Air pollution has been a major job throughout history [ 1,2 ] . The beginnings of air pollution are both natural and human based. The naturals are fire, air current, volcanic eruptions and vaporization of organic constituents. The combustion of fossil fuels is the key of the buildup C dioxide ( CO2 ) in environment, traffic and fluke gas depending on industrial beginning are the major beginnings of air pollution of the consequences of human activity. Air pollutants such as C monoxides ( COx ) , sulfur dioxide ( SO2 ) , nitrogen oxides ( NOx ) and other atoms or hint sums of heavy metals besides have harmful effects on natural ecosystem. As a consequence ; air pollution is a planetary job and both authoritiess and scientist survey to cut down effects of this job.
The engineering for cut downing CO2 and other pollutants from environment has been based on biological assimilation. Plants can cut down the atmospheric CO2 concentration by absorbing [ 3 ] . The most common renewable energy resource has a microalgal beginning [ 4,5 ] . Microalgae have the answer to many of the planetary jobs confronting us today and they use CO2, the major Global Warming Gas, with sunshine as their unlimited energy beginning, produce O2 and change over it many of the utile merchandises and energy [ 6 ] . The truth is that, without microalgae environment would be poorer and energy crisis would be a more and more of import job. This was a major ground why there is involvement in microalgae in different countries. It is besides good known that microalgae grow faster than tellurian workss and the efficiency of energy merchandise is much higher [ 7 ] .
Depending on the increasing sums of environmental pollution, several surveies have been published on the linkages between microalgae and toxic constituents [ 8-12, 24-26, 30, 35 ] and all these surveies indicated that a considerable attending in environmental pollution can be reduced by microalgal biomass.
On the other manus high sums of SO2 and NO2 cause toxic effects on microalgal growing. The present paper is proposed to get the better of the suppression of microalgal growing by SO2 and NO2 with the aid of the growing stimulators. To heighten the microalgal biomass and keeping to photosynthesis some of the growing stimulators added to the growing media. The first of them is Triacontanol ( TRIA ) and the 2nd is sodium hydrogen carbonate ( NaHCO3 ) . First, TRIA is a natural works growing endocrine, a long 3 concatenation 30-carbon primary intoxicant ( C30H61OH ) and stimulate photosynthesis and growing [ 13,14 ] . On the other manus Na hydrogen carbonate ( NaHCO3 ) , a type of inorganic C, can be fixed as a CO2 beginning by microalgae as Wang et Al. [ 15 ] reviewed.In the present survey two strains of microscopic microalgae were tested their ability to digest environmental pollutants. The choice of Chlorella sp. , was based on old plants to digest to high concentrations of CO2 [ 24, 26 ] and SOx, NOx [ 9, 30 ] . Synechococcus sp. was selected because of it is known to play an of import function in the planetary C rhythm [ 27 ] and to our cognition has non been studied to prove its ability to cut down of environmental pollutants.
The range of the current hunt was to find the synergistic effects of extremely toxic environmental pollutants and growing stimulators on different microalgal civilizations and to compare the tolerance of them.
2. Materials and Methods
Stock solution of TRIA ( 96 % , w/v ; Aldrich ) was prepared by fade outing of 0.5 g of the chemical in trichloromethane ( Merck ) . Sodium bicarbonate solution ( Merck ) was prepared by fade outing 17.2 g l-1 of the chemical in distilled H2O. Appropriate volumes of the solutions were added to the media.
The SO2 and NO2 gases were prepared by fade outing of Na2SO3 ( Merck ) and NaNO2 ( Merck ) in distilled H2O to a concluding concentration of 1 mol l-1 and by dropping into these solution 37 % HCl ( Merck ) for each flask sample.
2.2. Microorganisms and civilization conditions
A strain of unicellular cyanobacterium, Synechococcus sp. and unicellular green alga, Chlorella sp. were used in the survey, as provided by Ankara University, Faculty of Science Biotechnology Laboratory ‘s from the current civilization aggregation.
The micro-organisms were cultivated in BG11 medium [ 28 ] utilizing shake flask method. The medium pH was adjusted to 7.5 by 0.1 M H2SO4 and 0.1 M NaOH, before autoclaving. A micro-organism suspension of 20 milliliter was contacted with 80 milliliters of civilization media at 1 mg l-1 TRIA and 34 mg l-1 NaHCO3 for Synechococcus sp. and at 1 mg l-1 TRIA and 43 mg l-1 NaHCO3 for Chlorella sp. in an Erlenmeyer flask. Once inoculated, undaunted flasks were incubated under uninterrupted light at 2400 sixty light strength provided by cool white fluorescent lamps at 25 ± 2 & A ; deg ; C on 10 yearss.
