Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Year IX, No2 - 2010
RESEARCH AND ACHIEVEMENTS FOR NEW ELECTROCHEMICAL BIOSENSORS
Gheorghe GUTT1, Yarema TEVTUL2, Andrei GUTT1, Alina PSIBILSCHI1
1 Faculty of Food Engineering, Stefan cel Mare University, Street. Universitatii no.13, 720229, Suceava, Romania, [email protected] 2Yuriy Fedkovych National University of Chernivtsi, Kotsiubynsky St., 2, Chernivtsi, 58012 Ukraine
Abstract. The paper presents results and research of a team involved in instrumental analysis from Faculty of Food Engineering of Suceava University in biosensors field, for food, health and environment issues. Starting from previous achievements were developed electrochemical performance biosensors, extensive use, for analysis both in situ as well as in the laboratory, mainly in pursuing the universal use of such equipment in all reactions using as catalyst type oxidase enzymes. Another aim was to increase sensitivity of measurement and accuracy of such equipment, also by proposed solutions were removed single-use kits, commonly used in the realization of biosensors by using watertight vials containing oxidase which is enough for hundreds of tests. Combined biosensor described in this paper has the great advantage of using in the amperometric and conductometric methods only their advantages since the two electrochemical methods are complementary, also the dosing system of oxidazes is simple and accurate ensuring a good reproducibility of experimental data. By the avant-garde research and achievements of the team itis opened the way for development of new types of biosensors.
Keywords: amperometric and conductometric biosensor, universal biosensor, dual biosensor
Introduction. such as: electric charge, absorption / photon emission, temperature, refractive Biosensors are biological-selective index, layer thickness, values that are electronic integrated systems, consisting converted by transducers type of a biological active receiver, a amperometric, pH meters, photometers, transducer and an electronic amplification, thermal, refractive, piezo oscillator, surface processing and display data. Biological plasmon resonance, in proportional active receptor provides specific analytical electrical signals to the concentration of information enabling recognition of certain the species sought in the matter under biological or chemical species in a review. complex mixture that consists matter under Basic applications for biosensors are in review, followed by quantitative or semi- medicine, environmental quality control quantitative determination of it. issue, pharmaceutical industry and Following the interference of biological cosmetics, food industry and in active receptor and analytes resulted biotechnology. Experimental measurement changes of raport between reactants and with biosensors are simple and require no products of the reaction that causes in turn, specialized advanced knowledge but only depending on active biological used the user browsing of short successive systems, proportional evolution of stages in time. physico-chemical measurements values
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New Methods for achieving detection is a portable glucose biosensors electrochemical amperometric used to determine in situ blood sugar for biosensors diabetics and athletes. Glucose sensor has as an active biological material a Amperometric biosensors are used for the glucosoxidase enzyme used as biocatalysts detection of reaction products or in oxidation reaction of glucose with metabolites that oxidizes or shall slightly oxygen in the air with obtaining reduces. Amperometric method consists in gluconolactone and hydrogen peroxide measuring the electrical current of a (H2O2 -hydrogen peroxide) as products of miniature electrolysis cell electrolysis (this reaction. Biologically active receptor for is a biosensors system), fed at a constant rapid analysis of blood is in the form of voltage. single-use plastic strips that has at one end According to Faraday's law, a mass m of a two miniature plate electrodes between chemical species discharged from an which is deposited glucoseoxidase in a electrode of an electrochemical cell at a form of dry gel that containes a conductive time t is proportional to current I of polymer and at the other end the strip has electrolysis and with electrochemical electrical pine connection with the constant k: electronic system. When submitting a drop of blood on the area covered by the m k I t (1) glucoseoxidase the folowing reaction occurs: Currently, one of the most popular applications in biosensors amperometric
D Glu cose O Glucozoxidase Gluconolactone H O 2 2 2 (2)
Hydrogen peroxide is highly reactive and polyphenols, flavonoids and for the breaks down at the electrodes of development of substances with electrochemical cell, current intensity of antioxidant activity [3], [4] electrolysis is proportional to the amount Below are listed some typical examples of decomposed and proportional to the biosensors that use different enzymes from amount of glucose in analysed blood . oxidase type, as a catalyst, the result being Such biosensors systems can be used for also hydrogen peroxide obtaining: determining concentration of phenols,
Colesterol O ColesteroloxidaseColestenone H O 2 2 2 (3) Glutamate O DGlutamatoxidaseOxiglutarate NH H O 2 3 2 2 (4) Lactate O LactatoxidasePiruvat H O 2 2 2 (5) Piruvat HPO Piruvatoxidase Acetilfosfate CO H O 4 2 2 2 (6)
The concerns of analytical instrumental Engineering of Suceava University group from the Faculty of Food focused on improving the amperometric
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biosensors. Research focused on the In this issue is shown that using following directions: amperometric method, in which 1. achievement a conductometric electrolysis current is also adder, enables biosensor as a superior alternative to reducing the error by limiting electrolysis amperometric biosensors voltage of hydrogen peroxide at the Nernst 2. design and implementation a voltage. conductivity-amperometric biosensor To combine the advantages of both combined with concurrent analysis by both methods and to eliminate the methods, biosensor that can be used in disadvantages of each of them the team has reactions 1-6 using oxidase enzyme as developed a combined biosensor type catalyst conductometric - amperometric [3], [4] for 3. extending limits of concentrations of a the clinical laboratory and analytical food new type of biosensors from maximum In addition to those mentioned this 500mg/dl, as they are encountered in biosensor has a great advantage of not blood, to limit of 20% glucose, the latter working with expensive kits of single-use concentration being specific to analytical but with glucoseoxidase and pure liquid food, particular applications in colesteroloxidase which is precisely fermentative processes dosed[5], [6] using the equipment designed 4. elimination of chemical kit for single by collective use from typical biosensors Starting from the results of theoretical and Research have focused initially only on applied research has been developed a glucose biosensors [1], [2], [4], [6] and portable dual biosensor [7], [8] that allows were mainly intended to find an alternative concurrent determination of both glucose to low reproducibility of typical portable and cholesterol from a single drop of biosensors used to determine blood blood, but it is not the subject of this paper. glucose This low reproducibility is due to Contributions to conception, design and electrolysis phenomenology, electrolysis implementation of combined current, so the indication of glucometer, electrochemical biosensor with superior depending on the addition of hydrogen performance peroxide concentration and other parameters relating primarily to ionic Research team has designed and developed transport phenomena that lead to advanced a prototype of enzyme biosensor that uses electric polarization at electrodes that alter oxidase type enzymes as a catalyst in current yield. liquid form, biosensors being capable of Very promising results obtained in rapid determinations and in situ of experimental research [1], [2] have chemical or biological concentration of confirmed that using conductometric species without using of single-use method with frequency of 5 kHz eliminates biological kits [4 ], [4]. The advantages of the phenomenon of polarization and also using this measurement system are: allows determination both of glucose low - obtaining an universal and reliable concentration, as they are present in blood, biosensor a long-term usable in all types of as well as the concentration at high levels reactions catalyzed by enzymes type up to 20% , specific to analitical food. oxidase Using conductometric method presents - eliminating single-use kits and thereby also a disadvantage, namely that the value significantly lowering of cost price of the of conductivity is an adder for all analysis electrolytical species of blood. - combined use of both amperometric
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and conductometric method in the same of measurements determinations, it ensure a high a precision
Fig.1.Amperometric and conductometric scheme of measurement principle with enzyme biosensor. 2- acquisition electronics unit, data processing and display, 3-amperometric electronic unit, 4- conductometric electronic unit, 10- silicone hose, 14,15-platinum electrodes, 17-cup, 21- pipette or manually electronic unit.
In Figures 1.3 are represented elements of principle, construction and function of combined biosensors [4].
Fig.2.Sectional view of enzyme biosensors. 1-body 2- electronics unit of acquisition, data processing and display, 3-electronic amperometric unit, 4-conductometric electronic unit, 5-keyboard, 6- plastic capsule, o- oxidase solution, 7-bolt, 8-marker, p-drill, f-strip, c-con, 9-nut, 10-silicone hose, 11- supported eccentric cylinder l-conical seat, 12- ratchet, 13-bar 14.15-platinum electrodes, 16-con, 17-cup, 18.19- electric contacts, e-slot 20-ratchet ball, 21- pipette or manually hopper.
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The procedure to work with biosensors is system, of required volume for oxidase in the folowing: it is maximum screwed up 7 cup 17, with the stop of cylinder 11 cav screw then returned device to 1800 eccentric supported on full shuttered over normal working position, place the position of silicone hose, position indicated cap with oxidase in cavity of the screw 7 by arrow signs on the click swing button and after it is screwed by hand until it is but also by the ball's ratchet 12. sensitive mechanical strength which means It follows the dosage in cup 17 of the that strip of capsule 6 was pierced by the prescribed volume for examined species by knife p of the marker 8 and capsule neck 6 a dosing pipette 21 or manually hoppers sealing made with the marker 8, then after the device is keeped upright and returned again biosensors in working digital display is intended. Triggering of position and screw up removable cup 17 catalyzed reaction lead to the emergence of on rod 13, it is running several full first quantities of hydrogen peroxide, a rotations to the right of the cylinder 11 threshold of its concentration causes eccentric supported until on the perforated initiation amperometric and cone 16 it is a drop of oxidase conductometric measurements, these are After these operations the cup 17 is alternatively carrying by an automatic screwed and it is executed a full rotation of electronic switching. eccentric cylinder to dosing, by peristaltic
Fig.3.Exterior front and side of enzyme biosensors, 1-body, 2- electronic unit acquisition, data processing and display, 3-amperometric electronic unit, 4- conductometric electronic unit, 5-keyboard, 6- plastic capsules, o- oxidase solution, 7-bolt, 8-marker, p-drill, f-strip, c-con, 9-nut, 10-silicone hose, 11-cylinder eccentric supported, l-conical seat, 13-rod 14.15 platinum electrodes, 16-con, 17-cup, 21- pipette or manually hoppers.
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After 10 seconds is displayed the achievement of combined performance concentration value resulting from the biosensor that combines the advantages of average of dozens of amperometric and both electrochemical methods of conductometric measurements statistically measurement. processed by biosensors microprocessor. Devising a simple and accurate dosing The volume of oxidase capsule reaches system for catalytic enzymes type oxidase hundreds of determinations, between two and its implementation on the biosensor determinations with the same oxidase is allowed elimination of single-use kits that not required than washing under running determine the decisive price of analysis. At water of cup 17. On oxidase changing the the same time other design and whole route is washed using for this construction solutions embedded in the purpose a capsule with double distilled architecture of biosensors allow its water which is fixed, as already was universal use in all catalyzed reactions by described, on the body 1 of biosensors and oxidase run more full rotation of cylinder 11 Theoretical and applied research carried supported eccentric supported for pumping out allowed the design of new biosensors water through the circuit. In some one being a dual portable biosensor that situations it is necessary to replace the allows co-determination both of glucose silicone hose 10, these cases occur in and cholesterol in blood using for this specific applications where the volume of purpose a single drop of blood. This type dosing oxidase is different from the of biosensor is not the subject of this work, previous application. This requires research and prototype implementation are replacement of silicone hose with another still pending. with an greater inside diameter, that smaller, the operations arising in a few References seconds. The replacement of silicone hose 10 is required and its wastage in 1. GUTT G., SUMUSCHE I., GUTT A., Amperometric biosensors for glucose compression zone-stretching as a result of determination, Anals of the Suceava University- numerous pumping of eccentric supported Food Engineering, Year VII, No. 1, 2009, p 5-11 cylinder 11. At hose replacement it unwind 2. GUTT G., GUTT A., SUMUSCHE I., Research on achievement an conductometric enzyme biosensor, the nut on the opposite side of the Anals of the Suceava University-Food Engineering, actuating button of cylinder 11, after which Year VII, No. 1, 2009, p 11-18 it is extracted from the body 1 of 3. GUTT S., GUTT G, GUTT A.- Biosenzor , 0 Propunere de Inventie , Dosar OSIM A/00285/2008 biosensors, it follows the return with 180 4. GUTT A, GUTT S., GUTT G, - Biosenzor of the device from the working position, is enzimatic, Propunere de Inventie , Dosar OSIM unscrew the screw 7, extract of capsule 6 A/00857/2009 5. GUTT S., GUTT G, GUTT A, - Biosenzor and marker 8 from its slot, full unscrew of miniatural, Propunere de Inventie , Dosar OSIM nut 9, extraction of silicone hose on the A/00 854/2009 marker 8, followed by extraction of the con 6. GUTT A, GUTT S., GUTT G, - Biosenzor de glucoza, Propunere de Inventie , Dosar OSIM, 16 from rod 13, retrieval of hose 10 from A/00848/2009 cone 16, at the mounting of new siliconic 7. GUTT S., GUTT G, GUTT A, -Biosenzor de hose operations are repeated in reverse of laborator pentru glucoză şi colesterol, Propunere de Inventie , Dosar OSIM, A/00161/2010 of the dismantling of 10 silicone hose. 8. GUTT S., GUTT G, GUTT A, PSIBILSCHI A.- Biosenzor portabil pentru glucoză şi colesterol , Conclusions Propunere de Inventie , Dosar OSIM, A/00162/2010
Using both amperometric and conductometric principle it made possible
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MONITORING OF CONTAMINATION OF THE ENVIRONMENT IN BUKOVYNA WITH HEAVY METALS
Yarema TEVTUL1, Olga NECHYPORENKO1
1 Yuriy Fedkovych Chernivtsi National University, Kotsiubinsky St., 2, Chernivtsi 58012, Ukraine. E-mail: [email protected]
Abstract. The main sources of soils contamination with heavy metals have been analyzed. The results of monitoring agricultural soils contamination with heavy metals and the content of metals in the plant tissues in Chernivtsi region have also been registered. The average content of heavy metals in the agricultural soils of Chernivtsi region complies with the corresponding background values. An average content of the following heavy metals in a 0-5 cm horizon of the agricultural soils of Chernivtsi region was registered (mg/kg): 9,6 (Pb), 0,26 (Cd), 30,4 (Cu), 30,5 (Ni); 72,0 (Cr); 78,3 (Zn); 973 (Mn). Similar values for the urban soils within Chernivtsi were noticed as well: 51,3 (Pb), 0,3 (Cd), 79,2 (Cu), 25,3 (Ni); 41,0 (Cr); 73,4 (Zn); 654 (Mn). Heavy metals were arranged in a sequence according to their toxicity, which also varies for different organisms and content of free ions. The dependence of the metal ions content in food and the plants growing and cooking conditions has been analyzed. Some ways of heavy metals accumulation in plants are described. Local areas with higher concentration of heavy metals have been identified near roads and within some gardens. No such areas were found near industrial objects. The content of heavy metals in the soils of Prykarpattya and Prut-Dnister upland area remains below the background level, which ensures secure conditions for agricultural activities. A comparison between the heavy metals content in Bukovyna and other regions of Ukraine has also been made.
Keywords: monitoring of soils, accumulation of heavy metals, plants, human body, food chain.
Contamination of the environment with the study of various controllable and heavy metals compounds is very uncontrollable parameters’ effect on the dangerous since they can get into the transformation of pollutants. In this way human trophic sequence. The more stages we can simulate and forecast possible consists the sequence, the higher results of eco-catastrophes and massive accumulation ratio of toxic agents can be pollution of the environment and promote reached at the end. It is known that every better natural self cleaning by using plants, new stage of the sequence results in about which can absorb pollution agents. ten times higher concentration of the toxic Chemical compounds of heavy metals agents. cause structural changes in the soils Investigation of the environmental microcenosis. Sensitive species are being contamination with heavy metals is quite a gradually substituted by more metal- topical issue, which includes the study of resistant ones and rehabilitation of the pollutants toxic action on organisms, metal-polluted soils requires fixation of possible accommodation of the organisms heavy metals compounds in the indelible for functioning in a polluted environment, organomineral complexes.
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Heavy metals can be allocated in the decomposition inside the engine and then following descending toxicity sequence: this dust is being released together with mercury; silver; copper; cadmium; zinc; other combustion products. Long lead; chromium; nickel; cobalt. However, exploitation of TEL resulted in significant this sequence can be rearranged for accumulation of various lead compounds different organisms and depending on the along many roads. Car-related emission of nature of the acting compounds (free ions, lead made about 80 % of the total inorganic or organic compounds) [1]. anthropogenic lead emission. Hard cars Heavy metals ions oppress the functions of traffic also results in emission of some organisms by the ferment systems other toxic agents: compounds of blocking, strong bonding with the sulfide cadmium, zinc, copper, oxides of nitrogen, groups and the cell walls crippling [1]. carbon (II), hydrocarbons and aldehydes. Some microbes normally require ions of Cadmium can be emitted as a result of car cobalt, copper, iron and others, which can tires wearing, destruction of asphalt road be part of bioactive compounds: ferments, cover, leakage of engine oils and, slightly, vitamins and pigments. These ions are from the old and worn-out cadmium-nickel important agents, which ensure normal batteries with destroyed bodies. redox processes, which occur in a cell. Many compounds of manganese are used Some ions are necessary to ensure normal in engineering and medicine. The structure and functioning of the microbial manganese (IV) oxide can be used as ribosome. doping for some special steels, for Micromycets, yeasts, some thionic discoloration of the glass mass, for microbes and pathogenic microorganisms production of linoleum, some dyes and are the most metal-resistant. This lacquers. Potassium permanganate can be resistance can be caused by their ability to used in medicine as a disinfectant [2]. accumulate and fix the heavy metal ions in Ferrous and nonferrous metallurgy, the cells. Therefore, the intracellular thermoelectric power plants, mineral content of heavy metals can become fertilizers and pesticides production, hundreds times higher comparing with the irrigation and melioration equipment, corresponding concentration in the industrial and municipal wastes and environment. Mycobacteria and wastewaters can also emit significant actinomycetes are less metal-resistant. amounts of heavy metals compounds. Therefore, it is important to know the The transport (annual emission over 28000 possible ways how heavy metals ingress in t), industry (about 600 t), wastewaters the human and animal trophic sequence discharge and disposal of pesticides are the and their concentrations in the soils and main sources of environmental pollution environment. In this way we can with heavy metals in the region of effectively counteract the metals Bukovyna [3]. accumulation. We briefly describe some of The regional center for soils’ fertility these ways below. protection and control of the production Some compounds of lead were used to quality carries out regular control of heavy prevent detonation in the internal metals content in the soils of Bukovyna. combustion engines for a long time. The analysis results have proved that this Tetraethyl lead (TEL) was the most widely content complies with current requirements used detonation preventer. The fine dust of (see Table 1). lead is formed as a result of TEL
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Table 1 Total content of heavy metals ions in the agricultural soils of Chernivtsi region, mg/kg Upper layer (0 ÷5 cm) Pb Cd Cu Ni Cr Zn Mn Average content in the 9.6 0.36 30.4 30.5 72.0 78.3 973 region Range of concentrations 3.0 ÷ 0.29 ÷ 12.0 ÷ 12.0 ÷ 32.0 ÷ 20.0 ÷ 500 ÷ within the region 32.0 0.82 40.0 50.0 200.0 100.0 2000 Average content in the city 51.3 0.3 79.2 25.3 41.0 73.4 654 of Chernivtsi Range of concentrations 3.0 ÷ 0.29 ÷ 5.9 ÷ 12.0 ÷ 32.0 ÷ 3.6 ÷ 500 ÷ within Chernivtsi 32.0 1.4 40.0 50.0 200.0 100.0 2000
Higher content of lead has been identified excessive throat/mouth washing with for some spots near main roads. Higher potassium permanganate solution may content of copper has been determined in cause edema of vocal cords and mucous the soil samples taken from orchards. This coat of mouth and throat. The ingestion of can be explained by the influence of plants’ concentrated potassium permanganate treatment with some pesticides. solution may result in the stomach wall No local areas with excessive heavy metals perforation. The ingress of manganese pollution resulting from the industrial compounds into the urinary bladder or activity have been identified. The heavy uterine cavity causes peritonitis [2]. metals content in the soils of Prykarpattya The highest accumulation of manganese region and Prut-Dnister upland complies compounds in the human body is with the background values and can not registered in liver. Urine and digestive threaten normal quality of the agricultural system are the main ways of the products [4]. manganese compounds excretion. Massive It should be emphasized that the content of burns of gullet and degenerative changes in manganese in the soils of Bukovyna is the parenchymatous organs have been higher comparing to other metals. reported after autopsy of the manganese This is an important feature because the pollution victims [2]. compounds of manganese are widely used Plants accumulate manganese mostly in in engineering and medicine and their roots. The accumulation coefficient is sometimes may have poisoning effect. 62,0÷86,0 % comparing to the content in Manganese can often be produced from its the soil (see Table 2). ores and minerals: pyrolusite, manganese As seen from Table 1, the distribution of (IV) oxide. The technology of pyrolusite manganese among parts of the corn can be processing requires its grinding, which represented as follows: stems contain produces a lot of fine dust. Inhalation of 1,1÷4,5 %, leaves – 6,5÷30,8 %, coats – the dust may lead to serious poisoning. The 2,1÷3,8 % and grains – 0,7÷1,4 %. normal content of manganese in the cell Therefore, we can arrange the parts in the tissues is very low [2]. descending line based on the content of Manganese compounds have strong manganese: protoplasmatic toxic effect and affect the roots>leaves>coats>steams>grains [5]. central nervous system, kidneys, lungs and the blood circulation organs. Even
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Table 2 Content of manganese in different parts of corn and for growing at different distances from the roads Distance from Content of Mn2+, g/g (for the dry weight) the road, m soil root stems leaves coats grains >5 360 144 8 21 5 2 25 375 165 6 12 4 1 50 345 139 3 54 5 1 100 300 116 3 54 5 2 150 285 175 2 58 5 1 200 225 120 2 59 7 2 lowest. The wastes were mainly formed in Plants require manganese to ensure normal the city of Chernivtsi, then in the districts processes of growing and cell breathing. of Novoseytsya (12,3 t) and Khotyn (11,5 This element also plays an important role t). in photosynthesis. The ion of manganese The total amount of the most works in many ferments such as in environmentally dangerous wastes is oxydase. It acts as a reducing agent in the gradually decreasing. This amount was process of nitrate feeding and as a strong 74,2 t at the beginning of 2008 or 22,1 % oxidizer in the process of ammonium lower compared to the beginning of 2007. feeding. Only 17 t or 23 % of this amount is Manganese toxicity has been reported for classified as the first risk class materials. the plants, which grow on acid soils [6]. Again, most part (69,9 %) of the waste High concentrations of manganese affect materials are collected in the city of the normal development of roots and Chernivtsi, then in Novodnistrovsk (9,5 t) decrease the productivity of synthesis of and in the district of Putyla (9,2 t). chlorophyll. High concentration of starch, Novodnistrovsk is an area of construction phenolic compounds and nitrates can of the Dnister hydropower and energy worsen nitrogen metabolism efficiency for accumulation plant and the local wastes the manganese-affected plants. resulted from this activity. The district of Incorrect management with dangerous Putyla is located in mountain area. Local industrial waste materials can bring anthropogenic activity mostly deals with another potential threat for plants’ growth the primary wood processing. Therefore, and soils’ quality. 225,7 t of such the wastes mainly consist of wood chips dangerous materials have been formed at and other wood processing byproducts, industrial units in the region of Chernivtsi which can not bring any serious in 2007 [7]. This is more than it was environment threat. formed in 2006 (180,4 t). All environmental dangerous materials There were 40,1 t or 17,8 % of the first risk have been removed from the storage class materials (threshold limit facilities in the agricultural districts concentration – 0,1 mg/m3); 81,6 t or 36,1 (Vyzhnytsya, Hertsa, Kelmentsi) of the % of the second class materials (0,1÷1 region in 2008. mg/m3) and 104 t or 46,1 % of the third The decrease in amount of the class materials (1,1÷10 mg/m3) among all environmental dangerous wastes of I-III the wastes collected in 2007. As seen from classes collected in various industrial this distribution, an amount of the most storages is shown in environmentally dangerous wastes was the Fig. 1.
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food. On the other hand, different plants can uptake different amounts of heavy metals from the environment. The redistribution of heavy metals among different plant organs can also influence their final content in the tissues. Some cultivated plants can be placed into the following sequence according to the increasing ability to accumulate heavy Figure 1. Changes in amount of the I-III classes metals: beans < oats and rye < barley < wastes collected in the region of Chernivtsi in corn << cabbage < melons < potato < 2000 – 2007. radish < onion < carrot < red beat <<
squash < cucumber < tomato << parsley < There is a significant part of metal dill < onion leaves < spinach < lettuce [8]. compounds among the waste materials There are two ways of heavy metals’ salts collected in the Chernivtsi region. The total ingression into plants: apoplasmatic and amount of metals is estimated at 17,6 t and symplasmatic. This leads in general to the lead and chromium compounds constitute tendency of decreasing of heavy metals 86,2 % of all metal wastes. Lead is present content in the row: root > stem > leaves > mainly in old car and other batteries. There seeds [8]. are also 2,2 tons of old luminescent lamps, The size of vegetables is an important which contain mercury. Such dangerous factor influencing the content of heavy wastes can be utilized only outside the metals. The larger is the vegetable, the region. higher is heavy metals accumulation value The changes in the amount of the most [8]. dangerous 1 class wastes are shown in Biocleaning of soils by planting some Fig.2. agricultural species is very interesting and The amount of stored prohibited and old an important field. For example, corn can pesticides and other agricultural chemicals actively accumulate heavy metals [5] and is continuously decreasing. In the year this plant can facilitate in the soils 2008 this amount became 18,2 times lower biocleaning. than in 2007. There are only 1,4 tons of Lead is mainly accumulating in roots and these chemicals remaining in the region. this tissue can accumulate up to 63,0-81,0
% of the total lead in the plant. The lead content in stem is much lower (3,8-10,1 %), in the leaves – 4,8-19,0 % and in the coats – 6,2-13,2 %. It is needful to emphasize that lead can ingress into leaves and coats mainly from the air. Only traces of lead were found in the corn grains. An average concentration of lead in the corn leaves was 4,9 mg/kg [5]. This is lower than the threshold limit value for the cattle Figure 2. Changes in amount of the 1 class feed (5 mg/kg). wastes during 2000÷2007 years. Contamination with lead is irreversible, i.e. Many various factors such as growing this pollutant is gradually accumulating in conditions, methods of processing can the surface soil layer even at very low and influence the content of heavy metals in gradual ingress. The dependence of lead
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contamination on the distance from roads plants can become dangerous even before is nonlinear. Roadside forest shelter belts having affected plants by poisoning and can capture some amount of the metal and drop in the crop yield should be noticed. prevent its spreading in the nearest Natural accumulation of heavy metals in roadside area. On the other hand, air fluxes some plants can be used for the phyto- can spread lead-containing dust over the rehabilitation of the contaminated soils belts at a longer range, which causes the [11, 12]. This problem has been highest content of lead in the soils at about extensively investigated by Galiulin et al 100 m from roads [9]. A content of lead in [11], who proposed the use of chelate 100 m from the road was found 2,2 times compounds of etylendiamintetraacetate exceeding its limit threshold value, in 50 m (EDTA). Chen [13] proposed to use citic – 1,2 times higher. Related exceeding over acid. Kravets [14] reported higher mobility the background content were 26,8 and 14,4 and accumulation ratio of the mineral times. Similar spatial distribution has also pollutants for the plants exposed to the been determined for cadmium. Its content non-specific stresses (presowing in 100 m from the road was 1,5 mg/kg or irradiation, deviations from the optimal 15 times over its limit threshold value [9]. growing conditions; too dense sowing). The high content of cadmium has also This effect can be useful for more intense been determined in the corn roots. Its phyto-rehabilitation. concentration was several times higher Biomass of the phyto-rehabilitation plants than in the soil. The root barrier prevents can be utilized for the bio-fuel production further distribution of the metal towards [15]. Some non-ferrous metals can even be the vegetation and reproductive organs. cost-effectively extracted from this That is why its content in the stem and biomass [16]. leaves is much lower and in the grains it is It is recommended to keep growing the only at the trace level [10]. food vegetables not closer than 150-200 m The accumulation of copper in the plants from the roads. Highly accumulating plants tissues is about 34,0-42,0 of the are not recommended for growth on concentration in the soil. The stem of corn metals-contaminated soils. Regional accumulates 8,4-13,5 % of the total copper educational programs should promote accumulated in the plant; leaves – 20,3- more active testing of the metals content in 28,6 %; coats – 12,7-21,1 % and grains – some domestic cultivated plants in 9,9-18,4 % [5]. specialized laboratories. Special legislation Copper is less labile element as compared regulations should prevent the growth of to other ones. The most part of copper food plants at dangerous distances from stays in the root and leave tissues until they roads while the increase of the ecologically die off and only very small part of the clean products should be encouraged and metal can get into new tissues. Copper is a stimulated by various means. Ecolabeling vitally needful microelement. Unlikely to standards should be widely introduced. In lead and cadmium it is required for normal this way a potential consumer would be plants’ growth. On the other hand, the able to see more adequate picture of the exceeding concentrations of copper cause foods ecosafety and can make a well- hard intoxications which result in chlorosis grounded decision of buying needful and defects of the root system. foods. Special “green” shops, which work It is hard to foresee, which concentration only with ecologically safe and clean of copper can cause toxic effect on plants. products, can be recommended in a more It should be remembered that the food distinct prospect. manufactured from metals-contaminated
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References
1. KUSHKEVYCH I., GNATUSH S., GUDZ 10. STRAND V., ZOLOTAREVA B, S. Influence of heavy metals on microbial cells. LISOVSKY A. Dependence of adding the water Bulletin of Lviv University. Biology. (2007) vol. soluble salts of lead, cadmium and copper on on 45, P. 3-28. their ingress and crop yields for some agricultural 2. KRAMARENKO V. Toxicological plants. Agrochemistry. (1991) P. 76-83. chemistry. (1989). Vyscha Schkola, Kyiv. 447 p. 11. GALIULIN R, GALIULINA R. 3. Agrochemical services of Bukovyna. Phytoextraction of heavy metals from contaminated Development and current fertility of the soils. soils. Agrochemistry (2003) P. 77-85 (2004). Chernivtsi. 75 p. 12. GOLOVACH O, DEMKIV O, 4. DMYTRUK YU. M. Ecological and Phytorehabilitation – an effective method of geochemical analysis of the soils in cleaning heavy metals-contaminated soils. Bulletin agroecosystems. (2006). Ruta. Chernivtsi. 328 p. of LvivVeterinary Academy. (2003). Vol. 5. P. 21- 5. GOLOVACH O., KOZLOVSKY V., 27. DEMKIV O. Pollution of the agricultural soils with 13. CHEN Y. X., LIN Q., LUO Y. M. et al. heavy metals and their redistribution in the corn. The role of citric acid on phytoremediation of Bulletin of Lviv University. Biology. (2004). Vol. heavymetal contaminated soil. Chemosphere. 38. P. 205-211. (2003). vol. 50. – p. 807-811. 6. KABATA-PENDIAS A., PENDIAS A. 14. KRAVETS O., Current conditions and Microelements in the soils and plants. (1989) Mir. problems of phytocleaning the soils from Moscow. 439 p. radionuclides and heavy metals. Physiology and 7. Environment of Bukovyna-2007. biochemistry of cultivated plants. (2002). vol. 34. P. Statistical volume. (2008). 144 p. 377-386. 8. DUBININA A., MALYUK L., 15. SHVED O. Ecological biotechnology. SELYUTYNA G. Toxic compounds in foods and Lvivska Polytechnica. (2010). 424 p. methods of their determination. (2007). Lipski S. Ekopaliwo z kukurydzy. Kukurydza. Professional. Kiev. 384 p. (2002). vol. 2, P. 25-27 9. GLADKIH E., Details of accumulation of heavy metals in the soils depending on their allocation. Bulletin of Lviv University. Biology. (2004). vol. 38. P. 205-211.
