantum yields of Photosystem II (Fv/Fm,Fm/Fo) indicated a decrease of these ratios from optimal growth conditions to stresspollution conditions (Table 2). Non photochemical quenching coefficient qN have been decreased in growth pollution conditions (Table 2). Increase of the standard deviation values were observed in all calculated fluorescence ratios to pollution growth conditions in compare to optimal conditions. Observe increase can be illustrated the raise of the variability through the leaves as the effect to pollution exposure. 4.CONCLUSIONS Fluorescence images measured during induction kinetics, shape of the Chl fluorescence kinetics and the values of the fluorescence parameters in the leaves of endemic plant Cerciussiliquastrum grown in optimal conditions (Dajti area - site 1) exhibited a high photosynthetic activity as is demonstrated by the values of fluorescence ratios which evaluate the plant vitality and quantum yield of photosynthetic apparatus: Rfd=1.63, Fm/Fo=3.15. Activity of photosynthetic apparatus of leaves grown in plant steel air pollution conditions (site 2).was generally lower than activity of plants grown in optimal conditions: Rfd=1.26, Fm/Fo=3.1. Activity of photosynthetic apparatus of leaves grown in plant steel air pollution conditions (site 3) -Elbasan area) demonstrated reduction compare to other areas as is expressed by the lowest values of fluorescence decline ratio (Rfd=1.14,Fm/Fo=2.81); increased non-uniformity distribution and heterogeneity of signal of fluorescence over the leaf area and shape of induction kinetics. Observed differences on imaging of chlorophyll fluorescence signature and photosynthetic pigment content of spontaneous plants allowed to characterize the photosynthetic performance in order to evaluate the damage by plant steel air pollution related to the distance of the source of pollution. REFERENCES Babani F. and Lichtenthaler H.K. (1996). Light-induced and age-dependent development of chloroplasts in etiolated barley leaves as visualized by determination of photosynthetic pigments, CO2 assimilation rates and different kinds of chlorophyll fluorescence ratios. J Plant Physiol 148: 555-566 Buschmann C. and Lichtenthaler H.K. (1998). Principles and characteristics of multi-colour fluorescence imaging of plants. - J. Plant Physiol. 152, 297-314 Govindjee (1995): Sixty-three years since Kautsky: Chlorophyll a fluorescence. Australian Journal of Plant Physiology 22: 131-160 Govindjee (2004): Chlorophyll a fluorescence: A bit of basics and history. In: Papageorgiou G, Govindjee (eds) Chlorophyll fluorescence: A signature of Photosynthesis. Kluwer Academic Publishers, Dordrecht, The Netherlands. pp 1-42 93 �3rd International Symposium on Sustainable Development, May 31 - June 01 2012, Sarajevo Krause G.H. and Weis E. (1991). Chlorophyll fluorescence and photosynthesis: the basics. Ann Rev Plant Physiol Plant Mol. Biol 42: 313-349 Lichtenthaler H.K. (1987). Chlorophylls and carotenoids, the pigments of photosynthetic biomembranes. In: Douce R, Packer L (eds) Methods Enzymol 148, pp. 350-382. Academic Press Inc, New York. Lichtenthaler H.K. and Babani F. (2000). Detection of photosynthetic activity and water stress by imaging the red chlorophyll fluorescence. Plant Physiology Biochemistry 38: 889-895 Lichtenthaler H.K., Babani F. (2004). Light adaptation and senescence of the photosynthetic apparatus. Changes in pigment composition, chlorophyll fluorescence parameters and photosynthetic activity during light adaptation, In George Papageorgiou G, Govindjee (eds): Chlorophyll Fluorescence: A Signature of Photosynthesis, Kluwer Academic Publishers. Dordrecht,The Netherlands, pp 713-736. Lichtenthaler H.K. and Buschmann C. (2001). Chlorophylls and carotenoids–Measurement and characterisation by UV-VIS. Current Protocols in Food Analytical Chemistry (CPFA), (Supplement 1), pp. F4.3.1 - F 4.3.8. John Wiley, New York Lichtenthaler H.K. and Miehe J.A. (1997) Fluorescence imaging as a diagnostic tool for plant stress. Trends Plant Sci 2: 316-320. Lichtenthaler H.K., Babani F., Langsdorf G. (2007). Chlorophyll fluorescence imaging of photosynthetic activity in sun and shade leaves of trees. Photosynthesis Research, 93: 235244. Lichtenthaler H.K., Babani F., Langsdorf G., Buschmann C. (2000). Measurement of differences in red chlorophyll fluorescence and photosynthetic activity between sun and shade leaves by fluorescence imaging. Photosynthetica38: 521-529. Krause, G.H., Weis, E. (1991): Chlorophyll fluorescence and photosynthesis: the basics. Ann. Rev. Plant Physiol. Plant Mol. Biol. 42: 313-349. 94 � Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Extent The size or duration of the resource. 