EXTERIOR CHARACTERISTICS OF DUBROVNIK SHEEP ...multilingual.bionetsyst.com/images/docs/...sampling...

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RESEARCH PEOPLE AND ACTUAL TASKS ON MULTIDISCIPLINARY SCIENCES 8 10 JUNE 2011, LOZENEC, BULGARIA 1 EXTERIOR CHARACTERISTICS OF DUBROVNIK SHEEP - CROATIAN ENDANGERED BREED Z. Antunović, I. Marić, Đ. Senčić, J. Novoselec Abstract: The aim of this work is to show exterior characteristics of the Dubrovnik sheep - Croatian endangered sheep breed and to compare her with other Croatian sheep breeds. In the Republic of Croatia is grown 584 breeding heads of Dubrovnik sheep, which is only 0.14% of total breeding sheep. Breeding are stationed in Dubrovnik Neretva County, distributed in 27 registered breeders with the average herd size of 22 breeding head. The study was conducted with 30 Dubrovnik sheep during the summer feeding season. Sheep were fed on pasture, and upon returning to the barn were given hay (ad libitum). Weighing and sampling of exterior characteristics of sheep has been carried out in the morning before going out to pasture. Recorded an average body weight of Dubrovnik sheep was 45.80 kg, 62.58 cm height at withers and body length 65.07 cm. Compared with other Croatian sheep breeds Dubrovnik sheep is by the exterior characteristics medium-developed breeds. Indices of physical development of Dubrovnik sheep indicate good physical condition and sufficient nutrition. Although it can be said that the introduction in the meal some concentrated feed was necessary with the aim to improve productivity, especially in drought periods of the year. Key words: Dubrovnik sheep, Exterior characteristic, Indices of physical measures. INTRODUCTION Dubrovnik sheep, known as the Dubrovnik ruda, was created in random crossing domestic sheep-Pramenka with imported Spanish, French and Italian Merinos with aim to improve the quality of wool and body wool overgrown. However, the development of tourism and the loss of the importance of wool, breedings in this breed were significantly decreased. In recent years, has slowly working on its conservation. Dubrovnik sheep is most endangered Croatian native breeds of sheep. According to the report of the Croatian Agricultural agencies [9] in the Republic of Croatia has grown 584 breeding head of Dubrovnik sheep, which is only 0.14% of the total number of breeding ewes. Breeding are stationed in Dubrovnik Neretva County, distributed in 27 registered breeders with the average herd size of 22 breeding head. Given the vulnerability of breeding Dubrovnik sheep, arises question how to preserve and encourage its spread, not only in traditional areas where it is cultivated for centuries, but also in areas with similar geographical and climatic conditions. First of all, it is necessary to determine the productivity, nutritional and health status of the existing population of Dubrovnik sheep [2,3]. There are a very small number of papers which talk about breeding of Dubrovnik sheep [13, 11, 1, 4]. Therefore, the aim of this study are to show exterior and production characteristics of Dubrovnik sheep and compare them with other Croatian sheep breeds. MATERIAL AND METHODS The study was carried out on family farms in the Dubrovnik-Neretva County as part of project Ministry of Science, Technology and Sports "Nutritional aspects of modeling productivity and metabolic profile of sheep.” The experiment included 30 Dubrovnik sheep an average age 4 years. The sheep were healthy and in satisfactory physical condition. The research was conducted during the summer feeding season when the sheep stayed in the natural pasture, and upon his return to barn they received hay (ad libitum). Weighing and sampling of exterior characteristic of sheep has been carried out in the morning before going out to the pasture. Body condition score of sheep, with scores of 1-5, was determined by Russel [14]. Anamorphosis and body proportions indices of sheep were calculated by Chiofalo et al. [7]. Body measures of sheep (withers height, chest

Transcript of EXTERIOR CHARACTERISTICS OF DUBROVNIK SHEEP ...multilingual.bionetsyst.com/images/docs/...sampling...