17 Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Year IX, No2 - 2010
MIXTURES WITH GRADIENT OF MOBILE PHASES UTILIZED IN HPLC SEPARATIONS OF 2.4-DINITROPHENYLHIDRAZONES PROVIDED BY INFERIOR CARBONYL COMPOUNDS
Gheorghe ZGHEREA1, Cristina STOIAN1, Sandu PERETZ2
1University “Dunarea de Jos” of Galati, Department of Chemistry, 111 Domneasca Street, 800201 Galati, Romania, [email protected], [email protected], 2 Institute of Physical Chemistry “I. G. Murgulescu”, Department of Colloids, 202 Spl. Independentei, 060021, Bucharest, Romania, [email protected]
Abstract. Many mixtures of small quantities of carbonyl compounds are present in foods, concerning sensorial qualities (aroma and fragrance). The inferior carbonyl compounds (C2-C4, boiling point <100°C) – mono and dicarbonyl – can be identified and their concentrations can be measured, after being separated by distillation on water bath. They are transferred into a strongly acid solution of 2.4- dinitrophenylhidrazine (2.4-DNPH), generating a mixture of insoluble 2.4-dinitrophenylhidrazones (2.4-DNPH-ones). The 2.4-DNPH-ones are organic compounds with weak polarity, solids, crystallized, yellow and water insoluble, but soluble in organic solvents. The mixture of 2.4-DNPH- ones may be separated by liquid chromatography, using HPLC the reverse phase mechanism [1-3]. This paper contains experimental and theoretical considerations on the means of separation through liquid chromatography of two models and a natural mixture containing 2.4-DNPH-ones provided by inferior carbonyl compounds; to obtain decisive results, in the model mixtures 2.4-DNPH-ones provided by carbonyl compounds having three (acetone and propanal) and four atoms of carbon (isobutylaldehyde) were introduced.
Keywords: acetaldehyde, diacetyl, 2.4-dinitrophenylhidrazone, reverse phase, low polarity, gradient of mobile phase
Introduction In many cases, for the foods obtained by properly physical and chemical behavior, fermentation, it is very important to know assuring a good separation [4]. The the concentration of diacetyl and acetal- difficulty appears for the natural mixtures dehyde. According to literature, beer concerning acetaldehyde and diacetyl; the contains diacetyl and acetaldehyde in the two carbonyl compounds – and 2.4- ratio 1:100, in mass units. The interest on DNPH-ones – have similar physical diacetyl concentration requires especially behavior. In addition, their mass ratio analyticcal conditions. It is possible to creates great problems in liquid-chroma- solve that problem by HPLC for the tographic separation. To have a very good mixtures of 2.4-DNPH-ones provided by analytical performance, it is necessary to inferior carbonyl compounds. HPLC can use liquid chromatography separation with make a good separation only for the model gradient of mobile phase [5]; in addition, mixtures of 2.4-DNPH-ones; the molecules separation columns with gradient of of inferior carbonyl compounds have stationary phase are used in this paper.
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Materials and methods Solvents and mixtures of mobile phases The acetonitrile was utilized to solve the according with the programs abbreviated pure 2.4-DNPH-ones. For liquid- MGMPI and MGMPII, containing bidisti- chromatographic separations, two similar llated water (2 S·cm-1) and methanol pro mixtures with gradient of mobile phase, chromatography (Merck) were utilized.
The model mixtures of 2.4-DNPH-ones The pure 2.4-DNPH-ones – yellow pow- liquid chromatography), 5·10-4 M solutions ders – were obtained in our laboratory, u- were obtained. By controlled mixing we sing a strongly acid solution of 2.4-DNPH obtained two model mixtures, abbreviated and chemical pure carbonyl compound; by MMI and MMII, considered as approximate synthesizing: 2.4-DNPHAA (acetaldehy- models of natural mixtures; the ratio bet- de), 2.4-DNPHD (diacetyl), 2.4-DNPHA ween the quantities 2.4-DNPHAA and 2.4- (acetone) and 2.4-DNPHiBA (isobutyl- DNPHD is higher to one, in each mixture. aldehyde). In acetonitrile (Merck, pro
The beer’s mixtures of 2.4-DNPH-ones The carbonyl compounds from beer were mixture of 2.4-DNPH-ones. They are separated by distillation on water bath and isolated by filtration, washed with pure transferred into a strongly acid solution of water, dried and solved in acetonitrile. 2.4-DNPH; the precipitates form a natural
Apparatus For the separations we utilized Pye Uni- of 0-99 minutes. Every moment of cam Philips liquid chromatograph, equi- analytical separation, the value of B, the pped with: an installation for degasing of percent of the second component in the mobile phase (by refluxing) [7], gradient mixture of mobile phase (A + B=100%), is programmer for mobile phase (type LC- %B k t n (1) XPD, able to mix two different liquids), where: separation columns with gradient of - t – dimension of timing segment stationary phase, installation for column (minutes), thermostat control, electronic integrator - k – slope of curve (describes the (type DP101, Spectra-Physics) and evolution of B value on the t segment), potentiometer recorder (type PM8251, - n – exponent, with values 0.0-9.9 Philips). The mixing program of liquids (describes the geometry of mixing curve). has nine segments of time, g=1-9; each timing segment has independent dimension
Conditions of chromatographic separations - sample volume: 10 L; - LC UV detector, = 365 nm; - separation column: L = 25 cm, = 4.6 - flow rate: 1mL·min-1; mm; - eluate: a controlled mixture of A-metha- - stationary phase: Spherisorb 5 ODS, with nol and B-water, accordingly to two pro- gradient of stationary phase; grams – MGMPI and MGMPII – achieved - 37.5ºC, the temperature of separation co- by the LC-XPD chromatographic module. lumn:
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The liquid-chromatographic separations Both categories of liquid-chromatographic only the program of mixing liquids in separations were carried out on the same binary mixture of mobile phase. column, at the same temperature, changing
Results and Discussion The model mixtures (MMI and MMII) and The first program of binary mobile phase, a natural mixture from beer were separated MGMPI (32 minutes, figure 1), contains by liquid-chromatography, using a mecha- four time segments; figure 2 shows the nism of reverse phase. chromatogram of model mixture MMI, The mixtures of mobile phases have the sa- obtained with MGMPI, with the retention me initially composition, with 45% water. times (seconds, in brackets). In this instance, the 2.4-DNPHAA will be According to MGMPI, the separation eluted before the 2.4-DNPHD. In this way begins with a mobile phase having 45% we guaranteed the maximum difference water; the mixture of mobile phase has a between the values of retention times for higher polarity. This mixture is hold during the mentioned 2.4-DNPH-ones and a pre- the first time segment, g = 1 (t = 12 liminary separation of the 2.4-DNPH-ones minutes), assuring a better resolution provided by aliphatic carbonyl compounds between the peak of 2.4-DNPHAA (844 s) with three and four atoms of carbon. The and the peak of 2.4-DNPHD (918 s). The model mixture MMI contains four 2.4- weak polar molecules of two 2.4-DNPH- DNPH-ones, in the following ratio of vo- ones are strongly retained at the no polar lume: stationary phase. Therefore, the 2.4-DNPHAA: 2.4-DNPHD: 2.4-DNPHA: longitudinal diffusion of concentrated zone 2.4-DNPHiBA = 2:1:1:1. is higher.
Figure 1. The program of MGMPI Figure 2. The chromatogram of model
Simultaneously, in the stationary phase be reduced at 20%, thus the polarity of mo- 2.4-DNPH-ones provided by acetone and bile phase subsides; the 2.4-DNPHAA and isobuthanal are strongly retained. 2.4-DNPHD (more soluble in organic sol- On the second timing segment, g = 2 (t = vent) diffuse in the mobile phase and will 10 min, n = 0.1), the percent of water will be transported to the end of separation
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column. On the third timing segment, g = 3 model mixture MMII contains the same (t = 5 min, n = 1), the percent of water is 2.4-DNPH-ones as MMI, in the ratio of vo- constant (20%), to assure the best separa- lume: tion of 2.4-DNPHA and 2.4-DNPHiBA. 2.4-DNFHAA: 2.4-DNFHD:2.4-DNFHA: The last timing segment, g = 9 (t = 5 min, 2.4-DNFHiBA = 4:1:5:5. n = 5.5), is meant to recondition stationary phase, for a new separation; the interme- diate timing segments, g = 4-8, are inac- tive. The second program of binary mobile pha- se, MGMPII (40 minutes, figure 3), con- tains four time segments too; figure 4 shows the chromatogram of model mix- ture MMII, obtained with MGMPII. The
Figure 4. The chromatogram of model mixture abbreviated MMII
Figure 3. The program of MGMPII
As in the previous case, the first timing a better separation for the 2.4-DNPHAA segment, g = 1 (t = 15 min), the mixing and 2.4-DNPHD; for the 2.4-DNPHA and program assures an eluate with 45% water, 2.4-DNPHiBA, the resolutions being for the best separation between 2.4- comparable. DNPHAA and 2.4-DNPHD; the polarity of Figure 5 shoes a chromatogram of a na- mobile phase is higher (the molecules of tural mixture of 2.4-DNPH-ones, for the 2.4-DNPH-ones are strongly retained on inferior carbonyl compounds of beer. The the slow polar stationary phase). liquid-chromatographic separation is on The second timing segment, g = 2 (t = 10 the same column and the eluting program min, n = 9.9), the percent of water is redu- is MGMPI. The peaks for 2.4-DNPHAA ced at 20%; this value is constant and du- and 2.4-DNPHD are in the central zone of ring the third timing segment, g = 3 (t = 15 chromatogram. min). Subside of mobile phase polarity produces a desorptive process in the statio- nary phase. The timing segments, g = 4-8 have not got a specific content. In the last timing seg- ment, g = 9 (t = 5 min, n = 0.1) the initial mixture of mobile phase is quickly rebuilt; at cessation, the analytical system is completely ready for a new separation. By comparing the chromatograms from figures 2 and 4 we have drawn the following conclusions: the MGMPI assures
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Figure 5. The chromatogram of natural mixture accordingly to literature, namely 15 mg·L-1 of 2.4-DNFH-ones acetaldehyde [6]. In the same chromatogram, the peak of 2.4-DNPHD is Using the external standard quantitative the second, but it is on the tailing peak of method, for the chromatographic peak of -1 the first one. As for the natural mixture of 2.4-DNPHAA, we obtained 14.75 mg·L 2.4-DNPH-ones one may notice the as concentration of acetaldehyde in beer; following aspect: if literature offers real the value complies with literature data [6], concentration values for the two carbonyl assuring a higher satisfaction to the analyst compounds, the accuracy of analytical sys- operator. tem will be justified by high value, ~100, Using the same quantitative method, the of the ratio between the quantities of acet- surface of chromatographic peak of 2.4- -1 aldehyde and diacetyl. In this instance, the DNPHD gives a value of 2.5 mg·L as low peak of 2.4-DNPHD appears as a tai- concentration of diacetyl in beer; this value ling peak on the high peak of 2.4- is higher, bringing no satisfaction to the DNPHAA, thus, its surface is higher than analyst operator. In beer, the normal value the normal one; the mistake value of surfa- of diacetyl concentration is 0.01-0.2 mg·L- 1 -1 ce chromatographic peak became a source ; 0.15 mg·L is the threshold value [6]. of mistake for concentration value. 4. Each chromatogram – figures 2, 4, and 5 Conclusions – contains any peaks with lower values of On the basis of these experiments, the retention time; it is the peaks for solvent. following conclusions may be drawn: 1. We established two optimal programs References for providing mixtures of binary eluate; ac- 1. SIMION GOCAN, Cromatografie de Înaltă cording to them, two mixtures of binary Performantă, Partea a II-a, Cromatografia de phase that assure liquid-chromatographic Lichide pe Coloane, Editura RISOPRINT, Cluj- separation, with a good resolution are Napoca, Romania, 2002, 173-183 pp 2. DOUGLAS A. SKOOG, Principles of obtained for the etalon mixtures of 2.4- Instrumental Analysis, Third edition, Saunders DNPH-ones provided by the aliphatic College Publishing, 1985, 801-815 pp carbonyl compounds with a number of 2-4 3. CANDIN LITEANU; SIMION GOCAN; T. atoms of carbon. HODISAN; H. NASCU, Cromatografia de Lichide, 2. The binary mixtures of mobile phase, Editura Stiintifică, Bucuresti, Romania, 1974, 156- 157 pp obtained by the programs MGMPI and 4. RADU BACALOGLU; CAROL MGMPII, assure a better separation of mo- CSUNDERLIK; LIVIUS COTARCĂ; HANS- del mixtures which contain derivate com- HORST GLATT, Structura si Proprietătile pounds of inferior carbonyl compounds; in Compusilor Organici, vol I, Editura Tehnică, these model mixtures the ratio between Bucuresti, Romania, 1985, 116-127 pp 5. GORT S.M., HOGENDOORN E.A.; DIJKMAN 2.4-DNPHAA and 2.4-DNPHD is higher E.; VAN ZOONEN P.; R. HOOGERBRUGGE, The than one (in mass units), but very low. Optimisation of Step-Gradient Elution Conditions 3. In the case of natural mixtures of 2.4- in Liquid Chromatography, Chromatographia, vol. DNPH-ones, similarly with model mixtu- 42, No ½, ian 96, 17-24 pp res, the binary mixtures of mobile phase 6. BIERANALYSE 9.11 Bestimmung der Gärungsneben produkte in Bier, 246-250 pp offer only partially analytical satisfaction. 7. GHEORGHE ZGHEREA, Installation to Thus, the figure 5 shows obviously the do- Continuously Degassation by Refluxing of Mobile minant peak of 2.4-DNPHAA. By using Phases for HPLC, Annals of West University of the surface value for quantitative Timisoara, Romania, series Chemistry 12 (3) appreciation by external standard method, (2003) 1157-1160 pp we obtained experimental values
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BENEFITS OF MULTILATERAL LIBERALIZATION FOR THE COUNTRIES WHICH ARE EXPORTING LIGHT INDUSTRY PRODUCTS IN A KNOWLEDGE BASED SOCIETY
Alina-Petronela HALLER
PhD. senior researcher III ICES “Gh. Zane" - Iassy Branch of Romanian Academy [email protected]
Abstract:The evolution of international economic processes, interdependencies between countries, between production and goods movements under the incidence and the impact of scientific and technical progress required, in the middle of the 20th century, the institutionalization of the economic liberalization process by laying the foundations of GATT (General Agreement for Tariffs and Trade - transformed later in WTO - World Trade Organization). Initially, GATT's rules were applied only for goods trade but gradually it had broadened its sphere of activity. Based on interpretative approach, in this paper we analyze the degree in which GATT - WTO activity can solve the business problems of the countries which are exporting industrial light products (we are talking especially about developing countries). WTO multilateral agreements concern among others agriculture, textiles, food products and industrial standards. In this paper, we want to show that exporting countries of light products can enjoy many advantages due to the signing of agreements in a multilateral frame. This type of benefits are directly reflected in the economic growth and development of the society based on knowledge.
Keywords: economic liberalization, knowledge society, export, light industry products, economic and social growth, development.
Methods and methodology international division of labour, under the incidence and influence of the technical- The theme is dwelt upon from a conceptual scientific progress. Under these standpoint, on the basis of varied, direct circumstances, the commercial policies and indirect methods, like analysis, promoted by different states acquire new synthesis, observation. forms, sizes and values. After World War II, many economists Introduction believed in the possibility of a restructuring crisis, regarding the world The multilateral commercial system’s expansion as a key for the “free world’s” tendency towards development is power1, so that the United States came determined by the evolution of the with a project of reconstruction of the international economic processes, of the interdependencies between the production and the circulation of goods within the 1 Irwin Douglas, Free trade under fire, Princeton University Press, Oxford, 2003, p. 34.
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international trade based upon the rounds of negotiations, the most important suppression of the preferential regional one being the Uruguay Round, from 1986 agreements(following the to 1994. Commonwealth’s model) and upon One of the results of the UruguayRound multilateral negotiations.2 was the creation of the WTO (World Trade The measures the Americans foresaw were Organization), on 1st of January 1995. relying on the abolishing of trade GATT, under whose auspices these discrimination, on the control of the negotiations were started, was included in depreciation policy, on the liberty of the the WTO. capital mobility, on the free convertibility WTO is responsible for the control of the of currency, on the multilateralization of multilateral trade system, which has exchanges and payments. For them, peace gradually evolved over the last decades and the return to the liberty of trade do not and it also represents a forum for the exclude each other but, on the contrary, continuation of negotiations on the they presume each other. Thus, the world liberalization of trade with goods and states started a wide action, aiming at services, by abolishing barriers and liberalizing the multilateral trade elaborating new rules in the trade related exchanges in order to reduce economic and fields. The WTO agreements stipulate a social gaps between the industrialized and common mechanism to regulate the the industrializing countries3. disputes by which the members defend their rights and regulate the divergences Results and Discussion between them. Currently, only few countries are not part The first major effort to adopt the rules of the WTO system, which were in the past meant to organize the international trade planned economies, and some of the relationships was made, as we mentioned developing countries. The countries with a above, by some countries immediately transition economy regard WTO as a after World War II, when emerged the structure that offers important necessity to create an organism that should opportunities to get integrated in the world watch, control and get directly involved in economy and to improve the national legal the development of the international trade. basis in the economic and commercial These efforts led to the 1948 adoption of fields. the General Agreement on Tariffs and WTO represents the institutional-legal Trade (GATT), whose rules were applied basis of the multilateral trade system, to the international trade of goods. In time, being the only international organization the GATT text has been modified, to that deals with the global rules of trade include new provisions and especially to between countries4. deal with the developing countries’ trade The organization particularly focuses on problems. Moreover, a number of the necessity to make special efforts to subsidiary agreements were adopted, support the participation of the detailing some of the main provisions of developing countries and especially of GATT. GATT and subsidiary agreements LDCs in the raising of the world trade5. were revised and improved over several The WTO agreements are voluminous and complex, referring to agriculture, textiles 2 Teulon Frederic, Comerţ internaţional, Editura Institutului European, Iaşi, 1996, p. 43 4 Dinuţă I., Relaţii Economice Multilaterale, 3 Haller A.P., Exporturile - factor de dezvoltare şi Editura Scrisul Românesc , Craiova, 1995, p.54 creştere economică, Editura Performantica, Iaşi, 5 Dinuţă I., Relaţii Economice Multilaterale, 2008, p. 41 Editura Scrisul Românesc , Craiova, 1995, p. 56
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and clothing industry, bank system, situations commonly admitted, on an industrial standards, intellectual property undiscriminating basis, given that the and many other fields. At the basis of these protection of national economy in front of documents there are principles the foreign competition is made only by representing the foundation of the means of customs taxes. multilateral trade system: An exception allows the countries a) mutual and unconditional granting of facing difficulties related to the balance of the most favoured nation clause. According payments to restrict imports in order to to the WTO agreements, the states are not safeguard the external financial position. allowed to show discrimination towards This exception grants a bigger flexibility to their trade partners. If a partner was developing countries as far as the offered a special favour (like a smaller utilization of the quantitative restrictions in customs tax for a product), this will be imports are concerned, if these restrictions applied to all the other WTO members. are necessary to prevent a serious This principle is the first GATT article, downward slope of the currency reserves. being a priority in GATS (General d) The interdiction of disloyal competition, Agreement on Trade in Services, art. II) represented by the application of and in TRIPS (Agreement on Trade- subventions and dumping prices in export. Related Aspects of Intellectual Property The non-discriminatory rules – the most Rights, art. IV). favoured nation clause and the national Yet, the GATT rules allow for the customs treatment – are elaborated in such a taxes and other barriers standing in the manner as to ensure loyal conditions for way of trade to be preferentially reduced trade. Many of the WTO agreements play within the regional arrangements. The the role of supporting loyal competition (in reduction of customs taxes, applicable in agriculture, for instance, or intellectual the relations between the members within property, services). regional arrangements should not be e) The utilization of a methodology of extended upon other countries. The customs evaluation that could reflect the regional preferential arrangements real value of goods, in order to fulfil the represent an important exception to the correct functioning of the customs taxes rule of the most favoured nation clause. resulted from the multilateral negotiations. Such arrangements could borrow the f) The fulfilling of obligations. Within customs unions or the free trade areas. WTO, if the states agree to open their b) The granting of the national markets for goods and services, they make treatment clause, a fact that involves equal some mandatory commitments, which can attitude towards the import and the be modified after negotiations with the domestic goods, after the import goods’ trade partners, involving the payment of penetration on the market. The same is true some compensation for the partners’ in the case of the services made by foreign prejudice. and national agents, in the case of national g) The parties’ engagement to negotiate at and foreign trademarks, of the copyright a multilateral level the reduction or the and of patents. This principle is provided abolishing of customs taxes and of other in the three main agreements of the WTO, hinders in the way of the international though it is differently approached in each trade. of them. h) Encouraging the development and the c) The elimination of quantitative economic reform. restrictions and their application among the WTO offers multiple benefits, some of member states only in the exceptional them well-known, others less obvious. The
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objective of this system based on rules is to reduction of prices for finished articles and ensure that the markets stay open and to services. avoid the interruption of access by sudden e) trade contributes to income and arbitrary imposition of restrictions raising. The analyses and evaluation of the upon imports. impact had by the Uruguay Round (and In some developing countries, the business therefore of the WTO foundation) or by the communities are still not fully aware of the creation of the EU common market advantages of WTO trade system. Yet, the demonstrated that these processes ended multilateral trade system does not only up with considerable supplementary offer benefits to the industrial producers incomes, thus stimulating the processes of and to enterprises, but it also creates rights economic development. Trade involves in their favour6. These advantages would challenges and some risks – domestic be: producers must offer products as a) it contributes to maintaining qualitative as the imported ones, but not international security. Particularly, everyone succeeds. The fact that security is a result of the application of the supplementary incomes are generated, trade system’s most important principles. demonstrates the existence of the b) it allows constructive solutions for necessary resources for the governments to disputes. It offers effective modalities to redistribute the available benefits, for peacefully and constructively settle the instance, to help the economic agents disagreements pertaining to trade become more competitive. activities. Moreover, trade means several f) the system encourages the governing. possibilities of disagreements. The fact that The governments often use the multilateral the countries can resort to WTO to regulate trade framework as internal restriction their dispute is a substantial contribution to useful in elaborating economic policies, the soothing the international tensions arguing that they could not proceed from the trade area. differently, not to contradict the WTO c) trade stimulates economic growth, agreements. For business, engaging entails representing a potentiality to create a greater certitude and clarity in the trade workplaces. Practice shows that the conditions, and for governments, it means diminution of the number of trade more discipline and correctness. obstacles positively influence the number The advantages for the exporters of goods the workplaces. A well-planned policy and services are: could make easier the employment a) the security of access. In goods trade, problems. almost all the customs taxes of the d) free trade reduces costs. The studies on developed countries and a big part of the the impact of protectionism and of free developing ones were consolidated within trade demonstrated that protectionism is WTO against their rise. The consolidation expensive, determining the price raising. ensures the fact that the improved access The multilateral trade system aims at on the market, obtained by tariff reductions reducing trade barriers through the included in the lists of concessions for commonly agreed rules and bases upon the each country, should not be interrupted by principle of non-discrimination. The result sudden tax rises or the imposition of other – a reduction of production costs and a restrictions by the importing countries. In the field of services, the countries engaged 6 Hoekman Bernard, The Political Economy of the not to restrict the access of products World Trading System, Oxford Univesity Press, beyond the conditions and limitations 2001, p. 87 mentioned in their national lists.
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b) the stability of access. The system also the same time, the rule provides the ceiling ensures the stability of access on the export of reduced tariffs, that is the exclusion of markets, requiring all the states to apply a their subsequent raising. uniform set of rules, elaborated within 3. The most favoured country clause. This different agreements. The countries have important GATT rule refers to the principle the obligation to ensure conformity to the of non-discrimination. Thus, if a member pertinent agreement provisions for the state grants to another country any tariff or rules regarding the value determination in any advantage for certain products, it must customs, the inspection of products with a immediately and unconditionally extend it view to establishing the mandatory on the similar products of other countries. standards of quality or the emission of This obligation is applied both to imports, import licenses. and to exports. If a countries takes taxes There are frequent situations when for export towards a certain destination, it companies are forces to acquire from must take the same tax for the export to all import raw materials, intermediary destinations. There are derogations to it as products and necessary services for the well. The exchanges between the General export production. The basic rule, Agreements of Trade members submitted requiring for the exports to be allowed to preferential customs taxes or are without further restrictions, after the exempted from these taxes are one of the payment of taxes and the obligation to exceptions. Another one is the Generalized ensure conformity of the other national System of Preferences granted to the rules applied in customs with uniform rules developing countries by the developed within the Agreements, facilitates imports. ones. The exporting industries are thus granted a 4. National treatment. According to this certain guarantee to procure the necessities principle, an imported product that passed without delay and at competitive prices. the frontier after having paid the customs Furthermore, the tariff consolidations serve taxes and other taxes benefits by a to assuring importers that import costs will treatment equal to the one applied to local not be raise by increased customs taxes. products. The whole GATT system is based on four GATT also includes annexes that refer to basic rules: agriculture, textile industry, state trade, 1. The protection of national industry by standard products, subventions and actions customs taxes. Though GATT aims at against the dumping practices. The gradually liberalizing the trade, it admits countries engage in categories of that the member countries could be in the merchandise, referring to the reduction and situation to protect their domestic limitation of customs tariffs for the import production in front of the foreign of products. In certain cases, tariffs are competition. Yet, it asks the countries to reduced to zero, in other cases, they are ensure protection through customs taxes. just limited. Therefore, engagements are The utilization of quantitative restrictions taken to WTO, for a product to be applied is forbidden, with the exception of a one rate that will not be raised in the limited number of situations. future. 2. Ceiling of tariffs. The countries are The Uruguay Round offers some required, every time this is possible, to advantages for the developing countries. reduce or to eliminate the protection of the On the one hand, many of these states domestic production by reducing customs resisted, especially at the end of taxes and removing the other trade barriers negotiations, the American demands, that within multilateral trade negotiations. At their domestic policies, the industry of
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services, the foreign investments and the The agreement on agriculture allows the industrial property should become an developing countries to reduce protection object of negotiations within GATT. On the and internal support for a period of ten other hand, the perspective of a significant years. Article 15 stipulates that the normal reduction of agricultural protection and for period of implementation for the a big share of the textiles and clothing developing countries should be six years. industry, offers the possibility of real gains The effects of liberalization established in for many of the developing countries. the agreement for agriculture on the The Final Act of the Uruguay Round developing countries are different. Like reflects a numbers of compromises Winters suggests, the “exporters of between the developed countries’ products from the temperate areas, whose requirements and the developing countries’ prices are the most affected by the possibilities7. For instance, in the case of liberalization of industry, have direct and TRIPS, the licensing of patented strong interests in starting protection; their investments is allowed, but under certain gains will significantly raise”. The conditions, including the compensations importers will be submitted to less for the patent owner. This is a significant desirable consequences. The fact that the compromise of the American standpoint, liberalization of commerce in agriculture mainly shared by the UE and Japan, that can have negative outcomes as well, the mandatory licensing will not be especially for the developed countries, is allowed for a period of twenty years of taken into consideration in the decision patent protection. regarding the measures taken for possible In the case of TRIMS, the Final Act shortly consequences on the reform programmes characterizes the demands for the in the less developed countries, importers technological transfer that violates the of farm products. The ministers of trade rules and allows significant scope for Commerce should make sure that the investments with a view to make sure that LDCs benefit by sufficient quantities of the foreign companies will contribute in food products and technical assistance to the host-country’s developing. Again, USA develop their own farm sectors. made big pressures for a more restrictive When the Agreement on Textiles and variant as far as the investments measures Clothing Industry comes into force, the are concerned. members are to remove the restrictions While the general provisions of the Final based upon the MultiFiber Agreement or Act reflect the developing countries’ on the bilateral agreements upon the interests in several fields, the tendency was products summing up at least 16% of the not to grant them wider exceptions. total amount of their imports in the In certain cases, the developing countries following groups of goods: fibres, textile can benefit by a longer period to follow, at processed products, ready-made clothes. the domestic level, the rules, but the result Considering the restrictions remaining of the Uruguay Round demonstrates that after this initial phase of liberalization, they would be required to make mutual over three years, the members have to arrangements of trade liberalization8. remove the restrictions upon the products for a supplementary percentage of 17% of the total import amount in the terms of the 7 Jackson John, Word Trading System Law and Policy of International Economic Relations, 1990s. The elimination of restrictions for Princeton University Press, Oxford, 2003, p. 144 products holding at least 18% of the total 8 Jackson John, Word Trading System Law and Policy of International Economic Relations, Princeton University Press, Oxford, 2003, p. 145
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imports amount is necessary after seven transformation of the international trade years. All restrictions remained will be into a powerful engine of growth and into eliminated over ten years since the an important mechanism for the integration agreement’s enforcement. These of countries in the global economy. A big agreements of liberalization are only number of developing countries took applied to the restrictions imposed within advantage from the existing opportunities, the MultiFiber Agreement or the bilateral obtaining a rapid growth of their arrangements beyond the legal framework economies. Yet, not all states were able to of GATT. The agreement on the technical benefit from these opportunities. There is a barriers stipulates that the developing real risk for them to be still marginalized. countries will be offered, upon request, At the same time, it is well known that technical assistance and consultancy by their integration and full participation in other members, in order to facilitate the the global economy significantly process of technical standardization. They intensifies widening of international will occur under the “commonly agreed exchanges. terms and conditions”9. The agreement on Undoubtedly, the WTO developing the subventions forbids the subsidizing of countries can effectively capitalize on the exports after a transition period of eight market the results of their own farm and years (art. 27/1). Once a country reached industrial activities, especially the food and the “competitiveness of export” for a textile industry ones. certain product, it must give up Presently, WTO is an organization subventions (art. 27/4). Competitiveness of disposing of a multilateral trade legal and export supposes reaching the 3.25% of the institutional framework, which makes the world trade for two consecutive years (art. globalization process, with all of its 27/5). minuses and pluses, an unavoidable one, becoming a natural phase of the general Conclusions process of political, economic and cultural We could first of all conclude that the development. WTO enforcing represents a particularly important moment, but not important Acknowledgements: enough to ensure the effectiveness of the This paper is supported by the Sectorial new multilateral trade system. From the Operational Programme Human Resources practice of the first functioning months of Development (SOP HRD), financed by the the organization, one could infer that a European Social Fund and by the permanent “wakeful state” is necessary for Romanian Government under the contract all member states, meant to ensure the number POSDRU ID 56815” functioning of all engagements of this system in the requested rhythm. References: Secondly, the efforts meant to establish the 1. DINUŢĂ I, Relaţii Economice Multilaterale, Editura Scrisul Românesc, Craiova, 1995 trade multilateralism and the tendencies to 2. HALLER A.P., Exporturile - factor de dezvoltare şi solve the major problems in restricted creştere economică, Editura Performantica, Iaşi, circles should also be continued. 2008 3. HOEKMAN BERNARD, The Political Economy of The processes of globalization and the World Trading System, Oxford Univesity Press, liberalization determined the 2001; 4. IRWIN DOUGLAS, Free trade under fire, 9 Princeton University Press, Oxford, 2003; Mansfield E, World Trade Organisation – Facts 5. MANSFIELD E, World Trade Organisation – Facts and Policies, INC White Lains, New York, 1999, p. and Policies, INC White Lains, New York, 1999; 107 6. TEULON FREDERIC, Comerţ internaţional, Editura Institutului European, Iaşi, 1996
29 Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Year IX, No2 - 2010
HIGH-FIBER WHEAT BREAD PRODUCED WITH FERMENTED BRAN
Gabriela POP1, Adriana DABIJA1, Amelia BUCULEI1
1 Ştefan cel Mare University of Suceava, Faculty of Food Engineering, Str. Universitatii, no. 13, 720229, Suceava, Romania,, e-mail: [email protected]
Abstract: Dietary fiber is a common and important ingredient of a new generation of healthy food products demanded more each day by the consumer. One of the main sources of dietary fibre is cereals and cereal products. This study is dedicated to investigate the application of new sources of dietary fiber such as fermented bran for increasing the nutritional value of wheat bread. The objectives of the present investigation were to compare the effects of spontaneous fermentation of the bran fraction and fermentation with added yeast or added yeast and lactic aceid bacteria (Lactobacillus delbrueckii subsp.bulgaricus) on the quality of wheat bread supplemented with bran. Prefermentation with yeast and lactic acid bacteria had the greatest effect on the general quality of obtainig breads: improved the loaf volume, crumb struture and the shelf-life of bread supplemented with bran. Fermentation of wheat bran with yeast or with yeast and lactic acid bacteria improved the loaf volume, crumb structure and shelf-life of bread supplemented with bran. Even though we used bran instead flour for the fermentation, the results are similar to the effect of sourdough on bread quality. Spontaneous fermentation of the bran fraction did not have the same positive effects on bread quality.
