r r jid ji uaruuD www.institut-isi.si ERNATIONAL JOURNAL OF SANITARY GINEERING RESEARCH umber 1 | October, 2010 9771854067006 WT11 \M111J Editorial lllim;;/"/ §fm him ■ This year the publishing of the first issue is rather late. World economic crisis is evidently influencing the writing of the contributions to the International Journal of Sanitary Engineering Research. Hopefully the issue is completed and published with several interesting papers. Slovenia is known as intensive wine producing country. The area covered with the wine yards is approx. 22.000 ha or 3.9 % of the total Slovenian agriculture area. Therefore precautionary principle at using the phytop-harmaceutical substances is very important in order to decrease negative impacts of the heavy metals and chemicals on the environment, especially to the groundwater and to the sediments in the rivers. V. Weingerl carried out interesting study on the influence of the use the organometal-lic phytopharmaceutical fertilizers containing copper and zinc on the distribution those metals in the wine yard soil and in the Drava river sediments. Several wine producers in Slovenia and abroad are introducing the biodynamic methods in wine yards protection. This is evidently interesting topics for the one of the next issues of the Journal of the Sanitary Engineering Research. Small and Medium Sized Enterprises (SMEs) are generating substantial amounts of waste. The authors of the paper Waste Treatment Within Small and Medium Sized Enterprise and Proposals for Improvement found out that Slovenian SMEs have no enough knowledge and resources to deal with the wastes. Several years ago almost all EU countries established the National Cleaner Production Centers which have been helping companies with the waste of production and introducing the methods and technologies for the waste prevention at the source. Slovenia has not established the National Cleaner Production Center and it seems that it will never be established. So far companies themselves are obliged to find resources at solving problems related with the waste and how they are successful is evident from the presented study. Next paper deals with the analysis of the freezing time according to the composition of the product. The food composition of food product has significant effect on freezing time considering that the products have the same shape, mass and relief. The main influence on the freezing time is the water quantity and water soluble components in the product. Predicting the freezing time is therefore logical result of this study and authors are now obliged to develop mathematical model for predicting freezing time according to the product composition and temperatures for the particular type of product. The genetically modified organisms are in the centre of the interests of the general public, non governmental organisations, professional organisations, scientists and politics. On the one hand the world is faced with the lack of the food in less developed countries and on the other hand the developed countries have the surplus of food. In the near future almost all non renewable sources will be spent, the energy and raw materials will be produced from the biomass. The agriculture area will be divided between raw material production, energy production and food production. The prices of the food will rise and it seems that one of the answers for this challenge is the use of genetically modified organisms. Threads and opportunities of this organisms are presented in the paper Health Risk and Genetically Modified Organisms. Sincerely, Janez Petek Editor-in-Chief SANITARNO INTERNATIONAL JOURNAL N/ OF SANITARY ENGINEERING INZENIRSTVO RESEARCH Izdaja - Published by: INŠTITUT ZA SANITARNO INŽENIRSTVO THE INSTITUTE OF OCCUPATIONAL, FOOD AND ENVIRONMENTAL HYGIENE ZBORNICA SANITARNIH INŽENIRJEV SLOVENIJE THE CHAMBER OF SANITARY ENGINEERS OF SLOVENIA GLAVNI UREDNIK / Editor-in-Chief Janez PETEK Inštitut za sanitarno inženirstvo / Institute of Occupational, Food and Environmental Hygiene, Ljubljana, Slovenia TEHNIČNI UREDNIK / Technical Editor Aleš KRULEC Inštitut za sanitarno inženirstvo / Institut of Occupational, Food and Environmental Hygiene, Ljubljana, Slovenia NASLOV REDAKCIJE IN UPRAVE / Zaloška cesta 155, SI-1000 Ljubljana, Slovenija Address of the Editorial Board and Administration Tel: (+386)-1-5468-393, Fax: (+386)-1-5468-394 E-mail: info@institut-isi.si www.institut-isi.si UREDNIŠKI ODBOR / Editorial Board Borut POLJŠAK Univerza v Ljubljani, Zdravstvena fakulteta / University of Ljubljana, Faculty of Health Science, Ljubljana, Slovenia Matej GREGORIČ Inštitut za varovanje zdravja Republike Slovenije / Institute of Public Health of the Republic of Slovenia Mojca JEVŠNIK Univerza v Ljubljani, Zdravstvena fakulteta / University of Ljubljana, Faculty of Health Science, Ljubljana, Slovenia Dražen LUŠIC Sveučilište u Rijeci, Medicinski fakultet Rijeka / University of Rijeka, School of Medicine, Rijeka, Croatia Darko DREV Inštitut za vode Republike Slovenije / Institute for Water of the Republic of Slovenia, Slovenia Marina ŠANTIC Sveučilište u Rijeci, Medicinski fakultet Rijeka / University of Rijeka, School of Medicine, Rijeka, Croatia Martin BAUER Univerza v Ljubljani, Zdravstvena fakulteta / University of Ljubljana, Faculty of Health Science, Ljubljana, Slovenia Zoran KOVAČ Inštitut za sanitarno inženirstvo / Institut of Occupational, Food and Environmental Hygiene, Ljubljana, Slovenia Mile DAKIČ Univerza v Ljubljani, Zdravstvena fakulteta / University of Ljubljana, Faculty of Health Science, Ljubljana, Slovenia SVETOVALNI UREDNIŠKI ODBOR / Advisory Board Mateja ČEBULAR Inštitut za sanitarno inženirstvo / Institut of Occupational, Food and Environmental Hygiene, Ljubljana, Slovenia Eneja ZADRAVEC Inštitut za sanitarno inženirstvo / Institut of Occupational, Food and Environmental Hygiene, Ljubljana, Slovenia © Inštitut za sanitarno inženirstvo, 2010. © Copyright by Institute of Occuptional, Food and Environmental Hygiene. Vse pravice pridržane. Reproduciranje in razmnoževanje po zakonu o avtorskih pravicah ni dovoljeno! ■ © Inštitut za sanitarno inženirstvo, 2010. Content Influence of Food Composition on Freezing Time Sebastjan FILIP, Rok FINK, Mojca JEVŠNIK..................................4 Copper and Zinc Accumulation in Vineyard Soils Treated with Cu and Zn Containing Phytopharmaceuticals Vesna WEINGERL......................................................................14 Waste Treatment within Small and Medium Sized Enterprises and Proposals for Improvement Katarina ZELEZNIK LOGAR.......................................................25 Gensko spremenjeni organizmi in tveganja za zdravje/ Health Risk and Genetically Modified Organisms Sebastjan FILIP, Rok FINK.........................................................36 © Inštitut za sanitarno inženirstvo, 2010. Original scientific paper Influence of Food Composition on Freezing Time Sebastjan FILIP1, Rok FINK1, Mojca jEVŠNIK2* abstract Freezing is still the best way to preserve food when is carried out properly and allows production of food without any chemical preservatives. It is important to accurately predict the freezing time of different kinds of food to assess the quality and safety of the foodstuffs, processing requirements, and economical aspects of food freezing. The aim of this research was to investigate the influence of food product composition on freezing time. The research based on hypothesis: the composition of food product has significant effect on freezing time considering that the products have the same shape, mass and relief. In our research we recorded four batches of standard product with three different fillings (chocolate, vanilla and forest fruit) at the same freezing conditions. The products from puff pastry were always made from the same ingredients and also mass, shape, relief, slab and surface were constant. The core temperature after 41 minutes of freezing at air temperature - 32 oC ± 2 oC and constant velocity reached in chocolate product - 24.4 oC ± 0.2 oC, meanwhile in forest fruit - 17.0 oC ± 0.2 oC and vanilla only - 12.8 oC ± 0.2 oC; thus confirmed our hypothesis. For SMEs (small and medium size food enterprisers) which represent 99 % of all enterprises in the EU, it is essential to establish their own model for predicting freezing time which is more applicable that general theoretical models that require a lot of data which are not common accessible Received: 12. 1. 2010 for SMEs. Accepted: 10. 7. 2010 KEY WoRDs: Freezing, Food composition, Freezing time, Freezing point, Food quality, Food safety. 