1231 Title A name given to the resource Effect of steel plant pollution on photosynthetic apparatus of some spontaneous plants by chlorophyll fluorescence imaging Author Author Eriola, Zhuri Hida Abstract A summary of the resource. Environmental pollution by the steel plant in Elbasan, the largest metallurgical complex in Albania causes serious problems. Although the blast furnaces and basic oxygen converters closed in 1991, small scale steel production from scrap metal continued. This steel plant releasing chemicals and particulates into the atmosphere is considered a source of air pollution. Industrial siderurgic air pollution as other stressors affects growth and physiological functions of plants. Pollution has effects on the photosynthetic performance of leaves and can modify their optical and fluorescence properties either directly or indirectly. The chlorophyll (Chl) fluorescence provides ample information on the performance of photosynthetic apparatus. Chl fluorescence signatures of leaves as an efficient tool and a nondestructive method for the in vivo analysis of plant stress is applying to describe and investigate the photosynthetic light processes and quantum conversion at physiological conditions as well as to detect stress on the photosynthetic apparatus. The chlorophyll fluorescence images were measured on leaves using the FluorCam 700MF imaging system (Photon Systems Instrument) as a techniques that offer the possibility to study the distribution and patchiness of fluorescence signatures over the whole leaf area. Chl fluorescence images were measured on the leaves of some spontaneous plants grown in sites with different level of steel plant air pollution assessed on base of different distances and different directions from the source of the pollution. Efficiency of photosynthetic apparatus of analyzed plants was evaluated via chlorophyll fluorescence images during induction kinetics and various fluorescence ratio images. The images of Chl fluorescence ratios, acquired by pixel to pixel arithmetic operations performed by FluorCam software, were aplied as indicators of the functional state or the damage of the photosynthetic apparatus. The potential and effective quantum yields of photosynthetic electron transport were estimated too. Observed differences on imaging of chlorophyll fluorescence signature and photosynthetic pigment content of some spontaneous plants allowed characterizing of the photosynthetic performance in order to evaluate the damage by plant steel air pollution. Keywords: chlorophyll fluorescence imaging, induction kinetics, plant steel air pollution, photosynthetic apparatus, potential/effective quantum yield, spontaneous plant. Date A point or period of time associated with an event in the lifecycle of the resource 2012-05-31 Keywords Keywords. Conference or Workshop Item PeerReviewed H Social Sciences (General) https://omeka.ibu.edu.ba/files/original/99a99d901044cbf225748456c317163f.pdf 0a4f5df5dce467ff54885c5f8ac6a47c PDF Text Text Effect Of Trichoderma Harzianum And An Arbuscular Mycorrhizal Fungus Glomus Mosseae On Fusarium Crown Rot (Fusarium Culmorum) In Wheat (Cv Altay 2000) Ş.Evrim Arıcı University of Suleyman Demirel, Faculty of Agriculture, Department of Plant Protection Isparta/TURKEY evrima@ziraat.sdu.edu.tr Đlknur Eser University of Suleyman Demirel, Faculty of Agriculture, Department of Plant Protection Isparta/TURKEY evrima@ziraat.sdu.edu.tr Hulya Özgönen University of Suleyman Demirel, Faculty of Agriculture, Department of Plant Protection Isparta/TURKEY hozgonen@ziraat.sdu.edu.tr Abstract: The necrotrophic fungal pathogen Fusarium culmorum causes crown rot disease (CR) in wheat. The experiments were designed to compare the effectiveness of Trichoderma harzianum and Arbuscular Mycorrhizal Fungus Glomus mossea on reduction of F. culmorum in wheat (cv Altay 2000) in pots tests in this research. Three weeks after sowing, percentage of diseased emerging seedlings were evaluated. In pot trials, Trichoderma harzianum, Glomus mosseae were initially evaluated for reducing disease on wheat cultivars Altay 2000 planted in sterilized soil artificially infested with the pathogen. Wheat plants growing from T. harzianum, and G. mosseae showed the average disease severity of 82.68% for 30-40% while plants from untreated soil had the disease severity of 75% and 98%, respectively Both of them gave also a good control in wheat cultivars. The ability of these isolates to affect the infection of wheat seedlings by F. culmorum may be of potential value in field trials Introduction Fusarium culmorum (W.G.Smith ) is important soilborne pathogens that cause seedling, foot and root rot and head blight of wheat. There are no chemical means to control effectively both of fungi and resistant cultivars are not avaliable. Therefore, the use of microorganisms as a biological control for these diseases is of interest. However, there are few reports of a biocontrol agents against F. culmorum. Many studies have proved the potential of Trichoderma spp.and Arbuscular Mycorrhizal Fungus as biological agents antagonistic to several soil borne plant pathogens (Grondona et al. 1997; De Meyer et al. 1998; Altomare et al. 1999; Jacobs et al. 2000; Ahonen-Jonnart et al. 2000; Rosling et al. 2004; Kucuk & Kıvanc, 2005; Ozgonen & Erkilic, 2007; Askar & Rashad, 2010). Strains of Trichoderma can produce antifungal metabolites. They may also be competitors with fungal pathogens, which promotes plant growth. In addition, a number of Trichoderma strains are able to secrete lytic enzymes such as chitinases and β-(1,3)glucanases when grown in liquid media supplemented with either polymers such as laminarin or chitin or with fungal cell walls. The beneficial effects of arbuscular mycorrhizal fungion plant growth depend in part on the members of the symbiosis and their interactions with other organisms present in the rhizosphere (Ocampo, 1993). 