Keywords: yeast, lactic acid bacteria, fermentation
Introduction fundamental importance. The make of no refined based – products is Cereals (wheat, rye, oat, corn, sorghum, highly necessary especially that lately it millet and rice) have been an all times and has been demonstrated that weight increase will always be the most important plant is directly related to food ratio from group for the human existence and activity. refined cereals [1]. This show how Some researchers estimated, even from the important is for us to make the difference beginning of this century, the potential between the products of whole grain and advantages of food fibres in preventing those of refined cereals to help weight some chronic affection. For a healthy control. One way to obtain high-fiber person, the recommended fibre ratio is wheat bread is to add bran in the bread placed at medium value of 30grams formulas. [6] fibres/day, half of which must derive from A major disadvantage could be the fact that cereals. when we used bran in order to supplement Due to the main humankind concerns the wheat bread, ussualy we could observe regarding the increase of population health a wakenes of structure and baking quality state in general and obesity control in of wheat dough and, also, a decrease of particular, we may say that the volume and elasticity of the crumb. The development of some cereal and pseudo- effect has been atributed to the dilution of cereal processing technologies becomes of gluten wich would affest the gas-holding
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capacity of the dough [2] fermentation and fermentation with yeast The increased concentration of insoluble or with yeast and lactic acid bacteria on the and soluble cell wall material may also quality and the crumb structure of bread have detrimental effects on dough supplemented with 20% wheat bran. structure. Bran particles can machanically disrupt the structure of the gluten network. Materials and methods The baking quality of bran could be influence also by the size of particle or by In order to obtain some available the chemical components like soluble experimental data we used commercial pentosane and ß-glucan. [4] white wheat flour and wheat bran, As the specific volume of bread is one of provided by local producers. The chemical the important quality characteristics, composition of the material is given in differnt bran pretreatmens have been used table 1. to improve the volume of breads The analytical flour quality was supplemented with bran. determined according to the international The aim of the present study was to standard methods (ash content – ICC104/1, improve the quality of wheat bread wet gluten – ICC105/2, protein content – supplemented with bran by ICC106/2). The moisture content of the prefermentation of the bread fraction. We wheat flour and bran were determined by compared the effects of spontaneous oven drying at 1300C for 1 hour. [3] Table 1. Analytical Parameters of the Raw Materials Component Wheat flour Wheat bran 80% Flour +20% Bran Moisture 13.42 14.3 13.86 Protein (N x 6,25 8.5 8.2 8.5 Total dietary fiber 2.7 52.0 12.6 Total pentosan 1.9 26.0 7.3 Starch 78.8 12.0 64.2 Wet gluten 27.11 - 27.05 Ash 0.49 0.54 0.51
Starch content was determined by Constanţa. Instant active dry yeast from Megazyme colorimetric method and total Rompak S.A. Pascani, was used for bran dietary fiber byenzymatic-gravimetric pretreatment and baking. method of Douglas. [3] In order to obtain some available The starter culture of Lactobacillus experimental data our breads we use delbrueckii subsp.bulgaricus (JOINTEC) formulas which are given in table 2. was obtained from S.C.Liliput S.A. Table 2. Bread Formulas Component, g Control bread Bread with bran addition Wheat flour 500 400 Wheat bran - 100 Yeast 7.5 7.5 Salt 7.5 7.5 Sugar 7.5 7.5 Fat 7.5 7.5 Water 345 400
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The doughs were mixed with a spiral of Suceava, Faculty of Food Engineering. mixer for 8 min. After a floor time of 20 min at 280C, the dough was divided into Results and discussions 400g loaves. The loaves were proofed in pans (55 min at 350C) and baked at 2000C In order to obtain some available data we for 25 min. [5] made the following samples: After baking, the samples were cooled 6-8 hours in controlled atmosphere (UV P1: Bread with 20% bran without lamps). In order to be scoring (after 24 pretreatment hours), the samples were sliced for packed P2: Bread with spontaneus bran in plastic bags. Crumb structure was fermentation, 4hour determined after 24 hours of cooling by P3: Bread with spontaneus bran means of a Stereomicroscope fermentation, 16hour In order to obtain the fermented bran we P4: Bread with prefermentation bran with used an adapted method (M.Salmenkallio- yeast, 4hour Marttila, K.Katina, K.Autio, 2001): 100g P5: Bread with prefermentation bran with bran was mixed with 350 g of water and yeast and lactic bacteria, 16 hour dry micoroganisms (1.25g of yeast ore P6: Control bread without bran 1.25g of yeast and 0.162 of L. Bulgaricus Effect of bran fermentation on JOINTEC) in a large bowl; the bowl was the loaves volume covered with aluminum foil and incubated Baking experiments showed that wheat in temperature room. After tretment, the bran at a substitution level of 20% pH an total titrable acidity (TTA) were decreased loaf volume by 19% in measured. The TTA was expressed as cm3 comparision to white control bread (figure of 0.1 M NaOH/10g of bran suspension. 1) The experiments are made in the research laboratory of “Ştefan cel Mare” University
4,5
4 3,5
3 2,5
2 1,5
Specific volume,Specific mg/g 1
0,5 0 P1 P2 P3 P4 P5 P6
Figure 1. Effect of bran fermentation on loaf volume off breads supplemented
As it could see in figure 1, bran treated by The 4 hour spontaneous fermentation of short fermentation with yeast (P4) and long bran (P2) slightly improved specific fermentation with yeast and lactic acid volume, whereas the 16-hours spontaneous bacteria (P5) improved the specific bread fermentation (P3)had no effect on bred volume over untreated bran by 10-15%. volume.
32 Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Year IX, No2 - 2010
Effect of bran fermentation on good buffering capacity, that’s why the the pH and TTA of Bran-Water Slurries pretreatment had only a slight effect on the As it could see in table 3, bran possessed a pH of the bran slurries.
Table 3. Effect of bran fermentation on the pH and TTA of Bran-Water Slurries P2 P3 P4 P5 Yeast no no yes yes Lactobacillus delbrueki subsp.bulgaricus no no yes yes Fermentation time, hour 4 16 4 16 Temperature, 0C 28 25 28 25 Initial pH 6.6 6.6 6.6 6.6 Final pH 6.6 6.5 6.6 5.8 Initial TTA 3.3 3.3 3.3 3.3 Final TTA 3.3 4.4 5.9 8.7
The bran fermented by lactic acid bacteria slow the souring. Bran contains a much and yeast was the most acidic (pH=5.8, higher number of microbes than flour. So TTA=8.7), whereas spontaneous we could notice that adding starter cultures fermentation of the bran for 16 hours had a to the bran instead of spontaneous souring smaller effect, pH= 6.5 TTA = 4.4. Wheat is advisable for the production of bread sourdoughs usally have value of pH=3.5 to with an even repeatable quality. 4.5 and TTA values from 8 to 22, depending on the ash content of the flour Effect of bran fermentation on and the starter culture used. Higher ash the crumb firming. content of the flour gives more acidic pH and higher TTA values. In order to analyze the crumb firmness If we compare the pH and TTA values it were made tests on breads from each of presented in the literature we can see that the dough with Voland –Stevens crumb the lactic acid bacteria-yeast fermented firmness analyzer in baking day and after bran was not very acidic. each half of next three days.The results are Addition of baker’s yeast to the sourdough given in figure 2.
600
P6 500 P5 400 P4 300 P3 200
Hardness/g P2
100 P1
0 0 0.5 1 1.5 2 2.5 3 3.5 Storage time, days
Figure 2. Effect of bran fermentation on crumb firmness of bread supplemented
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Four– hours fermentation with yeast (P4) subsp.bulgaricus instead of Lactobacillus and 4-hours spontaneous fermentation of brevis (usualy used for flour fermentation the bran (P2) slowed the crumb firming in bakery). slightly during three days of storage in Endogenous enzymes of flour, comparision with white control bread (P6). especialy amylases and proteases, as well The bread baking with bran treted by 4- as the enzymes produced by yeast and hours fermentation with yeast (P4) was Lactobacillus delbrueckii subsp.bulgaricus slightly more firm than white control bread during fermentation seem to have a (P6) after storage for three days, but the positive effect on the dough properties and firming rate (the changes in firmness from the structure of bread. day0 to day 3) was much smaller. Acid produced during the fermentation The 16-hours fermentation with yeast and lower the pH level of the dough, thereby lactoc acid bacteria (P5) improved the affecting the enzyme activity and gluten crumb softness and keeping qualities most. characteristics. On the day of baking, all breads If we use bran fermented for repalcing supplemented with bran were firmer than 20% of flour and by optimizing the baking the white control bread (P6), but after process it is possible to produce good storage for only one day, the crumb consumer quality wheat bread containing firmnes of the white control bread (P6) up to 10% dietary fiber. was the same as that of the bread baking with fermented bran. After 3 days of storage the bread baking with yeast and lactic acid bacteria References (P5) was softer (20%) than the white control bread (P6). 1.C.S LAI, A.B DAVIS and R.C. HOSENEY, Production of whole wheat bread with good loaf volume. 1989, Cereal Chemistry, 66:224-227 2. D.ROGERS, Variety Whole Grain Ingredients, Conclusions International Whole Grains seminar, 2006, AIB International, Manhattan, KS, USA Fermentation of wheat bran with yeast 3. AMERICAN ASSOCIATION OF CEREAL or with yeast and lactic acid bacteria CHEMISTS, 2000, APPROVED METHODS OF THE AACC, 10TH ED. METHOD 74-09. The improved the loaf volume, crumb structure Association: St.Paul, MN, U.S.A and shelf-life of bread supplemented with 4. S. HEGENBART, Understanding Enzymes bran. Function in Bakery Foods,2006, Cereals Chemistry Even though we used bran instead flour 5. W. ATWELL, Wheat flour, Egan Press for the fermentation, the results are similar Handbooks Series, St. Paul, Minnesota, 2006, USA, 6. P. PYLER, Baking Science & Technology, 3rd to the effect of sourdough on bread quality. edition, vol. 1 Sosland Publishing Co., 1998, This a very interesting result especialy Kansas City MO pp. 30-32, 132-151. if we take into consideration the fact that we used Lactobacillus delbrueckii
34 Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Year IX, No2 - 2010
USE OF SOME CLAY MATRICES IN BIOTECHNOLOGY OF ACID DAIRY PRODUCTS
Alisa Vasilica ARUS1, Ana Maria GEORGESCU 1, Iuliana Mihaela LAZAR 1, Ileana Denisa NISTOR 1, Neculai Doru MIRON 1, Azzouz ABDELKRIM 2, Andrea – Eliza FARKAS 1, Ramona Mihaela COJOCARU 1
1Food Engineering Department, „Vasile Alecsandri” University of Bacau, [email protected] 2Département de Chimie, Université du Québec à Montréal
Abstract: In this paper we analyze the use of some clay matrices in the cultivation process of lactic acid bacteria, in dairy industry. The object of this paper is the use of anionic clays - hydrotalcite type, with food purity grade, that are synthesized in laboratory, and can be conditioned in porous tablets or pellets form. These are introduced into the culture medium and help optimal growth parameters achievement of lactic acid bacteria by temporary retention of lactic acid. After multiplication and cellular growth, the tablet or pellet insoluble clays, can be removed, washed and recovered. The lifetime (using) of such materials and devices is practically unlimited. According to this study, the cultivation process of lactic acid bacteria includes the immersion in culture medium of a hydrotalcite clay matrix, with anions exchange role, cultivated through continuous cycle of hydrotalcite matrices immersion, removing and recovering, the lactic anion is continuously fixed, achieving at the same time the proton neutralization of lactic acid, with an aqueous phase, separated from the culture medium, with subsequent extraction and separation of lactic acid. The advantages of the proposed process are: about four time decrease of achieved period of lactic bacteria cultivation phase; 2 - 3 time decrease of final cultures productivity growth; increase with 50-70% rate of bacteria growth in fermentative process; decrease with 30-60% of lactic bacteria cost for industrial fabrication of acid dairy products.
Keywords: hydrotalcites, lactic fermentation, lactic cultures, lactic acid
Introduction microorganisms eat nutrients and after metabolic formation of new substances. The growing process for the production of The classical growing process of lactic lactic acid is, generally, the same for all of bacteria which serves as the nearest these products, based on the classical solution consists in sterilization method, varying only the conditions of (pasteurization) of milk at the temperature control phase fermentation culture medium of 63…72ºC, for 0,5 h, cooling milk to the depending on the desired culture seeding temperature (40...48ºC) for microbiological component to be selected 0,25…0,5 h, adding the milk to seed [1]. powder culture, cultivation of lactic The disadvantages of these methods of bacteria at temperature 36…40ºC and their cultivating lactic bacteria are the difficulty separation [2]. Technologically the of maintaining the culture medium growing of lactic acid bacteria is inhibited composition optimum and stable. The by two processes: composition is changing as 1) gradual increase of v pH in the culture
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medium, followed by lactose conversion aqueous phase, which is separated from into lactic acid; matrix by the immobilized culture medium 2) accumulation of lactic groups. and after separation the lactic acid from These two processes combined have a bad aqueous phase returns to the culture influence on cellular growth and medium. development. The cultivation process of bacteria under In known processes the lactic acid anion is the second variant includes the not removed, the principal inhibitor of introduction in the culture medium of an cellular development. Thus, the classic alkaline agent with food purity, insoluble, drawbacks are large losses of bacteria in which in cultivation time sets continuously the fermentative process and low the lactic acid formed in the culture productivity of lactic bacteria, which leads medium to achieve continuous cycle entry, to reduced growth rate of bacteria and discharge and regeneration of basic agent growth in the cost of selected final and extraction and separation of lactic cultures. Therefore, according to statistical acid. laboratory data, growth of lactic bacteria The cultivating lactic bacteria according to from initial concentration of 5…50 cells/ the third variant includes immersion in the cm3 up to final concentration of 20…30 culture medium of a solid insoluble matrix millions cells/ cm3 takes 8…12h. of anion exchange resin, filled with groups We propose to solve this problem, by of OH- exchangeable, R+OH- type, during developing a lactic bacteria cultivation the cultivation period by continuous technology, to ensure their growth rate immersion cycle, discharge and during the fermentative process and to regeneration of resins into a solution of reduce considerable the cost of final sodium hydroxide, lactic anion is fixed selected cultures. continuously and with free OH- group to achieve neutralization of the proton which Materials and methods is made simultaneously with the formation of aqueous phase, which is separated from The problem presented above can be the culture medium by extraction and solved by making a cultivating lactic subsequent separation of lactic acid. bacteria procedure, which includes the In all these variants, the fermentation preparation of the culture medium, processes lasted for 2…3 hours. containing sterilized and cooled milk to the The common element for all the variants of seeding temperature, addition of the lactic bacteria cultivation processes is milk culture medium prepared by lactic acid sterilization (pasteurization) at the bacteria, cultivation and their separation. temperature of 63…72ºC for 0,5 h, cooling Lactic acid bacteria are used for seeding milk to the seeding temperature increase on a nutrient medium consisting (40…48ºC) for 0,25…0,5 h, addition in of yogurt: fresh pasteurized cow milk, milk of the culture medium seeding volumetrically taken 1:10 and powdered, powder, their cultivations for 2…3 h by which is added by agitation in the culture continuous agitation at the temperature of medium at constant temperature; then as 36…40ºC and lactic bacteria separation. the first choice the culture medium is As initial products we used fresh milk and immobilized in the porous matrix. Lactic one of the traditional culture used in milk bacteria cultivation is achieved at the industry, such as: Lactobacillus bulgaricus temperature of 36…40ºC for 2…3 hours (for milk fermentation), Lactobacillus with the elimination of lactic acid formed acidophilus (for acid fermentation of by continuous elution of the mobile milk), Lactobacillus longum (for milk
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coagulation), Lactobacillus casei (for for example, by treating with calcium fermentation and maturation of cheese) etc. hydroxide to obtain calcium lactate. After Culture medium composition includes: filling the precipitate, the liquid was - yogurt, buttermilk or another decanted and the sediment was passed medium in which lactic bacteria were through the filter. The liquid was treated grown – 1 ml; with sulfuric acid until the reaction became - pasteurized milk or cow milk fresh acid for the calcium decomposition. After pasteurized – 10 ml; the filtration of calcium, sulfate was - lactic cultures specific to the group formed; the diluted acid solution listed above. concentration was of approximately 50%, For the first variant of proposed thus obtaining a concentrated solution processes, as initial products we used fresh named „ lactic acid for technical use”. By milk, water and one of the cultures listed keeping the concentration of lactic acid at above. As porous matrices we used solid low pressure until the concentration matrixes from organic polymers and reaches 50...80% we obtained raw lactic mineral categories, for example acid, which can be used in industry, being montmorillonite clay type, or semi porous purified by re-crystallization of calcium matrix of cellulose and gels categories. lactate and subsequent treatment with For the second variant we used as base sulfuric acid, resulting free lactic acid. agent: Given the complexity of the technical a) a disperse aqueous phase, formed by an solutions adopted and proposed in this organic substance from chelates’ category, article, we made a statistical study of the dissolved in comestible liquid fat, from process of cultivating lactic bacteria using vegetables oil class and animal fat. The a factorial program, which took into continuous fixation of lactic acid is made account the individual effect of all by organization of a relative movement parameters, and their interactions can between disperse phase and culture potentially be synergistic. medium, and before evacuation is made, Such a study required a small number of the separation of disperse phase from experimental attempts made in short time culture medium takes place. and with negligible costs. b) an insoluble solid matrix, for example a The experimental testing methodology clay from the category used in medicine includes the performance of two series of such as gastric bandages, which is experiments. regenerated by washing and impregnation In the first of them we carried out a lactic with a sodium hydroxide solution. bacteria cultivation process. For seeding The carrying out of the process of the pasteurized milk (producer SC cultivating lactic bacteria on the proposed Prolabac SA Bacau, acidity 14…16ºT and variations is conditioned by three essential 1.8% fat) or fresh pasteurized cow milk factors: we used bacterial cultures grown in yogurt 1) Attachment velocity of the lactate and buttermilk (producer MAMY’S Dairy group; Sibiu) medium, the volumetric proportion 2) Elimination and regeneration speed of of „yogurt/ pasteurized milk” or the lactate group; “buttermilk/ fresh pasteurized cow milk ” 3) Attachment capacity of the lactate was 1:10. group. In “buttermilk/fresh pasteurized cow milk” Lactic acid separation from de case the composition of milk-culture regeneration medium or from aqueous medium contains: solution was achieved by a known process, - buttermilk MAMY’S Lactate, Sibiu
37 Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Year IX, No2 - 2010
– 1 ml; procedure consisted in very small values of - fresh pasteurized cow milk – 10 ml; efficiency, because of the relatively slow - specific lactic cultures listed above. kinetics of the diffusion process of lactic The fresh pasteurized cow milk acid during passage through the composition (energy value 64 kcal/ 100g) membrane. The main problem arises from was: nutrients – 100 g; the concern to ensure a good transfer of the fats – 2 g; lactic acid into the mobile aqueous phase carbohydrates – 3,9 g; that must be separated. proteins – 3,2 g; The third alternative method proved to be calcium – 125 mg; the most judicious that was adopted as dried substances – 11 g. optimal technical solution for the first After some time, the lactic bacteria variant of the proposed process of started to develop, the lactic acid formed in formation of lactic acid bacteria. The pursuing one of the proposed variants of problem was reduced by creating a porous the process of cultivating lactic bacteria matrix of organic polymers, cellulose, gels was partially and gradually removed from and mineral polymers type milk - culture – fermentation I medium. montmorillonite, in which the pore size Under the same conditions of temperature allows the development of bacteria mass. and acidity, we determined the rate of In the second series of experiments, yeast growth and development of the lactic (grown culture medium), obtained in the bacteria population that do not depend on fermentation process we used clot the milk-culture medium used, and thus we pasteurized milk (producer SC Prolabac presented below the results for both milk- SA)- fermentation II, to determine the culture media. degree of efficiency of the lactic ferment We studied three alternative ways of obtained, which is estimated in range of achieving continuous elution process of the the increase in acidity of inoculated milk in lactic acid. fermentation process II. 1. the culture medium was In this variant the culture fluid is not fixed, immobilized in a fix bed, the but has a mobile fluid phase. As insoluble conditions of the lactic acid were alkaline agent we used a liquid dispersed not disturbed by any of the phases, phase, which comes across milk-culture the milk-culture medium medium, and before discharge it was maintaining constant its separated from the culture medium. The composition testing of various lactic acid binding agents 2. the culture medium was proved that in this purpose the organic continuously dialyzed through a substances of chelate type (a complexing dialysis membrane agents class) that have a great affinity for 3. Immobilization of milk-culture acids are better suited. These compounds, medium in a solid or semi-solid before being added to the culture medium, matrix and continuous elution of were dissolved in a fat food fluid the lactic acid in the aqueous (vegetable oil or animal fat). mobile phase. The fermentation period depends on the We have noticed that the first way raises dose of reducing agent of lactic acid the problem of excessive consumption of concentration, which in this variant was culture medium and the necessity of dosed by varying the amount of insoluble recycling, which leads to the increasing of alkaline agent. As a criterion to start the technology spending. fermentation period, we chose the initial The major inconvenience of the second concentration of cells in culture medium –
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milk 20 cells/ m3, and as a finalization the amount of hydrocalcit. As criteria for criterion of the fermentation period we stating fermentation period we chose the chose the final concentration of bacterial initial concentration in milk-culture population of 25 million cells/ m3. medium 20 cells / m3 and quality criteria After selection of lactic bacteria grown for for finalization fermentation period, we each sample from fermentation I, we used chose the final concentration of bacteria’s in fermentation II, the efficiency of the population of 25 millions cells/ m3. culture bacteria grown in fermentation I After the selection of grown lactic bacteria testing at 360C for 7 hours and determining for each sample from fermentation I, these the average speed increase of acidity of ones were used in second fermentation of milk. efficiency testing of growth culture in first Alternatively, cultivation of lactic bacteria fermentation, achieved at 36ºC for 7 h and was carried out by stirring at the constant determining medium growing velocity of temperature of 37 C. In this variant the milk acidity. technical solution proposed supposes that the culture medium remains fluid and the Results and discussion lactic acid formed is fixed on some movable or immovable matrices (surfaces) The results of the three different methods with basic character. of cultivating lactic acid bacteria consist Surfaces were chosen to satisfy the of: following conditions: - reduction of four times the period of - have sufficient physical adsorption implementation of lactic acid bacteria capacity for each contact cycle in growing phase; milk-culture medium - increase of 2…3 times the growth - have no cation exchange properties, productivity of the final culture otherwise, the lactate group will not be - increased with 50…70% the evacuated growing process rate of bacterial - anion exchanger to allow elimination fermentation; of lactate group - reduction with 30…60% of lactic - to have a renewable surface bacteria cost for industrial production of - to have a long life lactic-acid products (buttermilk, kefir, sour - to present inertia in relation to other milk, cheese etc) depending on the constituents of culture medium procedure used; - to have a low cost. - use of cheap, affordable, virtually Such a surface can be achieved relatively unlimited lifetime materials, which do not easily, for example a classical anion resin affect the properties of dairy and food type. It was found an advantageous variant products and are not a health risk for of such a varied hydrocalcit similar to that consumers either; used for the treatment of stomach - obtain supplementary lactic acid - hyperacidity (clay, used in medicine as an additional valuable product for food, gastric bandages). chemical, leather industries and medicine. The surface can be easily made in laboratory according to standard Conclusion procedure. Hydrocalcits are also much cheaper than ion exchange resins. The result obtained is due to the following The fermentation period depends on the issues: dose reducing agent concentration of lactic - the lactic acid inhibiting cell acid; which in this variant was assayed by growth and proliferation of lactic acid
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bacteria, used to obtain cheese and lactic - after multiplication and cell growth, acid-products was removed from the the pills or insoluble clay plates can be culture medium; extracted, washed and regenerated with - contrary to known methods, which NaOH; do not eliminate lactic anion, the main - life (use) of such materials and inhibitor of cell growth, the technology devices is virtually unlimited. proposed expects the fixation of this ion by anion-exchange materials; References: - the lactic acid elimination which represents the main reason of the present 1. CHINTESCU G. Îndrumător pentru tehnologia study, is achieved by using insoluble produselor lactate, Bucureşti, ed. Tehnică, 1982, p. 175-177 materials with a high food purity grade and 2. CONSTANTIN I. MILICĂ, Biotehnologiile with basic characteristics; viitorului, Iaşi, ed. Ion Ionescu de la Brad, 1999, p. - insoluble material technologies and 232 – 233. technological interest, which correspond to 3. NISTOR D.I., AZZOUZ A., MIRON N.D the above, can be obtained by ion Ingineria proceselor chimice şi biochimice, ed. Tehnica-Info, Chişinău 2006; exchange resins; 4. NISTOR I.D., JINESCU C., AZZOUZ A., - insoluble material of technological Ingineria proceselor biotehnologice şi alimentare, interest, corresponding to the above vol. I, ed. Alma-Mater, Bacău, 2008. mentioned requirements, can be also 5. NISTOR I.D., JINESCU C., AZZOUZ A., obtained on anionic clay of hydrocalcit, Ingineria proceselor biotehnologice şi alimentare, vol. II, ed. Alma-Mater, Bacău, 2008. having a similarly structure with gastric bandages used in hyperacidity treatment of people; - anionic clay of hydrocalcit type can be synthesized in laboratory based on calcium, magnesium carbonate and sodium aluminates, after a 6 h of maturation period; - anionic clays can be conditioned under pills form or as a porous plate that is inserted into the culture medium for lactic acid removal;
40 Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Year IX, No2 - 2010
DISPERSION AND ASSESSMENT OF FORMALDEHYDE EMISSIONS BY AUTOMOBILES AND THEIR INFLUENCE ON THE AIR QUALITY
Boris SKIP 1, Sergiy ZELINSKYI 1
1 Chernivtsi National University by Yuriy Fedkovich, Chemistry Department, Ukraine e-mail: [email protected], [email protected]
Abstract: The paper deals with dispersion of formaldehyde, which is emitted by cars. According to our study of statistical data for Chernivtsi city, formaldehyde is the most dangerous component of the exhaust fumes.The intensity of the traffic as well as formaldehyde emission in city’s streets net according to a mathematical model of the chosen part of the Chernivtsi has been studied. Such modeling enables us to define the level of air contamination in any part of the chosen area. In the result of our research we defined the contour maps of concentration of formaldehyde.The objective of the study is to improve mathematical model, calculate 2D concentration profile of formaldehyde in surface air of the area under study with availability of wind, and conduct virtual experiment with different boundary conditions in the contours of the buildings and streets. The obtained results of the modeling are more accurate and more suitable for description of dispersion processes over crossroads. Under windy weather conditions, in particular during strong winds, we can observe considerable shift of concentration profiles of formaldehyde, and the highest pollution levels from motor vehicles that is observed and proven by the model in Holovna, Chervonoarmiyska, Prospekt Nezalezhnosti, Storozhynetska, Vynnychenka, Chapayeva, Stasiuka Streets. Realistic concentration profile is reached not only above the road but also over surrounding area and within the residential area. . Keywords: environmental pollution, fumes exhaust, convection-diffusive equation, mathematical model.
Introduction In city is necessary to take into consideration building (housing system). Global ecological security is very Because the building may be such so street important problem nowadays. This is ventilation process are absent or very global question and not problem of one small. In such situation to take place country or region. Preservation of increased (higher) concentration of atmosphere is actual task. Because, pollution. everyday we listen about ozone hole, In EU widely use a number of models for greenhouse effect, smog etc. environmental impact begins the built has This is actual problem for Ukraine too. In some region are considerable difference variable quality environment from standard been started. Such modelling permits to including atmospheric air. According to foresee and prevent ecological problem modern research, cars are one of the area and helps safe dispersion of pollutants biggest agents of air pollution especially in [1– 3]. big cities. Basic pollution substances are Chernivtsi city is comparatively clean and carbon oxide, carbon dioxide, nitrogen ecologically favorable region centre of oxides, compounds of sulfur, Ukraine. Motor transports are the biggest formaldehyde, volatile organic compounds. source of air pollution there. The motor
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transport emission made in mean value 29 According to statistics of thousands ton per year from 1999 till 2006. Environmental Protection Authority in There are 85,6% from total quantity of Chernivtsi region on 2008 was been emission. excelling of maximum permissible Tendency of increase of quantity of concentration of formaldehyde and emission has been starting from 2007. nitrogen dioxide. Environmental pollution Seeing the quantity of motor transport was by formaldehyde cause by increasing increased. Introduction in exploitation new quantity furniture manufactures big roundabout way for region centre can building shop, warehouse with materials of helped the quality of atmospheric air of timber industry, and quantity of Chernivtsi to do something better. automobiles
Table 1 Greatest mean and maximum concentration of pollution substances in city’s air in 2007 [5]
Maximum permissible Maximum of concentration Average Pollution 20–30 minutes Daily 20–30 minutes concentration* substances concentration* average, concentration, 3 3 mg/m mg/m Suspended 0.15 0.5 0.3 0.4 particles Sulfur 0.05 0.5 0.04 0.2 dioxide Carbon 3.0 5.0 0.5 1.4 oxide Nitrogen 0.04 0.085 0.7 2.8 dioxide Nitrogen 0.06 0.4 0.2 0.3 oxide Phenol 0.03 0.01 0.7 2.5
Fluoric 0.005 0.02 1.3 1.7 hydrogen Chlorine 0.2 0.2 0.9 4.9 hydrogen Formaldehyde 0.003 0.035 1.7 1.5
* - in deal of maximum permissible concentration
about oncological properties of Nitrogen dioxide is present in fumes formaldehyde. In last time formaldehyde exhaust. According to our study of called carcinogen in mass media but World statistical data for Chernivtsi city, Health Huard Organization in official formaldehyde is the most dangerous documents tells only about some component of the exhaust fumes [4]. carcinogenic effect on experimental Therefore formaldehyde emissions were animals [6]. taken by us for study of its dispersion and influence on environment. Practical application convection- International agency for studying cancer, diffusive model of dispersion of which is a part of World Health Huard admixture Organization, admitted that are many facts
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Scientific researches in sphere assessment of points which used in calculation were of fumes exhaust by motor transport in about 40 000. Scheme of streets and city’s atmospheric air basin are actual building is necessary for correct boundary problem nowadays. Especially dangerous conditions description for the convective- situations are when come into being traffic diffusion equation solving. Value of jam and slowly traffic in hot hours. concentration had received from own The quantity of emission create less if are observing and data of Research Institute of high speed traffic, new cars, high-quality Medico Ecological Problems. fuel. But in hot hours we locked many On boundary street - housing system traffic jams, where motor transport went constraints 3rd kind proceeding from very slowly. On pedestrian transit, traffic Schukarev’s law have been used: lights, crossroads the motor transport must C D (C C) . slow down. In downtown city’s parts x 0 streets are narrow with bad level of On boundary building’s geometry ventilation of air but with high traffic constraints 2nd kind by Neyman was used: intensity, because, above such streets we C observe high concentration of pollutant [7, 0 . x 8]. We used mathematical modeling on Nowadays literature has materials with convective-diffusion equation base because attempt of monitoring air pollution by the OND-86 and ISCLT2 methods have motor transport in Chernivtsi. But this limitation for calculating dispersion of papers pertain pollution only over roadway pollutant for objects with complicated and cannot answer the question about level geometry. of contamination on dwelling area or near the streets.