1 University of Ljubljana, Biotechnical Faculty, Department of Food Science and Technology, Jamnikarjeva 101, SI-1000 Ljubljana 2 University of Ljubljana, Faculty of Health Sciences, Department of Sanitary Engineering, Zdravstvena pot 5, SI-1000 Ljubljana * corresponding author: Mojca Jevsnik Tel.: ++38613001149 Fax: + +38613001119 e-mail: mojca.jevsnik@zf.uni-lj.si INTRODUCTION Like most food preservation methods, the freezing process for food has evolved over a significant period of time. As an extension of cooling, the primary goal of food freezing has been the reduction of microbial growth and/or control of reactions causing spoilage of the food, as well as the corresponding extension of product shelf-life. In addition, the process has extended the availability of many food commodities for consumption at any time during the year. Frozen food can be transported for longer distances and the process has contributed to making foods available on a worldwide basis. For most applications, the food products are held in storage for some period of time before thaned, and prepared for consumption. More recently, the process has evolved to include a variety of products that are consumed in a frozen state [1]. The principal objective of the new general and specific hygiene rules is to ensure a high level of consumer protection with regard to food safety. Food business operators shall ensure that all stages of production, processing and distribution of food under their control, satisfy the relevant hygiene requirements, laid down in Regulation (EC) No. 852/2004. This regulation emphasizes the importance of cold chain throughout HACCP (Hazard Analysis and Critical Control Point) system for all foodstuff that require cooling or freezing regime [2]. Kassianenko and co-workers [3] emphasis the importance of cold chain, which is a vital part of modern global trade as it has impact on all food commodities. In today's modern society, refrigerated storage is one of the most widely practiced methods of preserving perishable food. Improper use of this process increases the potential risk and microbial hazards will advance, thus leading to the food borne illnesses. The purposes of food freezing is preservation of food, reducing the activity of enzymes and microorganisms, reducing the amount of liquid water for microbial growth and reducing water activity of foods [2,4,5,6]. Freezing is still the best way to preserve food when is carried out properly and allows production of food without any chemical preservatives. Thermo physical properties of frozen food are used to estimate the rate of heat transfer and to calculate the heat load in process such as freezing and thawing [7]. The early calculations and analyses associated with freezing and thawing, primary used constant and uniform thermo physical properties [8]. The calculations and analyses were typically oversimplified and inaccurate. Numerical analyses such as finite difference methods were used widely to analyze thermal food processes [9]. The large number of food products, available today, create a great demand for knowledge on thermo physical properties. Since foodstuff are composite materials, the thermo physical properties are clearly a function of the components [10,11]. The core temperature for frozen food are prescribed by the Regulation upon safety of frozen food, 63/2002 [12] (the Council directive of 21 December 1988 on the approximation of the laws of the Member States relating to quick-frozen foodstuffs for human consumption (89/108/EEC)) and so it is an obligation to release the products on market with proper core temperature which has to be below - 18 °C. Nevertheless the monitoring of tempera- The principal objective of the new general and specific hygiene rules is to ensure a high level of consumer protection with regard to food safety. Freezing is still the best way to preserve food when is carried out properly and allows production of food without any chemical preservatives. The large number of food products, available today, create a great demand for knowledge on thermo physical properties. It is important to accurately predict the freezing time of foods to assess the quality and safety, processing requirements, and economical aspects of food freezing. The freezing time of a food of any shape can be calculated by quite a lot of different equations and are quite difficult for use in industry because of the wide range of variables. The most often mistake is overlaying the parameters to other similar products. ture during the transport is regulated with the Commission Regulation (EC) No. 37/2005 [13] of 12 January 2005 on the monitoring of temperatures in the means of transport; warehousing and storage of quick-frozen foodstuffs intended for human consumption and allowed short time deviation of temperature should not exceed 3 °C. It is important to accurately predict the freezing time of foods to assess the quality and safety, processing requirements, and economical aspects of food freezing [14]. But freezing process has a problem with moving boundaries [15]. Food, undergoing freezing, release latent heat over a range of temperatures. Freezing does not occur at unique temperature. In addition, food do not have constant thermal properties during freezing. As result, no exact mathematical model exists for predicting the freezing time of foods [16,17]. As proposed by Cleland and co-workers in successive papers [4], one of the ways to determine the freezing time of a multi-dimensional product is to know both the freezing time of a slab under the same operating conditions and a geometric shape factor [18]. Authors initially noted such a factor, such as EHTD (the equivalent heat transfer dimensionality). Therefore, the freezing time of a food of any shape can be calculated by quite a lot of different equations and are quite difficult for use in industry because of the wide range of variables. The experimental or testing method which has a lot of process traps in often used in practice. The most often mistake is overlaying the parameters to other similar products (mass, shape, relief, slab, etc.). The freezing time requirement for each entity is estimated by the simulation model using the Plank's equation. Plank developed an equation, based on the unique phase change model, for estimating freezing time for different geometrical shapes, and allowing for varying film coefficients. The equation, derived from one-dimensional infinite slab geometry and has been analytically extended for infinite cylinders, spheres and for finite parallelepiped geometries. Plank's equation is common worldwide used and can be expressed as follows [19], equation 1. Where the tf is time required for freezing (in min), p product density, AH latent heat of freezing, R characteristic half thickness of the food object, h, kf are heat transfer coefficient and thermal conductivity (before freezing) respectively 0.f, 0a denote the freezing point and ambient temperatures. p-AH tf =■ f -ea R R —+ - h 2k f (1) if P AH R h, kf 0f, 0 if' a time for freezing (min) product density (kg/m3) latent heat of freezing (kJ/kg) half thickness of the food object (m) heat transfer coefficient (W/(m2C)) thermal conductivity (W/(mC)) Carefully planned and efficiently provided education of employees is of the great importance. Lectures should be carried out by competent ex© Institut za sanitarno inzenirstvo, 2010. perts with relevant experience, who will provide specific information to every employee according to their assigned task. Employees should be personally responsible for their work [20]. The consumer is also a source of hazard for itself and that should be incorporated in the risk assessment. The results from research carried out by Ovca and Jevšnik [21] confirmed that the term ''cold chain" is not well known among consumers. They also confirmed that Slovenian and European consumers place the responsibility of maintaining a cold chain into other parts of the food chain. One of the reasons for poor knowledge of the cold chain could be due to the unsatisfactory efforts of governmental and nongovernmental organizations, which are responsible for educating and informing consumers. The role of the consumer in maintaining a cold chain is more important than is currently thought, and is greater than the importance attached to other parts of the food chain. MATERIALS AND METHODS Samples were prepared as part of daily production on continuous line at food enterpriser according to its production capacity. The product core temperature and time were measured during shock freezing using RT-DTEMP101; SN: M32551, for the range from -100 °C to +600 °C (MadgeTech, Contoocook, New Hampshire, USA), calibrated with measurement error 0.2 °C and evaluated using program MadgeTech Data Recorder Software, version 2.00.63. with the statistical parameters included. We recorded core temperature every 6 seconds and thus we got for each measure 410 thermal points for describing the decrease of temperature in core of product. Water activity was measure using Testo 650 with electrode 0628-0044 (910) at 25 °C, meanwhile the compositions were analyzed in Slovenian National Public Health Institute. The results of analyses are stated in Table 1. The puff pastry was always made from the same ingredients; also mass, shape, relief and surface were constant. Times of freezing, air temperature, air speed in continuous shock freezer were also constant and were not changed during the process. Freezing medium was always cooled air with temperature of -32.0 °C ± 2.0 °C, constant velocity and time of freezing for 41 minutes. Selected fills were vanilla, chocolate and forest fruits. Each piece of product was weighted on continuous packaging equipment (n = 9000; 100.00 g ± 2.8 g). We measured the temperature of each type of product in for parallel batch at the same conditions: (vanilla, n = 4; chocolate, n = 4; forest fruits, n = 4). Predicted freezing time for mass 100 g was 40 min - 45 min at - 32.0 °C ± 2.0 °C, constant velocity, as stated in producer's instructions. The time of freezing is managed by belt speed. At one time in freezer were 820 kg of product or 8,200 pieces. The results of core temperature after 41 minutes of freezing were processed by the XLSTAT 2009.6.01 Comparison of samples using