2.3. Gas readying system
In research lab experiments a design footing was devised for bring forthing of SO2 and NO2 gases. The theoretical account was besides used to reassign these acidic pollutants to the civilization media. The diagram of the experimental system is comprehensively given in Fig 1. System is composed of 500 and 250 milliliters Erlenmeyer flasks, magnetic heat scaremonger and heat immune hosiery. The 500 milliliter flask contains a gum elastic joggle with two holes that named acid-inlet ( 9.5 centimeter ) and gas-exit ( 4.5 centimeter ) and HCl drips into the acid-inlet hole with a aid of 5.0 milliliter enemas to Na2SO3 or NaNO2 chemical solutions, as above mentioned. As a consequence of HCl add-on, SO2 and NO2 gases is being produced in the 500 milliliter flasks with the aid of stirrer magnet and these gases are transferred with 23.5 centimeters length heat immune hosiery in 250 milliliter flask that contains microalgal civilization solution. The heat is provided by the magnetic heat scaremonger ( Velp Scientifica ARE Heating magnetic scaremonger ) to formation of the reaction where under the 500 milliliter flask. The 250 milliliter flask that contains of 100 milliliter of civilization solution includes a single-hole gum elastic joggle with a 19.0 centimeter length metal acerate leaf. The gases that are produced in 500 milliliter flask are transferred to the 250 milliliter flask by the aid of the metal acerate leaf that enters up to 1.0 centimeter into the civilization solution.
& A ; lt ; Here Fig.1 & A ; gt ;
Figure 1. The diagram of experimental set-up
1: Magnetic heat scaremonger
2: Erlenmeyer flask ( 500 milliliter )
3: Gum elastic joggle with acerb recess and gas issue holes
5: Heat resistant hosiery
6: Gum elastic joggle with cotton
7: Erlenmeyer flask ( 250 milliliter )
2.4. Consequence of different parametric quantities on microalgal gas tolerance
2.4.1. Consequence of pH
The consequence of median pH value on acidic gas tolerance of microalgal civilizations was determined in the samples that one time aerated with 10 ‘ exposing clip of SO2 and NO2 at the terminal of the 10 yearss of incubation period. The experiments were performed at pH 6.5, 7.5, 8.5 and 9.5 for each microalgal civilization and the samples were taken at the terminal of the 11, 13 and 15 yearss.
2.4.2. Consequence of gas exposing clip
To find the gas exposing clip to consequence on microalgal suppression, civilizations were one time aerated with 10 ‘ , 15 ‘ and 20 ‘ exposing clip of each SO2 and NO2 at optimal pH values for both microalgal civilizations after the 10 yearss of incubation period. Samples were taken at the terminal of the 11, 13 and 15 yearss for each experiment.
2.5. Analytic methods
The chlorophyll a, chlorophyll B and entire chlorophyll concentrations were determined for each strain by entering optical soaking up at 646.6 and 663.6 nanometer at the terminal of 11, 13 and 15 yearss with the method that developed by Porra et Al. [ 29 ] . The chlorophyll concentrations expressed in µg of chlorophyll per millilitre.
The OD600nm was measured with a Shimadzu UV 2001 theoretical account spectrophotometer at the same frequence. Cell dry weight was obtained by the measuring of the pellets that centrifuged at 5000 revolutions per minute for 10 min and after dried at 80 & A ; deg ; C overnight. Centrifugation and drying were performed utilizing a Hettich EBA 12 theoretical account extractor and N & A ; uuml ; ve FN 400 theoretical account sterilisation, severally. The settlement figure was determined by the numeration of the settlements by microscopic scrutiny. All the measurings were performed in triplicate.
2.6. Statistical analysis
The experiments were set in a wholly randomized design up with three replicates. The informations were subjected to analysis of discrepancy utilizing important differences among interventions agencies were compared by descriptive statistics ( ±S.E. ) .