Experimental and methods
The work purpose – to carry out an observation the intensity of the street traffic in some streets of Chernivtsi, to calculate 2D profile of concentration of formaldehyde in a near ground layer of atmosphere in the presence of air exchange processes, to make virtual experiment with different boundary conditions for geometry of streets and buildings. The task of study is to make mathematical model of a dispersion of formaldehyde in atmospheric air of a part of a city
Chernivtsi for possibility of an assessment Figure 1. Chosen part of Chernivtsi of a dispersion of emissions of motor transport including with presence of a network of streets and building. For attainment aim had chosen part of city which are shown on figure 1, in electronic version are represent on figure 2, with housing system a) and without b). Quantity
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high difference intensity traffic, are high difference concentration of formaldehyde. It cannot be in really. This situation was cause peculiarity of geometry record and boundary conditions of modeling. It had incited for next model upgrade, especially working about constrains for crossroads’ regions. We had attempted to write group
а) b) of streets as one continuous polygon. But such experiment hadn’t has positive result. Figure 2. Electronic version of selected In next studies this problem had been part: decided by writing 3rd boundary conditions a) with housing system; b) without constrains for streets’ geometry. In result housing system we have obtained the next contour maps, Results of study which are shown on figure 4a. We can see that such results are more As results of modeling calculation for two correct and describe dispersion process in variants: with housing system and without crossroads’ region more precise. Beside housing system on chosen part, we have that in given work has been writing more calculated contour maps of concentration correctly conditions of dispersion for of formaldehyde, which are shown on boundaries of selected area, that is figure 3. concentration in outside wasn’t as background impurity concentration, this is not absolutely right and had been described by 2nd boundary conditions later. In preceding models dispersion pollutant with housing system (picture 3, 4a), it had been use assumption that concentration on building-air boundary is constant and makes 10% from maximum level of
pollution. But air permeates into the a) b) building, opening and chink, that is process of replacement of air to takes place. It had Figure 3. Result of calculation: been written and 2nd kind’s boundary a) with housing system; b) without conditions for building geometry was housing system writing. After this it has been calculating.
We have obtained results, which are shown We can see on fig.3, contour maps are very on picture 4b. different. In the presence of housing If it is seen on picture 4b, when we used system takes place higher pollution over 2nd kind boundary conditions pollution roadways and in near laying territories, permeate deep inside dwelling area. This including over pavements. Such situation really describes real dispersion process and is dangerous for apartments which have mixing pollutant with air. If it is compared ventilation processes near the roadway. results, which are shown on figures 4a and As contour maps of concentration are so 4b, we can see that average concentration different next research will implement with of formaldehyde into the housing system housing system for trustworse results. -2 -2 makes 0.9·10 and 2.4·10 accordingly. It If we can see on figure 3, on the streets makes difference is approximately 2,5 - 3 crossroads, especially if across streets with
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times. According to statistics of Environmental Protection Authority in Chernivtsi region during 2009 for most of months the excelling of maximum concentration limit of formaldehyde level was been.
a) b) Figure 5. Dispersion calculation in cases of calm and the western wind.a) windless conditions; b) wind along an axis of abscissa of 3 m/s. So, apparently from results of study the greatest levels of pollution by
a) b) formaldehyde from motor transport Figure 4. Results modeling dispersion of emissions are observed and prove to be formaldehyde: а) the 1st kind’s boundary conditions true model oversuch streets: Golovna, have writing for building geometry (concentration of Chervonoarmiiska, Prospect Nezalezhnosti formaldehyde is constant on building-air boundary). b) the 2nd kind’s boundary conditions have writing for Storozhenetcka, high enough level of building geometry (concentration of formaldehyde formaldehyde also are over streets inside buildings is equal as concentration on building- Vynnechenka, Chapaeva, Stasyuka, air boundary) All results gained above are made for Chkalova, Gertcena and Karmelyuka. The windless cases. The given model allows realistic profile of concentration is making calculations of a dispersion of observed not only over road, but also over formaldehyde in the presence of a wind of the adjoining territories and in the heart of the set force and direction. For example if an inhabited area. to set the western wind force of 3 m/s then it is possible to obtain following results Conclusions which are presented on the figure 5. That is, apparently from results of The designed mathematical model modeling in cases windy weather, adequately describes substance dispersion especially at strong winds considerable in a near ground layer of atmosphere. displacement contour maps of It was obtained results of dispersion of formaldehyde concentration is observed. formaldehyde. It follows that in majority of housing system’s areas concentration of Building presence changes symmetry of -2 diffusive profile of substance dispersion. If formaldehyde at level 2,5-4,5·10 was to consider in the given model force and a observed, that does not exceed of 1,2 direction of winds which most typical for maximum permissible concentration. Chernivtsi throughout all year or for some However near the streets with high intensity of traffic concentration at level 5- season then it is possible to obtain results – -2 contour maps of formaldehyde 5,5·10 was observed or near 1,5 concentration and dispersion, that will be maximum permissible concentration. characteristic and are most probable at hot When calculation performs for cases of hour a current of year. presence of a wind it is observed more penetration into building area of a wind direction.
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References: 4. Website of Environmental Protection Authority in Chernivtsi region: www.ecology.cv.ua. [In Ukrainian]. 1. SKIP B. Influence of air quality and seasonal 5. Statement about environment situation on change in temperature and humidity on human Chernivtsi region in 2007. Official report. organism: modeling. In Environment Ecology and Chernivtsi Statistical Authrity. 2008. [In Ukrainian]. Life Safety. # 3, 2004, P. 87-91. In [Ukrainan]. 6. Medocoportal: http://www.health.gov.ua/ 2. GRAUR A., PURICH S., TURCU C., POPA V., 7. KIRICHEK Y.O. Ecology aspects problem of URSUL G., HOLOBICA M., OSEAN V., COCRIS roundabout ways’ projection and building // Motor I., NIKOLAYEV A., KYNAL O., SERGEEVA T., ways and road building. 2001. # 62.. [In Ukrainian]. RUDUSH B., SKIP B., Nehadailova T., Orlenko V., 8. Stepanchuk A.V. City’s ecology passport // Town Kartavina T. Ghid de actiuni comune pentru planning and territorial design: К.: Scientific – protectia calitatii aerului in context transfrontarier technical collection. – 2002 – number. 11 p.32 -34. (Manual of common action about air quality [In Russian]. protection in transboundary context) Suceava, 2008. – 149 p. 3. SKIP B., ZORINA T., STASHKEVYCH O. Influence of automobile’s air pollution on immune kinetic responce // 1st scientific practical conference «Computer modeling in chemistry and technology». Cherkasy: «Cherkasy CSCI», 12-16, May 2008 – p.114-118. [In Ukrainian].
46 Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Year IX, No2 - 2010
CONCERNS ON THE DEVELOPMENT OF ECUMENICAL TOURISM IN BUCOVINA
Elisabeta BOTEZ1,Constantina SĂLCEANU1, Oana – Viorela NISTOR1, Aura DARABĂ1
1 Bioingineering Department, Food Science and Engineering Faculty, „Dunarea de Jos” University, 111 Domneasca Street, 800201, Galati, Phone/Fax +40 236 460165, [email protected]
Abstract: The purpose of the study was to investigate the possibilities of the ecumenical tourism in Northern Moldavian monasteries’ area. The monasteries of Putna, Sucevita, Moldovita, Humor and Voronet are great touristic attractions of the beautiful Bucovina lands and being visited by a large number of foreign and Romanian tourists. Ecumenical tourism is a matter that already exists for centuries. It keeps some of the characteristics of pilgrimage, but it has greatly evolved over years. Today, ecumenical tourism asks for a higher level of culture and conditions for tourists. Thus, if in this context the tourists could assure their own necessities, they would be able to appreciate all the cultural objectives as well as the architecture, constructions, values, spiritual connotation and art. There are only two programs in Romania for this kind of tourism: “Know Bucovina” and “Easter in Bucovina”, both promoted by ANAT and by the Council of Suceava County. The accommodation facilities for the monasteries’ visitors have been studied as well as transportation and advertising points of view in order to enlarge the sizes of the actual tourism which converge to a higher number of tourists. In order to improve the touristic activity it was developed a ten-question questionnaire which had been distributed to a sample of 200 tourists.
Key words: pilgrimage, “Know Bucovina”, “Easter in Bucovina”, questionnaire
1. Introduction awarded by the International Federation of Journalists and Tourism Writers. The Tourism is defined as “the word to ancient architectural monuments with describe the activity of traveling” (Mill, internal and external frescoes from this 1990) or “tourism includes any activity part of the country were listed by that concerns the temporary movement on UNESCO as monuments of universal art. short term of people towards destinations (http://www.turisminbucovina.ro) far from their daily places and habits, Types of tourism: along with the activities developed during a. Cultural and ecumenical tourism: If their staying” (Middleton, 1993). Tourism we talk about the main touristic motivation means hiking or organizing and developing from Bucovina, the starting point is pleasure and recreation trips. Starting from represented by the unique and wonderful these definitions and from the ending of monasteries, subscribed to UNESCO’s the word, “-ism”, we can conclude that it is patrimony: Moldovita, Sucevita, Humor, “an action or process” (Theobald, 1994). Voronet, Putna, Arbore. The patrimony Bucovina is one of the most attractive and also enlists the museums, the ethnographic visited touristic regions from Romania. collection “Ion Gramada” and the black This area, well known nowadays ceramic of Marginea (pottery: expo sales worldwide, was awarded in 1975 with the show). This type of tourism can be international prize “Pomme d’Or”, developed without considering the fact that
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the region is on the land of Bucovina green are the main colors. The monastery’s monasteries. museum shelters embroideries, silver - Voronet Monastery pieces, popular paintings, manuscripts, It is an historical architectural monument, ancient books. These treasures confirm the the best representation of the Moldavian cultural role that this monastery had. architectural style. It was built between - Putna Monastery 26.05 – 14.09.1488 by Stephen the Great. The most important monument of Stephen Voronet became famous all around the the Great is situated at 80.5 km from world for its exterior paintings: Doomsday, Pojorata. Its construction started in 1466 Ieseu’s Tree, St. John the New’s Prayer for but it was burnt in 1484. The renovation the dead. Another peculiarity is the usage was made by the founder in 1498. The of a certain type of blue, so called “blue of monastery’s church shelters the graves of Voronet”. This color was made by locals the prince, his two wives and his sons. The including in the composition “lazurit” museum holds embroideries, fabrics, silver paint. pieces, valuable manuscripts, authentic - Humor Monastery medieval manuscripts. It is a valuable medieval architectural - Suceava Citadel monument, built in 1530 by the great The central part, “Musatin Fort”, belongs chancellor Teodor Bubuiog. The first to Petru Musat the1st. He was the one that element that became famous was the open gave up using Scheia. Alexander the Good veranda with its unique exterior painting. over-fortified Scheia and the one who These were made in 1536 by Toma from made it legendary was Stephen the Great. Suceava. The main piece that caught the He built the exterior walls, the defensive eye was “Constantinople Siege”. Red is the ditch and the main entry. Well defended, it dominant color and the founder’s portrait was able to withhold the ottoman siege is settled inside the crypt. from 1476, when Mahomed the Second - Moldovita Monastery had to admit himself defeated. It was built in 1532 and painted in 1537. It (http://www.romanianmonasteries.org/ro/b is one of the most valuable medieval ucovina) Romanian art monuments from Petru b. The hunting tourism Rares reign. The monastery appears to be a Its destination is for hunters and fishermen. fortress composed from the actual church, There is a rich hunting fund: bears, boars, the wall surrounding it with three massive mountain and birch rooster, stags and towers on each side and the royal dears, rabbits etc. Also, the fishing fund residency. The painting is of a great artistic has its resources: trout, dace, barbell, value, similar with the one from Voronet, boisten. The main species from the being special through the figure mountain area is the Carpathian stag. humanization tendency and their c. Hiking – it has as an advantage: presentation in movement. Green and blue beautiful marked routes (Rarau – are combined in a very original way, being Giumalau, Calimani, Rodnei etc.). It similar with the grass green and the blue reminds us of the old “Tartars’ Way” that sky. ends in Carlibaba. These routes can be run - Sucevita Monastery by foot or riding a local’s horse. Built at the end of the 16th century, it is the d. Recreation tourism has all the means most fortified monastery complex from to be put in practice, considering the Moldavia. The inner yard was made to positive effects of the climate and the hide Movilesti family and their huge beautiful scenery. fortune. In the exterior painting, red and
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e. Pleasure tourism: river rafting, a) Yes b) No c) paragliding, paintball, mountain biking and May be d) I don’t know climbing, archery, riding classes, long 7. What measures should ANAT take? walks through the mountains and a) The organization of a touristic tour of practicing winter sports. the monasteries The purpose of the study was to investigate b) The assuring of the roads’ infrastructure the possibilities of the ecumenical tourism to the monasteries gates in Northern Moldavian monasteries’ area. c) The monasteries and the places of worship promotion 2. Materials and methods: 8. Which do you think is the efficient 2.1. Materials method to promote the monasteries? a) By mass-media - Visiting Bucovina monasteries. b) By churches - Collecting data from the talks with the c) By authorities - from national and monks. international information programs - Interviewing the tourists. d) By tourism agencies - A questionnaire regarding the 9. How do you think Bucovina monasteries infrastructure quality and the local should improve their facilities? ecumenical tourism. The questionnaire a) By EU funds was made of 10 items, multiple b) By the City Councils choices. c) By the monasteries own funds The questionnaire d) From sponsorships 1. Which is the best known ecumenical 10. Would it be good for the monasteries to area from Romania? apply to European funds in order to a) Moldova b) Bucovina c) improve their facilities? Dobrogea d) Muntenia a) Yes b) No c) 2. Would you be interested in the May be d) I don’t know ecumenical tourism in Romania? a) Yes b) No c) 2.2. Methods May be d) I don’t know 3. What do you think would be the main - Graphical data interpretation – pie obstacles for a tourist not to visit Bucovina graphics. monasteries? - SWOT analysis based on the a) The lack of transportation comparison between the data collected b) The lack of accommodation place into on the spot and the solutions adopted the monasteries or in the neighborhood for improving ecumenical tourism. c) The impossibility of ensuring a meal SWOT analysis is the most important during the day managerial technique used for comparing 4. Would you like to receive a meal during and solving the improvement strategies of the monasteries visiting? a certain situation. The analysis starts with a) Yes b) No c) enumerating strong and weak points May be d) I don’t know concerning the actual state of the objective. 5. Would you be excited that the offered SWOT is an acronym, the letters coming meal to contain a specific monk menu? from: a) Yes b) No c) Strengths: strong points of the location, May be d) I don’t know the resources it had, its advantages, 6. Would you like to be accommodated to distinctive components of the personnel, the monasteries? managerial activities.
49 Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Year IX, No2 - 2010
Weaknesses: weak points, vulnerable areas, poor resources. Would you be interested in the ecumenical Opportunities: the ways to improve the tourism in Romania? purpose, ways to exploit the strengths and eliminate the weaknesses. Threats: points of danger for achieving the goal, negative aspects coming from faulty Yes resource management, generally, any type May be I don't know of risk. No
3. Results and discussion 3.1. Questionnaire graphic interpretation: Figure 2 Interest for ecumenical tourism We interviewed a number of 200 persons, Concerning the interest for ecumenical 75% women and 25% men. From these, tourism, 80% answered “Yes”, 8,88% 45% were undergraduates, 40% graduates, “Maybe”, 5,55% “I don’t know” and only 10% postgraduates and 5% had 2,22% said “No”. gymnasium studies. The Figure 3 presents the possible The interview had been attained at obstacles in monastery visiting. Sucevita and Voronet monasteries and the What do you think would be the main obstacles for a assigned time for this purpose was 3 days. tourist not to visit Bucovina monasteries? The interpretation of the principal questions from the questionnaire is presented in the following graphics, as the Lack of transportation area with the highest ecumenical potential Lack of accomodation is illustrated in Figure 1. Not having a meal insured during the visit The area with the biggest ecumenical potential
Figure 3 Possible obstacles in monastery visiting
Bucovina At this question, 85.55% considered that Moldova
Dobrogea the biggest problem for the ecumenical Muntenia tourism in Bucovina is the lack of means of transportation. At present, the access is possible only by car. The second problem for 40% of the tourists is the lack of Figure 1 The area with the highest ecumenical accommodation inside the monasteries potential (only Putna disposes of 120 places for accommodation). Only 5.55% considered The conclusion was that 93,33% from the that not insuring one meal per day when interviewed people indicated Bucovina as visiting a monastery is a problem. the area with the highest ecumenical When they had to choose between a meal potential, 21,11% Moldavia, 1,11% during the visit and accommodation, Dobrogea and 1,11% Muntenia. 55.55% chose the meal and 63.33% chose The interest for the ecumenical tourism is the accommodation. remarked in Figure 2. Figure 4 presents the possible measures
50 Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Year IX, No2 - 2010
that have to be taken by ANAT for the sponsorships, 21,11% public money given development of Bucovina ecumenical by the authorities and only 3,33% chose tourism. monasteries’ own funds.
What measures should ANAT takes? The possibility of applying for European funds/ European grants is the subject of Figure 6.
Would it be good for the monasteries to Organizing a monasteries' tour apply to European funds in order to Insuring transportation improve their facilities? Promoting the monasteries
Yes May be I don't know Figure 4 Measures that have to be taken by No ANAT
Concerning the measures that ANAT has to take, 56,66% thought that there should be Figure 6 The possibility of applying for some funds allocated for the infrastructure, European funds/ European grants 48,88% considered that the promotion of the monasteries is necessary and 15,55% When answering about Bucovina would organize a tour of the monasteries. monasteries applying for European funds, At the last question, 63,33% answered that the interviewed people 56,66% said “Yes”, mass-media advertising would be the most 25,55% “Maybe”, 12,22% “I don’t know” efficient, 38,88% chose promoting through and 2,22% thought that there is no need of national/international information European funds. programs, 31,11% would promote only inside the churches and 5,55% would use 4. Conclusions tourism agencies. The funds related to the improving of the Applying SWOT analysis demonstrated its accommodation inside the monasteries are efficiency in establishing the common represented in Figure 5. interest of sustainable development How do you think Bucovina monasteries should regarding the ecumenical tourism in improve their facilities? Bucovina. This analysis has conclusive results when having a significant number Using European funds of subjects with different interests and
With the help of City different ways of achieving the final goal. Council The main criterion is that all of them have Using monasteries funds to participate at the follow up and From sponsorships implementing possible solutions. From the visit and the talks with the monasteries’ management, it has been Figure 5 Improving accommodation inside concluded that their involvement in monasteries promoting and improving the conditions is From the interviewed people 64,44% minimal. considered that the monasteries from The monks consider that promoting the Bucovina should insure themselves with monasteries is not a task for them to do. funds for developing ecumenical tourism, European funds, 52,22% chose
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Table 1 SWOT analysis interpretation STRENGHTS WEAKNESSES - receiving ‘Pomme d’Or’ international award in - reduced number of national/international 1975, given by the International Federation of information programs Journalists and Tourism Writers - defective promotion of the monasteries - having the possibility to visit unique places - lack of transportation (Putna, Sucevita, Moldovita, Humor, Voronet) - lack of accommodation inside the monasteries - practicing hunting, recreation and pleasure (excepting Putna) tourism as well as hiking - not having a meal included during the visit - the possibility of visiting any time of the year - limited possibilities of applying for European - history and unique customs funds - the ‘1000 churches’ - memorial houses and museums OPPORTUNITIES THREATS - restoring historical monuments, especially the - no trademark for ‘Bucovina’ as a brand monasteries - defacement of art and architecture monuments - encouraging new forms of tourism (e.g.: - faulty collaboration between local councils scientific, adventure etc.) - PHARE funds - including agro-touristic mansions in the national network of tourism agencies - developing some resorts: Campulung Moldovenesc, Cacica, Botus (ethnographical resort) - expansions and renovation of infrastructure
Analyzing the present situation, it has been 3. http://www.turisminbucovina.ro discovered that Arbore Church and Voronet 4.http://www.scritube.com/geografie/turism/ETIM OLOGIA-CUVANTULUI- Monastery can not be the beneficiaries of a ECOTURIS133522615.php rehabilitation program from PHARE 5. S.C.E., SARINO, Tourism Principles, quote funds, because of the neglect of the City (Mill, 1990), (Middleton, 1993), 2009 Council and Suceava and Radauti 6. T., Filipas, Analiza strategica SWOT a unei Archiepiscopate. intreprinderi/organizatii, 2005 7. C., Băbăiţă, Impactul factorilor organizaþionali A PHARE program, started in 2003, asupra satisfacþiei consumatorului de servicii worthing 1,5 € millions, having as main turistice, 2006 beneficiary Suceava City Council, should 8. Agenţia de monitorizare a presei – Active Watch, have already modernized 13 monasteries Transparenţa fondurilor europene în and churches so far, some of them România – Studiu de caz, 2009 9. http://www.afaceripublice.ro/investigatie belonging to UNESCO patrimony. This 10. Commission of the European Communities, project was a failure because of the Basic orientations for the sustainability of European common neglect. tourism, Communication from the Commission, Although the solutions for reviving the Brussels, 2003 ecumenical tourism are pretty complicated 11. Commission of the European Communities, A renewed EU Tourism Policy: Towards a stronger at first sight, they can be solved from a partnership for European Tourism, Communication source of European funding, Regio from the Commission, Brussels, 2006 Projects, Espon Projects etc. 12. A., Holden, Environment and Tourism, (http://www.afaceripublice.ro/investigatie) Routledge, London, 2000 13. A., Ispas, Imaginea României ca destinaţie turistică, în Revista Convorbiri Economice, nr. 5. References 4/aprilie 2007, p.48-53. 1.http://www.romanianmonasteries.org/ro/bucovina 14. www.wto, Tourism Vision 2020 2.http://www.turismbucovina.ro/ghid%20bucovinea 15. www.mturism.ro/studii/studii internaţionale n.htm
52 Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Year IX, No2 - 2010
EVALUATION OF MINERAL ELEMENT CONTENT IN GRAPE SEED AND DEFATTED GRAPE SEED
Silvia MIRONEASA1, Costel MIRONEASA2, Georgiana Gabriela CODINĂ1
1Ştefan cel Mare University, Faculty of Food Engineering, 13th University Street, 720229, Suceava – Romania, e-mail address: [email protected]
2 Ştefan cel Mare” University, Faculty of Mechanical Engineering, Mechatronic and Management, Suceava – Romania 13th University Street, 720229, Suceava – Romania, e-mail address: [email protected]
Abstract: A study of the grape seed and of the defatted grape seed has been made in order to evaluate the content of mineral elements, with the purpose of investigating the possibility of using them for human/animal consumption. The research was made individually for each grape seed sample, defatted grape seed respectively. The total ash content was determined according to SR ISO 2171:2009, as a percent of dry substance. The determination of mineral content was performed by the energy dispersive spectrometer, using Röntgen radiation, Shimadzu type (EDX 900HS) with high resolution and sensibility. The results obtained show that the grape seed and defatted grape seed contain macro elements such as calcium (Ca), phosphorus (P), potassium (K) and sulphur (S) and trace elements such as: zinc (Zn), copper (Cu) and manganese (Mn). The seeds of the studied grape varieties and the defatted grape seed have proven to be good sources of mineral elements. Both varieties of seeds can be considered an important source used for fortifying aliments in order to obtain food products with higher nutritional characteristics. At the same time, these products can be valued to obtain new products which result in diminishing waste. The data regarding the mineral content of the grape seeds are useful for the formulation of new diets and food processing techniques.
Keywords: cakes, X-ray, Fluorescence spectrometer
Introduction Mineral elements play an important role in the human metabolism and the interest for The study of the physical and mechanical investigating them is rising [3, 4]. Although, properties of grape seeds, including the they do not produce energy, the mineral chemical composition and obtaining new elements have an important role in many products through the active natural activities of the human body [5, 6] and are components are the source of many necessary in order to maintain certain researches. The grape seed contains: oil, chemo-physical processes which are protein, cellulose, mineral elements and essential for life. Mineral elements represent other components, with benefits for the inorganic components which are vital human health [1]. In larger or smaller because they contribute to the normal quantities, such as salts (cations and ions), development of vital activities and to the especially in the solid part of the grape–peel, development of organism. As ions are linked seeds, the cellular walls of the core, soluble to vitamins, minerals play important elements from the soil can be found [2]. functional roles of metabolic nature,
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activating numerous enzymatic systems, of there is not enough evidence that could chemo-physical nature, controlling the pH, indicate their essential role in the organism electrical neutrality, and the gradients of the [15]. chemical potential [7]. The mineral The diversity of mineral elements which substances from vegetables and fruit help the exist in the seed constitutes an important ossification of the skeleton, growth, source of macro and microelements which maintaining different functions of the are necessary in covering the needs of the internal secretion glands and the forming of human body [16] and contributes to the blood. increase of food value of the products The mineral elements are a part of the obtained from these. An exception is made chemical composition of enzymes, pigments, by the contaminating materials (lead, nucleic acids, peptic substances. These have cadmium), which can appear in seeds due to an important role in nutrition because they the pollution of soil, water, in contact with are necessary in the development of the processing equipment and are toxic for the metabolic processes in the human body. No organism. matter of quantity existing in the human Analyzed macro elements and their role in body, mineral elements are essential because the organism the body cannot synthesize or replace them. Calcium (Ca) is present in the organism in For this reason, they must be consumed larger quantities than other mineral together with nourishment [8]. In accordance substances. Together with phosphor, calcium with the quantity that exists in the organism, contributes to the formation of the skeleton mineral elements are classified into three and teeth and insures their growth. Ca is categories: macro elements, microelements important in the formation and consolidation and ultra microelements. Macro elements of the cellular walls, and maintains the include: calcium, phosphorus, sodium and structure and permeability of the membrane, chloride and microelements include iron, activating the enzymes [17]. It facilitates the copper, cobalt, potassium, magnesium, normal activity of the neuromuscular system, iodine, zinc, manganese, molybdenum, of the heart, it intervenes in the blood fluoride, chromium, selenium and sulphur coagulation process, it increases the water in [6]. Macro-minerals are necessary for the protoplasm and the possibility of organism in larger quantity than 100 mg/day synthesizing cellular albumin. Ca decreases and micro-minerals, in smaller quantities the size of allergic processes, it increases the than 100 mg/day [9]. The lack and the excess resistance of organism to infectious diseases, of macro elements produce metabolic activates an important number of metabolic disorders. Trace elements are essential in the processes, stimulates the enzymatic maturation, activation and functions of host equipment of the organism, regulates defence mechanism [10; 11]. Essential energetic processes and tissue regeneration, components of the enzymatic systems, participates in the stimulation of gastric biological and of structural portions of the secretion, the use of iron and the absorption biologically active components [12] are used of vitamin B12 [18]. by the immune system in order to face Potassium (K) is an essential nutrient for the infectious agents [13, 14]. Ultra micro life and the human health. In the organism, K elements can be found as traces and include participates in the water circulation process, boron, silicon, arsenic, nickel, cadmium, intervenes in the muscular and myocardial lead, tin, lithium, vanadium, uranium, contraction, participates in the protein radium, thorium and a part of these have an synthesis and it is a diuretic. Potassium ions essential metabolic effect. As regards participate in the forming of acetylcholine cadmium, lead, tin, lithium and vanadium and in the transmission of nervous excitation
54 Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Year IX, No2 - 2010
to the muscles. The increase of K in the the organism organism will result in lower sodium and it Iron (Fe) is a component of vegetal and animal emphasizes its elimination because there is a organisms and it participates in metabolic conflict between the potassium and sodium oxidation. This element is a part of the chemical metabolism. The lack or insufficiency of K composition of some enzymes and participates determines the growth of enzymes which in the substance exchange of the organism; it is distorts the nutritional balance, especially an oxygen carrier and plays an important role in through the collapse of the phosphorylation cellular respiration. Fe balances the blood process [8]. This maintains the acid-basic forming processes. The value of a food product balance of the living cell, activating a series as a Fe source is influenced more by the of enzymes. The components of K influence chemical state of this micro element than the the tissue colloids. Through the reduction of total content of Fe. Solubility, slight ionization the hydration level of proteins from tissues, and the Fe balance state are properties which these determine the elimination of liquid determine the absorption of Fe. Ascorbic acid, from the organism [7]. sulphide groups and other reduction substances, Phosphorus (P) is a part of the nucleic and normal gastric acidity facilitate the amino acids structure, of the phospholipids absorption of integrated iron [7]. and participates in the division and Zinc (Zn) participates in the multiplication process of cells. In living metabolism of sugar and, especially, of matter, phosphorous has an important role in protein, it stimulates the activity of some growth. In medicine, P is used to treat enzymes, and is involved in digestion. Also, rickets, functional disorder of the nerve Zn has a specific role in the metabolism of system [19]. P is a partner of the majority of essential fatty acids [7]. In the human oxidation stages of sugar and lipids. These organism, Zn is a part of insulin, stimulating are a part of adenosine triphosphate (ATP) its secretion, of some enzymes, it takes part structure, made of a high quantity of energy, in the synthesis of vitamins C, D and P [20], which plays an essential role in the release of it facilitates the normal absorption and the energy from food products and supplies the vitamin action, especially those in the B1 energy necessary for the organism to complex and provitamin A. Zn participates in function. Moreover, P has an important role the biosynthesis of nucleic acids and of in the activity of vitamins from group B, the proteins while being an essential nutrient for majority of these become active only after the growth and development of children. Zn combining with phosphoric acid [7]. influences the memory and the behaviour of Sulphur (S) is present in all the cells of the individual. Zn has an important role in the organism. It is a component of: amino the metabolism of sugar and lipids acids with sulphur, vitamin B1, biotin, influencing the concentration of cholesterol. coenzyme A and the insulin hormone. This One of the first effects of Zn deficiency is mineral element has an important role in the loss of appetite which is established very process of substance exchange of the early. Along with this, growth stops and the organism and especially in detoxification. S individual can even experience weight loss. participates in expiration, has a favourable Copper (Cu) is present in all the fruits in action in a balanced diet. Through the variable quantity and has an essential role in bridges S-S, sulphur is one of the major the carbohydrate metabolism. In the human elements of the protein tertiary structure, and organism, Cu participates in the formation of the radicals -SH are involved in the activity haemoglobin and of erythrocytes, facilitating of some enzymes. S represents an integrated the absorption of Fe. part of the food protein. Manganese (Mn) together with iron, zinc and Analyzed micro elements and their role in copper stimulates the activity of some
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enzymes, takes part in the synthesis of some separation was made by eliminating the vitamins, the synthesis of proteins, lipids and impurities from seeds by sifting and free carbohydrates. At the same time, Mn falling in air. The seed were minced participates in the vitamin activity, improves mechanically as powder. the activity of the central nervous system and The defatted grape seed was also a study contributes to the forming of urea and of material in the present paper. blood, of milk, intervenes in the Chemical analysis. Determination of ash and development of the skeleton; also in the mineral content. In order to remove carbon, processes of reproduction contributes at every 2 g of powdered seeds of each variety maintaining the production rhythm of sexual were weighed and put into a crucible and hormones. The lack of Mn can delay the then incinerated in oven at 550 Celsius child’s growth; it can lead to deafness and degrees for 6 hours. The total ash content, blindness, and in adults it can produce ear determined according to SR ISO 2171:2009, noises or hearing loss. is the percentage of dry matter. The Cobalt (Co) is found in the organism in the determination of mineral content, from each smallest quantity, but it has a very intensive seed sample, and defatted grape seed activity. It is a component of vitamin B12 and it respectively, was performed by an energy participates in the proper functioning of red dispersive spectrometer using Röntgen blood cells. Recent studies have provided radiation, Shimadzu EDX type 900HS, of information on the biochemistry and high sensitivity and resolution. Before the bioinorganic chemistry of several proteins analysis, the homogenization of the samples containing cobalt in a form other than that in was taken into consideration so that the the corrin ring of vitamin B12 [21]. The level of radiation could penetrate each cell of the cobalt in the organism is dependent on the sample. hexogen, greater than in the case of other Analytical methods. All analytical nutrients, considering that there are no tissues determinations were performed at least three that can accumulate it directly. times and the value of each determined Nickel (Ni) takes part in blood formation parameter is the average of the three values, and plays an important biological role, which is added with ± the standard deviation generally similar to that of Co. Ni activates values. The veracities of the obtained results several enzymes [7] and is involved in and the ones presented in the paper are preserving the integrity of cell membranes [18]. confirmed at the signification level of 0,95. A comprehensive synthesis of the influence of minerals on the organism has been presented by Soetan [17]. Results and Discussion Materials and methods The results obtained at the determination of Seeds – study material. The grape seeds the total mineral elements (in %, dry matter come from grapes picked in 2009, from basis) from the grape seed, and from the different geographical regions of the country, defatted grape seed respectively are given in during the wine making season. The seed of Table 1. the grape varieties have been separated from the dry grape pomace (Mixtures of white varieties of Aligoté and Frăguţă b) and from wet grape pomace (white Nohan, Frăguţă a) which were dried in open air at that moment, at temperatures between 20oC and 25oC. The
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Table 1 0,8 milligrams is ingested by humans on a The total mineral elements from grape seed and daily basis, but the majority passes through defatted grape seed the organism without being absorbed [22]. Total mineral elements Grape varieties grape seeds defatted grape The content of mineral elements from the [% s.u.] seed [% s.u.] grape seed and defatted grape seed, from the White variety 3.18 2.73 Aligoté variety is given in Figure 2. Mix In the case of seeds from the Aligoté variety, Aligoté 3.35 2.84 the mineral elements Ca, K, S are found in Hybrid variety 2.14 1.80 Nohan white larger quantities in the defatted grape seed Hybrid variety than in seeds; only P is in a smaller 3.20 2.86 Frăguţă a percentage in the defatted grape seed Hybrid variety 3.28 2.80 (13.12%) than in the seeds (16.632%). The Frăguţă b concentration of Ca from the defatted grape seed (53.477%) does not differ significantly The way in which the content of mineral from the concentration of Ca in grape seed elements from the grape seed varies in report (54.11%). with the defatted grape seed for the white Mix The variation of mineral elements from the variety is given in the graphic from Figure 1. grape seed and defatted grape seed of Nohan For the white Mix varieties, a larger variety is given in Figure 3. The predominant concentration of calcium in seeds is obtained mineral elements Ca, K, S from the seeds of (55.74%) than from the defatted grape seed Nohan hybrid variety can be found in (49.51%), and the other predominant mineral smaller concentration than in the defatted elements, K, P and S are found in larger grape seed. In seeds, just P is found in large concentration in the defatted grape seed than concentration (18,853%) than in the defatted in seeds. Also, in the defatted grape seed grape seed (14,364%). from the white Mix variety the element Ti The content of mineral elements from grape (0,090%±0,006) is present. Ti is non-toxic seed and defatted grape seed for Fraguţă even in larger quantities and has no natural hybrid variety is presented in Figure 4 and role in the human organism. A quantity of for Fraguţă b in Figure 5.