  • RESEARCH PEOPLE AND ACTUAL TASKS ON MULTIDISCIPLINARY SCIENCES 8 – 10 JUNE 2011, LOZENEC, BULGARIA

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    EXTERIOR CHARACTERISTICS OF DUBROVNIK SHEEP - CROATIAN

    ENDANGERED BREED

    Z. Antunović, I. Marić, Đ. Senčić, J. Novoselec

    Abstract: The aim of this work is to show exterior characteristics of the Dubrovnik sheep - Croatian endangered sheep breed and to compare her with other Croatian sheep breeds. In the Republic of Croatia is grown 584 breeding heads of Dubrovnik sheep, which is only 0.14% of total breeding sheep. Breeding are stationed in Dubrovnik Neretva County, distributed in 27 registered breeders with the average herd size of 22 breeding head. The study was conducted with 30 Dubrovnik sheep during the summer feeding season. Sheep were fed on pasture, and upon returning to the barn were given hay (ad libitum). Weighing and sampling of exterior characteristics of sheep has been carried out in the morning before going out to pasture. Recorded an average body weight of Dubrovnik sheep was 45.80 kg, 62.58 cm height at withers and body length 65.07 cm. Compared with other Croatian sheep breeds Dubrovnik sheep is by the exterior characteristics medium-developed breeds. Indices of physical development of Dubrovnik sheep indicate good physical condition and sufficient nutrition. Although it can be said that the introduction in the meal some concentrated feed was necessary with the aim to improve productivity, especially in drought periods of the year.

    Key words: Dubrovnik sheep, Exterior characteristic, Indices of physical measures.

    INTRODUCTION Dubrovnik sheep, known as the Dubrovnik ruda, was created in random crossing

    domestic sheep-Pramenka with imported Spanish, French and Italian Merinos with aim to improve the quality of wool and body wool overgrown. However, the development of tourism and the loss of the importance of wool, breedings in this breed were significantly decreased. In recent years, has slowly working on its conservation. Dubrovnik sheep is most endangered Croatian native breeds of sheep. According to the report of the Croatian Agricultural agencies [9] in the Republic of Croatia has grown 584 breeding head of Dubrovnik sheep, which is only 0.14% of the total number of breeding ewes. Breeding are stationed in Dubrovnik Neretva County, distributed in 27 registered breeders with the average herd size of 22 breeding head. Given the vulnerability of breeding Dubrovnik sheep, arises question how to preserve and encourage its spread, not only in traditional areas where it is cultivated for centuries, but also in areas with similar geographical and climatic conditions. First of all, it is necessary to determine the productivity, nutritional and health status of the existing population of Dubrovnik sheep [2,3]. There are a very small number of papers which talk about breeding of Dubrovnik sheep [13, 11, 1, 4]. Therefore, the aim of this study are to show exterior and production characteristics of Dubrovnik sheep and compare them with other Croatian sheep breeds.

    MATERIAL AND METHODS

    The study was carried out on family farms in the Dubrovnik-Neretva County as part of project Ministry of Science, Technology and Sports "Nutritional aspects of modeling productivity and metabolic profile of sheep.” The experiment included 30 Dubrovnik sheep an average age 4 years. The sheep were healthy and in satisfactory physical condition. The research was conducted during the summer feeding season when the sheep stayed in the natural pasture, and upon his return to barn they received hay (ad libitum). Weighing and sampling of exterior characteristic of sheep has been carried out in the morning before going out to the pasture. Body condition score of sheep, with scores of 1-5, was determined by Russel [14]. Anamorphosis and body proportions indices of sheep were calculated by Chiofalo et al. [7]. Body measures of sheep (withers height, chest

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    circumference, carcass length, shin-bone circumference, chest width, chest depth, pelvis width, pelvis length) were performed by the Lydtin’s stick or stock band. Indices of chest, chest depth, body compactness, massines, body weight, length and bony of sheep were determined by Ćinkulov et al. [8]. The research results were analyzed by descriptive statistics in the computer program Statistica Stat Soft Windows [15].