10 �The purpose of this study was to evaluate the interaction between the pathogen and the fungal antagonist and the possibility of mycoparasitism under the biological control of F. culmorum by T. harzianum and Glomus mosseae. Material and Methods T. harzianum was isolated from soil surrounding wheat roots infected by Fusarium culmorum. The isolated culture was kept on Potato Dextrose Agar (PDA). F. culmorum and T .harzianum was grown at 7 days on potato dextrose agar (PDA), Growing medium (115 gr sand, 35 g corn meal) were autoclaved twice on subsequent days at 121 0C for 60 min, and then were inoculated with one 6-mm-diameter PDA plug excised from an actively growing each F. culmorum and T. harzianum culture (Sneh et al. 1998). Flasks were then placed in a growth chamber at 24 0C for 21 days. Glomus moseae was bulked up on maize and used as mycorrhizal fungal inoculum. Mycorrhizal inoculation were performed by incorporating the inoculum including soil infested with spores mixed with root fragments, 2-3 cm below the seeds (Miles & Wilcoxson, 1984). Inoculum amount was determined as 1000 spores 10 g -1 for each plant as seed treatment. Wheat seed cv cv Altay 2000 were sterilized (%5 sodium hypocloride) and sown in pots. 1 seeds/cultivar in five pots was sown for each soil sample After 11 days 2% growing medium was added in steril soil in pot Prothioconazole +Tebuconazole was used as fungucide (Table 1). Treatment Control (-) Control (+) Glomuss mosseae +F.culmorum+T.harzianum F.culmorum+T.harzianum Glomuss mosseae +F.culmorum F.culmorum+Prothioconazole +Tebuconazole Table 1:Treatments on wheat plants. After sowing each pot received 100 ml of tap water. Pots were maintained in a greenhouse at 21 C and watered daily for 21 days. After plants had been harvested and the roots were washed free of soil, disease was assessed on roots and subcrown internodes. Severity of disease was evaluated on a scale of 0–5 in which 0: no lesion evident, 1: <25% roots and subcrowns with necrosis, 2 :26–50% roots and subcrowns with necrosis, 3: 51–75% roots and subcrowns with necrosis, 4: 76–100% roots and subcrowns with necrosis, and 5: no plant emergence. Obtained data from bioassays were analyzed by SPSS version 16.0.1. Data were analyzed by ANOVA and treatment means separated by TUKEY test (P <0:05) to select isolates for a new assay with several cultivars 0 Results and Discussion In greenhouse experiments, of T. harzianum, Glomus mosseae and Lamardor initially screened for ability to suppress damping-off of wheat when applied to wheat seeds in sterilized soils. The results obtained for the pot/soil experiment are given in Table 2. Lamardor caused a decrease in the percentage of diseased plants. Lamardor provided 60% inhibition rate for F.culmorum. Treatments with T. harzianum and Glomuss mosseae significantly reduced severity of symptoms of F. culmorum damping-off as compared with the non-treated controls. Among the antagonists were significantly better than the control, but it was not statistically different from each other. The results shown that T. harzianum was found to have a wide range of inhibitory effects against Fusarium culmorum. T. harzianum provided 65-70% inhibition rate for F.culmorum. T. harzianum gave also a good behaviour and growth in soil. It was observed colonization of Trichoderma harzianum (data not shown). T. harzianum is a potential agent for the biocontrol of plant pathogens. 11 �Treatment Control (-) Control (+) Glomuss mosseae +F.culmorum+T.harzianum F.culmorum+T.harzianum Glomuss mosseae +F.culmorum F.culmorum+Prothioconazole +Tebuconazole Scala Value 0±0,0 a 4,7±0,3 c Diseases severity % 0 95 1,3±0,3 b 30 1,6±0,5 b 35 1,6±0,3 b 2±0,3 b 35 40 Table 2 Cultivar reaction to the inhibition of F.culmorum after treatment with Glomuss mosseae , T.harzianum, Prothioconazole +Tebuconazole. Means with different letters are significantly different at p<0,05 using TUKEY test. Mycorrhizal fungi reduced the percentage of diseases severity in infected wheat plants. These result is in agreement with Askar & Rashid (2010). We found that Glomuss mosseae decreased root rot by 65-70%. Many researchers have reported that colonization of mycorrhizal fungi can reduce root diseases caused several soil born pathogens (Wehner et al. 2009; Hozgonen et al. 2010). Mycorrizal colonization was observed in the root of wheat and in soil (data not shown). Wheat with Mycorrizal fungi was observed to receive protect from F.culmorum relative to control plants in our experiment. Biological control is becoming an important component of plant disease management. It was reported that Trichoderma harzianum and Glomuss mosseae are the most effective agent for the biocontrol of fungal pathogens (Kucuk & Kıvanc, 2005; Ozgonen & Erkilic, 2007). Dunlop et al. (1989) showed that an isolate of T. koningii inhibited the saprophytic growth of G. graminis (Sacc.) Arx and Oliver var. tritici Walker (Ggt). In conclusion, our study showed that T. harzianum and Glomus mosseae were superior in suppressing the disease. Fron the obtained results that both of biological agents showed an antagonistic effect on plant pathogenic fungi and Glomus mosse colonization increased plant resistance against infection with F. culmorum as well as on their biochemical and physiological features. Thus they could be used in certain biological control studies References Ahonen-Jonnart U., Van Hees, P.A.W., Lundstrom, U.S., & Finlay, R.D. (2000). Organic acids produced by mycorrhizal Pinus sylvestris exposed to elevated aluminium and heavy metal concentrations. New Phytol. 146: 557– 567. Altomare, C., Norvell, W.A., Bjorkman, G., & Harman G. (1999). Solubilization of phosphates and micronutrients by the plantgrowth- promoting and biocontrol fungus T. harzianum Rifai 1295-22. Applied and Environmental Microbiology, Vol. 65, No. 7, 2926-2933. De Meyer, G., Bigirimana, J., Elad, Y. & Hofte, M. (1998). Induced system resistance in Trichoderma harzianum T39 biocontrol of Botrytis cinerea. Eur. J. Plant Pathol. 104: 279–286 Dunlop, R.W., Simon, A., & Sivasithamparam, K. (1989) An Antibiotic from Trichoderma koningii Active Against Soilborne Plant Pathogens. Journal of Natural Products 52: 67-74. Grondona, I., Hermosa, R., Tejada, M., Gomis, M.D., Mateos, P.F., BrIdge, P.D., Monte, E. & Garcia-Acha I. (1997). Physiological and biochemical characterization of Trichoderma harzianum, a biological control agent against soilborne fungal plant pathogens. Appl. Environ. Microbiol. 63: 3189–3198. Jacobs, H., Boswell, G.P., Ritz, K., Davidson, F.A. & Gadd, G.M. (2002). Solubilization of calcium phosphate as a consequence of carbon translocation by Rhizoctonia solani. FEMS Microbiol. Ecol. 40: 65–71. 12 �Kücük, K., & Kıvanc, M., (2003). Isolation of Trichoderma Spp. and Determination of Their Antifungal, Biochemical and Physiological Features .Turk J Biol 27, 247-253 Rosling, A., Lindahl, B.D., Taylor, A.F.S. & Finlay R.D. (2004). Mycelial growth and substrate acidification of ectomycorrhizal fungi in response to different minerals. FEMS Microbiol. Ecol. 47: 31–37. Sneh, B., Burpee, L., & Ogoshi, A. (1998). Identification of Rhizoctonia species. American Phytopathological Society Press. St Paul, 133 pp. 13 � Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Extent The size or duration of the resource. 321 Title A name given to the resource Effect Of Trichoderma Harzianum And An Arbuscular Mycorrhizal Fungus Glomus Mosseae On Fusarium Crown Rot (Fusarium Culmorum) In Wheat (Cv Altay 2000) Author Author Arıcı, S.Evrim Eser, İlknur Özgönen, Hulya Abstract A summary of the resource. The necrotrophic fungal pathogen Fusarium culmorum causes crown rot disease (CR) in wheat. The experiments were designed to compare the effectiveness of Trichoderma harzianum and Arbuscular Mycorrhizal Fungus Glomus mossea on reduction of F. culmorum in wheat (cv Altay 2000) in pots tests in this research. Three weeks after sowing, percentage of diseased emerging seedlings were evaluated. In pot trials, Trichoderma harzianum, Glomus mosseae were initially evaluated for reducing disease on wheat cultivars Altay 2000 planted in sterilized soil artificially infested with the pathogen. Wheat plants growing from T. harzianum, and G. mosseae showed the average disease severity of 82.68% for 30-40% while plants from untreated soil had the disease severity of 75% and 98%, respectively Both of them gave also a good control in wheat cultivars. The ability of these isolates to affect the infection of wheat seedlings by F. culmorum may be of potential value in field trials Date A point or period of time associated with an event in the lifecycle of the resource 2010-06 Keywords Keywords. Conference or Workshop Item PeerReviewed Q Science (General) https://omeka.ibu.edu.ba/files/original/0bed6a65d0b57cf646e0783e059b7814.pdf af8c307a067e56185e57c3d303df8cfc PDF Text Text Effect of Vaccinium vitis-idaea tea and Arctostaphylos uva-ursi tea on growth of causative agents of urinary tract infections Lamija Hafizović1, Selma Karup1, Almin Hadžialić 1 1 International Burch University, Sarajevo, Bosnia and Herzegovina lamija.hafizovic@stu.ibu.edu.ba selma.karup@stu.ibu.edu.ba almin.hadzialic@stu.ibu.edu.ba Abstract - Urinary