nonparametric test, multiple pair wise comparisons using the Nemenyi's procedure/Two-tailed test with significance level 5 %. Lectures should be carried out by competent experts with relevant experience, who will provide specific information to every employee according to their assigned task. The role of the consumer in maintaining a cold chain is more important than is currently thought, and is greater than the importance attached to other parts of the food chain. Figure 1. Core temperature in correlation to time of freezing for four batches. a. the first batch; b. the second batch; c. the third batch; d. the fourth batch. results and discussion According to our four batches we recorded core temperature every 6 seconds and thus we got for each measure 410 thermal points for describing the decrease of temperature in core of product. When we compare the data between four parallels within one type of product we ascertain no statistical difference (p < 0.001). Results from recorded data for all products are stated on the Figures 1 from a to d. Within one type of product there is no difference within type of product or according of the used fill. As Figure 1 shows that the time of freezing was constant 41 minutes and at this time only one product (chocolate filling) reached the temperature below -18 °C and the core temperature after 41 minutes of freezing reached -24.4 °C ± 0.2 °C. The core temperature for product with forest fruit filling almost reached the required temperature (-18 °C) but nevertheless it was still with its -17.0 °C ± 0.2 °C more than 1.0 °C to high. The highest core temperature we measured in product with vanilla filling and it was -12.8 °C ± 0.2 °C after 41 minutes of freezing. Summary data of four investigated batches are shown in the Table 2. Reason for such differences between similar products is in the specific heat and enthalpy. In general we can conclude that the food with higher content of water have higher specific heat and higher enthalpy (at 0 °C) thus need longer freezing time compare to similar prod- 30,00 Ï 5 -10,00 -30,00 30,00 Time (minutes) vanilla chocolade forest fruits E oj CD S vanilla chocolade forest fruits -30,00 Time (minutes) £ 8 30,00 ..... vanilla d 20,00 \\ - chocolade ....... forest fruits 10,00 \ V, p 2? 00,00 10 ( io'\. \-20 30 40 50 0.05 p >0.05 p <0.001 p <0.001 p <0.001 p <0.001 p <0.001 p <0.001 p <0.001 t: time of freezing in minutes, V: vanilla product, C: chocolate product, FF: forest fruit product, x : average values for four batches, Significant at p <0.001; significant at p <0.01; significant at p <0.05; p >0.05, not significant, Values followed by a different letter are significantly different along the column for the Duncan (0.05) test. Figure 2. Phase change from liquid to solid (ice). T: predicted temperature (oC); t: time (min) in range from 0 to -12 oC. Figure 3. Food composition in correlation to core temperature in sampled products. ucts with lover water content. Freezing time as correlation to water content as well as dry matter from our research is described with equation on the Figures 2 and 3. There are exceptions like cucumber which has water content 95.4 % and enthalpy 390 kJ/kg (at 0 °C), but low specific heat capacity (1.02 kJ/(kgK)) thus its water in 100 % freeze at -5 °C, meanwhile tomato pulp with water content 92.9 % and enthalpy 382 kJ/kg (at 0 °C), but higher specific heat capacity (4.02 kJ/(kgK)) at -5 °C have still 16 % of unfrozen water [11]. The time of cooling when the 0 °C was food composition (%) 20 40 60 80 100 120 -5 - water (w) dry matter (dm) -30 J T (°C) = 0.1797 w(%) - 24.653 R2= 0.9993 T (°C) = -0.1797 dm(%) - 6.6835 R2= 0.9993 achieved, was also different - 9 minutes for vanilla, 13 minutes for chocolate and 15 minutes for forest fruit. So the filling with the highest content of water cool down to 0 °C rapidly compare to other two. The cooling gradient from 24 °C to 0 °C for vanilla was 2.7 °C/min, 1.8 °C/min for chocolate and 1.6 °C/min for forest fruit filling. This temperature cooling gradient is also shown on the Figures 1 to 3. Although any solid food is not solution, the aqueous component in the frozen solid food can be considered as a mixture of ice and a solution of nonaqueous components in liquid water. As water in the food freezes into pure ice during the freezing, the remaining solution becomes more and more concentrated. Thus, depression of the freezing point of the food occurs and continues as concentration increases. The net effects of dynamic freezing point depression are that the initial freezing point of the food is below 0 °C. The freezing process occurs over a range of temperatures, which is different from the freezing process of pure water at unique temperature. The phase changes from liquid water to solid (ice) are shown in the Figure 2. When the 0 °C was reached after different time in each product, the temperature gradient in all products changed rapidly because of the changing of physical state from liquid to solid (ice). At this point the phenomena discussed above appear, so the products with higher content of water need more energy to convert all water to ice. The product with chocolate filling which has the highest content of fat continues to cool down at the quite the same temperature gradient. It can be assumed that the fat in the chocolate consist majority from saturated fatty acids which are solid at room temperature, thus there is less water than in other two cases that has to be converted from liquid to solid. CONCLUSIONS As expected, the product containing the highest level of water (vanilla filling) has not reached the target temperature and it was with the core temperature extremely above the -18 °C and thus can represent a food safety hazard as well as risk for consumers. There are data confirming microbial growth of some Gram-negative bacteria at -4 °C and growth of bacilli, that is, Gram-positive bacteria at -7 °C [4]. The problem of higher core temperature than -18 °C in frozen food products, which is also obligatory by the law, is not concerning only the primary production process or enterprisers but concerns all actors in food supply chain [2]. The data of food composition as well as shape, density, surface are necessary to predict freezing conditions. On one hand it is possible to reach target temperature by pre-cooling of raw material (where possible), while on the other hand we can prolong the time of freezing and lower the temperature of freezing media (air) and increase its velocity for better heat transfer (not always applicative). In general we need longer time of freezing for foodstuff with high water in the water soluble components. Because of the effect of lowering freezing point as result of concentrating the solution in food, as well as knowing the mean of the importance of specific heat. There are also some other methods to Although any solid food is not solution, the aqueous component in the frozen solid food can be considered as a mixture of ice and a solution of nonaqueous components in liquid water. The products with higher content of water need more energy to convert all water to ice. In general we need longer time of freezing for foodstuff with high water and the water soluble components. In practice every part of food supply chain has the manage system which pushes the action to limits like just in time management. decrease freezing time such as use of cryogenic freezing, where extreme low temperatures are used by liquid gasses (N2, He, CO2) or increasing the dry matter by adding hydrocolloids, proteins and sugars. Global food safety will be achieved, when every single link in the food chain will entirely (in its indoor and outdoor environment) become master of its particular area and will trust in activity of both previous and following link in the food safety circle "from farm to table", not ignoring consumer [22]. Usually in practice every part of food supply chain has the manage system which pushes the action to limits like just in time management. So there is possibility that frozen food with inadequate temperature from enterpriser over the logistics, reach the retailer shelves thus even consumer, who can storage such a frozen food according to expire date. For SMEs is essential to establish their own model for predicting freezing time which is more applicable than general theoretical models that require a lot of data which are not common accessible for SMEs. REFERENCES [1] Heldman DR. Nesvadba P. Food Freezing History. Encyclopedia of Agricultural, Food, and Biological Engineering, New York 2003: 350-352. [2] Likar K. Jevsnik M. Cold chain maintaining in food trade, Food Control 2006; 17: 108-113. [3] Kassianenko A. Wszol K. Oggel J. Jol S. The Cold Chain, One Link in Canada's Food Safety Initiatives. Food Control 2006; 6: 18-19. [4] Cleland DJ. Valentas KJ. Prediction of freezing time and design of food freezers. In: KJ Valentas, E. Rotstein, RP Singh (Eds). Handbook of Food Engineering Practice. Boca Raton, FL; CRC Press 1997; 71-124. [5] Zorrilla SE. Rubiolo AC. Mathematical modelling for immersion chilling and freezing of foods. Part I: Model development. Journal of food engineering 2005; 66: 329-338. [6] Goni SM. At al. Prediction of food freezing and thawing times: Artificial neural networks and genetic algorithm approach. Journal of food engineering 2008; 84: 164-178. [7] Delgado AE. Sun DW. Influence of surface water activity on freezing/ thawing times and weight loss prediction. Journal of Food Engineering 2007; 83: 23-30. [8] McKellar A. Paterson J. Pham QT. 2009. A comparison of two models for stress and strains during food freezing. Journal of food Engineering 95: 142-150. [9] Chin SW. Spotar SY. Freezing time prediction for film packed food. International journal of engineering and technology 2006; 3: 2,182-190. [10] Liu Y. Zahao Y. Feng X. Exergy analysis for freeze-drying process. Applied Thermal Engineering 2005; 28: 675-690. [11] Wang L. Weller CL. 2006. Thermophysical properties of frozen foods. In D.W. Sun Handbook of frozen food processing and packaging. Taylor&Francis, Boca Raton 101-127. [12] Uradni list Republike Slovenije. St. 63/2002. Datum izdaje 19.07.2002. Stran: 6721-6722. [13] Commission Regulation (EC) No 37/2005 of 12 January 2005 on the monitoring of temperatures in the means of transport, warehousing and storage of quick-frozen foodstuff intended for human consumption. 