60 55.74 Grape seed Defatted grape seed 49.251 50
40 28.374 30 26.447
% s.u. % 19.139 20 15.346
10 2.34 0.289 0.332 0.11 0.074 0.09 2.088 0.253 0 0.123 0.069 0 0 Ca K P S Mn Fe Zn Cu Ti
Figure 1. The content variety of mineral elements from grapes and defatted grape seed for the white variety Mix
57 Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Year IX, No2 - 2010
53.477 60 54.11 Grape seed Defatted grape seed
50
40 29.913 27.403 30 % % s.u. 16.632 20 13.12
10 2.53 0.227 0.115 0.083 1.942 0.314 0.149 0.082 0 Ca K P S Mn Zn Cu
Figure 2. The content of mineral elements form seeds and defatted grape seed – Aligoté variety
60 55.058 56.89 Grape seed Defatted grape seed
50
40
30 23.655 25.699 % s.u. 18.853 20 14.364
10 2.419 0.297 0.206 1.759 0.048 0.078 0 0.246 0.07 0.054 0 Ca K P S Mn Fe Zn Cu
Figure 3. The content of mineral elements form seeds and defatted grape seed – Nohan hybrid variety
60 52.153 Grape seed Defatted grape seed 53.285 50
40
30 23.918 % % s.u. 22.389 21.263 20 19.202
10 2.557 1.904 0.272 0.23 0.059 0.088 0.014 2.247 0 0.236 0 0.082 0.1 0 0 Ca K P S Sc Mn Fe Zn Cu Ni
Figure 4. The content of mineral elements form seeds and defatted grape seed – Frăguţa a hybrid variety
58 Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Year IX, No2 - 2010
60 52.697 Grape seed Defatted grape seed
50 50.912
40
30 23.051
% s.u. 22.298 21.676 20 21.512
2.778 10 1.725 0.329 0.284 0.069 0.058 0.035 2.238 0 0.236 0 0.097 0.068 0 0 Ca K P S Sc Mn Fe Zn Cu Co
Figure 5. The content of mineral elements form seeds and defatted grape seed – Frăguţa b hybrid variety
When compared with the other varieties - defatted grape seed from the Frăguţă a varieties from Vrancea, in the hybrid variety hybrid variety, variety gathered from humid Fraguţă – variety from the Suceava county, a marc. Its occurrence may be due to the new element appears – scandium (Sc). In the contact with processing machinery. Its defatted grape seed which results after the concentration is in the allowed limits. extraction of oil, the nickel (Ni) cation contamination can be found in concentration Conclusion of 0,014%. From the comparative analysis of the content The results of the mineral elements show of mineral elements of the Fraguţă hybrid that all the studied grape seed variety and the variety, it can be observed that seeds defatted grape seed contain considerable gathered from the dry pomace – Fraguţă b quantities of mineral elements: calcium, have a larger concentration of Ca (52,697%) potassium and phosphorus which can be when compared to the seeds gathered from used to improve the nutritional value of the the wet pomace – Fraguţă a (52,153%). human diet. The grape seed and the defatted Moreover, in the seeds gathered from the dry grape seed, due to their high content of pomace, Ni is not present but a new element mineral elements, can constitute an appears - Co. alternative in the manufacture of products Generally, the chemical composition of the which ensure benefits for health, following mineral elements of the tested samples is the acceptance of the idea that diet plays a comparable with the one presented in the decisive role in the prevention and treatment specialty literature [23, 24]. of disease. Such products, in addition to From the analysis of the obtained results for having appropriate nutritional effects - they the mineral content, it can be observed that improve health and/or reduce the risk of in the grape seed and also in the defatted disease - also have components which are grape seed, there is a predominant quantity, derived from natural ingredients that provide in the specified order, of calcium, potassium, additional health benefits. phosphor and sulphur. It has been found that cations such as arsenic, cadmium, selenium, mercury, etc. do not exist. The only cation that appears is nickel, in the case of the
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References
1. MIRONEASA, S., LEAHU, A., CODINĂ, G.G., 14. SHERMAN, R.A., Influence of iron on immunity STROE, S.G. and MIRONEASA, C. Grape Seed: and disease resistance, Ann. N.Y. Acad. Sci., 1990, physico-chemical, structural characteristics and oil 587, 123-139. content, Journal of Agroalimentary Processes and 15. ALBION RESEARCH NOTES. A compilation of Technologies 2010, 16 (1), 1-6. vital research updates on human nutrition, 5: 2, 2. POMOHACI, N., STOIAN, V., GHEORGHIŢĂ, Albion Laboratories, Inc. May, 1996. M., SÎRGHI, C., COTEA, V. V. and 16. FAUR, V. and GOIAN, M. Fitoterapie şi NĂMOLOŞANU, I., Oenologie vol. I, Prelucrarea fitoprotecţie alimentară, Editura Dacia, Europa Nova, strugurilor şi producerea vinurilor, Editura Ceres, Lugoj, 1998. Bucureşti, 2000. 17. SOETAN, K. O., OLAIYA, C. O. and 3. PELUS, E., ARNAUD, J., DUCROS, V., FAURE, OYEWOLE O. E., The importance of mineral H., FAVIER, A. and ROUSSEL, A.M., Trace element elements for humans, domestic animals and plants: A (Cu, Zn, Fe, Mn, Se) intakes of a group of French men review. African Journal of Food Science, 2010, 4(5), using the duplicate diet technique. Int. J. Food Sci. 200-222. Nutr. 1994, 45, 63 –70. 18. DUMBRAVĂ, D.G., Contribuţii la studiul, 4. FENNEMA, O.R., Food Chemistry. New York: izolarea şi purificarea pigmenţilor carotenoidici din Marcel Dekker, 2000. produsele naturale. Teze de doctorat ale UPT, Seria 4, 5. MALHOTRA, V.K., Biochemistry for Students, Nr. 11, Editura Politehnica, 2008. Tenth Edition, Jaypee Brothers Medical Publishers 19. SEGAL, B., COTRĂU, M. and SEGAL, R., (P) Ltd, New Delhi, India, 1998. Factori de protecţie prezenţi în produsele alimentare, 6. ERUVBETINE, D., Canine Nutrition and Health. Editura Junimea, Iaşi, 1996. A paper presented at the seminar organized by 20. ŢÂRDEA, C., Chimia şi analiza vinului, Editura Kensington Pharmaceuticals Nig. Ltd., Lagos on „Ion Ionescu de la Brad”, Iaşi, 2007. August 21, 2003. 21. KOBAYASHI, M. and SHIMIZU, S., 1999, 7. SEGAL, R., Principiile nutriţiei, Galaţi, Editura Cobalt proteins, European Journal of Biochemistry, Academica, 2002. 1999, 261 (1), 1–9. 8. JAMBA, A. and CARABULEA, B. Tehnologia 22. EMSLEY, J., "Titanium". Nature's Building păstrării şi industrializării produselor horticole, Blocks: An A-Z Guide to the Elements. Oxford, Chişinău, Editura Cartea Moldovei, 2002. England, UK: Oxford University Press, 2001. 9. MURRAY, R.K., GRANNER, D.K., MAYES, P.A. 23. GURIN, A.A., Razrabotka tehnologhii and RODWELL, V.W., Harper’s Biochemistry, 25th proizvodstva ăkstaktov, konţentratov i vin iz sladkoi Edition, McGraw-Hill, Health Profession Division, vinogradnoi vîjimki. Avtoreferat kand. Disertaţii, USA, 2000. Moskva, 1991. 10. BENDICH, A., Antioxidant micronutrients and 24. GASANOV, D.H.A., Razrabotka tehnologhii immune responses, Ann. N.Y. Acad. Sci., 1990, 587, proizvodstva kormovogo belkovogo producta iz 180. vinogradnîh bîjimok. Ealta, Avtoreferat kad.diss, 11. ROMERO, R., Intra-uterine infection, premature 1990. birth and the Fetal Inflammatory Response Syndrome. J. Nutr. 2003, 33, 1668-1673. 12. ARINOLA, O.G., NWOZO, S.O., AJIBOYE, J.A. and ONIYE, A.H. Evaluation of trace elements and total antioxidant status in Nigerian cassava processors, Pak. J. Nutr., 2008, 7(6), 770-772. 13. SPALLHOLZ, E.J., BOYLAN, M.L. and LARSEN, S.H., Advances in understanding selenium’s role in the immune system, Ann. N.Y. Acad. Sci., 1990, 587, 123-139.
60 Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Year IX, No2 - 2010
FRESHNESS EVALUATION OF CHICKEN MEAT USING MICROBIOTA AND BIOGENIC AMINE INDEX.
Octavian BASTON1, Octavian BARNA1, Aida VASILE1
1 Dunarea de Jos University of Galati, Faculty of Food Science and Engineering, 111, Domeasca St., Tel./Fax: +40 236 460165, e-mail: [email protected].
Abstract: The purpose of our study was to evaluate freshness of raw chicken meat using microbial and biogenic amines content. The objectives were to determine the variation of microbiota (total viable count, psychrotrophic and Pseudomonas spp.), to study the variation of some important biogenic amines and to calculate a biogenic amine index for refrigerated chicken carcasses aerobically stored for one week. Hereby, we used microbial analysis and HPLC determination for the following biogenic amines: tryptamine, β-phenylethylamine, putrescine, cadaverine, histamine, serotonin, tyramine, spermine and spermidine. Our determinations showed that total viable count increased in value from the first to the seventh day (the last day of storage), psychrotrophic microorganisms also increased in number and Pseudomonas spp. increased from the first day to the seventh day of storage. Regarding biogenic amines variation, tryptamine had a low initial content and after a week of storage the content was below 5mg/kg, β-phenylethylamine had also a small initial value and after a week of storage its value is slightly higher than 5 mg/kg, serotonin had a similar comportment with β-phenylethylamine and tyramine the same with β-phenylethylamine only that the final value was slightly higher. Cadaverine and putrescine were detected beginning with the third day of storage and they had the highest values after a week refrigeration of chicken meat. After one week of refrigerated storage, spermine had a particular allure because it decreased permanently. Spermidine had a very slow increase, from 4.8 mg/kg to 6 mg/kg. Biogenic amines index were calculated according to the mathematical relation proposed by researchers from Barcelona University.
Keywords: meat, freshness, microbiota, Pseudomonas, biogenic amine index
Introduction Microorganisms’ activity is slowed down by using refrigeration temperatures for The spoilage of refrigerated chicken meat meat conservation purpose. In order to when stored for a long period is due to the obtain products with high conservation microorganisms’ action and biochemical durability and to increase the refrigeration transformations inside the product. If the effect, it is necessary to have as less initial refrigerating chain from producer to microbial load as possible [1]. consumer is not ensured, or if the seller Initially, chicken meat quality was overpasses shelf life, the consumer can evaluated by determination of have an unpleasant surprise of buying an microbiological and sensorial attributes. altered product. After chicken slaughter, For the identification of the early signs of the muscular tissue suffers irreversible meat alteration, some chemical indices physical, chemical and biochemical were proposed: volatile nitrogen basis, transformations which determine the composites resulted after breaking the muscle to convert into meat. The microbial nucleotides, volatile acidity and the spoilage processes occur later. biogenic amine content [2]. The biogenic
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amine occurrence is a consequence of the Romanian slaughterhouse. The meat was enzymatic decarboxylation of the precursor analyzed after cooling, packaging and amino acids because of the transportation from the plant the first day microorganisms’ activities. Polyamines: after slaughter. All the carcasses were spermine and spermidine are natural stored aerobically, without package foil, amines produced by the body. The for 7 days, at the temperature of 4±1°C in biogenic amines: putrescine, cadaverine, the refrigerator type Electrolux histamine, tyramine, tryptamine, β- ENB43691S. The carcasses weight varied phenylethylamine can be formed when between 1.2÷1.5 kg. The samples were storing the chicken meat due to analyzed the first day when the meat was microorganisms’ action. The biogenic received, recorded as day 1, then at the 3rd, amine determination is important not only 5th and 7th day. because of their toxicity but also their The dry matter determination was made potential use as freshness indicators [3]. according to Romanian STAS 9065/3-73. The occurrence of these amines is Pieces of raw chicken meat with skin (16 dependant on different factors that vary in cm2 in area) were aseptically excised from time. The microbial population influences carcasses and each piece was homogenized the profile of biogenic amines. Spoilage with 100 ml of saline water (0,8% NaCl) responsible microorganisms might not by using a homogenizer model Bagmixer have the capacity of amine forming. It is 400. Duplicate 0,1 ml aliquots of suitable difficult to establish quality limits dilutions of each skin homogenate were universally accepted based on the biogenic spread on the surface of nutrient agar amine content. The above mentioned plates. Inoculated plates were incubated in reasons are partially justifying the relative ATICH 9082 Incubator in aerobically dispersal of biogenic amine values for condition at 4ºC for 14 days for meat, in various researches. From a psychrotrophic microorganisms and at practical point of view, the relative 30ºC for 2 or 3 days for total viable count. simplicity and quickness identification and Pseudomonas were determined on quantification of the biogenic amines centrimide-fusidin-cephaloridine agar (compared to the microbiological supplemented with SR 103 for the measurement) besides the economical selective isolation of Pseudomonas spp. advantages (for example the quick test for generally, after incubation at 25ºC for 1 or determining the diamines described by 2 days, and oxidase-positive colonies were Hall et all. [4], are the reasons for using enumerated. [5] After that viable colonies these substances as chemical indices for were counted using Automatical Colony animal origin product freshness. Counter SC6. The purposes of the study are: The measurement of biogenic amines - determination of microbiota variation content using high performance liquid from chicken meat, concerning total viable chromatography was performed according counts, psychotrophic microorganisms and to the method proposed by Food Research Pseudomonas spp at refrigerated raw Institute from Helsinki, Finland [6]. The chicken meat storage; method principle is as follows: - evaluation of freshness of the refrigerated -Bioactive amines are extracted from a chicken meat using biogenic amine index. homogenized sample with diluted perchloric acid; Materials and methods -An aliquot of the extract is derivated with dansyl chloride reagent; Chicken carcasses were purchased from a -Separation and quantification of
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dansylated amines is performed by mean ± standard deviation (SD). The reversed phase liquid chromatography with standard deviation is a measure of the ultraviolet detection at 254 nm. dispersion of outcomes around the mean. All the reagents used were analytic pure, The differences among means were for HPLC use. Te water used was determined using the method of the deionised. The necessary reagents were smallest squares and the significance level purchased from the Merck and Sigma- was p< 0.05. Aldrich companies. Installations and equipment used for biogenic amine Results and discussion determination: Philips 7768 food processor, homogenization device 7011S, After the chicken carcasses were Kern 770-60 analytical balance, Silent refrigerated for three weeks at 4ºC, we CrusherM homogenization device, determined the variation in time of total centrifuge EBA 21, filter paper for quick viable count and psychotroph count. In filtering with 55 mm diameter, syringe figure no. 1 we present the variation of filters with porosity of 0,45 µm and 13 mm refrigerated storage of total viable count. diameter, Heidolph REAX control agitator, The average initial contaminations of the ultrasonic water tank Aquawave TM, chicken meat were 5.11 log CFU/cm2 incubator BMT INCUCELL 55, water (CFU – colony forming unit), it increase at deionising system EASY pure RoDi, the third day at 5.95 log CFU/cm2 and in filtering assembly with vacuum pump. The the 5th day of storage at 7.1 log CFU/cm2. device for the HPLC determination was a The shelf life of carcass, as stated by the liquid chromatograph model SURVEYOR food manufacturer, was the fifth day of produced by Thermo Electron company, storage. In the 7th day of refrigeration the configured with detector model PDA total viable count were 7.56 log CFU/cm2. PLUS DETECTOR, auto-sampler model This slow increase of microbiota is due to AUTOSAMPLER PLUS, pump model LC refrigeration temperatures that do not allow PUMP PLUS and detector UV-VIS. a quick spoilage of chicken carcasses and Chromatography column is type BDS preserving the freshness for longer period Hipersyl C18. The biogenic amines of time. quantification: quantitative measurement Psychrotrophic count is characteristic for was performed depending on the internal food that is preserved at refrigeration standard using peaks for each biogenic temperatures. We determined amine. The 254nm wavelength absorbance psychrotrophic microbiota after seven days was measured and the resulted peaks were of storage. The variations of integrated with CromQuest software. The microorganisms are presented in figure 2. concentration of each biogenic amine was The initial load were 4.00 log CFU/cm2, expressed in mg/kg d.m. (d.m. = dry increasing permanently to 6.9 log matter). Our determinations refer to the CFU/cm2 for the 7th day of storage at following amines: tryptamine, β- refrigerated temperature. Comparing to phenylethylamine, putrescine, cadaverine, total viable count, on the first day of histamine, serotonin, tyramine, spermine storage we have a difference of more than and spermidine. 1 log CFU/cm2, at the expiration date of The statistical analysis of the obtained data chicken carcass (the 5th day), the difference was made using Microsoft Excel features was higher than 1.5 log CFU/cm2 for the for 10 samples in each of the storage days. total viable count. In the 7th day of Each sample was analyzed in triplicates. storage, the difference begins to decrease, The results obtained are presented as the being only of 0.6 log CFU/cm2. It appears
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9 8 7
6 (CFU/cm2) 5
Total Total viable count 4 0 2 4 6 8 Storage time (days)
Figure 1. Total viable count variation for refrigerated chicken meat that after the 5th day of storage, the psychrotrophic microbiota begin to prevail
8
7
6
5 (CFU/cm2) 4
Psychrotrophic count count Psychrotrophic 3 0 2 4 6 8 Storage time (days)
Figure 2. Psychrotrophic variation for refrigerated chicken meat
Pseudomonas spp. is very important be mesophile or psychrotrophic microbiota that spoils chicken carcasses microorganisms, we can compare the during aerobically refrigeration storage. In values obtained with those of figure 3 we show the variation of psychrotrophic biota. Pseudomonas spp. from chicken carcasses Thus, the initial contamination of carcasses until the 7th day of storage at 4ºC. As it can is mostly by Pseudomonas spp., the trend be noticed, the number of Pseudomonas kept until the 5th day of storage. On the7th increase in time, from 3.8 log CFU/cm2 on day, besides Pseudomonas spp, it appears the first day of storage to 6.4 log CFU/cm2 that another biota become to increase. on the 7th day. Because Pseudomonas can
7
6
5
(CFU/cm2) 4
3 Pseudomonas spp. count count spp. Pseudomonas 0 2 4 6 8 Storage time (days)
Figure 3. Pseudomonas spp. variation for refrigerated chicken meat
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Biogenic amines can be an indicator of hystamine cadaverine putrescin BAI chicken meat freshness. Researchers from 1 spermine spermidine Barcelona University proposed from raw . Based on this index it can be determined foods a biogenic amine index that has a allergenic risk and freshness of raw foods. mathematical relation of: Our determined values for considered biogenic amines are presented in figure 4.
TRIP FEN PUT CAD HIS SER TIR SPD SPM
25
20
15
10
5 Biogenic amines values (mg/kg d.w.) 0 0 2 4 6 8 Storage time of carcasses at 4 grd C (days)
Figure 4. Biogenic amines variation for refrigerated chicken meat, were: d.w.-dry weight
As it can be seen, in time, the values of derivatives being also hormones (tyramine mostly biogenic amines increase, with one is in fact 4-hydroxi-phenylethylamine). exception, of spermine which decreases. Spermine and spermidine are two amines On the first day of storage, the biogenic involved in cellular metabolism, essential amine content is low, spermine having the for organism growth and for development highest value: 21.5 mg/kg d.w. followed by and proliferation of cells. Spermine can be spermidine, of 4.8 mg/kg d.w. Also, synthesized from spermidine. Histamine is putrescine and cadaverine have not been present in small amount (1.8 mg/kg d.w.). detected on the first day of storage. This is The third day of storage of raw chicken due to the fact that either our method is not carcasses is important to be highlighted as sensible enough to detect smaller amounts we detected small amounts of putrescine of those biogenic amines, or they do not and cadaverine. exist in chicken meat. Tryptamine and On the day of shelf life ending cadaverine tyramine presence is due to their action as and putrescine amounts are increasing, hormones, neurotransmitters, and serotonin cadaverine increase being over four times, being one of tryptamine derivative. β- putrescine increase being of 7 times, phenylethylamine action as spermine are decreasing. The spermine neuromodulator and neurotransmitter, its decrease can be due to its use by the
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microorganisms as nitrogen source. microorganisms that characterized best the Histamine has a particular variation chicken meat spoilage and being the because on the fifth day is slowly principal cause for freshness loss. Chicken decreasing, after a slow increase from the meat freshness decreases in time and this first day of chicken meat refrigeration leads to quality loss and shelf life storage. reduction. All biogenic amines that we determined Chicken freshness is good if IAB is below could be made as a result of microbiota 1.00. activity on amino acids chicken carcasses. It is well known that cadaverine and References putrescine are two amines with characteristic repugnant smell especially in 1.TOFAN, I., 2005, Lantul frigorific al produselor more advanced stadium of alteration. A alimentare perisabile (Refrigerating chain of the perishable food products), Ed. Agir, Bucuresti, p. 9, slight modified smell was distinguished on 20, 197-198. seventh day of storage, but we cannot be 2.HALASZ, A. BARATH, A. SIMON-SARKADI, sure if this was due to cadaverine or L. HOLZAPFEL, W., 1994, BAs and their putrescine. production by microrganisms in food, Trends of We calculated the BAI for the chicken Food Sci. Technol. 5, 42-49. 3.BALAMATSIA, C.C. PALEOLOGOS, EK. carcasses and we had the following results: KONTOMINAS, MG. SAVVAIDIS, IN., 2006, -IAB for the first day were 0.068 Correlation between microbial flora, sensory -IAB for the third day were 0.30 changes and biogenic amines formation in fresh -IAB for the fifth day were 0.66 chicken meat stored aerobically or under modified -IAB for the seventh day were 1.37 atmosphere packaging at 4C: possible role of biogenic amines as spoilage indicators, Antonie van Compared with the first day we can Leeuwenhoek, Springerlink, 89, 9-17. observe that IAB of the third day 4.HALL, M. SYKES, PA. FAIRCLOUGH, DL. increased 5 times and IAB for the fifth LUCCHESE, LJ., 1999, A test strip for diamines in day 10 times. We can also say that fresh tuna, Journal AOAC International, 82, 5, 1102- raw chicken meat has an IAB below 1108. 5.MEAD, G.C. AND ADAMS, B.W., 1977, A 1.00. selective medium for the rapid isolation of Pseudomonads associated with poultry meat Conclusion spoilage, Br. Poult. Sci., 18:661-670. 6.EEROLA, S., HINKKANEN, R., LINDORFS, E. An increase in total viable count number AND HIRVI, T., 1993, Liquid chromatographic determination of biogenic amines in dry sausages, leads to spoilage, loss of freshness and Journal AOACInternational, vol 76, nr. 3, p. 575- shelf life of raw chicken meat. 577. Pseudomonas spp. was the prevalent
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INFLUENCE OF FERTILIZATION TREATMENTS ON NITRATES CONTENT OF SOME VEGETABLE SPECIES CULTIVATED IN THE FIELD
Enuţa IORGA1, Monica CATANĂ1, Luminiţa CATANĂ1, Mioara NEGOIŢĂ1, Alina BÂLEA1, Gabriela LILIOS2
1National Institute of Research&Development for Food Bioresources – IBA Bucharest, 021102, Bucharest 2, 6 Dinu Vintila Street, Romania, e-mail: [email protected] 2Ovidius University, 900527, Constanţa, 124 Mamaia Street, Romania, e-mail: [email protected]
Abstract: The existing amount of nitrates in plant at a given time is the result of balance between the amount absorbed and used in protein-genesis. A main cause of nitrate accumulation in vegetables is the use of nitrogenous fertilizers on crop land. High concentration of nitrates in plants (especially in vegetables) means a hazard for human and animal body, for two reasons: possibility of methemoglobin appearance at children and nitrates conversion to nitrites in saliva and formation of cancerigen nitrosamines in the intestinal tract. This paper presents the results of researches made in order to establish the influence of fertilization treatments on nitrates content of some vegetable species, cultivated in the field: egg plants, green peppers, bell peppers and carrots. In the culture technology of these vegetables, the following fertilization variants were applied (fertilizer being ammonium nitrate with 33% nitrogen): V1 = 0 kg nitrogen/ha; V2 = 100 kg nitrogen/ha; V3 = 200 kg nitrogen/ha; V4 = 400 kg nitrogen/ha; V5 = 800 kg nitrogen/ha. In order to establish the accumulation potential of nitrates in vegetables cultivated in the field, an enzymatic method was used. When no fertilizer was added in the culture, bell peppers had the minimum value of the nitrates - - content (11.53 mg NO3 /kg), and carrots the maximum one (90.42 mg NO3 /kg). - In the case of the highest fertilization level, carrots had the highest nitrates content (391.20 mg NO3 - /kg), and bell peppers the lowest one (63.08 mg NO3 /kg).
Key words: egg plants, green peppers, bell peppers, carrots, nitrates
Introduction quantities can pollute soil, groundwater and end products. When determining the dose Achieving a sustainable soil fertility of fertilizer, the following aspects should be reintegration involves the mineral, plant taken into consideration: nutrients in the and animal in its natural flow. Under natural soil, the degree of assurance of water conditions, soil organic matter is plants, pre-plant, the planned production on maintained in terms of intake of animal and cultivated plot [2]. vegetable, devoid of life, which falls on the Naturally, between nitrates and nitrites in surface of the land mass of dead roots, soil soil, water and plants equilibrium is microorganisms that die, all incorporated by established, which can be disrupted by biological cycles characteristic of intensive use in agriculture of natural biocenoses [1]. organic fertilizers and of those synthetic Fertilizer application is made on the basis nitrates, especially. Their degradation of careful analysis for each crop separately products enrich the soil and they can be and for each type of soil, taking into accumulated in the cultivated plants until account that their use in uncontrolled deleterious levels for consumers.