tract infections pose a serious problem to people, both in the hospital environment and outside world. They are characterized by high mortality and ability to cause health problems in areas of the human body other than the urinary tract. It has been long clinical practice to treat these infections with antibiotics, a tactic made very ineffective with the advent of antibiotic-resistant microbial strains. The research has turned to alternative modes of treatment, such as use of herbal remedies to combat urinary tract infections. Effect of two types of herbal teas was observed through use of broth microdilution assay, to test varying concentrations of teas on the growth of selected microorganisms. Results were verified by assessment of colony growth on Mueller Hinton Agar plates. Tested microorganisms exhibited very dense colony growth. Similarity of conditions between urinary retention and conditions under which microorganisms were cultured in 96-well plates possible reason for density of growth. Methods with higher degree of confidence in treatment of urinary tract infections could likely be the combination of antibiotics with herbal teas. Keywords: antibiotic resistance, Arctostaphylos uva-ursi, broth microdilution assay, urinary tract infections, Vaccinium vitis-idaea 1. Introduction Urinary tract infections (UTIs) are, by definition, categorized as diseases according to clinical symptoms, laboratory indicators and microbiological findings, and are most often caused by various bacterial species. They are designated as cystitis or as infections affecting lower urinary tract, or as prostatitis. Based on clinical factors, they are classified in different groups: acute uncomplicated cystitis, urinary tract infections caused by indwelling catheters, recurrent cystitis in young women, urinary tract infections in men, complicated urinary tract infections and asymptomatic bacteriuria [1,2]. In the cases of often repeated and inadequately treated urinary tract infections, these infections may become permanent or chronic diseases [3], which can lead to development of other types of diseases, and further increase the already high mortality rate exhibited by urinary tract infections [4]. Antibiotics are the most common method of treatment for urinary tract infections. Some of the antibiotics have been highly effective in treatment of UTIs, while others had little to no effect. However, the misuse of antibiotics has led to development of bacterial resistance, where increasing numbers of antibiotics have no effect in treatment of urinary tract infections [5,6]. This has led to discovery of alternative methods of treatment for UTIs, such as use of herbal products to combat these infections. Certain herbs have �exhibited ability to prevent bacterial invasion into the urinary tract (Agropyron repens), to impede adhesion of bacteria to bladder walls (Urtica spp., Betula spp.), and to inhibit formation of bacterial colonies [7,8]. Bioactive compounds of Vaccinium vitis-idaea (lingonberry, mountain cranberry) have demonstrated antimicrobial, anti-inflammatory and antioxidative activity [9,10], while Arctostaphylos uva-ursi (bearberry) was also noted to have anti-inflammatory effect in lower urinary tract [11]. The objective of this experiment was to determine the effect of Vaccinium vitis-idaea tea and Artctostaphylos uva-ursi tea on growth of primarily bacteria which are responsible for the development of various urinary tract infections. 2. Materials and methods Microorganisms used in this experiment were Escherichia coli ATCC 14169, Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923, Staphylococcus aureus ATCC 6538, Staphylococcus aureus ATCC 12493, Enterococcus faecalis ATCC 29212, Candida albicans ATCC 10231 and Pseudomonas aeruginosa ATCC 27853. They were cultured and kept in Tryptic Soy Broth (TSB) containing 50% glycerol, since it was necessary to store them at -80˚C. The herbal teas chosen for this experiment were the Vaccinium vitis-idaea tea and Arctostaphylos uva-ursi tea, both of which were acquired from herbal pharmacy ″Faveda″ in Sarajevo. Teas were made and tested in two different concentrations: in concentration recommended on tea packaging and in concentration two times stronger than the recommended one. Since the objective of this experiment was to study the effect of teas on bacterial growth, it was necessary to detect minimal inhibitory concentration (MIC) for both teas. Broth microdilution assay was used for detection of minimal inhibitory concentrations for each of the tested microorganisms. Tested microorganisms were first grown in the medium Tryptic Soy Broth until desired growth phase. In each well of microtiter plate a 100μl of medium was pipetted. Tea, also made in TSB medium, was added in column 3 (100μl) and mixed using micropipette to suck the liquid up and down a few times. Then, a 100μl of dilution was transferred from column 3 to column 4 and mixed using micropipette. The procedure was repeated until the last column, so the concentration of tea was lessened by half in each subsequent well. Following that, a 10 μl of microorganism was added (each tested microorganism to an individual plate) in columns 2 to 12. Column 1 did not contain microbes, as it represented negative control. Plates were incubated overnight at 37˚C. After incubation, microbes from wells containing different concentrations were inoculated onto Mueller Hinton Agar plates and incubated overnight at 37˚C. Bacterial growth was observed from the plates and it was visually assessed and numerically classified by the density of formed colonies, as shown in Table 1. 