2005. Official Journal of the European Union. L 10: 18-19. [14] Jafari M. Alavi P. Analysis of food freezing by slot jet impingement. Journal of applied sciences 2008; 8: 1188-1196. [15] Fricke BA. Becker BR. Sensitivity of freezing time estimation methods to heat transfer coefficient error. Applied Thermal Engineering 2006; 26: 350-362. [16] Rolfe EJ. The chilling and freezing of foods. In: N Blakebrought, Ed. Biochemical and Biological Engineering Science 1968; 2: 137-208. [17] Pham QT. Simplified equation for predicting the freezing time of foodstuffs. International Journal of Food Science &Technology 2007; 21: 209-219. [18] Salvadori VO. De Michelis A. Mascheroni RH. Prediction of freezing times for regular multi-dimensional foods using simple formulae. Lebensmittel Wissenshaft und Technologie 1997; 30: 30-35. [19] Ramakrishnan S. Wysk RA. Prabhu VV. Prediction of process parameters for intelligent control of tunnel freezers using simulation. Journal of Food Engineering 2004; 65: 23-31. [20] Jevsnik M. Ovca A. Likar K. Maintaining a cold chain in retail: does it work? 13th World Congress of Food Science & Technology. 17 - 21 September 2006. Nantes. Food is life. Nantes: IUFoST, 2006: 35-45. [21] Ovca A. Jevsnik M. Maintaining a cold chain from purchase to the home and at home: Consumer opinion. Food Control 2009; 20: 167-172. [22] Raspor P. Jevsnik M. Good nutritional practice from producer to consumer. Critical Review in Food Science and Nutrition 2008: 48(3): 276-292. © Inštitut za sanitarno inženirstvo, 2010. Original scientific paper Copper and Zinc Accumulation in Vineyard Soils Treated with Cu and Zn Containing Phytopharmaceuticals Vesna WEINGERL ABSTRACT Long term use of copper and zinc containing phytopharmaceuticals in vineyard areas causes increased accumulation of total Cu and Zn in the surface layer of soil. Copper and zinc accumulation and distribution in vineyard soils is discussed as a step towards understanding Cu and Zn existence in soils and their availability to plants. Several single extractions were performed to the determination of total Cu and Zn content after aqua regia digestion. The use of Cu containing phytopharmaceuticals has increased the total Cu concentration in analyzed vineyard soils to 97 mg/kg compared to background levels of approximately 30 mg/kg. The amounts of plant available Cu and Zn extracted by the EDTA ranged as follows, when expressed relative to the total Cu or Zn content in the soils: 13.6 % - 40.4 % for Cu and 3.5 % - 11.6 % for Zn. The study shows that copper stays active in soils longer than ten years, and may result in leaching and can transfer to deeper soil layers and consequently to ground or the surface waters. KEYWORDS: Vineyard, Soil, Copper, Zinc, Phytopharmaceuticals. Received: 4. 12. 2009 Accepted: 10. 7. 2010 University of Maribor, Faculty of Agriculture and Lifesciences, Pivola 10, 2311 Hoce, Slovenia e-mail: vesna.weingerl@uni-mb.si introduction In order to control vine (Vitis vinifera L.) diseases, such as downy mildew caused by Plasmopara viticola, different phytopharmaceuticals, especially Cu-based ones, have been applied extensively to vineyards. The application of these phytopharmaceuticals (CuSO4 + Ca(OH)2); Cu2O), including the Bordeaux mixture CuSO4-3Cu(OH)2<3CaSO4 and more recently copper oxychloride CuCl2-3Cu(OH)2 and hydroxide (Cu(OH)2), resulted into increased Cu concentrations in vineyard soils. The content of Cu significantly increases with a vineyard's age. Heavy metals are the main pollutants which accumulate in vineyards' ecosystems. Because of viticulture intensive practice, the phytopharmaceuticals and fertilizers are the main source of metal pollution. While the soil purifies slowly and partially, metal pollutants tend to accumulate [1]. In many regions of the world there is a concern about the risks that the accumulation of copper in agricultural soils may pose to human health, ecological health, and the long-term sustainability of agricultural lands. The phytotoxic effects of copper to plants have been widely studied [2,3], but it is not expected that copper causes phytotoxicity in mature grapevines as they tend to have deep root systems that extend below the surface soils where most of the applied copper tends to remain. The impacts on soil biology and implications for soil fertility are more of the concern in established vineyards. Several studies have shown the elevated copper concentrations to adversely affect the fertility of vineyard soils. Increased Cu concentrations in soil (more than 80 mg/kg) reduce earthworm abundance; therefore, any harm to earthworm populations could affect the productivity and efficiency of vineyards [4]. In response to environmental concerns over the use of copper fungicides such as the accumulation of copper in agricultural soils and the potential impact on soil ecology, regulators in some European countries have imposed restrictions on the use of copper-based fungicides. For example, copper use has been banned in The Netherlands, and Switzerland has restricted the amount of copper that can be applied per hectare [2]. Micronutrient elements like Cu and Zn form insoluble compounds and also bond strongly to mineral particles and organic matter. Considering availability to vines, only a nutrient in the soil solution is immediately available to the vines. Deficiency occurs when a nutrients supply is un-sufficient to meet a vines demand; conversely, when supply exceeds demand or a nonnutrient element accumulates in the root zone, toxicity may occur. In previous research main focus has been pointed out to Zn determination in vineyard soils. Zinc additions in the Orlica vineyard soil in Mer-anovo were much larger than uptake; the consequence was accumulation of zinc in the soil. The analysis of the Meranovo stream sediment which springs in the Pohorje territory and runs below the vineyard to the river Drava showed much lower zinc concentrations in the stream Heavy metals are the main pollutants which accumulate in vineyards' ecosystems. The impacts on soil biology and implications for soil fertility are more of the concern in established vineyards. Micronutrient elements like Cu and Zn form insoluble compounds and also bond strongly to mineral particles and organic matter. Zinc is hard to wash away, therefore the contents of zinc are the highest in the surface layer. High levels of phosphorous, zinc, iron, manganese and aluminum may also restrict copper absorption by roots. sediment before the stream reaches the vineyard and than after it leaves the vineyard. The increasing zinc content in the sediment is caused by the use of zinc containing phytopharmaceutical substances and mineral fertilizers [5]. Mobility of copper and zinc in soil is closely connected to soil reaction. With increased acidity of the soil there is also the increase of the quantity of element available to plants. The content of zinc is considerably lower in acid than in alkaline soil. Zinc is hard to wash away, therefore the contents of zinc are the highest in the surface layer [6,7]. Contents of copper, zinc and other heavy metals in Slovene soil on the country area were researched by Andjelov and Šajn [8,9]. Increased contents of copper and zinc were established in the Carinthia area (the Mežica mine), in Celje area (the Celje zinc processing plant) and in the areas of intensive plantation production of fruit and grape (organometal-lic phytopharmaceutical pesticides and fertilizers). Micronutrients present the limitation of growth, development and consequently production of vine. Considerable deficiency as well as larger surplus of specific nutrients can be detected on vine by visual inspection. Zinc deficiency in vine occurs in the situation where soil is rich with phosphorus, phosphate ions surrounded the roots and cause the change of zinc ions to insoluble zinc phosphate when contacting the roots. The surplus of phosphate ions deactivates zinc ions into insoluble zinc phosphate as well. High phosphorus content in vineyard soil samples is caused by use of mineral fertilizers containing phosphorus (superphosphate, Thomas phosphate, complex NPK fertilisers). Reports of copper deficiency in vineyards are rare. More common are reports of copper toxicity. Copper is an essential element for living organisms, as a constituent of several enzymes and as a redox catalyst in a variety of metabolic pathways. At high concentrations, however, copper inhibits growth and interferes with several cellular processes, including