    RESULTS AND DISCUSSION Tables 1 and 2 shows exterior characteristics and indices of physical measurement

    Dubrovnik sheep. Table 1. Exterior characteristic of Dubrovnik sheep

    Traits, cm Statistical values

    mean sd min max SE

    Body weight, kg 45.80 9.31 29.00 63.50 1.70

    Withers height 62.58 3.69 49.00 67.00 0.67

    Chest circumference 83.98 6.17 70.00 96.00 1.13

    Carcass length 65.07 2.68 59.00 70.00 0.49

    Shin-bone circumference 7.21 0.21 6.80 7.60 0.04

    Chest width 19.42 2.17 15.00 24.00 0.40

    Chest depth 30.15 1.37 27.00 33.00 0.25

    Pelvis width 18.60 1.50 14.50 21.00 0.27

    Pelvis length 19.53 1.87 15.00 23.00 0.34

    sd- standard deviation; SE- standard error

    From Table 1 it is visible that the Dubrovnik sheep achieved an average weight of 45.80 kg and height of the withers of 62.58 cm.

    Mentioned indicates that Dubrovnik sheep are medium size breed in compare to other Croatian sheep breeds (Table 3). Research in the past with Dubrovnik sheep, although they are very rare, indicate considerable variability in body weight of sheep. Namely, in the research Rako [13] determined the average body weight was 32.94 kg, and in the research Mioč et al. [11] 47.93 kg. The reasons for these variations primarily we can see in the quality of feeding and rearing of Dubrovnik sheep.

    Table 2. Physical measures indices of Dubrovnik sheep

    Index Statistical values

    mean sd min max SE

    Body condition score 3.37 0.51 2.5 4.0 0.09

    Anamorphosis index 113.54 17.21 83.05 153.60 3.14

    Body proportion index 73.18 14.29 49.15 101.67 2.61

    Index of chest 64.37 6.31 53.57 77.42 1.15

    Index of chest depth 48.30 3.04 43.65 57.14 0.56

    Index of body compactness 129.11 8.57 115.70 148.39 1.56

    Index of massiness 134.60 12.05 118.64 161.23 2.19

    Index of body weight 73.18 14.29 49.15 101.67 2.61

    Index of leg length 51.70 3.04 42.86 56.35 0.55

    Index of bony 11.55 0.70 10.30 14.29 0.13

    sd- standard deviation; SE- standard error

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    Shown physical development indices of Dubrovnik sheep indicate good physical condition and sufficient nutrition. Although it can be said that the introduction in the meal some concentrated feed (corn, oats, barley, wheat bran) was necessary in order to improve productivity, especially in drought periods of the year (hot summer months). It is known to be very simple and convenient monitoring sheep nutritional status can be done by establishing the body mass and body condition scores [16, 5]. The research Ćinkulov [7] conducted on Tsigai shows that the indices of physical development were higher, which is reasonable because the Tsigai sheep physically developed breeds in relation to Dubrovnik sheep, which is confirmed by research Antunović et al. [6]. Possible direction of growing Dubrovnik sheep in the future are in the function of tourism through the production of lamb meat and sheep cheese.

    Table 3. Exterior characteristics of Croatian sheep breeds

    BM Dubrovnik sheep1

    Tsigai2 Istrian sheep3

    Lika pramenka4

    Pag sheep3

    Cres sheep3

    Dalmatian pramenka3

    Krk sheep3

    BW 47.93 74.64 67.38 49.25 43.05 41.58 39.43 35.44 WH 60.12 81.20 73.51 60.75 56.14 59.97 59.43 54.64 CL 65.05 91.21 77.33 67.35 64.27 67.26 65.78 61.36 CD 30.32 32.11 32.98 29.28 27.91 29.34 28.55 28.14 CW 19.81 22.75 21.71 16.64 17.11 17.75 17.75 16.20 SBC 7.54 9.12 9.02 7.48 7.04 7.93 7.45 6.96 CC 86.45 111.7 96.69 83.83 83.26 83.1 82.39 76.39

    BM- body measures; BW- body weight, kg; WH- Withers height, cm; CL- carcass length, cm; CD- Chest depth, cm; CW - Chest width, cm; SBC - Shin-bone circumference, cm; CC- Chest circumference, cm; 1 Mioč et al. [11]; 2 Antunović et al. [6]; 3 Mioč et al. [12], 4 Mioč et al. [10]

    CONCLUSIONS AND FUTURE WORK

    Based on the results obtained in this study we can conclude that Dubrovnik sheep belongs to a group of medium developed Croatian sheep breeds and that it must work on the quality of nutrition in order to repair their production properties and indices of physical development.