3. Results Bacterial growth was assessed from Mueller Hinton Agar plates. The objective was to find minimal inhibitory concentration, so microorganisms were inoculated only from certain wells of 96-well plates (where the probability of finding minimal inhibitory concentrations seemed the highest). Starting concentrations of tea were the recommended concentration (RC) on the tea packaging (Table 2 and 3), �adjusted for the volume of the well, and the second concentration that was twice as strong as the recommended one (Tables 4 and 5). Using broth microdilution assay as described in the previous section, the concentration of tea was lowered by half in each subsequent well. Recommended concentration on the tea packaging was one coffee spoon (10 g) per 200 ml of liquid (TSB, in this case). Table 1. Evaluation of microbial growth Evaluation Area of plate covered by microbial growth 10^7 100% 10^6 85%-90% 10^5 70%-75% 10^4 50% 10^3 35% 10^2 15% 10^1 <15% Sterile 0% Table 2. Effect of recommended concentration of Vaccinium vitis-idaea tea on bacterial growth 0,5*RC 0,25*RC 0,125*RC 0,0625*RC E. faecalis 29212 10ˆ7 10ˆ7 / / C. albicans 10231 10ˆ7 10ˆ7 / / P. aeruginosa 27853 10ˆ7 10ˆ7 / / E. coli 25922 10ˆ7 10ˆ7 / / E. coli 14169 10ˆ7 10ˆ7 10ˆ7 10ˆ7 S. aureus 25923 10ˆ7 10ˆ7 / / S. aureus 12693 10ˆ7 10ˆ7 / / S. aureus 6538 10ˆ7 10ˆ7 / / Table 3. Effect of recommended concentration of Arctostaphylos uva-ursi tea on bacterial growth 0,5*RC 0,25*RC 0,125*RC 0,0625*RC E. faecalis 29212 10ˆ7 10ˆ6 / / C. albicans 10231 10ˆ6 10ˆ7 10ˆ7 10ˆ7 �P. aeruginosa 27853 10ˆ6 10ˆ7 10ˆ7 / E. coli 25922 10ˆ7 10ˆ7 / / E. coli 14169 10ˆ7 10ˆ7 / / S. aureus 25923 10ˆ7 10ˆ7 / / S. aureus 12693 10ˆ6 10ˆ6 10ˆ6 10ˆ6 S. aureus 6538 / 10ˆ7 10ˆ7 10ˆ7 Table 4. Effect of second concentration of Vaccinium vitis-idaea tea on bacterial growth RC 0,5*RC 0,25*RC E. faecalis 29212 10ˆ7 10ˆ7 10ˆ7 C. albicans 10231 10ˆ7 / / P. aeruginosa 27853 10ˆ7 10ˆ7 / E. coli 25922 10ˆ7 / / E. coli 14169 10ˆ7 10ˆ7 / S. aureus 25923 10ˆ7 10ˆ7 / S. aureus 12693 10ˆ7 10ˆ7 / S. aureus 6538 10ˆ7 10ˆ7 / Table 5. Effect of second concentration of Arctostaphylos uva-ursi tea on bacterial growth RC 0,5*RC 0,25*RC 0,125*RC E. faecalis 29212 / 10ˆ7 10ˆ7 10ˆ7 C. albicans 10231 10ˆ7 10ˆ7 / / P. aeruginosa 27853 10ˆ7 10ˆ7 / / E. coli 25922 10ˆ7 10ˆ7 / / E. coli 14169 10ˆ6 10ˆ7 10ˆ7 / S. aureus 25923 10ˆ7 10ˆ7 / / �S. aureus 12693 10ˆ7 10ˆ7 10ˆ7 / S. aureus 6538 10ˆ7 10ˆ7 / / As observed from results presented in tables above, microorganisms have demonstrated formation of highly dense bacterial colonies, covering the entirety of inoculated surface of agar plates, both in case of recommended tea concentrations and the second tested concentration. 4. Discussion Urinary tract infections present a great issue, since they are capable of causing a multitude of other health complications, as well as possessing a high mortality rate, which is 3% in women and 1% in men [4]. A fact which further exacerbates this problem is the ability of microorganisms to form structures such as biofilm and to produce antibiotic-negating enzymes, resulting in bacteria being capable of withstanding and surviving antibiotic activity, which is a characteristic termed antibiotic resistance. Microbial strains resistant to a variety of different antibiotics, designated as multidrug-resistant strains, have also emerged [6]. It is likewise highly concerning that the discovery of new types of antibiotics capable of successfully combating UTIs has substantially decreased, which has led researchers to turn their investigations to alternative methods of treatment for UTIs. One of these alternative methods is the use of herbal products, since some of their compounds are capable of mitigating or eliminating the symptoms of UTIs. In this experiment, we observed the growth of microorganisms under influence of herbal tea. One of the objectives was to determine whether tea alone has the ability to impede the growth of bacteria. Broth microdilution assay was used to test the effect of different concentrations of tea on microbial growth, because it provides quantitative data and it is possible to use this method in any laboratory. Inoculation onto Mueller Hinton Agar plates was used to verify the results. After overnight incubation at 37˚C, it was observed