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- + - + Vegetables and fruits can also accumulate NO3 +NADPH + H NO2 + NADP + nitrogen as nitrates and nitrites form. H2O Mineral nitrogen amount in tissues of The amount of oxidized NADPH is vegetable and fruit is higher in the case of stoechiometrically equal to the nitrate one. species at which nitrates reductions is made Decreasing of NADPH amount is measured .nm 340 = ג in leaves and when light intensity and by absorbance at temperature are lower [3]. The factors determining the nitrates accumulation in cultivated horticultural products within protected environments (greenhouses, solar) or in the open field are [4]: genetical potential of species high doses of mineral fertilizers with nitric nitrogen, applied to cultures low temperature reduced light intensity Figure 1. Kit for nitrates determination through maintenance system of soil enzymatic method
Experimental Results and Discussion
We carried out a study to determine the In the case of experimental variant V1 influence of fertilization treatments on (without fertilizer addition), the nitrates nitrates content of some vegetable species, content of egg plants is in the range 41.15 – cultivated in open field: egg plants (Zaraza - 44.55 mg NO3 /kg. The average value of variety), green peppers (Buzău 10 variety), accumulation potential of nitrates, in this bell peppers (Splendens variety), and - case is 42.61 mg NO3 /kg. carrots (Nabuco variety). In the case of fertilizer application, the The samples were supplied by the nitrates content of egg plants increases with Vegetable Research Development Plant for fertilizer dose (ammonium nitrate with 33% Horticulture Buzau. In the culture nitrogen). Thus, in the case of fertilization technology of these vegetables, the variant V2 (100 kg nitrogen/ha) the egg following fertilization variants were applied plants have the nitrates content in the range (fertilizer being ammonium nitrate with - 81.95 – 84.67 mg NO3 /kg, and the average 33% nitrogen): V1 = 0 kg nitrogen/ha; V2 = accumulation potential of nitrates is 83.22 100 kg nitrogen/ha; V3 = 200 kg - mg NO3 /kg, with 95.30% much higher than nitrogen/ha; V4 = 400 kg nitrogen/ha; V5 = in the case when no fertilizer is applied. 800 kg nitrogen/ha. For each fertilization In the case of fertilization variant V3 (200 variant, we made 4 repetitions. kg nitrogen/ha) the egg plants have a To determine the accumulation potential of nitrates content in the range 137.59 – nitrates in vegetables cultivated in open - 143.25 mg NO3 /kg, and the average field, an enzymatic method was used. In accumulation potential of nitrates is 140.56 this method, nitrate is reduced by the - mg NO3 /kg, approximately 3.3 times reduced nicotinamide adenine dinucleotide higher than in the case when no fertilizer is (NADPH), to nitrite, in the presence of applied. nitrate-reductase (NR) [5]: The application of fertilizer dose of 400 kg NR
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nitrogen/ha (fertilization variant V4), Correlation between fertilization level with nitrogen and accumulation determines an increase of 4.83 times of the potential of nitrates in egg plants, cultivated in the field 350 average accumulation potential of nitrates 300 in egg plants cultivated under these 250 conditions, in comparison with those non- 200 y = 0,3216x + 58,607 fertilized ones. In the case of this 150 R2 = 0,9732 fertilization variant, egg plants have nitrates 100 Nitrates (mg NO3-/kg) content in the range 203.55 – 209.23 mg 50 - 0 0 200 400 600 800 1000 NO3 /kg, and the average accumulation Fertilization variants potential of nitrates is 206 mg NO -/kg. 3 Figure 3. Correlation between fertilization levels The use within culture technology of a with nitrogen and accumulation potential fertilizer dose of 800 kg nitrogen/ha of nitrates in egg plants, cultivated in the field (fertilization variant V5), determines an increase of 7.11 times of the average In the case of experimental variant V1 accumulation potential of nitrates within the (without fertilizer addition), the nitrates egg plants cultivated under these content of green peppers is in the range 19.8 - conditions, in comparison with those non- – 21.57mg NO3 /kg. And the average value fertilized ones. In the case of this of accumulation potential of nitrates, in this - fertilization variant, the egg plants have case, is 20.79 mg NO3 /kg. nitrates content in the range 295.37 – In the case when fertilizer is applied, the - 308.19 mg NO3 /kg, and the average nitrates content of green peppers increases accumulation potential of nitrates is 303.05 with fertilizer dose (ammonium nitrate with - mg NO3 /kg. 33% nitrogen). Thus, in the case of According to the achieved results, egg fertilization variant V2 (100 kg nitrogen/ha) plants are a vegetable species with an green peppers have the nitrates content in the - average accumulation potential of nitrates. range 32.28 – 33.05 mg NO3 /kg, and average accumulation potential of nitrates is - Acumulation potential of nitrates in eggplants cultivated in field, 32.58 mg NO3 /kg, with 56.71% much depending on fertilization level higher than in the case when no fertilizer is 350 300 applied. V1 - 0 kg nitrogen/ha /kg)
- V2 - 100 kg nitrogen/ha 3 250 In the case of fertilization variant V3 (200 kg V3 - 200 kg nitrogen/ha 200 V 4 - 400 kg nitrogen/ha V 5 - 800 kg nitrogen/ha nitrogen/ha) green peppers have the nitrates 150 - Nitrates content in the range 46.24 – 49.77 mg NO3 100 content (mg Nitrates (mg NO 50 /kg, and the average accumulation potential - 0 of nitrates is 48.02 mg NO3 /kg, V1 V2 V3 V4 V5 Fertilization variants approximately 2.31 times higher than in case
Figure 2. Accumulation potential of nitrates when no fertilizer is applied. in egg plants cultivated in the field, depending on The application of fertilizer dose of 400 kg fertilization level nitrogen/ha (fertilization variant V4), determines an increase 3.23 times of the Between the fertilization level with nitrogen average accumulation potential of nitrates in of egg plants culture and accumulation green peppers under these conditions, in potential of nitrates in egg plants, a linear comparison with those non-fertilized ones. In correlation is established, described by the the case of this fertilization variant green equation peppers have nitrates content in the range - y = 0.3216x + 58.607, linearity coefficient 65.74 – 68.80 mg NO3 /kg, and the average 2 (R ) being 0.9732. accumulation potential of nitrates is 67.14 - mg NO3 /kg.
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Acumulation potential of nitrates in green peppers Within the fertilization variant V1 (without cultivated in field, depending on fertilization level fertilizer addition), the nitrates content of 100 V1 - 0 kg nitrogen/ha
/kg) bell peppers is in the range 11.53 – 14.04 - 80 V2 - 100 kg nitrogen/ha 3 V3 - 200 kg nitrogen/ha - 60 V 4 - 400 kg nitrogen/ha mg NO3 /kg. The average value of the V 5 - 800 kg nitrogen/ha 40 accumulation potential of nitrates in this Nitrates - 20 content case is 12.63 mg NO3 /kg.
Nitrates (mg NO Nitrates (mg 0 When fertilization is applied, nitrates V1 V2 V3 V4 V5 Fertilization variants content of bell peppers increases once with increasing of fertilizer dose (ammonium Figure 4. Accumulation potential of nitrates in green peppers cultivated in the field, depending nitrate with 33% nitrogen). Thus, in the on fertilization level case of fertilization variant V2 (100 kg nitrogen/ha) bell peppers have a nitrates - The use within the culture technology of content in the range 17.03 – 19.25 mg NO3 fertilizer dose of 800 kg nitrogen/ha /kg, and the average accumulation potential - (fertilization variant V5), determines an of nitrates is 18.17 mg NO3 /kg, with increase 4.59 times of the average 43.86% much higher than in the case when accumulation potential of nitrates in green no fertilizer is applied. peppers cultivated under these conditions, Within the fertilization variant V3 (200 kg in comparison with those non-fertilized nitrogen/ha) bell peppers have a nitrates - ones. In the case of this fertilization variant, content in the range 27.10 – 29.35 mg NO3 the achieved green peppers have nitrates /kg, and the average accumulation potential - - content in the range 93.56 – 98.34 mg NO3 of nitrates is 28.08 mg NO3 /kg, of 2.22 /kg, and the average accumulation potential times higher, than in the case when no - of nitrates is 95.41 mg NO3 /kg. fertilizer is applied. Between the fertilization level with nitrogen The application of a fertilizer dose of 400 of green peppers culture in field and kg nitrogen/ha (fertilization variant V4) accumulation potential of nitrates in green determines an increase of 3.22 times of the peppers, a linear correlation is established, average accumulation potential of nitrates described by the equation y = 0.0922x + in bell peppers cultivated under these 25.14, linearity coefficient (R2) being conditions, in comparison with those non- 0.9769. fertilized ones. In the case of this According to the achieved results, green fertilization variant bell peppers have peppers are a vegetable species with low nitrates content in the range 39.67 – 41.55 - accumulation potential of nitrates. mg NO3 /kg, and the average accumulation - potential of nitrates is 40.67 mg NO3 /kg. Correlation between fertilization level with nitrogen and accumulation The use in the culture technology of a potential of nitrates in green peppers, cultivated in the field 120 fertilizer dose of 800 kg nitrogen/ha 100 (fertilization variant V5) determines an -/kg) 3 80 increase of 4.88 times of the average
60 y = 0,0922x + 25,14 accumulation potential of nitrates in bell 2 40 R = 0,9769 peppers cultivated under these conditions,
Nitrates (mg NO 20 in comparison with non-fertilized ones. In
0 the case of this fertilization variant, bell 0 100 200 300 400 500 600 700 800 900 Fertilization variants peppers have the nitrates content in the - Figure 5. Correlation between fertilization level range 60.85 – 63.08 mg NO3 /kg, and the with nitrogen and accumulation potential of average accumulation potential of nitrates is - nitrates in green peppers, cultivated in the field 61.70 mg NO3 /kg.
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nitrate with 33% nitrogen). Thus, in the
Acumulation potential of nitrates in bell peppers cultivated case of fertilization variant V2 (100 kg in field, depending on fertilization level nitrogen/ha) carrots have a nitrates content - 70 V1 - 0 kg nitrogen/ha in the range 159.55 – 161.93 mg NO3 /kg, 60 V2 - 100 kg nitrogen/ha V3 - 200 kg nitrogen/ha and the average accumulation potential of /kg) -
3 50 V 4 - 400 kg nitrogen/ha - 40 V 5 - 800 kg nitrogen/ha nitrates is 160.98 mg NO3 /kg, with 77.19% 30 much higher than in the case when no 20 Nitrates content 10 fertilizer is applied. (mg
Nitrates (mg NO 0 Within the fertilization variant V3 (200 kg V1 V2 V3 V4 V5 nitrogen/ha) carrots have a nitrates content Fertilization variants - in the range 199.85 – 201.85 mg NO3 /kg, Figure 6. Accumulation potential of nitrates in and the average accumulation potential of bell peppers cultivated in the field, depending on - fertilization level nitrates is 200.83 mg NO3 /kg, of 2.21 times higher than in the case when no fertilizer is Between the fertilization level with nitrogen applied. of culture of bell peppers in the field and The application of a fertilizer dose of 400 accumulation potential of nitrates in bell kg nitrogen/ha (fertilization variant V4) peppers, a linear correlation is established, determines an increase of 2.87 times of the described by the equation y = 0.0617x + average accumulation potential of nitrates 13.736, linearity coefficient (R2) being in carrots cultivated under these conditions, 0.9901. in comparison with those non-fertilized ones. In the case of this fertilization variant Correlation between fertilization level with nitrogen and accumulation carrots have nitrates content in the range potential of nitrates in bell peppers, cultivated in the field - 70 260.53 – 261.44 mg NO3 /kg, and the
60 average accumulation potential of nitrates is - 50 260.62 mg NO3 /kg. /kg ) - 3 40 The use in the culture technology of a
30 y = 0,0617x + 13,736 fertilizer dose of 800 kg nitrogen/ha R2 = 0,9901 20 (fertilization variant V5) determines an Nitrates (m g NO 10 increase of 4.3 times of the average
0 accumulation potential of nitrates in carrots 0 100 200 300 400 500 600 700 800 900 Fertilization variants cultivated under these conditions, in
Figure 7. Correlation between the fertilization comparison with non-fertilized ones. In the level with nitrogen and accumulation potential of case of this fertilization variant, carrots nitrates in bell peppers, cultivated in the field have the nitrates content in the range - 389.72 – 391.50 mg NO3 /kg, and the According to the obtained results, bell average accumulation potential of nitrates is - peppers are a vegetable species with low 390.49 mg NO3 /kg. potential of nitrates accumulation. Between the fertilization level with nitrogen Within the fertilization variant V1 (without of culture of carrots in field and fertilizer addition), the nitrates content of accumulation potential of nitrates in carrots carrots is in the range 90.18 – 91.88 mg a linear correlation is established, described - NO3 /kg. The average value of the by the equation y = 0.3544x + 114.42, accumulation potential of nitrates in this linearity coefficient (R2) being 0.9806. - case is of 90.85 mg NO3 /kg. According to the obtained results we have When fertilizer is applied, the nitrates drawn the conclusion that carrots are a content of carrots increases once with vegetable species with an average increasing the fertilizer dose (ammonium accumulation potential of nitrates.
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3. Bell pepper samples obtained in the field Acumulation potential of nitrates in carrots cultivated in field, depending on fertilization level culture, through application of 5
400 fertilization variants, have an accumulation V1 - 0 kg nitrogen/ha potential of nitrates in the range: 12.63 mg /kg) - 300 V2 - 100 kg nitrogen/ha 3 - - V3 - 200 kg nitrogen/ha NO3 /kg – 61.70 mg NO3 /kg. The V 4 - 400 kg nitrogen/ha 200 V 5 - 800 kg nitrogen/ha minimum value of nitrates content is for Nitrates non-fertilized bell peppers, and the 100 content (mg NO3- Nitrates (mg NO maximum value is for fertilized bell 0 V1 V2 V3 V4 V5 peppers with 800 kg nitrogen/ha. Fertilization variants 4. Carrot samples obtained in the field
Figure 8. Accumulation potential of nitrates in culture, through application of 5 carrots cultivated in the field, depending on fertilization variants, have an accumulation fertilization level potential of nitrates in the range: 90.85 mg - - NO3 /kg – 390.49 mg NO3 /kg. The Correlation between fertilization level with nitrogen and minimum value of nitrates content is for accumulation potential of nitrates in carrots, cultivated in the field 450 non-fertilized carrots, and the maximum 400 value is for fertilized carrots with 800 kg
/kg) 350 - 3 300 nitrogen/ha. 250 200 y = 0,3544x + 114,42 2 150 R = 0,9806 Acknowledgments 100 Nitrates (mg NO 50 0 The experiments were performed within the 0 200 400 600 800 1000 contract no. 51-050/18.09.2007, financed Fertilization variants through Programme 4 “Partnerships in Figure 9. Correlation between the fertilization priority S&T Domains” 2007 – 2013 – level with nitrogen and accumulation potential of nitrates in carrots, cultivated in the field National Centre for Projects Management.
Conclusions References
1. Egg plant samples achieved within 1. LUCHIAN V., Legumicultură generală şi culture in the field, through application of 5 specială, Ed. Elisavaros, Bucureşti, 2007 2. BORLAN Z., DORNEANU A., Nutriţia minerală fertilization variants, have an accumulation a plantelor în sisteme durabile de protecţie vegetală potential of nitrates in the range: 42.61 mg şi cu referire specială la fertilizarea cu fosfor, Ed. - - NO3 /kg – 303.05 mg NO3 /kg. The Europa Nova, Bucureşti, 1999 minimum value of nitrates content is for 3. GHERGHI A., BURZO I., MIRUNA BIBICU, non-fertilized egg plants, and the maximum LIANA MĂRGINEANU, LILIANA BĂDULESCU and all, Biochimia şi fiziologia legumelor şi value is for fertilized egg plants with 800 fructelor, Ed. Academiei Române, Bucureşti, 2001 kg nitrogen/ha. 4. BIBICU M., Cercetări metodologice privind 2. Green pepper samples achieved within determinarea nitraţilor şi nitriţilor din ţesuturi culture in the field, through application of 5 vegetale şi nivelului de acumulare în produsele fertilization variants, have an accumulation horticole, Bucureşti, 1994 5. *** Protocol de lucru pentru metoda enzimatică - potential of nitrates in the range: 20.79 mg UV de determinare a nitraţilor din produse - - NO3 /kg – 95.41 mg NO3 /kg. The alimentare, R – Biopharm, 2008 minimum value of nitrates content is for non-fertilized green peppers, and the maximum value is for fertilized green peppers with 800 kg nitrogen/ha.
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INFLUENCE OF TOTAL SOLUBLE CONTENT, STARTER CULTURE AND TIME PERIOD ON RHEOLOGICAL BEHAVIOUR OF CULTURED BUTTERMILK
Mircea-Adrian OROIAN1, Gheorghe GUTT1
1 Ştefan cel Mare University of Suceava, Faculty of Food Engineering, Universităţii Street, 720229, Suceava, Romania, e-mail: [email protected], [email protected]
Abstract: Casein gels are part of the dairy products, hard or soft products, as cheese and yogurt, cultured buttermilk, and their rheological behaviour can be measured by various instrumental techniques. For this study we used three types of milk (7.5%, 10%, and 12.5% respectively TSS) and starter culture (Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. cremoris) in different concentrations (2 g/100 l, 3 g/100 l, 4 g/100 l). The rheological properties of the cultured buttermilk obtained from milk were investigated by a Brookfield RV Pro II+ viscometer at different shear rates, during the time period keeping (from the 1st day – the obtaining day, till the 20th day – the last day when the product is good for human consumption). The samples with the highest TSS (total soluble content) present the highest viscosity, while the samples with 7.5% TSS content present the smallest viscosity. During the rheological tests, the samples with 3 g/100 l starter culture presented the best rheological properties (the samples had the highest viscosity), while the samples with 4 g/100 l had the smallest viscosity due to the proteolysis generated by the starter culture activity. The flow index behaviour was influenced more by the TSS content rather than by the starter culture dose. It can be seen that the change in the apparent viscosity was not linear with solids concentration, where a 5% (from 7.5% to 12.5%) increase in the solids concentration led to triplicate in the apparent viscosity.
Keywords: viscometer, shear stress, shear rate, flow index
1. Introduction
Rheology is the study of deformation and deformation responses of foods when flow of matter. The science of rheology subjected to normal and tangential stresses grew considerably due to research work [2]. In food research, the term is often used done on synthetic polymers and their interchangeably with texture, which refers solutions in different solvents that in turn to the flow, deformation, and disintegration was necessary due to the many uses of the of a sample under force. Strictly speaking, polymers in day-to-day and industrial texture relates to solid foods, and applications. Nevertheless, because of the viscosity—the tendency to resist flow— biological nature of foods, food rheology relates to fluid foods. Food can exhibit offers many unique opportunities of study. both solid and liquid characteristics, and Many foods are composed mainly of rheology can identify the properties of biopolymers and aqueous solutions such foods [3]. containing dissolved sugar and ions. The Typically, milk is fortified with dairy former ones are large molecules, often ingredients to produce a milk base which is called macromolecules, such as proteins, then submitted to a drastic heat treatment, polysaccharides, and lipids from a wide which results in a high level of thermal range of plant and animal sources. [1] denaturation of the whey proteins and their Rheological properties are based on flow partial fixation on the casein micelles. As a
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consequence, aggregation is promoted, which can be oval. The cells, which can be giving stronger gels and decreasing the associated in pairs or chains, have a extent of acidification required to allow diameter between 0,5-1,5 µm and do not association. Finally, the lactic acid present mobility. Lactococcus lactis lactis production during the fermentation step and Lactococcus lactis cremoris are results in the destabilization of the micellar mesopilic, and are characterised by their system and associated gelation of the potential to produce lactic acid. They have proteins. As the isoelectric point of the capacity to ferment lactose under 0,5% denatured proteins (pH 5.2), and casein and have a reduce acidotolerance (are (pH 4.6) are reached, low-energy bonds, inhibited at pH<4.5) [9]. mainly hydrophobic, are progressively established between the proteins [4]. 2. Materials and methods Casein gels are responsible for many 2.1. Materials rheological properties of cheese, yogurt, Milk with a TSS 7,5 %, 10%, 12,5% buttermilk, cream and other dairy products reconstitute, Lactococcus lactis subsp. that gel, stretch, and fracture. Rheological lactis and subsp. cremoris pure starter studies are performed as a quality control culture (Enzymes & Derivates S.A. Costisa method in dairy plants and as a technique Neamt); orbital shaker; thermostat; for scientists to study the structure of the Brookfield viscometer Model RV- DV II product [5]. Pro, with disk spindle, RV2 type. The dairy products (yogurt, cultured buttermilk, sana cultured buttermilk) Tab. 1. presents the thixotropy phenomena. Milk properties Thixotropy is the property of certain gels 7,5% 10% 12,5% or fluids that are thick (viscous) under Fat, g/100g 2.1 2.8 3.5 normal conditions, but flow (become thin, Protein, g/100 g 1.92 2.56 3.2 less viscous) over time when shaken, Sugar, g/100g 2.7 3.6 4.5 agitated, or otherwise stressed. Thixotropy Ash, % 0.43 0.57 0.72 0 should be defined as: the continuous Acidity, T 18 18 18 decrease of viscosity with time when flow is applied to a sample that has been 2.2. Sample preparation previously at rest and the subsequent All the samples were made using recovery of viscosity in time when the reconstituted milk. The starter culture is flow is discontinued. This is consistent extracted from the freezer and thawed. with the IUPAC terminology [6]. It should The milk was heated at 90 0C for 5 min, be noted that various general scientific afterwards the samples are cold to 28 0C. dictionaries and encyclopedias still give The starter culture was inoculated at 28 0C different definitions, often more closely in (2 g, 3 g and respectively 4 g starter culture line with Freundlich's original definition /100 l milk with 7.5%, 10% and [7]. The essential elements of the definition respectively 12.5% TSS). For a good used nowadays are that: it is based on activation of the starter culture, the viscosity; it implies a time-dependent samples are shaken for 15 min on a orbital decrease of the viscosity induced by flow; shaker at 150 rpm. The starter culture was the effect is reversible when the flow is inoculated at 28 0C. All the samples were decreased or arrested [8]. maintained at 28 0C for 12 h in the The members of Lactococcus gen are thermostat. The samples were kept in the Gram positive lactic bacteria, having a refrigerator at 4 0C for 20 days. coccus form or swerved form from it,
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2.3. Determination of rheological was indicative of the thixotropy nature of properties of cultured buttermilk culture buttermilk. The power law equation Viscosity measurements were carried was found to be an adequate model to describe the flow behaviour of the samples out on the cultured buttermilk samples at 2 room temperature (200C), with a in this study. The R values of the Brookfield viscometer (Brookfield equations log shear rate versus log Engineering Inc, Model RV- DV II Pro+) rotational speed were found to vary from at 0,5; 1; 2; 2,5; 4; 5, 10 and 20 rpm with 0.950 to 0.998. The flow index behaviour RV spindle (RV2). The spindle nos (n) of the power law model varied between (spindle nos: 2) was used in accordance 0.208 to 0.255. Although n does not have a with the sample nature to get all readings strong dependence on the concentration of within the scale. starter culture, while the TSS concentration The samples in 300 ml of beaker with a has a great impact on the index flow 8,56 cm diameter (according to the behaviour. Brookfield requests) were kept in a According to Mitschka, P., Steffe, J.& thermostatically controlled water bath for Daubert, C. [10, 11] the Brookfield about 10 min before measurements in viscometer with disk spindles represents a order to attain the desirable temperature of good method to determine non-Newtonian 250C. fluid properties. First measurements were taken 2 min One of the manifestations of thixotropy after the spindle was immersed in each is the hysteresis loop. The hysteresis sample, so as to allow thermal equilibrium technique was introduced by Green and in the sample, and to eliminate the effect of Weltmann [12]. It consists in immediate time dependence. systematically increasing and decreasing All data were then taken after 40 s in the shear rate between zero and a each sample. Each measurement was maximum value.
duplicated on the sample. ]] The obtained empirical data were 2 converted using the Mitschka relationships dyne·cm to shear rate and shear stress. The shear [ rate versus shear stress data were 10 · interpreted using the power law expression σ=k·γn, where σ – shear stress (N/m2), γ is -1 the shear rate (s ), n is the flow behaviour Shearstress index, k is the consistency index (Nsn/m2). The values for the flow behaviour Shear rate [s-1] index n, were obtained from plots of log shear stress versus log rotational speed; the Fig.1. Hysteresis loop of culture buttermilk slope of the line (if the dependence is sufficiently close to a linear one) is simply The change can be a continuous ramp or equal to the flow index of the fluid, n. a series of small steps. When the transient data are plotted as shear stress versus shear 3. Results and discussion rate, a thixotropic sample will describe a hysteresis loop (fig.1) because the stress Viscosity data showed that all samples will lag behind the shear rate [8]. The gap under examination were non-Newtonian between the two signals leads from the fluids, since the values for the flow breakdown of the casein chain during the behaviour index, n were under 1, which strain application.
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The empirical data were converted to next equation: shear rate and shear stress. τi=kτ·αi·C [1] The shear stress is calculated using the
Apparentviscosity [Pa·s] Apparentviscosity [Pa·s] Apparentviscosity [Pa·s]
Shear rate [s-1] Shear rate [s-1] -1 Shear rate [s ] Fig. 3 Changes in the apparent viscosity with shear rate in cultured buttermilk: A. 7.5% TSS; B. 10% TSS; C. 12.5% TSS: 2 g starter culture / 100 l milk; 3g starter culture/100 l milk; 4 g starter culture / 100 l milk *The apparent viscosity measuring were made for the 1st day
The influence of TSS on the rheology of culture buttermilk is a positive one, the viscosity increases with the increasing of TSS percent, due to the quantity of protein in the sample. The samples with 12.5% TSS had the highest viscosity, while the samples with 7.5% TSS the lowest viscosity (fig. 2), the shear stress had the same evolution like the viscosity did. The flow index behaviour varies between 0.211 and 0.255 due to the increasing of protein Fig. 2. Changes in the viscosity in cultured content of the samples. buttermilk (1st day) It can be seen that the change in the apparent viscosity was not linear with Where solids concentration, the same observation 2 τi – shear stress [dyne/cm ] was obtained by Hazim et al. [13], where a kτ= 0.119, this constant is for the 5% (from 7.5% to 12.5%) increase in the spindle nos 2 solids concentration led to triplicate in the αi – torque dial, % apparent viscosity (from 11.14 to 30.05 C – 7,187 dyne/cm for RV viscometer Pa·s at 0.5 rpm). This is an indication that The shear rate is calculated using the the control of the solids concentration is an next equation: important quality factor and it may affect γi = kγ(n)·Ni [2] the final acceptance of cultured buttermilk -1 γi – shear rate, s by the consumer. kγ(n) – constant, depends by the value of The changes of the viscosity with the n starter culture dose is not a linear one, the Ni – rotational speed, rpm increasing of the dose from 2 g to 3 g
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increased the viscosity and the shear rate viscosity and the shear rate decreased. The (tab.2, fig.4), but from 3 g to 4 g the maximum of rheological behaviour
] ] 2 ] 2 2
Shearstress [dyne/cm hearstress [dyne/cm Shearstress [dyne/cm S
Shear rate [s-1] Shear rate [s-1] Shear rate [s-1] Fig. 4 Changes in the shear stress with the shear rate in the cultured buttermilk: 2 g starter culture /100 l milk 1st day; 3g starter culture/100 l milk 1st day; 4 g starter culture /100l milk 1st day; 2 g starter culture /100 l milk 10th day; 3g starter culture /100 l milk 10th day; 4 g starter culture / 100 l milk 10th day; 2 g starter culture /100 l milk 20th day ; 3g starter culture /100 l milk 20th day ; 4 g starter culture / 100 l milk 20th day
( viscosity, shear rates, flow index) is of the cell increased in all the samples and observed at 3 g starter culture as is led to the decreasing of shear stress for all presented by the producers on the starter the samples due to the break of the casein culture description sheet. The decreasing chains by the activity of cells (fig.4). of the rheological properties can be generated by the activity of the cells of Conclusions Lactococcus lactis subsp. lactis and subsp. The rheological properties of cultered Cremoris, which is a power full proteolysis buttermilk are influeced by the TSS, starter activity. culture and time period keeping, leading The starter culture activity increased form st th to the conclusions that the samples with the 1 day to the 20 day, and the viability 12.5% TSS and 3 g starter culture/100 l milk have the best rheology (viscosity and Tab. 2. Changes in the rheological properties of the shear stress functions are the highest for cultured buttermilk prepared in three different these samples). formulations The Brookfield viscometer Model RV- DV Apparent Flow index II Pro+, represents a cheap method to Sample viscosity [n]* determine rheological properties of non- [Pa·s]** 2 g/100 l 0.211 8.49 Newtonian fluids, providing information 7.5% 3 g/100 l 0.216 11.14 about viscosity, flow index behaviour, TSS 4 g/100 l 0.208 9.57 shear rate and shear stress of fluids. 2 g/100 l 0.229 15.41 10 % The flow index behaviour (n) varies 3 g/100 l 0.234 18.89 TSS between 0.211 and 0.255, the variation is 4 g/100 l 0.227 17.13 greater with the increasing of the TSS 2 g/100 l 0.238 27.07 12.5% content compared with the starter culture 3 g/100 l 0.255 30.05 TSS 4 g/100 l 0.235 23.53 dose. It can be seen that the change in the *The flow index is for the samples with 3 g/100l starter culture, apparent viscosity was not linear with in the 1st day solids concentration, where a 5% (from **The apparent viscosity was measured at 0.5 rpm 7.5% to 12.5%) increase in the solids
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concentration led to triplicate in the 4. LUCEY, J. A., & SINGH, H.. Formation and apparent viscosity. This is an indication physical properties of acid milk gels: a review. Food Research International (1998), 30(7), 529– that the control of the solids concentration 542. is an important quality factor and it may 5. TUNICK, M.H. Rheology of Dairy Foods the affect the final acceptance of cultured Gel, Stretch, and Fracture, Journal Dairy Science buttermilk by the consumer. 83:1892-189 6. IUPAC. Compendium of chemical terminology, electronic version http://goldbook. iupac.org. Acknowledgment W06691.html. 7. BARNES HA. J Non-Newton Fluid Mech This paper was supported by the project 1997;70:1. "Knowledge provocation and development 8. MEWIS, J., WAGNER, N.J. Thixotropy, through doctoral research PRO-DOCT - Advances in Colloid and Interface Science 147-148 (2009) 214-227 Contract no. POSDRU/88/1.5/S/52946 ", 9. COSTIN, G.M., Produse lactate fermentate, project co-funded by European Social Editura Academica, 2005 Fund through Sectorial Operational 10. MITSCHKA, P. A simple conversion of Program Human Resources 2007-2013. Brookfield R.V.T. readings into viscosity functions, Rheologica Acta, 21, 207-209, SpringerLink, 1982 11. STEFFE, J., DAUBERT, C., Bioprocessing References pipelines: Rheology and Analysis, Freeman Press. 1. RAO, M.A. Phase transitions, food texture and USA, 2006 structure, in Texture in Food, Volume 1: Semi-Solid 12. GREEN H, WELTMANN RN. Ind Eng Chem Food, ed. B.M.McKenna, pp. 36-62, Woodhead Anal Ed 1943;15:201 Publishing Ltd., Cambridge, UK. 13. HAZIM, A.M., BASIM, A.-J., ALI, Al-S., 2. RAO, M.A., Rheology of Fluid and Semisolid Effect of solids concentration on the rheology of Foods, Principles and Applications, Second Edition, labneh (concentrated yougurt) produced from sheep Food Engineering Series, Springer, 2007 milk, Journal of Food Engineering 61 (2004) 347- 3. GIESE, J.. Measuring physical properties of 352 food. Food Technol. (1995), 19:54–63.
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GENETIC RESOURCES AND THE PROGRESS REACHED BY WINTER WHEAT BREEDING
Constantin DROBOTĂ 1 , Ioan GONTARIU2
1 Agricultural Research and Development Station of Suceava, B-dul 1 Decembrie 1918 no.15, Suceava, e-mail: [email protected] 2 Stefan cel Mare University of Suceava, Faculty of Food Engineering, 13 Universitaţii St., 720229, Suceava, Romania, e-mail: [email protected]
Abstract: This paper shows the contribution in the development of the winter wheat crop, from the beginning of the breeding activity, through the testing and identification of the most adequate autochthonous and foreign wheat varieties, and later, through creating, introducing and extending in production of eight varieties: Bucovina, Suceava 84, Aniversar, Gaşparom, Magistral, Esenţial, Drobeta and Voroneţ. The cultivars Aniversar, Gaşparom, Magistral and Esenţial were cultivated during period 1996-2005. These cultivars registered an average yield of 6367 kg/ha, reaching a yield rise of 197% in comparison with the average yield registered during the period 1951-1955. Thus, during the last 60 years an ascending dynamic progress, materialized through an yearly increasing rate up to 1,4% was registered, in comparison with the average yield obtained at the standard variety Flamura 85, during 1996-2005. The superiority of the eight varieties created at the Agricultural and Development Research Station of Suceava, from the productivity and yield stability point of view, was statistically demonstrated, through the distribution of the correlations and variability of productions, in comparison with the standard varieties from the above mentioned periods. An important progress concerning the physiological breeding characteristics such as: winter, foliar diseases and lodging resistances, was registered, too.