    REFERENCES

    [1]. Antunović, Z. et al. 2007. Exterior, productive and metabolic characteristics of Dubrovnik lambs. 1st Conference on Native Breeds and Varieties as part Natural and cultural Heritage. Šibenik, Croatia, Novemeber 13-16 2007, pp. 6-10. [2]. Antunović, Z. et al. 2009. Blood metabolic profile of Dubrovnik sheep - Croatian endangered breed. Proceedings of IV International Symposium of Livestock Production, Struga, Macedonia, 9-12. 09. 2009., p. 46. [3]. Antunović Z. et al. 2010. Blood haematological and biochemical profil of Dubrovnik sheep. 2nd Conference on Native Breeds and Varieties as part Natural and cultural Heritage. Poreč, Croatia, September 22-25 2010, pp. 10-11. [4]. Antunović Z. et al. 2010.:Primjena manan-oligosaharida u hranidbi sisajuće janjadi u ekstenzivnom uzgoju. Krmiva 52, 2, 71-76. [5]. Antunović Z. et al. 2010. Praćenje hranidbenog statusa janjadi iz ekološkog uzgoja. Krmiva 52, 1, 27-34.

    http://bib.irb.hr/prikazi-rad?&rad=477804http://bib.irb.hr/prikazi-rad?&rad=477804

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    [6]. Antunović et al. 2011. Fenotipske odlike cigaje u ekološkom uzgoju. Zbornik radova 46. hrvatskog & 6. međunarodnog znanstvenog simpozija agronoma. 14.-17. 02. 2011.(in press). [7]. Chiofalo, V. et al. 2004. Effects of the administration of lactobacilli on body growth and on the metabolic profile in growing Maltese goat kids. Reprod. Nutr. Dev. 44, 449-457. [8]. Ćinkulov, M. et al. 2003. Phenotypic differences between two types of Tsigai breed of sheep. Lucrai stiintifice Zootehnie si Biotehnologii, vol XXXVI, Timisoara, Romania, pp.1-6. [9]. HPA (2010.): Godišnje izvješće. Ovčarstvo, kozarstvo i male životinje. Izvješće za 2009. godinu. Križevci. [10]. Mioč, B. et al. (1997). Odlike eksterijera ličke pramenke. Stočarstvo 52, 1, 93-98. [11]. Mioč, B. et al. 2003. Odlike eskterijera i polimorfizmi proteina dubrovačke ovce. Stočarstvo 57, 1, 3-11. [12]. Mioč, B. et al. 2007. Exterior characteristics and production traits of some Croatian autochtonous sheep breeds. 1st Conference on Native Breeds and Varieties as part Natural and cultural Heritage. Šibenik, Novemeber 13-16 2007, pp. 190-193. [13]. Rako, A. 1949. Dubrovačka ovca. Veterinarski arhiv XIX, 3/6, 63-122. [14]. Russel, A. 1991. Body condition scoring of sheep. In: Sheep and goat practice. Boden E. (ed.). p. 3. Bailliere Tindall, Philadelphia. [15]. STATISTICA-Stat Soft, Inc. version 8.1, 2008, www.statsoft.com. [16]. Whitney, T.R. et al. 2009. Evaluating Nutritional Status of dorper and rambouillet ewes in range sheep production. Sheep and Goat Research Journal, 24, 10-16.

    Acknowledgements This article is a part of a research project “Nutritional aspects of modeling productivity and metabolic profile of sheep” financed by the Ministry of Science, Education and Sports of the Republic of Croatia.