photosynthesis, respiration, enzyme activity, pigment and protein synthesis and cell division [22]. Copper is strongly bound to organic matter. Peat soils and mineral soils with high levels of organic matter (6-10%) are most likely to be deficient in plant available copper. Copper availability is reduced as pH increases to 7 and above. High levels of phosphorous, zinc, iron, manganese and aluminum may also restrict copper absorption by roots. Copper is an important constituent of many proteins like ascorbic acid oxidase, cytochrome oxidase, diamine oxidase, and polyphenol oxidase. Copper is an important nutrient for many microbes. It controls molds and often alleviates perceived zinc deficiencies. Copper interacts with iron and manganese [23]. Predicting the mobility of heavy metals in soils requires the mathematical models that accurately describe metal adsorption in the presence of competing cations. A reaction-based sorption model was developed using a combination of nonspecific cation exchange reactions and competitive sorption reactions to sites with high affinity to heavy metals. The proposed model describes competitive sorption and transport of Cd, Zn, and Ni in pH range between pH 4.6 and 6.5 [8]. Heavy metal extraction efficiency depends on the combination of many factors, such as the liability of heavy metals in soil, the concentration and nature of the chelant, pH and the properties of the soil matrix. Raising soil pH is a prevention mechanism for elution of heavy metals. Chelant assisted soil washing has been widely studied for remediation of contaminated soil. A number of chelants have been tested. For soils contaminated with Pb, Zn, Cu and Cd, ethylenediamine tetraacetate (EDTA) has often been found to be the most effective [13]. The study of transition metals (Cd, Co, Cu, Mn, Mo, Pb and Zn) in the soils of the sub-Mediterranean winegrowing areas in Slovenia confirmed that the main source of transition metals pollutant in the region is intensive viticulture practice, especially the use of copper substances [10,11]. The object of this research was to investigate the influence of organo-metallic phytopharmaceutical fertilizers use on copper and zinc dynamic and distribution in vineyard soil. With the use of the control soil samples, differentiation of natural and anthropogenic soil enrichment in heavy metals in wineyard areas, was possible. Content of Cu and Zn in a control sample gives natural soil profile of these two elements in soil, including the atmospheric influence. Total element contents, EDTA- and AL- (ammonium lactate) extractable (plant available) Cu and Zn with flame atomic absorption spectrometry (FAAS) method was determined. Contents of Cu and Zn were determined in wine, produced on vineyard areas, included in research. Weight balance was calculated from the data on copper content in wine (Cu uptake) and content of copper in used phytopharmaceutical substances (Cu addition). In addition, content of Cu in the sediment of the stream at a foothill of the vineyard Orlica was measured. The sediment of the stream was sampled on several sites from the spring to the river Drava. Increased copper concentration in the stream sediment was expected as the evidence of the long term use of the phytopharmaceutical substances containing copper. METHODS Soils of two vineyards from both riverbanks of the river Drava near Maribor, Slovenia, were sampled. Surface of the Orlica vineyard in Mer-anovo is 50,000 m2 with 17,800 vines. Surface of the Kalvarija vineyard is 45,000 m2 with 15,000 vines. Upper part of the vineyard Kalvarija (soil sample named Kalvarija 2) and lower part of the vineyard Kalvarija (soil sample named Kalvarija 3) were sampled and analysed separately. The particular vineyards were chosen because of their diversity in acidity of soil - vineyard Orlica possesses acid soil whereas Kalvarija possesses alkaline soil. Quite different content of free carbonates in vineyardsoil Kalvarija was the reason to separate this vineyard in two parts regarding to the altitude. Control soil sample was sampled above the two vineyards on uncultivated surfaces. The distance between sampling sites was approximately 10 m. In all vineyards, soil samples were collected as close to the vine lines as possible and in the middle of the row, between two sampled lines. The samples were sampled with metal probe (2 cm diameter) at depth of 0-20 cm and 20-40 cm. Two layInternational Journal of Sanitary Engineering Research The study of transition metals in the soils of the sub-Mediterranean winegrowing areas in Slovenia confirmed that the main source of transition metals pollutant in the region is intensive viticulture practice, especially the use of copper substances. The distance between sampling sites was approximately 10 m. We sampled according to ONORM L1057 which regulates soil samples taken in vineyards and orchards. Soil samples were first dried in the air to the constant weight. Roots and larger stones were removed from the dried samples. Samples were ground in the mill and passed through 1 mm sieve. ers were sampled to assess the distribution of Cu and Zn with depth. We sampled according to ONORM L1057 which regulates soil samples taken in vineyards and orchards [14]. Soil samples were first dried in the air to the constant weight. Roots and larger stones were removed from the dried samples. Samples were ground in the mill and passed through 1 mm sieve. Humidity was determined by weighing 10 g of air-dried sample into paper bag and dryed at 60 °C in a drying machine to the constant weight. Contents of Cu and Zn were calculated according to the dry weight of the sample. Copper and zinc were determined by flame atomic absorption spectrometer, SpectrAA-10 (Varian, Mulgrave, Australia) in air/acetylene flame. Zinc absorption was measured with the use of deuterium lamp for background correction at 213.9 nm and copper absorption at 324.7 nm wavelength. Soil samples were digested in PFTE (polytetrafluor-oethylene) containers in MDS 2000 microwave oven produced by CEM (CEM, Matthews's n. c., USA). Phosphorus concentration was determined by spectrophotometric ammonium molybdate method on Varian Cary 1E spectrophotometer (Varian, Mulgrave, Australia) at 659 nm wavelength. For measuring pH in solution, Iskra MA 5750 pH meter was used (Iskra, Horjul, Slovenia) with HEC 0101 combined glass electrode. Microwave-assisted digestion of soil samples with aqua regia: 0.5000 g ± 0.0001 g of air-dried soil sample were weighted in PTFE container and 16 mL of aqua regia (HNO3 and HCl, 1+3) were added. The containers were covered and puted in the microwave oven. Heating programme for 12 containers was following: 1 min at 180 W, 4 min at 480 W and 60 min at 650 W. Containers were cooled to room temperature. Contents were poured through wrinkled filter paper (MN 6191/4) into 50 mL volumetric flask, deionized water were added to the mark. Contents of Cu and Zn in filtrate were determined by the FAAS method in air/acetylene flame. Calibration curve was prepared with standard solutions, added the same amount of aqua regia as the samples. A blank sample was prepared for controlling acid purity. Total microwave-assisted digestion of soil samples: in PTFE container for digestion 1.0000 g ± 0.0001 g of air-dried soil sample were weighted, 10 mL of deionized water, 5 mL of HNO3, 2 mL of HCl and 3 mL of HF were added. The containers were covered and put in the microwave oven. The pressure control was programmed on 6.9-105 Pa. Twelve containers were heated for 60 min at 650 W. The containers were cooled to room temperature and the content was poured through wrinkled filter paper into 50 mL PE volumetric flask. Deionized water was added to the mark. Cu and Zn contents in the filtrate were determined by FAAS method in air/acetylene flame. The calibration curve was prepared with standard solutions added the same amount of acids as to the samples. Blank sample was prepared for acid purity control. Extraction of soil samples in EDTA-solution: 10.000 g ± 0.001 g of fine air-dried soil sample were weighed to 250 mL plastic container and poured over by 100 mL concentration of EDTA-solution (0.05 mol/L). Preparation of EDTA-extraction solution is described in literature [5]. The plastic container was sealed and shaken on horizontal shaker for two hours with 180 min-1. After the shaking process the suspension was filtered through wrinkled filter paper. At first a few mL of the filtrate were discharged. Contents of Cu and Zn in the filtrate were determined by the FAAS method. soil sample extraction with AL-solution: into plastic container with cover 5.000 g ± 0.001 g of soil sample were weighted and 100 mL of AL-extraction solution was added. The container was covered and shooked for two hours on rotary shaker at 35 min-1. Suspension was filtered through dry wrinkled filter paper of medium porosity (white strip). The first few mL of the filtrate were discarded. Concentration of phosphorus in the filtrate was determined by measuring molecular absorption at 659 nm wavelength in air/acetylene flame. AL-extraction solution was used as a blank sample. Preparation of AL-solution is described in literature [5] Soil acidity was determined according to ONORM (16), free carbonates by Scheibler [15]. results and discussion In our research the main focus was pointed to the copper and zinc determination in vineyard soils. Copper addition in the Orlica vineyard soil in Meranovo is much