that tested microorganisms exhibited highly dense growth, under both normal and doubled concentrations. Conclusion is that this might have happened because the conditions in 96-well plates, during the performance of broth microdilution assay, were similar to those occurring in urinary retention. Urinary retention is an inability to completely remove urine from the bladder, which can lead to kidney and bladder damage, and development of urinary tract infections. Bacteria within urine, which are normally mostly harmless since they are removed with urine in healthy people, in case of urinary retention are able to accumulate and cause the development of various urinary tract infections [12]. Increased liquid intake was reported to have beneficial effects for the patients affected by UTIs, since it causes dilution of metabolic waste products which serve as nutrients for microorganisms [13]. In addition, diuresis that results from increased liquid intake yields the benefit of so-called mechanical ″flushing″, meaning that bacteria are physically removed from the urinary tract along with urine, denying them the time necessary to accumulate and cause further issues. As noted in introduction, some herbs possess compounds which exhibit the ability to prevent bacterial adhesion or inhibit bacterial growth [7,8]. ″Flushing″ the urinary tract with herbal tea used for alleviating the symptoms of UTIs can possibly exhibit higher effectiveness in treatment of these infections. �Herbal teas used in this experiment consisted of dried leaves (in case of A. uva-ursi) and dried fruits or baccae (in case of V. vitis-idaea). This might have contributed to their low effect on microbial growth. Extracts of fresh leaves and fruits could be more effective in treatment of urinary tract infections. Despite not being able to inhibit the growth of microbes on their own, both V. vitis-idaea and A. uva-ursi have properties which make them useful in treatment of UTIs, such as the anti-inflammatory activity and antioxidative activity, as well as other properties. Their use in treatment of UTIs is a subject of many studies [9-11,14-17]. Antibiotic resistance of microorganisms responsible for development of UTIs extends to some of the most commonly used antibiotics in treatment of UTIs. However, there are several antibiotics which still demonstrate significant effect in inhibiting the growth of UTI causative agents. There is a high possibility that the combined effect of antibiotics and herbal teas could result in greater effectiveness of treatment of urinary tract infections. Further research into combined use of antibiotics and herbal remedies should be done, with aim of reduction of unnecessary antibiotic use, which leads to the development of antibiotic resistance. 5. References [1] Grabe, M., Bjerklund-Johansen, T. E., Botto, H., Çek, M., Naber, K. G., Pickard, R. S., ... and Wullt, B. (2013). Guidelines on urological infections. European Association of Urology guidelines. [2] Stamm, W. E., and Hooton, T. M. (1993). Management of urinary tract infections in adults. New England journal of medicine, 329(18), 1328-1334. [3] Finer, G., and Landau, D. (2004). Pathogenesis of urinary tract infections with normal female anatomy. The Lancet infectious diseases, 4(10), 631-635. [4] Daswani, P. G. (2019). Non-antibiotic potential of medicinal plants to combat urinary tract infections. CURRENT SCIENCE, 117(9), 1459. [5] Sabir, N., Ikram, A., Zaman, G., Satti, L., Gardezi, A., Ahmed, A., and Ahmed, P. (2017). Bacterial biofilm-based catheter-associated urinary tract infections: Causative pathogens and antibiotic resistance. American Journal Of Infection Control, 45(10), 1101-1105. [6] Gopichand, P., Agarwal, G., Natarajan, M., Mandal, J., Deepanjali, S., Parameswaran, S., and Dorairajan, L. N. (2019). In vitro effect of fosfomycin on multi-drug resistant gram-negative bacteria causing urinary tract infections. Infection and drug resistance, 12, 2005. [7] Rafsanjany, N., Lechtenberg, M., Petereit, F., and Hensel, A. (2013). Antiadhesion as a functional concept for protection against uropathogenic Escherichia coli: In vitro studies with traditionally used plants with antiadhesive activity against uropathognic Escherichia coli. Journal Of Ethnopharmacology, 145(2), 591-597. [8] Beydokthi, S., Sendker, J., Brandt, S., and Hensel, A. (2017). Traditionally used medicinal plants against uncomplicated urinary tract infections: Hexadecyl coumaric acid ester from the rhizomes of Agropyron repens (L.) P. Beauv. with antiadhesive activity against uropathogenic E. coli. Fitoterapia, 117, 22-27. [9] Wang, S. Y., Feng, R., Bowman, L., Penhallegon, R., Ding, M., and Lu, Y. (2005). Antioxidant activity in lingonberries (Vaccinium vitis-idaea L.) and its inhibitory effect on activator �protein-1, nuclear factor-κB, and mitogen-activated protein kinases activation. Journal of agricultural and food chemistry, 53(8), 