3.1. Consequence of initial pH on microalgal gas tolerance
The pH is one of the most of import environmental factors greatly act uponing non merely growing of micro-organisms but besides solution chemical science. The optimal pH of the culturing media is rather of import to heightening the tolerances of the microalgae to SOx and NOx [ 30 ] . To find the pH consequence experiments were performed with 10 ‘ exposing clip of acidic pollutants ( Fig.2 ) . As shown in the Fig.2a, B, degree Celsius and vitamin D pH consequence were determined at four different pH values for both microalgal civilizations. Fig.2a and B shows the pH consequence on chlorophyll a content of Synechococcus sp. The maximal chlorophyll a content was 0.93 µg ml-1 at pH 7.5 for SO2 and was 1.50 µg ml-1 at pH 7.5 for NO2 at the terminal of 15 yearss. The maximal chlorophyll a contents were 0.28, 0.35 and 0.19 µg ml-1 at pH 6.5, 8.5 and 9.5, for SO2, severally. Synechococcus sp. had much more tolerance to NO2 than SO2. The chlorophyll a contents were 0.62, 0.91 and 0.99 µg ml-1 at pH 6.5, 8.5 and 9.5, severally.
& A ; lt ; Here Fig.2 & A ; gt ;
a ) B )
degree Celsius ) vitamin D )
Figure 2. The consequence of initial pH values on chlorophyll content of Synechococcus sp. and Chlorella sp. at 1 mol l-1 SO2 or 1 mol l-1 NO2 concentrations after 15 yearss ( TRIA concentration, 1 mg l-1 ; NaHCO3 concentration, 34 mg l-1 ; exposing clip 10 ‘ ; light, 2400 sixty ) . a ) Synechococcus sp. – SO2, B ) Synechococcus sp – NO2, degree Celsius ) Chlorella sp. – SO2, vitamin D ) Chlorella sp. – NO2.
The information given in Fig.2c and vitamin D shows the pH consequence of acidic pollutants on Chlorella sp. The civilization had much more tolerance to both of the acidic pollutants than Synechococcus sp. The chlorophyll ( a+b ) contents were 2.07, 2.72, 3.06 and 1.88 µg ml-1 at pH 6.5, 7.5, 8.5 and 9.5 for SO2, severally. The civilization besides showed much more tolerance to NO2 and chlorophyll ( a+b ) content increased from pH 6.5 to 8.5 and so decreased to pH 9.5. The maximal chlorophyll ( a+b ) content was 3.3 µg ml-1 at pH 8.5.
3.2. Consequence of gas exposing clip on microalgal gas tolerance
Exposing clip, has a significant impact on the finding of mark experiments, was determined at optimal pH values for both microalgal civilizations. Exposing clip may change from proceedingss to hours harmonizing to the biomass and mark analyses. In the visible radiation of the above information, a clip scope from 10 ‘ to 20 ‘ was tested for SO2 and NO2 gases. The exposing clip consequence on chlorophyll content, dry weight and settlement figure were comprehensively given above.
The acidic pollutants would do toxic effects on the microalgal growing or photosynthesis. To turn to the photosynthetic suppression or tolerance we besides researched chlorophyll contents. To happen the maximal gas exposing clip that the civilizations able to digest and were inhibited experiments were set up to 15 ‘ and 20 ‘ . The consequences were recorded on 10 ( without gas ) 11, 13 and 15 yearss ( with 10’,15 ‘ and 20 ‘ gas exposing clip intervals ) . It was recorded that when exposing clip increased, chlorophyll values of both microalgal civilizations decreased. Fig.3 and 4 nowadayss all of the exposing clip effects. As shown in Fig.3a, exposing clip of 10 ‘ did non suppress the increasing of microalgal chlorophyll content until the twenty-four hours 13. The chlorophyll content was 0.8 µg ml-1 on twenty-four hours 10 and increased to 1.03 µg ml-1 on twenty-four hours 13. The civilization began to be inhibited at the terminal of 15 yearss and chlorophyll content decreased to 0.93 µg ml-1.
There was merely a small addition in 11 yearss at 15 ‘ ( 0.49 µg ml-1 ) and 20 ‘ ( 0.39 µg ml-1 ) SO2 gas exposing clip harmonizing to the informations and after 11 yearss Synechococcus sp was quickly and well inhibited. The chlorophyll a content was 0.16 µg ml-1 at 15 ‘ and 0.13 µg ml-1 at 20 ‘ at the terminal of 15 yearss.