larger than uptake, the consequence being accumulation of Cu in soil. Vineyard soils polluted with heavy metals present a serious environmental concern. Copper and zinc originating from the intensive application of Cu- and Zn-based phytopharmaceuticals belongs to the most important contaminants of the vineyard soils. Both elements can migrate through the soil layers in vineyard and pose thus an important risk for groundwater quality. Detection and quantification limits for total and extractable copper determination methods in soil samples are shown in the Table 1. To verify the accuracy of total Cu, standard reference material 2709 (San Joaquin Soil, National Institute of Standards and Technology) was analysed. Comparison of our results with certified values in SRM is given in the Table 2. It is evident from the Table 1 and Table 2 that sensitivity, precision and accuracy of analytical values were satisfactory. Considering pH soil value, the particular vineyards were chosen for their diversity in acidity of soil. Vineyard Orlica possesses acid soil whereas vineyard Kalvarija possesses alkaline soil. Control soil samples were sampled above the two vineyards on uncultivated surfaces. Acid soil reaction is caused by the lack of exchanging metal ions. Cu and Zn con- Copper addition in the Orlica vineyard soil in Meranovo is much larger than uptake, the consequence being accumulation of Cu in soil. Detection and quantification limits for total and extractable copper determination methods in soil samples are shown in the Table 1. Table 1: Linear ranges, detection and quantisation limits for determination of Cu by FAAS and P2O5 by MAS. Linear range Detection limit Quantisation limit (mg/L) (mg/L) (mg/kg) Cu - total 0.2 - 1.0 0.1060 10.6 Cu - EDTA 0.5 - 5.0 0.0350 3.5 Cu - AL 0.5 - 5.0 0.0300 2.8 P2O5 0.1 - 4.0 0.0002 30.0 Table 2: Precision of total Cu content determination in SRM 2709. Mean conc.* rsd Certified value Measured value (mg/kg) (%) (mg/kg) (mg/kg) Cu 34.6 1.9 34.6 ± 0.7 34.6 ± 1.1 *Mean concentration, result of ten determinations of the same sample. Figure 1. Correlation between total and EDTA extractable Cu and total and EDTA extractable Zn content (amounts in mg/kg on primary axis) and pH soil value (amounts on secondary axis). tent therefore much lower in acid that in alkaline soil. Results of comparison between total copper and zinc content in the vineyard soil samples and control samples to pH value of the samples are shown in the Figure 1. The use of Cu containing phytopharmaceuticals has increased to total Cu concentration in analysed vineyard soil, 97 mg/kg, compared to background levels of approximately 30 mg/kg. Content of the Zn increases with soil pH value in all analysed vineyard soil samples and control samples. Content of the Cu trended increasing Orlica Control Orlica Kalvarija Control Kalvarija 2 Kalvarija 3 —A— Cu - total —O— Zn - total Cu EDTA Zn EDTA -o- pH with pH soil value with one exception - the control soil sample from Kalvarija vineyard area. Mobility of copper and zinc in soil is closely connected to soil reaction. With increased pH value of soil there is also an increase in the quantity of element available to the plants. Contents of copper and zinc are considerably lower in acid than in alkaline soil. Amounts of plant available Cu and Zn extracted by the EDTA ranged as follows, when expressed relative to the total Cu or Zn content in the soils: between 13.6 % and 40.4 % for copper and between 3.5 % and 11.6 % for zinc. Proportion between AL-extractable phosphorus and copper content in vineyard soil samples and control soil samples is shown in the Table 3 and graphically in the Figure 2. It is evident from the Figure 2 that there is obvious difference between amounts of EDTA extractable copper and phosphorus in vineyard soil samples (Orlica, Kalvarija 2, Kalvarija 3) and amounts of the same elements in control soil samples (Orlica control, Kalvarija control). Concentrations of AL-soluble phosphorus are up to nineteen times higher in vineyard soils than in control soil samples (uncultivated surfaces). Phosphorus and copper proportion is 2 to 4 times lower in control soil samples than in intensively cultivated soil samples. Addition and uptake of copper in the Orlica vineyard was also calculated. Annual copper addition was calculated from copper content data in phytopharmaceutical substances used for vineyard protection in one-year period. The data are shown in the Table 4. Annual copper addition with phitopharmaceuticals amounts to 8.5 kg per vineyard surface which is 1.7 kg of Cu per ha. Since 1996 the vine- Figure 2. Content of AL-soluble copper and phosphorus, and P/Cu proportion in soil samples. Table 3: Phosphorus and copper content in AL- extracts from soil samples and P/Cu proportion. soil sample P (AL-soluble) (mg/kg) Cu (AL-soluble) (mg/kg) P/Cu Mass proportion Orlica control 4.6 6.0 0.8 Orlica 40.6 11.5 3.5 Kalvarija control 5.9 4.2 1.4 Kalvarija 2 109.1 34.2 3.2 Kalvarija 3 115.4 40.4 2.9 Table 4: Content of Cu in phytopharmaceutical substances used for Orlica vineyard protection. Dosage (kg) Cu (%) Cu (kg) Cu - Antracol 40 17.5 7 Cu - Euparen 10 15 1.5 Determination of Cu in pomace and the rest of the vine may count significantly for total uptake of Cu from vineyard soil. yard Orlica is not treated with the mineral fertilizers. Annual Cu uptake was calculated from contents data in the Sauvignon wine and than the amount of wine produced in the Orlica vineyard. Content of Cu in wine was determined by flame atomic absorption method. The wine contained 0.05 mg Cu per liter. Estimated quantity of wine produced in the vineyard Orlica is 24,000 L meaning that our copper uptake with wine from this vineyard is 1.2 g per year. It is important to highlight that with wine analysis the total uptake of copper from soil could not be included. Determination of Cu in pomace and the rest of the vine (leaves, shoots ...) may count significantly for total uptake of Cu from vineyard soil. Another consideration is transfer of copper caused by rain to the nearest surface waters. Sediment of the Meranovo stream which springs in the Pohorje territory above the vineyard Orlica, and runs below the vineyard to the river Drava, was analysed. Cooper and zinc contents in the sediment are shown in the Table 5. Copper content in the Orlica vineyard soil was 48 mg/kg, zinc content was 99 mg/kg. Due to the fact that Cu is strongly immobilized by soil organic matter and Fe, Mn, Al-(hydr)oxides, elevated Cu concentrations originating from phytopharmaceutical applications were observed in superficial ho- Table 5: Copper and zinc content in the Meranovo stream sediment. sampling site Zn (mg/kg) Cu (mg/kg) Spring 96 37 Below the vineyard 95 51 Behind the vineyard 120,5 89 Mouth of the Drava 130 107 Table 6: Limit, warning and critical imission values for Cu and Zn in soil. Limit imission value Warning imission value Critical imission value (mg/kg) (mg/kg) (mg/kg) Cu 60 100 300 Zn 200 300 720 rizons of vineyard soils. The highest Cu concentrations were found in the surface layer (0 cm - 20 cm). Concentrations of Cu and Zn in the stream sediment are much lower before the stream reaches the vineyard than after it leaves it. Increased concentrations of both elements are a consequence of spraying the vineyard with zinc and copper containing phytopharmaceutical substances. With analysis of the stream sediment which springs in the Pohorje territory and runs below the vineyard Orlica to river Drava can be inferred that the increasing Cu and Zn contents in the sediment were caused by the use of phytopharmaceuticals containing those two elements. Rain washes away the vineyard soil and the silt gathers in streams. Amounts of copper in stream sediment behind the vineyard Orlica and in the mouth of the Drava River exceed the limit imission value for Cu in soil. Contents of zinc in soil and stream sediment do not exceed limit emissions. Regulation about limit, warning and critical imission values of hazardous substances in soil deals with heavy metals in soil in Slovenia (Official gazette of the Republic of Slovenia, Vol. 68/96). Limit, warning and critical imission values for copper and zinc in soil are set out in the Table 6. The copper amounts in vineyard Kalvarija exceed the limit imission value for Cu in soil for both parts of the vineyard. Zinc concentrations were not exceeded in discussed vineyards soils. Amounts of copper in stream sediment behind the vineyard Orlica with 89 mg/kg and in the mouth of the Drava River with 107 mg/kg exceed the limit imission value for Cu in soil. There is a much higher portion of available copper in the vineyard soils relative to the control soil samples; there is no overall relationship between total and available copper in soils. Concentrations of plant available copper in soils cannot be predicted on the basis of total copper concentration, which is important for developing environmental guidelines. references [1] Kabata-Pendias. A. Trace Elements in Soil and Plants. Boca Raton. Florida: CRC Press, 2001. pp. 128. [2] Wanga Q-Y, Zhoua D-M and Canga L. Microbial and enzyme properties of apple orchard soil as affected by long-term application of copper fungicide, Soil Biology and Biochemistry. 