Keywords: average yield, ecological factors, agricultural areas
Introduction: that are superior from both the quality and the quantity point of view to the varieties The presence of some risk ecological cultivated in time, as the wheat cultures factors with a limitative action not only on behave risks because of the biotic and the quantity but also on the yields quality abiotic factors, registering frequently big has determined the fact that during the 60 yield fluctuations [5, 6, 9]. years period since the foundation of In connection with that, at the A.R.D.S. of A.R.D.S. Suceava, a correct strategy of the Suceava, a research activity sustained in way of organization and development of the breeding field, following initial the breeding researches for winter wheat selection and identification of the most should be objectively approached. This appropriate varieties for winter wheat, in affirmation is also sustained through the our country and abroad, having in mind the results that have been registered [1, 2, 3, 6, introduction of them in the culture and 7]. replacing from the cultivated variety the The agricultural increasing rate on the one with poorer production performances, more poor lands in the agricultural areas has been carried on, since its foundation in with a wet and cold climate, from the north 1946 [1, 4, 7, 12]. and north west of Moldova have amplified The research organized on the basis of a the necessity to create some new varieties national experimentation system of the
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wheat varieties has developed only after future new more performing genotypes. 1950, increasing after 1970, when we The value dimensioning of the superiority initiated the special works of breeding in of the varieties created is presented during the creation of new varieties and wheat three significantly periods of time as lines more valuable and better adapted to follows: the ecological and cultural conditions in 1. Period 1946-1975 highlights the Moldova and in the northern part of the difference in behavior of varieties and the country [1, 2, 6]. wheat lines verified at Suceava having in Through the development of breeding consideration the aspect of physiological, researches, superior wheat genotypes have quality and production features. been created, this being one of the safest 2. Period 1976-1995 presents the ways and efficient ones to increase productive value of the varieties Bucovina, productivity, stability and yield quality. Suceava 84 and Aniversar, cultivated in Having this in mind, at present the this period of time, comparing with the appreciations made by the best researchers Fundulea 29 used as a standard in the in the field are unanimously accepted, as northern areas. they estimate a continuous genetic 3. Period 1996-2005 highlights the progress for the wheat varieties created in Gaşparom, Magistral, Esenţial, Drobeta our country, dimensioned to a annual rate and Voroneţ varieties, in comparison with of up to 1, 4 % and even more for other witness variety Fundulea 4 used in this agronomical characters, with a direct period of time. influence on the stability of yield [8, 9, 10, For the appreciation of the experimented 11, 12]. varieties value, from the point of view of This work aims at appreciating the the productive potential, the obtained contribution brought to wheat culture, results have been used, during the initially through the testing and experimentation years, in the comparative identification of the most appropriated national crop, comparing it with the national and foreign wheat varieties and witness varieties with the greatest spread in after that through the creation, introduction production: Fundulea 29 and Fundulea 4. and extension in the production of the The relation between the results of the varieties obtained at A.R.D.S. of Suceava varieties created in Suceava and those of in 60 years since its foundation. the standard variety Fundulea 4 in the period 1996-2005 has been described Biological Material and Research through the calculation of the linear Method regressions and the establishment of the production has been characterized through The breeding working method used so far the determination of the variances(s2).The at the A.R.D.S. of Suceava in the last 60 results of the phenological observations years has contributed in the first half of made during the vegetation are presented the period to the testing and generalization in the work through smaller values, and the in the culture of the most valorous interpretation of the production data has autochthonous and foreign wheat varieties, been made through statistic methods of and in the second half of the period to variance analyses creation of the 8 wheat varieties and also to The experimentation has been carried out obtaining of a valuable breeding material, in comparative crops, executed at the well adapted to specific conditions of beginning after the method of plots culture in the north part of the country, that position in Latin rectangle (with 5 offers the possibility of creating in the replications and 10 m2 the cropped
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surfaces of one plot) and then in the ammonium nitrate or nitro-limestone have balanced grating with reiteration of the been administered. basis scheme (6 replications and 7 m2 The sowing has been carried out, in the cropped plot) majority of the experimental years, in the Over the years the experiments have been optimum epoch for this area being assured placed after potato, in the leached 550 germinated grains/ m2. For the chernozem soil, reached in humus and total creation of the 8 winter wheat varieties to nitric, middle provided with phosphor and A.R.D.S. of Suceava, with a genealogy and mobile kalium, not being necessary the method obtained being presented in short phase fertilization in autumn, as the potato terms, valuable genetic resources have has received larger quantities of chemical been used, most of them being adapted to fertilizer. In spring, before the beginning of the northern and west northern part of vegetation period, 60-70 kg N s.a. /ha as Moldova (table 1).
Table 1 The genealogy and obtaining method of the winter wheat varieties created at the A.R.D.S. of Suceava Variety Parental forms Cross Homologation Obtaining year year. method Bucovina F.53-67 x F. 117-67 1968 1979 Suceava Bezostaia 1 x F.208 – 65 1969 1984 84 The individual Aniversar Lovrin 11 x F.53 – 67 1970 1986 selection Magistral 1502W4-1/154I1-1//Aniversar/Roxana 1986 1998 (Pedigree Gaşparom Sv. 9710-79 x Fundulea 4 1984 1998 method) Esenţial Sv. 2946 – 86 x Sv. 9710 – 79 1988 2001 Drobeta Sv. 9710 – 79 x F. 4141 – W 1 1991 2003 Voroneţ Sv. 9710 – 79 x Progres 1994 2004 Climatic conditions, appreciated from the have been closed, having a normal point of view of the annual precipitation density. quantity and the way of distribution of In the majority of the years the regime of these in the course of vegetation, have the rains has been richer in the intense been favorable in the majority of the vegetation period, May – June, being an experimentation years. Not only in the advantage for the growths and autumn period but also in the winter development of plants, exception making period, the climatic conditions have not the years: 1996, 1999, 2000, 2003 and influenced negatively the vegetation of 2004, when a humidity deficit of 116.3 mm plants , so that on spring arrival the crops has been registered (table 2). Table 2 The precipitation regimes at ARDS of Suceava during vegetation periods (1996 – 2005) Precipitation per years, mm: Multiyear Period 1995/ 1996/ 1997/ 1998/ 1999/ 2000/ 2001/ 2002/ 2003/ 2004/ average 1996 1997 1998 1999 2000 2001 2002 2003 2004 2005 August 51.7 136.0 88.1 21.1 68.2 39.1 73.2 112.3 58.0 52.9 69.7 September- 95.0 150.6 100.8 186.5 92.0 81.9 191.0 108.3 74.4 65.1 79.2 Oct. November 65.5 27.1 21.2 30.5 22.6 10.8 63.5 47.0 10.9 39.5 30.4 Dec. - March 126.5 84.5 104.4 68.3 77.3 89.1 76.6 67.8 90.8 101.4 92.8
April 38.3 52.8 63.0 87.6 50.9 60.5 6.3 10.8 19.0 100.6 45.6 May – June 93.3 153.2 262.9 90.1 54.0 123.9 119.0 46.4 61.6 225.4 163.2 July 53.8 111.9 101.6 65.8 115.0 91.4 238.0 252.5 160.7 45.3 89.3 Total 524.1 716.1 742.0 549.9 480.0 496.7 767.6 645.1 475.4 630.2 570.2
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The normal calculation of 61 years wheat , and the other plants taken into The wet and cold regime in the vegetation consideration in the area has registered period has positively influenced the an ascending line . appearance and development of foliar Beginning with the year 1946 and until diseases attack and contributed in the 2005 , during six decades , a number of same time to the amplification and 6391 varieties and wheat lines of sometimes early falling of plants from the perspective, have been verified, from the most sensitive varieties . point of view of production and Under the report of thermal regime, in the vegetation behavior of which 1750 majority of the experimental years , autochthonous and foreign varieties. The values relatively close to the multi- most valuable of these varieties have annual average have been registered , been introduced and generalized in without inducing negative influences over production , occupying at the level of the plants vegetation. different periods numerous surfaces.
RESULTS AND DISCUSSION The yield quality
A valuable contribution in the agricultural From the point of view of the physical progress in Bucovina is brought to the aspects of production it is easy to see that plants breeding research field, beginning one thousand kernel weight of the with the foundation of the ARDS in 1946 experimented varieties during 1970 -1975, through verifications of different has varied greatly from one variety to varieties and wheat populations’ behavior another, ranging values from 33,8g to 42,4 from the western part of Suceava Plateau g (table 3). It has been noticed that from area, the research activities being carried this point of view the Lovrin 10, Aurora out on the basis of a national and Kaukaz varieties have registered the experimentation system of wheat varieties highest values for one thousand kernel which have been developed only after weight (39,9 g –42,4 g) 1950 . It can be appreciated that winter
Table 3 The yield quality of wheat varieties and lines which were experimented during period 1970 -1975 at ARDS of Suceava One Total Gluten From 100g thousan protein Pelshenk Farinog flour d kernel % from e index raf note Bread Bread Variety or line weight d.s. N x volum weigh wet Dry -g- 5,7 e t -cm3- -g- Lovrin 10 42.4 15.4 32.9 10.3 59 45 508 152 Aurora 41.0 15.3 31.1 10.4 81 45 518 156 Kaukaz 39.9 16.1 33.6 10.9 75 46 531 154 Brucker 6111 35.5 15.4 31.6 10.4 94 48 530 151 T. 195 - 65 33.8 14.0 27.2 9.6 135 50 514 153 Dacia 36.8 16.1 31.6 10.9 101 49 540 153 Favorit 33.9 16.2 32.9 11.4 107 59 547 154 Bezostaia1-st. 33.8 14.4 29.0 9.8 114 51 496 151
According to protein content, represented noticed for the Favorit , Dacia and Kaukaz through values of 14,0-16,2%, it has been varieties, to have higher content of wet
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and dry gluten. values up to 33.3 % at wet gluten and From the point of view of gluten quality, 10.8 % for the dried one. Taking into which bread making features depend on consideration this aspect we have noticed ultimately, for the Favorit, Bezostaia 1, that, through the increased gluten content, Dacia, Brucker 6111 varieties and for the varieties Bucovina, inbred line T. 195 – 65, the following Aniversar, Suceava 84, Apullum, Ariesan, aspects have been analyzed: the highest Bezostaia 1 and Dropia have registered value for Pelshenke index, of 94 – 135, values up to 30% at wet gluten and 10.8% farinograf note, up to 48, the bread volume dried gluten. From the point of view of the calculated at 100 g flour between 496 and gluten quality, which finally determines 547 cm3, and its weight up to 151 g the bread making features of flour, we can observe that higher gluten content The yield quality varieties had also the higher glutenin index, from 54.5 at Bucovina, up to 60.5 From the point of view of the quality at Ariesan. features we can notice that between the The bread making features value of the experimented varieties in the period 1991- experimented varieties, appreciated 1995 there are no differences that could through bread volume and weight, from surprise us, these displaying good 100g of flour , demonstrates that from qualities of milling and bread making this point of view the advanced varieties features (table 4). are mostly varieties with good bread According to the protein content, making features qualities, the bread represented by average values from 12.2 volume registering from 441 cm3 in the % at Transilvania 1 and Turda 81, up to variety Apullum, up to 506 cm3 at Dropia, 13.8 %, in the varieties Aniversar, and their weight varying between 139 g at Fundulea 29, Dropia and Bezostaia 1, the variety Turda 81 and 144 g at varieties gluten content has registered higher Dropia, Aniversar and Fundulea 4.
Table 4. Quality traits of the wheat varieties experimented at ARDS of Suceava (1991-1995) Curre Variety % % gluten Glutenic Bread from 100 g flour nt No. protein wet dry index Volume -cm3 weight –g 1 Fundulea 29 13.8 28.4 9.7 52.8 477 143 2 Fundulea 4 12.4 25.3 8.5 54.4 443 144 3 Flamura 85 13.6 28.1 9.5 54.2 453 142 4 Rapid 13.6 29.0 9.5 58.6 457 143 5 Dropia 13.8 32.1 10.2 59.6 506 144 6 Bucovina 13.1 30.9 9.3 54.5 476 142 7 Suceava 84 12.8 33.0 9.5 54.7 480 143 8 Aniversar 13.8 33.3 10.1 55.3 479 144 9 Moldova 83 12.8 29.4 9.2 53.0 495 140 10 Turda 81 12.2 25.2 8.3 49.6 489 139 11 Apullum 13.4 30.2 9.5 50.2 441 142 12 Arieşan 13.3 30.5 9.4 60.5 437 140 13 Transilvania 1 12.2 26.9 8.6 53.9 502 141 14 F. 308 13.2 28.5 9.6 52.5 495 141 15 Bezostaia 1 13.7 33.3 10.8 57.1 496 141
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The yield quality requests regarding this matter (table 5). The chemical content of grains frequently The physical features of production, at registers high values, superior to the the varieties created at ARDS of Suceava Flamura standard, not only for the protein between 1996-2005, are highlighted by content from 13.8 % to 14.2 %, but also bigger and heavy grains, fact that confers for the wet gluten content (28.4 – 32.4 %) them higher values not only for one and dried gluten (9.5 – 10.5 %). The gluten thousand kernel weight, most often indicator between 85.0 and 97.9 ml, registered between 45-50 g, and the test superior to the Flamura 85, proves in a weight had values between 75-80 kg. special way that the quantity and quality Taking into consideration the bread values of the gluten from protein for the making features, the Gasparom, Magistral, newly varieties created at ARDS of Esential, Drobeta and Voronet varieties Suceava between 1996 and 2005, is frame themselves among the crops with obviously due to the lower deformation superior qualities, proving also to be indicator level (6 – 7 mm as against to 8 competitive in comparison to the actual mm at standard).
Table 5 The main indicators for bread making features quality for the created and experimented varieties at ARDS of Suceava during period 1996-2005 in comparison with the Flamura 85 variety.
Crt FLAMURA Specification GASPAROM MAGISTRAL ESENTIAL DROBETA VORONET No. 85 1 Protein content (%) 14.2 13.9 13.8 13.8 14.1 13.6 2 Wet gluten (%) 30.2 32.4 28.5 28.4 29.8 28.1 3 Dried gluten (%) 10.5 9.8 9.8 9.5 9.5 9.5 4 Glutenic index (ml) 85.0 89.6 95.8 97.9 91.8 84.2 The deformation 5 7 6 6 7 6 8 index (mm) FARINOGRAMA Hydration capacity 6 66.3 62.8 61.2 62.4 61.8 61.2 (%) 7 Developing (min.) 2.1 2.1 2.3 2.5 2.5 2.1 8 Stability (min) 2.3 2.2 2.4 2.6 2.6 2.1 9 Elasticity (uv) 130 128 126 133 131 125 10 Power 56.8 57.7 59 56.6 57.5 50.5 11 Value index 68.4 67.4 70.8 65.2 67.4 61.8 12 Bread volume (cm3) 510 512 510 505 510 500 13 Bread weight (g) 145 145 145 143 145 142
VALUE GROUP A2 –B1 A1 –B1 A2 –B1 A2 –B1 A1 –B1 B1 –B2
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7000
6000
5000
4000
6367 3000 5740 kg/ha kg/ha 4650 kg/ha 2000 3410 2680 kg/ha 2145 kg/ha 1000 kg/ha 1000 kg/ha 0 1946-1950 1951-1955 1956-1965 1966-1975 1976-1985 1986-1995 1996-2005
Fig. 1 The genetic progress made by winter wheat, in the period of time 1946-2005, through utilization in the crop of the most valuable varieties, under crop conditions offered by ARDS of Suceava
Analyzing the evolution of the genetic lines and afterwards creating new progress registered by wheat through the varieties and wheat lines more utilization in the crop production of the productive and better adapted to crops most valuable varieties, during the 60 conditions from the North Moldova , years since the foundation of the ARDS of have contributed to the rational zoning Suceava, we can notice that there is a of the best varieties, at the introduction continuous increase, quantified by average and generalization in the crop and productions from 2145 kg/ha, during the through this to progressive increase of period 1951 – 1955, to 6367 kg/ha, production. during the period 1996 – 2005, which in Through the researches carried out over relative values means an increase of 197 time at ARDS of Suceava, 8 common % (fig.1). winter wheat varieties have been created The genetic progress reflects especially (Bucovina, Suceava 84, Aniversar, the increase of production capacity, Gasparom, Magistral, Esential, Drobeta registered especially in the last 30 years and Voronet) more productive and better (1976-2005), when as a consequence of adapted to the specific crop conditions intensification of creating new varieties, for the wet and cold climatic areas. wheat productivity has increased a lot, The global analyses of the production reaching an average level of 4650 kg/ha, performances have proved that, through during the period 1976-1985, up to 6367 the creation and utilization in the crop kg/ha during the period 1996 – 2005 of these genotypes, in time, dynamic increasing progress has been registered, CONCLUSIONS dimensioned through an annual increasing rate of production, in The research activities carried out comparison with the annual average between 1946-2005 at ARDS of yield of the standard variety Flamura 85, Suceava, initially in the direction of identifying varieties and more valuable
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of up to 1.4 % during the period 1996- References 2005 Significant progresses concerning the 1. GAŞPĂR, I., 1968 – Soiuri şi linii de grâu de breeding of the physiological features toamnă de perspectivă pentru cultură în Podişul Sucevei. Probleme agricole, nr. 8; and quality features have been also 2. GAŞPĂR, I., 1976 – Soiuri şi linii de grâu şi registered: secară valoroase şi de perspectivă pentru - The winter resistance and resistance to judeţul Suceava. Volum omagial, 1946 – 1976: cryptogrammic diseases attack has 47-61; been increased from average forms to 3. GAŞPĂR,I., 1984 – Soiul de grâu de toamnă Suceava 84. Cercetări Agronomice în Moldova, resistant forms; vol. 3 (67): 39-44; - The lodging resistance has been 4. GAŞPĂR, I., 1987 – Caracterizarea improved, through the shortening of bioproductivă a soiului de grâu „Aniversar”, height and the thickening of stem, from creat la Staţiunea Agricolă Suceava. Cercetări low to very good; Agronomice în Moldova, Anul XX, vol. 4 (80):37-42; - Concerning the quality features, the 5. GAŞPĂR,I., 1993 – Rezultate experimentale progress obtained places the new privind comportarea unor soiuri de grâu, varieties, created at ARDS of Suceava, secară şi triticale la S.C.A. Suceava. Cercetări among the wheat varieties with superior Agronomice în Moldova, Anul XXVII, vol. 3-4 bread making features (classes A -B (102): 31-38; 1 1 6. GAŞPĂR,I., ZAMĂ, E., - 1996 – Rezultate and A2-B1). experimentale privind comportarea unor soiuri The winter wheat varieties that may be şi linii de grâu de perspectivă în condiţiile de found at present in Moldova, especially cultură din Podişul Sucevei. Volum omagial, in the northern and west-northern part of 1946-1996: 21-36; Moldova, are autochthonous varieties with 7. GAŞPĂR, I.,CRISTEA, M., POPOVICI,D., 1996 – Strategii şi obiective în ameliorarea multiple adaptability qualities to the plantelor la Staţiunea Suceava. Lucrări ecological conditions, and until the ştiinţifice- volum omagial, 1946-1996. creation of some more performing S.C.Agris-Red. Rev. Agricole S.A.: 11-21; varieties , we strongly recommend the 8. GAŞPĂR, I., ZAMĂ, E., POPESCU, following ones: Gasparom, Magistral, CONSTANŢA, MURARIU, M., 2000 – Gaşparom şi Magistral – soiuri noi de grâu de Esential, Drobeta and Voronet. toamnă, create la Staţiunea Suceava. Analele I.C.C.P.T. Fundulea, vol LXVII: 49-64; 9. GAŞPĂR, I., MURARIU, M., ZAMĂ, E., 2002 – Progresul realizat în ameliorarea grâului de toamnă la S.C.A. Suceava în ultimele trei decenii, Cercetări de genetică vegetală şi animală, vol.VII: 59-69.
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A REVIEW ON THE EFFECT OF FUNGI ON THE WHEAT GRAIN UNDER POST HARVEST STORAGE ECOLOGY
Mathew SHIJU1
1Ministry of Higher Education, Department of Natural Sciences, Aksum University, Ethiopia Contact No.: (+251) 920126043; e-mail: [email protected]
Abstract: The review paper is aimed to assess the effect of fungi on the wheat grain under the post harvest storage system. It has been observed that very high percentage of wheat kernels has been lost under post harvest storage. The grain quality greatly depends upon the storage conditions. The storage requirement violation can result in grain deterioration. The biotic and abiotic factors influence the various physical and chemical properties of wheat grains under post harvest storage. The fungal ecology present in the stored grain has an important role in spoilage and the production of mycotoxins. The mycoflora of stored wheat grains predominantly consisted of ubiquitous mould genera Aspergillus, Alternaria, Cladosporium, Fusarium, Mucor, Rhizopus and Penicillium possibly because of their omnipresence, capacity to grow on all possible substrates and a wide range of temperature and humidity. The most frequent species observed on the stored wheat grains of Aspergillus were A. niger, A. fumigatus, Alternaria alternata, Fusarium moniliformis, Rhizopus arrhizus and a few Pencillium species. Among these almost all have the capacity to produce mycotoxin which can contaminate and cause spoilage. The various physical, microbial and biochemical analyses of post harvest storage wheat grain is an essential component to evaluate the grain quality in various Indian wheat varieties and also important for facilitating the minimal post harvest food grain loss.
Keywords: loss, biotic factor, abiotic factor, mycotoxin, infestation.
1.1 Introduction traditionally regarded as an indicator of poor storage conditions. Mycotoxins may The grain production varies from year to already be present in grains coming into year and hence the grains should be stored storage or may be produced as a result of strategically from years of overproduction poor storage only if there is sufficient for the use in year of under production. moisture. Conversely, not all moulds that Stored grains can have losses in both grow in stored commodities produce quantity and quality. The wheat grain after mycotoxins. The metabolic activity of harvest is influenced by a wide variety of these pioneer species raises the moisture biotic and abiotic factors and has been content of the grain, which may allow studied as a stored grain ecosystem. Losses growth of mycotoxigenic species and occur when the grain is attacked by ultimately, the formation of mycotoxins. microorganisms and particularly by fungi. Alternaria, Aspergillus and Penicillium can The grain losses in quantity and quality act as pre or post-harvest pathogens of can be under the form of depletion in seed grain and may form mycotoxins. viability, hardness, colour, size and shape, grain weight and various biochemical 1.1.1 The origin and classification of parameters, protein, carbohydrate and wheat vitamins under post harvest storages. The presence of mycotoxins in grains is Wheat belongs to the genus Triticum of the
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grass family, Poaceae. This genus is chromosomes [1-9]. Globally, wheat is the originated in the tropical South west Asia, most-produced food among the cereal where it occurs in wild as well as in crops after rice. Table 1.1 shows the cultivated forms. Wheat is well adapted to taxonomic status of the genus Triticum . harsh environments and is mostly grown The development of a modern on wind swept areas that are too dry and classification depended on the discovery, too cold for the more tropically inclined in the 1918s, of the fact that wheat was rice and corn, which do best at classified according to three ploidy levels intermediate temperature levels. Wheat [10-16]. The classification of Van Slageren was first grown in the United States in (1994) is probably the most widely used 1602 on an island of the Massachusetts genetic-based classification at present. coast. Man has been dependant upon the Most species of wheat can be described in wheat plant for thousands of years. Wheat Latin binomials, e.g., Triticum aestivum, genetics is more complicated than that of rather than the trinomials necessary in the most other domesticated species. Some genetic system, e.g., Triticum aestivum wheat species are diploid, with two sets of subsp. aestivum. Both approaches are chromosomes, but many are stable equally valid and both are widely used polyploids, with four sets of chromosomes [17]. (tetraploid) or six (hexaploid) set of
Table 1.1: The Taxonomical status of Triticum Kingdom Plantae Plants Subkingdom Tracheobionta Vascular plants Superdivision Spermatophyta Seed plants Division Magnoliophyta Flowering plants Class Liliopsida Monocotyledons Subclass Commelinidae Order Cyperales Family Poaceae Grass family Genus Triticum Wheat Species aestivum Common wheat
material for roofing thatch. Although Wheat (the Triticum spp.) is cultivated worldwide. Globally, wheat is the most- wheat supplies much of the world's dietary produced food among the cereal crops after protein and food supply, as many as one in rice. Wheat grain is a staple food used to every 100 to 200 people suffers from make flour for leavened, flat and steamed Coeliac disease, a condition which results breads; cookies, cakes, breakfast cereal, from an immune system response to a pasta, noodles; and for fermentation to protein found in wheat: gluten [18-20]. make beer, alcohol, vodka or even biofuel. Grain fed to livestock whole or coarsely Durum is the most commonly used to ground. Starch is used for pastes and sizing make pasta. The most common wheat is textiles. Straw is turned into mats, carpets, the hexaploid wheat and includes spelt, baskets, and used for packing material, modern bread wheat and soft wheat used cattle bedding, and paper manufacturing. for cookies and cakes. Wheat is grown to a Some wheat is cut for hay. Wheat grown limited extent as a forage crop for for grain crop is also used for pasture livestock, and the straw can be used as before the stems elongate and as a fodder for livestock or as a construction temporary pasturage; it is nutritious and
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palatable. mycotoxins into the food chain. In some cases, pre-harvest decisions can 1.1.2 The effect of stored wheat grains significantly impact the capability for under post harvest storage subsequent post-harvest control. The propagates of mould fungi, frequently The spoilage of grain in storage is brought found in grain and its surroundings, start to about by two variables, the biotic and develop when favourable conditions abiotic [21]. Wheats are attacked by many emerge. Temperature and substratum fungi and other organisms. Some cultivars moisture are the most significant factors are resistant to the various rusts, smuts, that determine the intensity of and virus diseases. The most important micromycete development and grain fungal diseases of wheats are the extension damage. agents which are concerning diseases in an area before growing wheat. Also cvs 1.1.2.1 The effect of fungi on the odour should be selected for growing as resistant and discolouration of the stored wheat to various diseases. The biotic variables are grains different types of insects, mites, rodents, The fungal contamination of the grains is birds and microorganisms, i.e., bacteria responsible for emitting undesirable odours and fungi. The abiotic variables include [31] found that under high moisture temperature and moisture content of the condition cereals gave a musty odour. The grains; the latter in grains increases due to dark colouration of germs of the wheat seepage of moisture from soil or from grains are due to the damage caused by cemented floor [22]. The micro flora of fungi [32]. This is discoloration of the cereal grains normally includes fungi, germ end of the wheat grain. In the bacteria and actinomycetes [23-25]. A commercial grading system, black point number of fungi have been reported to be involves the germ end only and not associated with cereal grains and their extended into the grain crease. If more than milling fraction [26-27]. The majority of half the grain is damaged and well into the mycotoxins are chemically tough, resistant crease, it is classified as smudge. It has to temperature, conditions of storage and been also observed that if the wheat grains, processing technologies. Therefore, with moisture content of 13% or above is detoxication is not always effective. Based kept long enough at a temperature of 350C on the stages at which invasion and growth to 400C, the grains turn brown even in the occur, the fungi associated with grains can absence of storage fungi [33]. Grain itself conveniently be divided into two groups and the microbial contaminants respire i.e., Field fungi and Storage fungi [28]. slowly when stored dry. However, if the Spoilage of stored grain by fungi is water availability is increased to 15-19% determined by a range of factors which can moisture content (=0.75-0.85 water activity be classified into four main groups (aw), wheat) spoilage fungi, particularly including (a) intrinsic nutritional factors, Aspergillus and Penicillium species grow, (b) extrinsic factors (c) processing factors resulting in a significant increase in and (d) implicit microbial factors. The respiratory activity [34]. This can result in factors produce fungal colonization within an increase in temperature and sometimes the stored grains [29-30]. These reviews spontaneous heating from the colonization examined some important mycotoxins and by a succession of fungi resulting in the post-harvest control strategies which colonization by thermophilic fungi and have been developed for effective actinomycetes [35-37]. management to minimize entry of The quantitative grain contamination with
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micromycete propagules was established occurrence. During the course of their by a number of methods given by various development breakdown and utilization of scientists [38-44]. The grain colour the constituents of the seeds occur [88-89]. changes by some micromycetes produced The dark germs of grains are known to be and contaminate it with poisonous due to damage by storage fungi [90-94]. substances including mycotoxins [45-52]. The high moisture and temperature conditions for cereals gave a musty odour 1.1.3 The presence of micro flora due to fungi, instead of bacteria [95]. contaminating the stored wheat grains 1.1.4 Fungal infestation in post harvest Mould contamination of food and feed is a stored wheat grains constantly occurring phenomenon but the attitude towards it has been far from The wheat grains come in association with consistent [53]. It has been found that the fungi from the time of grain maturity fungal damage of grain is caused in and also at the time of storage. Some of various ways. They reduce the grain these fungi are in intimate association and viability, produce odour, grain are present as dormant mycelium under the discoloration and depletion in food added pericarp or dormant spores on the surface value (FAV) by the increase in FAV and of the kernel. However, there is a number also produce toxins injurious to the health of fungi which are only superficially of consumers. The fungi are responsible associated with stored grains. Mould for emitting of undesirable odour from growth in grains may cause deleterious stored grains [54]. A number of studies changes in addition to the formation of have been reported the loss of dry matter mycotoxins. Many spoilage content and visible moulding of stored fungi cause loss of germination in seed wheat grains [55-62]. This could be grains, discolouration and darkening of the considered as a subjective index of the safe grains, reduction in protein content, musty storability of grain. There are problems odours, and changes in fatty acid profiles with the use of visible moulding as a and other constituents of the grains. Mould criterion of deterioration [63-66]. Wheat development may also encourage mite and quality loss has been measured and models insect infestation. The association of fungi developed based on germination rates, with cereal grains starts from the field visible mould growth or respiration of itself. Shortly after the grain reaches to grain and microorganisms. The maximum size, the lemma and palea microscopic growth may be a more protecting it are pushed apart exposing the effective measurement of initial grain to infection by fungi [96] and their colonization than visible moulding [67-70] extensive studies have been carried out in An extensive micro flora has been found to the laboratory on these aspects [97-99]. be associated with stored wheat grains [71- Poor post harvest management can lead to 83]. Earlier in the laboratory a number of rapid deterioration in grain quality, cereals have been screened with respect to severely decreasing the germinability and micro flora associated in storage grains nutritional value of stored grains. Fungal [84-87]. activity can cause undesirable effects in The effect of microbial infestation on grains including discolouration, contribute decrease in the germination of grains is to heating and losses in nutritional value, now well known. It has been established in produce off-odours, losses in other cereals and especially in wheat. germinability, deterioration in baking and Discolouration of wheat is a common milling quality, and can result in
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contamination by mycotoxins [100-102]. to a loss of moisture) is a measure of food A number of studies have been reported on loss but the latter may be proportionately the loss of dry matter content and visible larger owing to selective feeding by the moulding of stored wheat grains [103- pests. Rodents and moth larvae may 110]. This could be considered as a preferentially attack the germ of the grain subjective index of the safe storability of thus removing a large percentage of the grain. There are problems with the use of protein and vitamin content, whereas visible moulding as a criterion of weevils feeding mainly on the endosperm deterioration [111-114]. Fungi seldom will reduce the carbohydrate content. occur on grains in isolation, but usually as Many pests may eat the bran of cereals a mixed consortium of bacteria, yeasts and reducing vitamins such as thiamin [120- filamentous fungi. It is thus inevitable that 123] interspecific and intraspecific interactions The quantity and quality of endosperm will occur depending on the nutritional proteins are the major factors responsible status of the grain and the prevailing for baking quality, and nutritional value of environmental conditions. Wheat quality wheat [140-142]. Various seed storage loss has been measured and models proteins in wheat are classified upon their developed based on germination rates, solubility in different solvents, such as visible mould growth or respiration of water, saline solution, 70% aqueous grain and microorganisms [114-118] The ethanol and diluted acid or alkali solutions microscopic growth may be a more and are named as albumins, globulins, effective measurement of initial gliadins, and glutenins, respectively [126- colonization than visible moulding. Some 128]. attempts have also been made to relate dry The other storage factors such as moisture matter losses to actual calorific losses due and fungal infection also lead to changes in to the activity of mycotoxigenic moulds. vitamin content. In beans in particular, loss of protein is very important where there is 1.1.5 Decrease in nutritional value as infestation, as up to 25% of the dry matter due to myctoxin production in stored may be raw protein. Fungi growth leads to wheat grains reduced nutritional and technical quality of cereal grains [130-132]. Grain storage During storage, the grains undergo some conditions affect its quality due to high biochemical changes like the increase in percentage of mycotoxins [133-137]. fatty acids. The increase in fatty acid value Wheat grains are also rich in pantothenic is chiefly due to storage fungi and not due acid, riboflavin and some minerals, sugars to the activity of seeds [119]. The stored etc [138]. The composition of wheat grain grain will have quantitative and qualitative and flour is given in Table 1.2. losses. Weight loss during storage (not due
Table 1.2: Composition of wheat grain and flour [139] Parameter Grain (%) Flour (%) Moisture 9 - 18 13 - 15.5 Starch 60-68 65-70 Protein 8-17 8-15 Cellulose 2-2.5 Trace Fat 1.5-2 0.8-1.5 Sugars 2-3 1.5-2 Mineral matters 1.5-2 3-6
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Storage temperature affects the keeping mycotoxin which can contaminate and quality of wheat. Increase in temperature cause spoilage. accelerates the rate of respiration until it is limited by oxygen supply; build up of carbon dioxide concentration, exhaustion References of substrate, and thermal inactivation of essential enzymes. Heat damages the 1. U.S. DEPARTMENT OF AGRICULTURE APPROPRIATIONS (1957). Hearings of 84th Congress. gluten proteins and even discolours the 2nd day Session in 1956. United States Congress. pp 242. kernels of the stored hard red winter wheat 2. SMITH, J.E. AND ALBERT, E. (1995). Handbook with moisture contents of 10.95% in sealed of Weed Management Systems. Marcel Dekker. pp. 411. 0 0 3. BELDEROK, B., HANS, M. AND DINGENA A.D. (2000). half gallon glass jars at -1 C and 24 C for Bread-Making Quality of Wheat. Springer. pp3. two years [140-142]. The grain losses in 4. PALMER, J. J. (2001). How to Brew. quantity and quality can be under the form Defenestrative Pub Co. pp. 233. 5. NEILL, R. (2002). Booze: The Drinks Bible for the of depletion as mentioned above like that 21st Century. Octopus Publishing Group - Cassell of seed viability, hardness, colour, size and Illustrated. pp.112. shape, grain weight and various 6. CAUVAIN, A., STANLEY, P. AND CAUVAIN, P. (2002). Bread Making. CRC Press. pp 540. biochemical parameters, protein, 7. COLLEDGE, S., CONOLLY, J. AND SHENNAN, carbohydrate and vitamins under post S. (2002). Archaeobotantical Evidence for the Spread of harvest storages. The storage fungi damage Farming in the Eastern Mediterranean. Curr. Anthropol. 45:S35-S58. the grains in several ways; they reduce the 8. U.S. DEPARTMENT OF AGRICULTURE (2003). Annual germinability, produce undesirable odour World Production Summary, Grains, and kernel discoloration, decrease the food http://www.usda.gov/Agriculture crop production. 9. HANCOCK AND JAMES, F. (2004). Plant value and also produce toxins injurious to Evolution and the Origin of Crop Species. CABI the health of consumers. Publishing. The effect of mycotoxigenic fungi in post 10. MAGAN, N. (1993a). Early detection of fungi in stored grain. Int. J. Biodeterior. Biodegradat. 32:145- harvested stored wheat grains must be 160. examined in the context of the ecology as a 11. MAGAN, N. (1993b). Use of sulphur dioxide to whole in order to understand the control fungi in stored grain. In: Proceedings of Int. Conference on Controlled Atmosphere and Fumigation in dominance of certain species of fungi Grain Storages (S. Navarro and E. Donahaye, eds.). under certain environmental conditions. Capital Press, Jerusalem, Israel. pp. 163-171 Interactions between these fungi and other 12. KABARITY, A. (1966). On the origin of the new cultivated wheat II. Cytogenetical studies on the contaminants are complex and are karyotypes of some T. macha varieties. Beitr. Biol. significantly affected by the prevailing and Pflanzen. 42: 339-346. changing environmental factors. As a 13. MORRIS, R. AND SEARS, E.R. (1967). The cytogenetics of wheat and its relatives. In: Wheat and conclusion the mycoflora of stored wheat wheat improvement. American society of Agronomy grains predominantly consisted of monograph (K.S. Quisenberry and L.P. Reitz, eds.). ubiquitous mould genera Aspergillus, Madison, WI. pp 18-97. 14. BOWMAN, C.M., BONNARD, G. AND DYER, Alternaria, Cladosporium, Fusarium, T.A. (1983). Chloroplast DNA variations between species Mucor, Rhizopus and Penicillium possibly of Triticum and Aegilops. Location of the variation on the because of their omnipresence, capacity to chloroplast genome and its relevance to the inheritance and classification of the cytoplasm. Theor. Appl. Genetics grow on all possible substrates and a wide 65: 247-262. range of temperature and humidity. The 15. KERBY, K. AND KUSPIRA, J. (1987). The most frequent species observed in the phylogeny of the polyploidy wheat Triticum aestivatum (bread wheat) and Triticum turgidum (macaroni wheat) stored wheat grains of Aspergillus were A. Genome. 29: 722-737. niger, A. fumigatus, Alternaria alternata, 16. BREIMAN, A. AND GRAURA, D. (1990). Wheat Fusarium moniliformis, Rhizopus arrhizus evolution. Israel J. Plant Sci. 43: 85-98. 17. WAINES, J.G. AND BARNHART, D. (1992). and a few Pencillium species and some of Biosystematic research in Aegilops and Triticum. the fungi have the capacity to produce Hereditas 116: 207-212.