Fig.3b shows the consequence of NO2 exposing clip on Synechococcus sp. As seen in the figure NO2 did non suppress the microalgal civilization at 10 ‘ on the contrary had a positive consequence. The making to chlorophyll values in a short clip could be explained that photosynthesis is much less sensitive to the presence of N oxides than other pollutants [ 34 ] . The chlorophyll a content had a rapid addition at 10 ‘ on 10 yearss ( 0.82 µg ml-1 ) and had a stable consequence at 15 ‘ . For illustration chlorophyll a content was 0.48 µg ml-1 on 10 yearss and was 0.48, 0.45 and 0.43 µg ml-1 at 15 ‘ on 11, 13 and 15 yearss, severally. There was a similar consequence between the 20 ‘ and 15 ‘ consequences until the twenty-four hours 13, but at the terminal of the 15 yearss chlorophyll a contents pronounced declined to 0.19 µg ml-1and photosynthetic suppression of Synechococcus sp. was occurred.
& A ; lt ; Here Fig.3 & A ; gt ;
a ) B )
Figure 3. The consequence of SO2 and NO2 exposure clip on chlorophyll a content of Synechococcus sp. after 15 yearss ( TRIA concentration, 1 mg l-1 ; NaHCO3 concentration, 34 mg l-1 ; light, 2400 sixty ) . a ) SO2 consequence, B ) NO2 consequence
The experiments showed that 20 ‘ exposing clip of both SO2 and NO2 could significantly suppress the growing of Synechococcus sp. cells. On the other manus we saw better consequences for Chlorella sp. than Synechococcus sp. when civilization aerated with SO2 during 10 ‘ civilization did non suppress ( Fig.4a ) . The photosynthetic suppression consequence of SO2 was more singular than NO2. Chlorophyll ( a+b ) content was 1.66 µg ml-1 at 0 ‘ on 10 yearss and increased to 3.06 µg ml-1 at 10 ‘ on 15 yearss. Culture was able to digest SO2 when was applied until 10 ‘ and than chlorophyll ( a+b ) content began to fall after 11 yearss and was 1.51 µg ml-1 on twenty-four hours 15 at 15 ‘ SO2 exposing clip. Chlorophyll ( a+b ) content decreased significantly at 20 ‘ SO2 exposing clip from 1.56 µg ml-1 on twenty-four hours 10 to 1.08 µg ml-1 on twenty-four hours 15.
& A ; lt ; Here Fig.4 & A ; gt ;
a ) B )
Figure 4. The consequence of SO2 and NO2 exposure clip on chlorophyll ( a+b ) content of Chlorella sp. after 15 yearss ( TRIA concentration, 1 mg l-1 ; NaHCO3 concentration, 43 mg l-1 ; light, 2400 sixty ) . a ) SO2 consequence, B ) NO2 consequence
As present in the Fig.4b, NO2 had rather a batch of positive consequence at 10 ‘ and partly at 15 ‘ . Chlorophyll ( a+b ) was 2.02 µg ml-1 on twenty-four hours 10 and increased to 3.14 µg ml-1 and 3.30 µg ml-1 on twenty-four hours 13 and 15, severally. Chlorophyll ( a+b ) decreased from 2.34 µg ml-1 to 2.23 µg ml-1 from twenty-four hours 10 to 15 at 15 ‘ NO2 exposing clip. There was important negative consequence on the civilization at 20 ‘ NO2 exposing clip, chlorophyll ( a+b ) decreased to 1.13 µg ml-1 on twenty-four hours 15. This might be explained that NO2 and SO2 foremost dissolves in aquaculture and at higher times react with microalgae. The decrease in NO2 occurred with in 10 minute and continued easy depending on TRIA and NaHCO3 in aquaculture.
It is good known that acidic pollutants had a negative consequence on microalgal biomass [ 9,11,12,30 ] . For that ground we wanted to look into how to impact SO2 and NO2 on microalgal biomass and determined dry weight and settlement figure after the gas application for each SO2 and NO2 gases and both microalgal civilizations. Table 1 is a sum-up of all the informations that was determined at the terminal of 15 yearss. As discovered in the chlorophyll contents, both dry weight and settlement figure were high in civilizations that were exposed to NO2.