2009; 41: 1504-1509. [3] McLaughlin MJ, Hamon RE, McLaren RG et al. Review: A bioavailabili-ty-based rationale for controlling metal and metalloid contamination of agricultural land in Australian and New Zealand. Aust. J. Soil Res. 2000; 38: 1037-1086. There is a much higher portion of available copper in the vineyard soils relative to the control soil samples; there is no overall relationship between total and available copper in soils. [4] Paoletti MG et al. Earthworms as useful bioindicators of agroecosytem sustainability in orchards and vineyards with different inputs. Appl. Soil Ecol. 1998; 1G: 137-15G. [5] Weingerl V, Kerin D. Distribution of zinc in vineyard areas treated with zinc containing phytopharmaceuticals. Acta Chim. Slov. 2GGG; 453467. [6] Arias M, López E, Fernández D, Soto B. Copper Distribution and Dynamics in Acid Vineyard Soils Treated With Copper-Based Fungicides. Soil Science. 2GG4; 169: 796-8G5. [7] Stoate C et al. Ecological impacts of arable intensification in Europe, Journal of Environmental Management. 2GG1; 63: 337-365. [8] Andjelov M. Rezultati radiometričnih in geokemičnih meritev za karto naravne radioaktivnosti Slovenije. Geologija. 1994; 34: 223-248. [9] Šajn R, Geokemične lastnosti urbanih sedimentov na ozemlju Slovenije, Doktorsko delo, Univerza v Ljubljani, Naravoslovno tehniška fakulteta. 1998. [1G] Rusjan D, Strlič M, Pucko D and Korošec-Koruza Z. Copper accumulation regarding the soil characteristics in Sub-Mediterranean vineyards of Slovenia. Geoderma. 2GG7; 141: 111-118. [11] Rusjan D, Strlič M, Pucko D et al. Vineyard soil characteristics related to content of transition metals in a Sub-Mediterranean winegrowing region of Slovenia. Geoderma. 2GG6; 136: 93G-936. [12] Voegelin A, Vulava VM, Kretzschmar R. Reaction-Based Model Describing Competitive Sorption and Transport of Cd, Zn, and Ni in an Acidic Soil, Environ. Sci. Technol. 2GG1; 35: 1651-1657. [13] Finžgar N and Leštan D. Multi-step leaching of Pb and Zn contaminated soils with EDTA. Chemosphere. 2GG7; 66: 824-832. [14] Önorm L 1G57 Probenahme von wein- und obstbaulich genutzten Böden und Böden von Baumschulen, Österreichisches Normungsinstitut, Wien, 1993. [15] Önorm L 1G84 Chemische Bodenuntersuchungen, Bestimmung von Carbonat, Österreichisches Normungsinstitut, Wien, 1989. [16] Önorm L 1G83 Chemische Bodenuntersuchungen, Bestimmung von Acidität, Österreichisches Normungsinstitut, Wien, 1989. [17] Flores-Velez LM, Ducaroir J, Jaunet AM, Robert M. Study of the distribution of copper in an acid sandy vineyard soil by three different methods. Eur. J. Soil Sci. 1996; 47: 523-532. [18] Brun LA, Maillet J, Richarte J, et al. Relations between extractable copper, soil properties and copper uptake by wild plants in vineyard soils, Environ. Pollut. 1998; 1G2: 151-161. [19] Besnard E, Chenu C and Robert M. Influence of organic amendments on copper distribution among particle-size and density fractions in Champagne vineyard soils, Environ. Pollut. 2GG1; 112: 329-337. [2G] Brun LA, Maillet J, Hinsinger P, and Pépin M. Evaluations of copper bioavailability to plants in copper-contaminated vineyard soils. Environ. Pol-lut. 2GG1; 111: 293-3G2. [21] Official gazette of the Republic of Slovenia. Directives about limit, warning and critical imission values of hazardous substances in soils. In: Official gazette of Republic of Slovenia, Ljubljana 1996; 68: 5773-5774. [22] Deveza A, Gomeza E, Gilbinb R, et al. Assessment of copper bioavailability and toxicity in vineyard runoff waters by DPASV and algal bio-assay, Science of The Total Environment 2GG5; 348: 82-92. [23] Andersen AB, Science in Agriculture, Acres USA, Kansas City, Missouri, 2GGG. p.p. 376. Professional paper ■ © Inštitut za sanitarno inženirstvo, 2010. Waste Treatment within Small and Medium Sized Enterprises and Proposals for Improvement Katarina ZELEZNIK LOGAR* ABSTRACT Small and medium-sized enterprises (SMEs) play an important role in many economies around the world and are vitally important for a health dynamic market economy. Collectively, SMEs also cause significant impacts on the environment by their activities, products and services. The present situation, when SMEs are unaware of their environmental impact, can pose an important threat not only to the environment but also can have a negative effect on worker's health and safety. The main research questions of our study were: do entrepreneurs performing craft activities, have the necessary knowledge to carry out their waste management activities and environmental performance in general, and what possibilities for better waste treatment, waste minimization and reduction can be offered to SMEs and how? The results supported this hypothesis are that the level of knowledge about their responsibilities according to the regulations among many SMEs is still low. In the future, it will be crucial that public authorities and other key players support SMEs in increasing their efforts for improvement. KEYWORDS: Craft, Environmental impact, Small and medium-sized enterprises (SMEs), Waste, Waste management. Received: 5. 7. 2010 Accepted: 10. 7. 2010 Chamber of Craft and Small Business of Slovenia Celovška c. 71, Ljubljana, Slovenia e-mail: katarina.zeleznik@ozs.si * A statement: The article sent is original, the manuscript or any part of it has not been sent to any other publisher and it is not in consideration for publication anywhere in any language. The total environmental impact of small firms is not known either at national or regional levels, but according to some surveys about 70 % of all industrial pollution is contributed by SMEs. SMEs are responsible for the waste generated within their business; from when it is produced until they have transferred it to an authorized person. introduction A great share of the businesses in Slovenia and European Union is small, representing some 99 % of all enterprises, 57 % of economic value added and about 66 % of total employment. Based on the definitions micro, small and medium-sized enterprises is made up of enterprise are which employ fewer than 250 persons and which have an annual turnover not exceeding 50 million Euro, and/or an annual balance sheet total not exceeding 43 million Euro [1]. The Law of Craft defines that a craft activity is a profit oriented activity being divided into craft activity, activity similar to craft and activity of home craft and applied arts being performed by self employed persons, an economic company or a craft collective. An individual person can perform a craft activity for which certain qualification repairements must be fulfilled and on the basis of craft permission and registration in the craft register [2]. The Slovenian craft with 52,109 craft enterprises and over 150,000 employees represents an important part of the Slovenian economy and the bulk of craft enterprises are SMEs, according to definition above. Special categories in the Slovenian craft are definitely craft enterprises without employees, which represent 55,4 % of all registered craft enterprises [3]. The total environmental impact of small firms is not known either at national or regional levels, but according to some surveys about 70 % of all industrial pollution is contributed by SMEs. The problem is not so much in individual firms, although in some cases there can be serious individual impacts on the local environments and communities, but rather in their combined total impact across sectors [4]. Over the years, worldwide concern for the environment is getting bigger attention. There are clear signals that wide industrial sectors are moving towards cleaner production processes, in order to respond, among other factors, to climate change imperatives. Industrial activities which cause an environmental pollution has been bounded to more comprehensive and effective environmental schemes or legislations (such as the IPPC Directive, the Directive on the Emission Trading Scheme), while SMEs often fall below the thresholds that trigger the application of instruments that concentrate on major individual sources of pollution. In cases, where environmental legislation is applicable to SMEs, they tend to presume that they are complying and, as a result, full compliance is often the result of external action after an inspection rather than an on-going process of checking that legal requirements are actually being met [5]. At the same time, SMEs often do not have the necessary legal and environmental expertise to cope with environmental legislation. SMEs are responsible for the waste generated within their business; from when it is produced until they have transferred it to an authorized person. The field of waste treatment within crafts is well regulated. Slovenian environmental policy has been based on the Waste Management Decree, and has been harmonized with the corresponding Euro© Institut za sanitarno inzenirstvo, 2010. pean Union's legislation. The amount of waste produced is usually lower than it is within the industry sector, but nevertheless the heterogeneous and diverse nature of the craft sector makes it difficult to generalize about the environmental impacts and strategies of the sector. Some of the empirical literature shows that the majority of SMEs have little awareness of the environmental impacts and how to manage them. Most of them (between 75 % and 90 % according to some surveys) think that their activities do not have any impact on the environment. Therefor, most of them have not introduced any practical measures to reduce their environmental impacts [6]. SMEs usually do not have any employees dealing with environmental issues, so this remains the duty of the managing director or