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18. HILL, I. D., HORVATH, K. AND FASANO, A. stored grain: a promising new approach. J. Stored (1995). Epidemiology of celiac disease. Am. J. Product Res. 38: 191-218. Gastroenterol. 90: 163-164. 36. MAGAN, N., SANCHIS, V. AND ALDRED, D. 19. FASANO, A. BERTI, I. AND GERARDUZZI, T. (2004). Role of spoilage fungi in seed deterioration. (2007). "Prevalence of celiac disease in at-risk and not- Chapter 28, In: Fungal Biotechnology in Agricultural, at-risk groups in the United States: a large multicenter Food and Environmetal Applications (D.K.Aurora, ed.). study". Arch. Int. Med. 163: 286-292. Marcell Dekker. pp. 311-323. 20. PRESUTTI, J., (2007). Celiac Disease. Am. Family 37. SHUEY, W.C. (1960). A wheat sizing technique for Physician 76: 196-1802. predicting flour milling yield. Cereal Sci. Today 5: 71-72 21. WHITE, N.D.G., SINHA, R.N. AND MUIR, W.E. (1982). 38. ELLIS, M. B. (1971). Dematiaceous Intergranular carbon dioxide as an indicator of biological Hyphomycetes. Kew: Commonwealth Mycological activity associated with the spoilage of stored wheat. J. Institute. pp. 608. Can. Agric. Eng. 24: 35-42. 39. ELLIS, M. B. (1976). More Dematiaceous 22. PINGALE, S.V. (1970). Prevention and losses in Hyphomycetes. Kew: Commonwealth Mycological storage. Bull. Grain Technol. 8: 3-13. Institute. pp. 785. 23. JAMES, N., WILSON, J. AND STARK, E. (1946). 40. DOMSCH, K. H., GAMS, W. AND ANDERSON, The microflora of stored wheat. Can. J. Res. 24: 224-233. T. (1980). Compendium of Soil Fungi. London: 24. GRAVES, R.R., ROGERS, R.F., LYONS, A.J. JR. Academic Press. Vol.1. pp. 405. AND HESSELTINE, C.W. (1967). Bacterial and 41. GERLACH, W., AND NIRENBERG, H. (1982). actinomycetes flora of Kansas – Nebraska and Pacific The Genus Fusarium: a pictorial atlas. Berlin–Dahlem: Northwest wheat and wheat flour. Cereal Chem. 44: 288- Institute of Microbiology. pp 406. 299. 42. RAMIREZ, C. (1982). Manual and Atlas of the 25. MEHROTHRA, B.S. AND BASU, M. (1975). Penicillia. Elsevier, Amsterdam. pp. 874. Survey of microorganisms associated with cereal grain 43. LUGAUSKAS, A., BRIDZIUVIENE, D., and their milling fraction in India. J. Imported wheat Int. LEVINSKAITE, L., PASKEVICIUS, A., PECIULYTE Biodegradation Bulletin 11. D., REPECKIENE, J., SALINA, O. AND VARNAITE, 26. MEHROTHRA, B.S. AND KUNWAR, I. K. R. (1997). Mikrobiologiniai medziagų pazeidimai. (1980). Survey of microorganisms associated with cereal Vilnius: Valstiecių laikrastis. pp 469. grain and their milling fraction in India II. Indigenous 44. LUGAUSKAS, A., PASKEVICIUS, A. AND wheat Int. Biodegradation Bull. ISSN0020 - 6164 spring. REPECKIENĖ, J. (2002). Pathological microorganism 27. Wicklow, D. T. (1995). The mycology of stored grain: an application Vilnius pp. 434. ecological perspective. In: Stored grain ecosystems 45. SCOT, P.M. (1994). Penicillium and Aspergillus toxins. In: (D. S. Jayas, N. D. G. White and W. E. Muir, eds.). New Mycotoxins in grain compounds other than aflatoxins (J. York, Marcel Dekker, pp. 197-249. O. Miller and H. L. Trewnholm, eds.). Eagan Press, St. 28. CHRISTENSEN, C.M. AND KAUFMANN, H.H. Paul, Minesota, USA. pp 261-268. (1969). Grain storage the role of fungi in quality loss. 46. MOSS, M. O. (1991). Mycology of cereal grain and University of Minnesoto Press, Minn U.S.A., pp. 153. cereal products. In: Cereal Grain: Mycotoxins, Fungi and 29. WALLACE, H.A.H. (1973). Fungi and other Quality in Drying and Storage (J. Chelkowski, ed.). organisms associated with stored grains in grain storage Elsevier, Amsterdam–London–New York–Tokyo. pp. 23- part of a system (R.N. Sinha and W.E. Muir, eds.). AVI 51. publishing Co., Connecticut Sec. II Capt. 4 : 71-78. 47. LACEY, L. AND MAGAN, N.(1991). Fungi in 30. WALLACE, H.A.H. AND SINHA, R.N. (1981). cereal grains: their occurrence and water and temperature Causal factors operative in distributional patterns and relationships. In: Cereal Grain: Mycotoxins, Fungi and abundance of fungi: a multivariate study. In: The Fungal Quality in Drying and Storage (J. Chelkowski, ed.). Community- Its Organisation and Role in Ecosystems Elsevier, Amsterdam–London– New York–Tokyo. pp. 77- (D.T. Wicklow and G.C. Carroll, eds.). Marcell Dekker 118. Inc., New York. pp. 233-247. 48. KUZMIENE, G., PAZARAUSKIENE, J. AND 31. SCHROEDER, H.W. AND HALICK, J.V. (1963). SIRTAUTAITE, S. (1991). Augalininkystės produktų Effect of moisture content, humidity and length of laikymas ir perdirbimas. Vilnius: Mokslas. pp.381. storage on maintenance of quality in rough rice. 49. MARIN, S., SANCHIS, V., RAMOS, A.G. AND Phytopathol. 54: 858-862. MAGAN, N. (1998a). Environmental factors, 32. GOLUMBIC, C. AND LAUNDANI, H. (1966). interspecific interactions, and niche overlap between Storage and warehousing. Year book of Agriculture (U.S. Fusarium moniliforme and F. proliferatum and Fusarium Department of Agriculture) pp. 139-152. graminearum, Aspergillus and Penicillium spp. isolated 33. GOLDBLATT, L.A. (1969). Aflatoxin scientific from maize. Mycol. Res. 102: 831-837. background control and implications. Academic Press, 50. ABRAMSON, D. (1998). Mycotoxin formation New York. pp. 472. and environmental factors. In: Mycotoxins in Agriculture 34. PETERSSON, S. AND SCHNURER, H. (1995). and Food Safety (K. K. Sinha and D. Bhatnagar, eds.). Biocontrol of Mold Growth in High-Moisture Wheat New York: Marcel Dekker Inc. pp. 255-277. Stored under Airtight Conditions by Pichia anomala, 51. Singh, H. Singh, N. Kaur, L., and Saxena, S.K. (2001). Pichia guilliermondii, and Saccharomyces cerevisiae. Effect of sprouting conditions on functional and dynamic Appl. Environ. Microbiol. pp. 1027-1032. rheological properties of wheat. J. Fd. Eng. 47: 23-29 35. FLEURAT-LESSARD, F. (2002). Qualitative reasoning and integrated management of the quality of
93 Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Year IX, No2 - 2010
52. ZVICEVICIUS, E., RAILA, A., NOVOSINSKAS, 67. ANJUM, F.M. AND WALKER, C.E. (1991). H. AND KRASAUSKAS, A. (2006). Mycotoxin Review on the significance of starch and protein to wheat producents in the grain layer. J. Ecologia. 3: 105-111. kernel hardness. J. Sci. Fd. Agric. 56: 1-13. 53. FORGACS, J. (1965). Stachybotrys toxicosis and 68. ARMOLIK, N., DICKSON, J.G., DICKSON, A.D. moulds corn toxicosem. In mycotoxins in food stuffs. (1956). Deterioration of Barley in storage by (G.N. Wogans, ed.). M.L.T. Press Cambridge, microorganisms. Phytopathol. 46: 457-461. Massachusetts. pp. 87- 104. 69. AUTRAN, J.C., HAMER, R.J., PLIJTER, J.J. AND 54. F. A. O. (1966). Grain of Life. pp. 193. POGNA, N.E. (1997). Exploring and improving the 55. KREYGER, J. (1972). Drying and storing grains, industrial use of wheats. Cereal Fd. World 42: 216-227. seeds and pulses in temperate climates. Institute for 70. BAKUTIS B. (2004). Microrganism grown on Storage and processing of Agricultural Products, Bulletin seeds. Kaunas: Terra Publication. pp. 81. 205, Wageningen, The Netherlands. 71. BALIUKONIENE, V., BAKUTIS, B. AND 56. SEITZ, L.M., SAUER, D.B., MOHR, H.E. (1982). STANKEVICIUS, H. (2003). Mycological and Storage of high moisture corn: Fungal growth and dry mycotoxicological evaluation of grain. Ann. Agric. matter loss. Cereal Chem. 59: 100-105. Environ. Med. 10: 223-227. 57. WHITE, N.D.G., SINHA, R.N. AND MUIR, W.E. 73. BASU (1974). Studies of microorganisms (1982). Intergranular carbon dioxide as an indicator of associated with stored wheat and its flour in India. biological activity associated with the spoilage of stored Research paper presentation at University of Lucknow, wheat. J. Can. Agric. Eng. 24: 35-42. India. 58. HOFSTEIN, R. S., DROBY, E., CHALUTZ, C., 74. BERG, S.O. (1947). Is the degree of grittiness of WILSON AND FRILENDER, B. (1990). Scaling-up the wheat flour mainly a varietal character? Cereal Chem. production for application of an antagonist-from basic 24: 274-283. research to R & D, In: Biological control of postharvest 75. BROOK, P.J. AND WHITE, E.P. (1966). Fungus diseases of fruits and vegetables, workshop proceedings. toxins affecting mammals. Ann. Rev. Phytopathol. 4: Agricultural Research Service document ARS-92. U.S. 171-194. Department of Agriculture, Washington, D.C. pp. 197- 76. BYERS, M., MIFFIN, B.J. AND SMITH, S.J. 210. (1983). A quantitative comparison of the extraction of 59. HAMER, A., LACEY, J. AND MAGAN, N. (1991). protein fractions from wheat grain by different solvents Use of an automatic electrolytic respirometer to study and of the polypeptide and amino acid composition of the respiration of stored grain. In: Proceedings of 5th Int. alcohol solution proteins. J. Sci. Fd. Agric. 34: 447-462. Working Conference of Stored Product Protection, (F. 77. CHELKOWSKI, J. (1991). Mycological quality of Fleurat-Lessard and P. Ducom eds.). Bordeaux, France, mixed feeds and ingredients. In: Cereal grain, Imprimerie dumedoc, 1: 321-329. mycotoxins, fungi and quality in drying and storage, (J. 60. KRASAUSKAS, A., STEPONAVICIENE A., Chelkowski, ed.): Elsevier, Amsterdam, London, New RAILIENE, M., LUGAUSKAS, A., RAILA, A. AND York, Tokyo, pp. 217-227. RAUDONIENE V. (2005). Impact of environmental 78. CHRISTENSEN, C.M. (1955). Grain storage conditions on the spread of micromycetes in grain during studies 21: Viability and Moldiness of Commercial wheat its harvesting and storage. Bot. Lithuanica. 11: 101- in relation to the incidence of germ damage. Cereal 109. Chem. 32: 507-518. 61. WAINES, J.G. AND BARNHART, D. (1992). 79. CHRISTENSEN, C.M. (1956). Deterioration of Biosystematic research in Aegilops and Triticum. stored grains by moulds. Wallerstein Lab. Commun. 19 : Hereditas 116: 207-212 31-48. 63. SINGH, H. SINGH, N. KAUR, L., AND 80. CHRISTENSEN, C.M. AND LINKO, P. (1963). SAXENA, S.K. (2001). Effect of sprouting conditions on Moisture content of hard red winter wheat as determined functional and dynamic rheological properties of wheat. by meter and by oven drying and influence of small J. Fd. Eng. 47: 23-29. differences in moisture content upon subsequent 64. LEELAVATHI, K., VETRIMANI, R. AND deterioration of the grain in storage. Cereal Chem. 40: HARIDAS-RAO, P. (1990). Changes in the functional 129-137. characteristics of wheat during soaking and subsequent 81. CHRISTENSEN, C.M., OLAFSON, J.H. AND germination. J. Fd. Sci. Technol. 27: 349-354. GEDDES, W.F. (1949). Grain storage studies 8: Relation 64. WOSNIAK, W. AND STYK, W. (1996). Internal damage of moulds in moist stored cotton seed to increase to wheat grain as a result of wetting and drying. J. Drying production of carbon dioxide, fatty acids and heat. Cereal Technol. 14: 349-365. Chem. 26 : 106-129. 65. LACEY, J., HAMER, A. AND MAGAN, N. 82. CIEGLER, A. (1978). Fungi that produce (1997). Respiration of wheat grain in different mycotoxins conditions and occurrence, Mycopathologia environments. (E.J. Donahaye and S.Navaro eds.). Proc. 65: 5-11. Int. Symp. on Controlled Atmospheres and Fumigation 83. COSKUNTUNA, L., KOÇ, F. AND COSKUN, F. (CAF), Caspit Press, Jerusalem, Israel. pp. 113-122. (2008). Effect of Chemical Content and Physical 66. ABRAMSON, D. (1997). Toxicans of genus Characteristics on Nutritional Value of Wheat Varieties Penicillium. In: Handbook of plant and fungal toxicants Collected from Thrace Region. Pak. J. Nutrition 7: 218- (J.P.F. D’Mello, eds.). CRC Press, Boca Raton, FL. pp. 221. ISSN 1680-5194 303-317. 84. DROCHNER, W. AND LAUBER, U. (2001). Occurrence of the three important Fusarium - toxins Deoxynivalenol, Nivalenol and Zearalenon in grains in
94 Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Year IX, No2 - 2010
Central Europe and effects in farm animals. Proc. Soc. of 104. KURASOVA, V.V., KOSTIN, V.V. AND Nutrition Physiol. pp 163-168 SALOVSKAJA (1945). Methodolgy of isolating 85. DUGGELI, M. (1904). The mycological vertanary microrganism, pp. 194 -195. florainfection. Zentr. Microbial Infection 12: 602-614. 105. LANGSETH, W., STENWING, H., SOGN, L. 86. ERIKSEN, G.S. AND ALEKSANDER, J. (1998). AND MO, E. (1993). Growth of moulds and production Fusarium toxins in cereals – a risk assessment. Tema of mycotoxins in wheat during drying and storage. Acta Nord. Nordic Council of Ministers, Copenhagen, Agric. Scand. [B] Soil Plant Sci. 1993, 43, 32-37 Denmark, 10-165. 106. LUGAUSKAS, A. (1997). Microbiological book 87. FIELD, R.W. AND KING, T.H. (1962). Influence for research: Vilnius, 35-390. of storage fungi on the deterioration of corn in storage a 106. MACHACEK, J.E. AND GREANEY, F.J. (1938). The review. Lowa Acad. Sci. 51: 265-280. block point of kernel smudge disease of cereals. Can. J. 88. FINNEY, K.F. AND BARMORE, M.A. (1948). Res. 16: 84-113. Loaf volume and protein content of hard red winter and 107. MAGAN, N., HOPE, R., CAIRNS, V. AND ALDRED, spring wheats. Cereal Chem. 25: 291-312. D. (2003). Post-harvest fungal ecology: impact of fungal 89. GOODING, M.J. AND DAVIS, W.P. (1997). Wheat growth and mycotoxin accumulation in stored grain. production and utilization, systems, quality and the European J. Plant Pathol. 109: 723-730. environment. Book CAB International. 108. MEHROTHRA, D.K. (1974). Studies on some 90. HALVERSON, J. AND ZELENY, L. (1988). microrganisms associated with rice in storage. Paper Criteria of wheat quality. Chapter 2. In: Wheat: presentation Abstract at University of Lucknow. Chemistry and Technology, Vol. 1. (Y. Pomeranz, ed.). 109. MILNER, Y. AND GEDDES, W.F. (1946). Grain Am. Assoc. Cereal Chem., Inc., St. Paul, MN. pp. 15. storage studies of influence of certain moulds inhibitors 91. HASAN, H.A.H. (1999). Mycoflora and changes of on respiration of moist wheat. Cereal Chem. 24 : 507- safflower, wheat and faba bean seed quality during the 517. storage. Rostl. Vyr. 45: 85-91. 110. MILNER, Y., CHRISTENSEN, C. M. AND 92. HESSELTINE, C. W. (1968). Flour and wheat: GEDESS, W.F. (1947). Grain storage studies 6: Wheat Research on their microbiological flora, Bakers Dig. 42: respiration in relation to moisture content, mould growth, 40-42, 66. chemical deterioration and heating. Cereal Chem. 24: 93. HESSELTINE, C. W. AND GRAVES, R.R. (1968). 182-199. Microbiology of flours. Econ. Bot. 20: 156-168. 111. MIS, A. AND GRUNDAS, S. (2002). Wheat grain 94. HOCKING, A.D. (2003). Stored grain in Australia hardness and quality. Int. Agrophysics 16: 283-288. 2003. Proceedings of the Australian Postharvest 112. MOHAMMADKHANI, A. (2005). Study of Technical Conference, Canberra. CSIRO Stored Grain Pentosans (Non Starch Polysaccharides), in Durum Research Laboratory, Canberra. pp. 55-58. Wheat and its Relation to the Quality of Protein and 95. HOSENEY, R.C. (1987). Wheat hardness. Cereal Grain Hardness Index (H.I.). Pak. J. Nutri. 4: 208-209. Fd. World 32: 320-322. 113. ORTH, R.A. AND SHELLENBERGER, J.A. 96. HUMPISCH, G. (2001). Gesundheitsvorsorge im (1988). Origin, production and utilization of wheat. In: Getreidelager. Neue Landwirtschaft. Vol. 2. pp. 44- Wheat: Chemistry and Technology (Y. Pomeranz, ed.). 48. Vol. I, AACC, St. Paul, MN, pp.1-14. 97. INAGASKI, N. AND IKEDA, M. (1959). Studies 114. PALNI, I.K. (1975). Studies on the microflora on the fungi isolated from foods II. Identification of associated with wheat grains and various milling fraction. Penicillia and Aspergilli from flour. Bull. Nat. Hygenic Paper presentation. Abstract, University of Lucknow. Lab. Tokyo 77: 347-366. 115. PAPAVIZAS, G.C. AND CHRITENSEN, C.M. 98. JAYAS, D. S. (1995). Stored-grain ecosystems. (1960). Grain storage studies 29: Effect of invasion by New York: Dekker, pp. 757. individual species and mixture of Aspergillus upon 99. JONES, D.B. AND GERSDORFF, C.E.F. (1941). germination and development of discoloured germs in The effect of storage on the protein of wheat, white flour, wheat, Cereal Chem. 27: 197-230. and whole wheat flour. Cereal Chem. 18: 417. 116. PELHATE, J. (1968). Inventaire de la mycoflora 100. JONSSON, N., PETTERSON, H. AND des bles de conservation (Inventory of mycoflora of SCHNURER, J. (2000). Study of the relationship stored wheat) Bull. Soc. Mycol. France 84: 127-143. between storage conditions and the growth of moulds and 117. POISSON, J. AND GUILBOT, M.A. (1956). production of Ochratoxin A in grain 24 preliminary Mycoflora des Farines Franscaises. Menuerie, France. pp results. Proc. Eur. Agricul. Eng. Sym. Warwick 42-52. University, U.K. pp.44. 118. POMERANZ, Y. AND WILLIAMS, P.C. (1990). 101. JORGENSEN, K. AND RASMUSEN, G. (1999). Wheat hardness: its genetic, structural, and biochemical Ochratoxin in Danish cereals 1986- 1992 and daily intake background, measurement, and significance. In: by Danish population. Fd. Additives Contaminants. 13: Advances in Cereal Science and Technology (Y. 95-104. Pomeranz ed.). American Association of Cereal 102. KENT- JONES, D.W. AND AMOS, A. J. (1930). Chemists, St. Paul, MN. 10: 471-548. Preliminary studies in bacteriology of wheat and flour. 119. PRESTON K.R., DEXTER J.E. AND KRUGER Analyst 55: 248-268. J.E. (1978). Relationship of exoproteolytic and 103. KOEKSEL, H., ATLI, A., OEZKAYA, H. AND endoproteolytic activity to storage protein hydrolysis in DEMIR, Z. (1993). Comparison of physical properties of germinating durum and hard red spring wheat. Cereal wheat and NIR spectroscopy hardness value for Chem. 55: 877-888. prediction of semolina yield. J. Agr. Forest. 17: 821-830.
95 Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Year IX, No2 - 2010
120. SAMSON, R.A. AND VAN REENEN, H. (1988). 137. MAGAN, N. (1993b). Use of sulphur dioxide to Introduction to Foodborne Fungi. 3rd ed. Centraalbureau control fungi in stored grain. In: Proceedings of Int. voor Schimmelcutures, Baar. Delff. Institute of the Royal Conference on Controlled Atmosphere and Fumigation in Netherland Academy of Arts and Sciences. Grain Storages (S. Navarro and E. Donahaye, eds.). 121. SANKARAN, R. (1976). Mould Problems in the Capital Press, Jerusalem, Israel. pp. 163-171 Storage of Food Grains. Symposium ‘Stratergies on 138. ORTH, R.A. AND SHELLENBERGER, J.A. Storage, Handling and Processing of Food Grains in the (1988). Origin, production and utilization of wheat. In: Present Context'. Bangalore Chapter of Association of Wheat: Chemistry and Technology (Y. Pomeranz, ed.). Food Scientists and Technologists, India. Vol. I, AACC, St. Paul, MN, pp.1-14. 122. SANKARAN, R., SEHGAL, D. D., PARIHAR, D. 139. GOODING, M.J. AND DAVIS, W.P. (1997). Wheat B. AND NATH, H. (1976). Cereal grain fungi and grain production and utilization, systems, quality and the quality. Def. Sci. 26: 161. environment. Book CAB International. 123. SANKARAN, R., MATTADA, R. R. AND 140. FINNEY, K.F. AND BARMORE, M.A. (1948). THANGAMANI. (1975). Mycoflora and changes of Loaf volume and protein content of hard red winter and wheat seed quality during storage. Itzd. Fd. Packer 29: 1. spring wheats. Cereal Chem. 25: 291-312. 124. SCHNURER, J. AND JONSSON, A. (1992). 141. POMERANZ, Y. AND WILLIAMS, P.C. (1990). Wheat Ergosterol levels and mould colony forming units in hardness: its genetic, structural, and biochemical Swedish grains of food and food grade. Acta Agric. background, measurement, and significance. In: Scand. [B] Soil Plant Sci. 42: 240-245. Advances in Cereal Science and Technology (Y. 125. SHUEY, W.C. AND GILLES, K.A. (1969). Pomeranz ed.). American Association of Experimental milling of sized wheat. Northwest Mill. 142. HALVERSON, J. AND ZELENY, L. (1988). 276: 9. Criteria of wheat quality. Chapter 2. In: Wheat: 126. SINGH, H.B., ANDERSON, E. AND PAL, B.P. Chemistry and Technology, Vol. 1. (Y. Pomeranz, ed.). (1957). Studies in the genetics of Triticum vavilovii Am. Assoc. Cereal Chem., Inc., St. Paul, MN. pp. 15. Jacob. Agron. J. 49: 4-11.
127. SMIRNOVA, A., KOSTROVA, O. (1989). Mikrobiologija zerna i produktov jego pererabotk. pp.195. 128. TATHAM, A.S., AND SHEWRY, P.R. (1985). The conformation of wheat glutenin proteins. The secondary structure and thermal stabilities of glutein and gliadins. J. Cereal Sci. 3: 103-113. 129. TUITE, J.F. AND CHRISTENSEN, C.M. (1957). Grain storage studies 24: Moisture content of wheat seed in relation to invasion of seed by species of the Aspergillus glaucus group and the effect of invasion upon germination of the seed. J. Phytopathol. 47: 323-327. 130. TURNBULL, K.M. AND RAHMAN, S. (2002). Endosperm texture in wheat. J. Cereal Sci., 36: 327-337. 131. WALL, J.S. (1979). The role of wheat proteins in determining baking quality. In: Recent Advances in the Biochemistry of Cereals, (D.L. Laidman, and R.W.Wyn- Jones, eds.). Phytochem. Soc. Eur. Symp. Ser. No. 16, Academic Press London NY and San Fransico. pp. 275- 311. 132. WICKLOW, D. T. (1995). The mycology of stored grain: an ecological perspective. In: Stored grain ecosystems (D. S. Jayas, N. D. G. White and W. E. Muir, eds.). New York, Marcel Dekker, pp. 197-249. 133. WORZELLA, W.W. (1942). Inhe ritance and interrelationship of components of quality, cold resistance, and morphological characters in wheat hybrids. J. Agric. Res. 65: 501-522. 134. WOSNIAK, W. (2001). Effect of moisture treatment of wheat grains on technological hardness (in Polish). Acta Agrophysics 46: 227-233. 135. WRIGLEY, C.W. AND BIETZ, J.A. (1988). Protein s and amino acids. In: Wheat: Chemistry and Technology (Y. Pomeranz, ed.). American Association of Cereal Chemists, Saint Paul, MN, 1: 159-275. 136. MAGAN, N. (1993a). Early detection of fungi in stored grain. Int. J. Biodeterior. Biodegradat. 32:145- 160.
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Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Year IX, No2 - 2010
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Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Year IX, No2 - 2010
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