& A ; lt ; Here Table 1 & A ; gt ;
Table 1. The consequence of SO2 and NO2 exposure clip on dry weight ( X ) ( g l-1 ) and colony figure ( x104 settlement ml-1 ) of Synechococcus sp. and Chlorella sp. after 15 yearss ( TRIA concentration, 1 mg l-1 ; NaHCO3 concentration, 34 milligram l-1 for Synechococcus sp. , 43 milligram l-1 for Chlorella sp. ; light, 2400 sixty ; SO2 and NO2 concentration, 1 mol l-1 )
Ten ( g l-1 )
Colony figure ( x104 settlement ml-1 )
2.79 ± 0.40
2.83 ± 1.29
0.48 ± 0.54
1.66 ± 1.36
0.39 ± 0.11
1.00 ± 0.58
4.32 ± 0.55
4.50 ± 0.86
1.29 ± 0.27
3.60 ± 0.58
0.57 ± 0.36
1.66 ± 1.37
2.65 ± 0.47
9.00 ± 0.89
1.31 ± 0.10
3.00 ± 1.13
0.94 ± 0.18
2.00 ± 1.03
2.86 ± 0.17
10.5 ± 1.03
1.93 ± 0.23
8.60 ± 0.52
0.98 ± 0.37
2.66 ± 0.89
The dry weight decreased from 5.01 g l-1 to 3.73 g l-1 and 2.38 g l-1 for Synechococcus sp. when SO2 exposing clip increased from 10 ‘ to 20 ‘ . In parallel with these consequences, colony figure was 2.83, 1.66 and 1.00 x104 settlement ml-1 at 10 ‘ , 15 ‘ and 20 ‘ , severally. The maximal dry weight was 6.44 g l-1 and colony figure was 4.5 x104 settlement ml-1 at 10 ‘ NO2 exposing clip.
The dry weight consequences of Chlorella sp. that exposed to SO2 were similar with the consequences of Synechococcus sp. that exposed to NO2. So that dry weight was 6.23 g l-1 at 10 ‘ and decreased to 3.70 g l-1 at 20 ‘ SO2 exposing clip. The highest consequences in table were NO2 experiments. The maximal dry weight was 6.63 g l-1, 4.85 g l-1 and 3.76 g l-1 and the settlement figure was 10.5, 8.60 and 2.66 x104 settlement ml-1 at 10 ‘ , 15 ‘ and 20 ‘ NO2 exposing clip, severally. These consequences indicated that 10 ‘ and 15 ‘ exposing clip was non repressive to microalgal biomass under these conditions.
There are two sorts of major of import jobs, environmental pollution and higher energy depletion jobs. Microalgae have the answer to many of the planetary jobs. The usage of microalgae to repair CO2 and digest other pollutants produce energy and aid to clean the environment, so microalgae may potentially work out all of our jobs.
The survey was designed to get a better apprehension of some basic microalgal growing and suppression responses under consequence of acidic pollutants and the possible physiological enhance over biomass production in a stirred civilization. The consequence of different parametric quantities was studied by the aid of the gas manufacturer and transportation system. The advantages of the system are ( 1 ) can be easy established and used in each research lab without any cost ; ( 2 ) allows the green goods and direct transportation of different gases through assortment of chemicals ; ( 3 ) the transportation of SO2, NO2 or other gases can be achieved by increasing gas exposing clip and this clip can be adjusted at the optimum scope of different gases.
Acidic pollutants in fluke gas, such as SO2 and NO2 might do negative effects on the growing and physiology including suppression of photosynthetic pigments, acidification of civilization and suppression of microalgal growing. Therefore, it may be concluded that adding growing stimulators to the civilization media was an effectual method to heighten the tolerances of microalgae to acidic pollutants. A inquiry is why these negative effects may be minimum at lower exposing clip degrees. Our experiments pointed out that lower unmasking degrees of acidic pollutants can be significantly tolerated by both of the microalgal civilization by the aid of growing foils. It is besides interesting to compare which species is better for this procedure. For comparing of pertinence of microalgal civilizations under straight SO2 and NO2 gases, Chlorella sp. reveals a better tolerance to acidic pollutants than Synechococcus sp. It could be explained by the response of different genus and spp. show differences in aquaculture as it can be expected. Therefore research of the pollutant tolerant species will be need for farther surveies.
Three chief decisions can be explained ; First, CO2 in NaHCO3 can significantly heighten biomass output with a growing stimulator TRIA. Second, acidic constituents may do important harm to microalgal chlorophyll content and biomass and therefore they need to be used to a safe clip degree and thirdly, the presence of NOx in microalgal civilization did non suppress in harmonizing with the findings of Doucha et Al. and Hauck et Al. [ 10,12 ] .