the business owner who also handles all other activities of the enterprise. It is hard for them to comply with environmental legislation. Actions are usually taken in order to avoid costs, but not at the expence of environmental protection [7]. Environmental planning, if any, in most cases is only of a short term and is applied as necessary. Sometimes companies have difficulties in understanding how the legislation affects them, which is accompanied by poor knowledge about their responsibilities according to the regulations; separating hazardous and special waste from others, false treatment and storing, keeping records and the awareness that these responsibilities have no time limit. If SMEs are unaware of the environmental impacts of their own activities, and of the environmental legislation that applies to them, their activities could present an important threat for the environment. The environmental impacts of SMEs which are associated with low awareness and low legal compliance could also have the effect on increasing health and safety risks of workers. And at the end they can miss the chance to reap the economic benefits of the opportunities presented by better environmental management and eco-innovation [7]. The main hypothesis of this research work was that the majority of SMEs have little awareness of their environmental impact, particularly within the waste management operations, which makes them problematic in terms of compliance with environmental legislation. Through the research work we wanted to find out what is the level of knowledge about environmental requirements, waste and hazardous waste, waste minimization, and waste treatment in general between SMEs. We also wanted to find out how to provide possibilities that are not connected to high costs and investments for better waste treatment? By the help of questionnaires, interviews and on-site visits it has been endeavored to analyze the current situation, regarding how craft shops are dealing with waste and hazardous waste generated within business. The study identifies some proposals for waste management improvements within SMEs starting with the prevention of waste generation in the first place. Also some other measures and tools to help SMEs to comply with the legislation and to improve their environmental attitude were identified. Actions are usually taken in order to avoid costs, but not at the expence of environmental protection. The environmental impacts of SMEs which are associated with a low awareness and low legal compliance could also have the effect of increasing health and safety risks to workers. The results consist of data collected through the analysis of returned questionnaires and of data collected from on-site interviews. METHODS The main objective of the study is to identify some proposals for waste management improvements within SMEs, starting with the prevention of waste generation, and other measures and tools to help SMEs to comply with the legislation and to improve their environmental attitude. The empirical part of presented work included a desktop research, which was in May 2008, continued with analyzing the questionnaires sent randomly using the data from the Craft Register [8], to 300 entrepreneurs who are members of the Chamber of Craft and Small Business of Slovenia from the section of maintenance of textiles and car services. The survey also included an on-site visit along with interviews conducted with some SMEs. The results consist of data collected through the analysis of returned questionnaires and of data collected from on-site interviews. The response rate was about 40 % and lower than had been anticipated, particularly from the car service section. There were 21 questions in the questionnaire; most of them were fixed-choice with two, three or more possible answers to select. In the introduction, it was asked about their core business and number of employees. It was continued with questions about quantities of waste and hazardous wastes produced per year, collecting and separating wastes at source, storing and recycling. It was foreseen that SMEs do not have full awareness of the legal requirements associated with their operation, so asking them about being aware of the demands of legislation. The questions following referred to specific waste treatment procedures which Table 1: Main themes of the interviews. Waste and waste management within business: Actions to meet legislation and complying with legislation. Waste treatment within the business. Waste minimization, recovery, recycling. Removal by authorized person, disposal. Keeping the necessary documentation. Awareness of environmental impact of the business: Is there any impact and what kind of? Responsibility towards the environment. Reducing environmental impacts. Environmental planning. Environmental inspector visits: Supervisions within last five years. Measurements set down by the inspector. Improvements after the visit? Need for further information and better advice: How, what? Guidance. Motives to build up en EMS: Improvements regarding environmental issues. Willingness for on-going efforts. Short term plan to implement ISO 14001, EMASeasy. Expected benefits, obstacles. are performed by each SME, such as: costs which appear yearly for hazardous and non hazardous waste treatment, having waste documentation as a waste management plan according to the Decree on waste management, who is involved in environmental issues, where do they usually get information and necessary knowledge and skills to comply with environmental issues, do they need additional education, do they have any controls and examinations by environmental inspectorate in their crafts units and finally, what measures were given, if any. Interviews and on-site visits were meant as continuation work after the survey. Interviews were performed in May 2009, in selected companies within two businesses: car section, and textile maintenance 2009. The interviewing method was not based on a formal schedule of questions but the researchers had a list of selected themes for the discussion. On-site visits have followed the interviews and helped to better understand the situation and problems appearing within SMEs. Main themes of the interviews with managing directors-owners as interviewee are listed in the Table 1. results and discusion The results of this study clearly showed that SMEs in general are very passive in environmental protection and that they primarily follow the development of tightening legislation. Actions taken are usually in order to avoid costs, not for sustainability or environmental protection. They often do not deal with environmental planning but if they do, planning is only for the short term. There are still some entrepreneurs who feel limited responsibility towards the environment. They often think they are small entrepreneurs and that their contribution towards environmental impacts is negligible. Sometimes, even if they were familiar with some options to improve their environmental attitude, there was no action taken because there was no willingness and positive aspiration to comply. The results of the study showed that we are dealing with very small entrepreneurs (average number of employees is 3.1 with owners included), which are generating wastes and hazardous waste. A high percentage of those asked - 34 % - do not assort waste and dispose of the municipal waste, and from the answers which followed, it was clear that the knowledge about waste treatment is generally low. The Figure 1 shows different types of wastes which were stated as most common by entrepreneurs. It is obvious from the results that the surveyed entrepreneurs do not know enough about municipal wastes they listed only a few types of waste, in spite of the previous question in which many of respondents - 62 % - affirmed that they know about the municipal waste. They were asked which hazardous wastes have been generated within their business? Answers given by participants performing textile processing showed that they know their hazardous waste very well, while answers received from participants of the section of car services showed that they know only few of the hazardous wastes generated The results of this study clearly showed that SMEs in general are very passive in environmental protection and that they primarily follow the development of tightening legislation. It is obvious from the results that the surveyed entrepreneurs do not know enough about municipal wastes they listed only a few types of waste. Knowing municipal waste > 60 0) u a 50 a> 40 ■2 30 M « o* 20 o S 10 -Q 0 I—I glass paper plastic metal biodegradable waste glass no answer types of wastes Figure 1. Specify the municipal wastes which are being collected and disposed separately. Figure 2. Which hazardous wastes are generated within your business? within their business (Figure 2). Mechanics are less aware of hazardous waste generated, even if there can be different types which can cause mismanagement with hazardous types of waste and pose a threat towards the environment and also to human health. Only 10 % of the entrepreneurs surveyed generate less than 200 kg/a of hazardous waste and are consequently not obliged to prepare the waste management plan. But the following answers showed that more than 30 % of respondents thought they are not obliged to have the Types of hazardous waste in your business 70 ■ Graphic Design & Layout: Tempora, Rijeka, HR