3156-3166. [10] Puišo, J., Jonkuvienė, D., Mačionienė, I., Šalomskienė, J., Jasutienė, I., and Kondrotas, R. (2014). Biosynthesis of silver nanoparticles using lingonberry and cranberry juices and their antimicrobial activity. Colloids and Surfaces B: Biointerfaces, 121, 214-221. [11] Schink, A., Neumann, J., Leifke, A. L., Ziegler, K., Fröhlich-Nowoisky, J., Cremer, C., ... and Lucas, K. (2018). Screening of herbal extracts for TLR2-and TLR4-dependent anti-inflammatory effects. PloS one, 13(10), e0203907. [12] Barrisford GW and Steele GS. Acute urinary retention. Post T, ed. UpToDate. Waltham, MA: UpToDate; 2018. [13] Tian, Y., Cai, X., Wazir, R., Wang, K., and Li, H. (2016). Water consumption and urinary tract infections: an in vitro study. International Urology and Nephrology, 48(6), 949-954. [14] Moore, M., Trill, J., Simpson, C., Webley, F., Radford, M., Stanton, L., ... & Griffiths, G. (2019). Uva-ursi extract and ibuprofen as alternative treatments for uncomplicated urinary tract infection in women (ATAFUTI): a factorial randomized trial. Clinical Microbiology and Infection, 25(8), 973-980. [15] Afshar, K., Fleischmann, N., Schmiemann, G., Bleidorn, J., Hummers-Pradier, E., Friede, T., ... & Gágyor, I. (2018). Reducing antibiotic use for uncomplicated urinary tract infection in general practice by treatment with uva-ursi (REGATTA)–a double-blind, randomized, controlled comparative effectiveness trial. BMC complementary and alternative medicine, 18(1), 1-8. [16] Ek, S., Kartimo, H., Mattila, S., & Tolonen, A. (2006). Characterization of phenolic compounds from lingonberry (Vaccinium vitis-idaea). Journal of agricultural and food chemistry, 54(26), 9834-9842. [17] Antolak, H., Czyzowska, A., Sakač, M., Mišan, A., Đuragić, O., and Kregiel, D. (2017). Phenolic compounds contained in little-known wild fruits as antiadhesive agents against the beverage-spoiling bacteria Asaia spp. Molecules, 22(8), 1256-1274. � Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Journal of Natural Sciences and Engineering Identifier An unambiguous reference to the resource within a given context 2637-2835 DOI Digital object identifier 10.14706 Publisher An entity responsible for making the resource available International Burch University Description An account of the resource Journal of Natural Sciences and Engineering (JONSAE) is a peer-reviewed, biannually published international journal focusing on empirical and theoretical research in all branches of Engineering and Natural Sciences. It is published on the behalf of Faculty of Engineering and Natural Sciences of International Burch University and aims to provide the best content regarding by publishing original research papers, review articles, special issues, feature articles, and book reviews. All manuscript submissions are subject to initial appraisal by the Editor, and, if found suitable for further consideration, to peer review by independent, anonymous referees. All peer review is double-blind and submission is online. The journal welcomes theoretical, applied, interdisciplinary and methodological work, with preference on empirical research, critical approach and problem-solving methods in manuscripts. Language A language of the resource English Dublin Core The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/. Title A name given to the resource Effect of Vaccinium vitis-idaea tea and Arctostaphylos uva-ursi tea on growth of causative agents of urinary tract infections Author Author Lamija Hafizović, Selma Karup, Almin Hadžialić Abstract A summary of the resource. Urinary tract infections pose a serious problem to people, both in the hospital environment and outside world. They are characterized by high mortality and ability to cause health problems in areas of the human body other than the urinary tract. It has been long clinical practice to treat these infections with antibiotics, a tactic made very ineffective with the advent of antibiotic-resistant microbial strains. The research has turned to alternative modes of treatment, such as use of herbal remedies to combat urinary tract infections. Effect of two types of herbal teas was observed through use of broth microdilution assay, to test varying concentrations of teas on the growth of selected microorganisms. Results were verified by assessment of colony growth on Mueller Hinton Agar plates. Tested microorganisms exhibited very dense colony growth. Similarity of conditions between urinary retention and conditions under which microorganisms were cultured in 96-well plates possible reason for density of growth. Methods with higher degree of confidence in treatment of urinary tract infections could likely be the combination of antibiotics with herbal teas. Keywords Keywords. antibiotic resistance, Arctostaphylos uva-ursi, broth microdilution assay, urinary tract infections, Vaccinium vitis-idaea Identifier An unambiguous reference to the resource within a given context 2637-2835 DOI Digital object identifier 10.14706/JONSAE2021321