Some of the of import parametric quantities such as temperature and pH besides were detected during the experiments. A important difference was non observed on the initial temperature of the civilization media after the transferring of SO2 and NO2 gases at different clip. The temperature was 25 ± 2 & A ; deg ; C before the acidic pollutants, and decreased to max 25.75 ± 2 & A ; deg ; C after the acidic constituents.
It was reported that the initial optimal pH degree was 7.5 for Synechococcus sp. and was 8.5 for Chlorella sp. into the SO2 and NO2 experiments. Depending on nature of the biomass, this happening optimal pH values for different surveies have been reported for blue-green algae and Chlorella sp. For illustration, Lawry and Jensen [ 31 ] reported that Synechococcus sp. showed its maximal phosphate consumption at pH 7.5-8.5 scope and Baebprasert et Al. [ 32 ] explained that Synechocystis sp. showed its maximal hydrogenase activity at pH 7.5. On the other manus Su & A ; aacute ; rez et Al. [ 33 ] discovered the pH consequence of the binding of metals in solution by Chlorella vulgaris with a scope of pH 4.0, 6.0 and 8.0 and they found the higher per centum of adhering at alkali pH 8.0.
The biomass productiveness of Chlorella sp. was higher than that of Synechococcus sp. , besides the entire chlorophyll content of Chlorella sp. was higher. The following singular topic is why Synechococcus and Chlorella sp. cells are inhibited under high transportation degrees of acidic pollutants? One of the possibilities is an acidification of the civilization media. The pH alteration is one of the significant differences between the two types of microalgae which the consequence of different initial pH degrees. It has been good documented that when the acidic pollutants are transferred into the civilization media, the pH of the media go down and this was a ground of the disintegration of acidic pollutants in civilization media [ 35 ] . In this survey the pH of the growing medium was about 3.0 and 4.8 for Chlorella sp. and 2.59 and 2.72 for Synechococcus sp. at 15 ‘ exposing clip of SO2 and NO2, severally and went down at approximately 2.50 when exposing clip increased to 20 ‘ . In a old study, pH markedly decreased from 7.3 to 2.3 for Chlorella vulgaris and the lessenings in the pH degrees did non inhibited the growing of Cyanidium caldarium [ 12 ] . It was besides possible to command the pH scope to be able to digest the SO2 and NO2 gases [ 30 ] and they reported that a important lessening in pH from 5.8 to 3.0 was observed after 12 h. for Chlorella sp. and they found that Chlorella sp. was wholly inhibited from the beginning if pH was non controlled hence, they controlled pH to 7.00 by adding NaOH solution. In this survey the pH of the Chlorella sp. was more alkali than Synechococcus sp. before the transportation of the acidic pollutants and more tolerated to these constituents. On the other manus the Chlorella cells did non wholly suppress. This was peculiarly encouraging because the presence of acidic pollutants in aquaculture could do wholly suppression of growth.These findings suggested that a big sum of microalgal biomass may be used for farther surveies. It is clear that all of the above consequences have provided many of the advantages and belongingss to microalgal tolerance of acidic pollutants under consequence of growing stimulators toward the future practical applications.
The use of Synechococcus sp. and Chlorella sp. civilizations on the tolerance of acidic constituents with growing stimulators was investigated for the first clip. The important consequences from the survey can be summarized as follows ;
TRIA and NaHCO3 helped to microalgal growing under consequence of SO2 and NO2,
Chlorella sp. dominant microalgal civilization was much more tolerance to acidic pollutants than Synechococcus sp. ,
NO2 did non merely do negative effects but besides behaved as an foil on the growing of both microalgal civilizations.
A different attacks to digest acidic gases by the usage of two different microalgal species was investigated. The survey suggested that use of this method is executable and an case for the farther probe into civilization physiology under consequence of acidic constituents and environmental surveies, with no major engineering and cost are needed.
The writers are thankful to the Scientific and Technological Research Council of Turkey ( T & A ; Uuml ; B & A ; deg ; TAK-B & A ; deg ; DEB ) for fiscal support and want to show their gratitude to Prof. Dr. Orhan ATAKOL and Prof. Dr. Mustafa TaAYtekin ( Ankara University, Faculty of Science, Department of Chemistry ) for their valuable remarks.