    ABOUT THE AUTHORS

    PhD. Zvonko Antunović, Full professor, Department of Animal Science, Faculty of Agriculture, University of J.J. Strossmayer, Trg sv. Trojstva 3, 31000 Osijek, Croatia, tel: 00385 31 224 220; Fax: 00385 31 224 220; E- mail : [email protected]

    PhD. Đuro Senčić – full professor; Department of Animal Science, Faculty of

    Agriculture, University of J.J. Strossmayer, Trg sv. Trojstva 3, 31000 Osijek, Croatia, tel: 00385 31 224 220; Fax: 00385 31 224 220; E- mail : [email protected]

    BSc. Josip Novoselec – scientific novice, Department of Animal Science, Faculty of

    Agriculture, University of J.J. Strossmayer, Trg sv. Trojstva 3, 31000 Osijek, Croatia; E-mail : [email protected]

    BSc. Ivica Marić – Croatia Agency of Agriculture, M. Marojevića 4, 20000 Dubrovnik,

    Croatia; E- mail: [email protected]

    http://www.statsoft.com/mailto:[email protected]:[email protected]:[email protected]:[email protected]

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    BENEFIC EFFECTS OF SEVERAL ESSENTIAL OILS TREATMENTS IN HEALTHY AND POTATO VIRUS Y INFECTED PLANTS Solanum

    tuberosum L. AND Nicotiana tabacum

    Carmen Liliana BĂDĂRĂU , Angela MĂRCULESCU, Nicoleta CHIRU, Florentina DAMŞA, Andreea NISTOR

    Abstract: Antioxidants,poliphenols presents in essential oils (Lamiaceae plants) and other compounds

    (hydrogen peroxide, ascorbic acid) are implicated in processus signaling against stress. The treatments of positive potato plants with Rosmarinus officinalis oils significantly reduced the number of minitubers, enhancing their weights, while leaf pigment content also increased. Concerning the antiviral effect of Thymus serpyllum, Lavandula officinalis oils,all the treated tobacco plants presented after PVY infection values of absorbances at 405nm signifficantly lower than the untreated and inoculated controls.

    Key words: potato virus Y, essential oils, Rosmarinus officinalis, Thymus serpyllum, Lavandula

    officinalis, tobacco Abbreviation: AA ascorbic acid; ROS reactive oxygen species; RA rosmarinic acid; RO Rosmarinus

    officinalis; PVY potato virus Y; OD optic density; SD standard deviation

    INTRODUCTION Over the past 20 years, potato virus Y (PVY) (Potyviride) has become a serious

    constraint to potato (Solanum tuberosum L.) production in the world [7],[16]. High virus level can cause stand loss, reduced yields and reduced quality [5]. Thus, efforts to control PVY are essential when producing potatoes [2],[3],[5],[16],[23]

    Being very susceptible to potyvirus infection, Nicotiana tabacum (family Solanaceae) is used usually like test plant for potato virus Y [5].

    Phenolic compounds and other constituents of Rosmarinus officinalis, Thymus serpyllum, Lavandula officinalis plants (Family Lamiaceae, order Lamiales) have antioxidant activity and pharmaceutical properties [4],[21]. They are also antimicrobial, antiviral wich protects the plants. Oils extracted from Rosmarinus officinalis, Thymus serpyllum, Lavandula officinalis introduced in healthy and infected potato plants could be implicated in the processus signaling against stress [24]. Plant cells have defensive responses to pathogen attack associated with changes in oxidative metabolism [13]. One of the consequences of stress is an increase in the cellular concentration of reactive oxygen species (ROS), which are subsequently converted to hydrogen peroxide (H2O2). These ROS, particularly H2O2, play versatile roles in normal plant physiological processes and in resistance to stresses. H2O2 produced in excess is harmful, but lower concentrations are beneficial [22], [15], [17] in microplants of Solanum tuberosum. Genetic and physiological evidence suggests that H2O2 mediates the acquisition of tolerance to biotic and abiotic stresses [1], [22]. Another molecule that participates in response to both biotic and abiotic stresses is ascorbic acid (AA), which acts as an antioxidant, protecting the cell against oxidative stress caused by environmental factors and pathogens [19],[20]. Considering that compounds from Lamiaceae plants oils have antiviral and antioxidant activity [4], [24] and that H2O2, AA have been implicated in signaling gene expression against biotic and abiotic stresses [10], [19], the objectives of this work were to evaluate the effects of treatments with Rosmarinus officinalis oils, hydrogen peroxide and AA on photosynthetic pigments in healthy and mechanical inoculated potato plants with potato virus Y (PVY). and to study the antiviral activity of Thymus serpyllum, Lavandula officinalis oils treatments on Nicotiana tabacum plants inoculated with PVY.

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    MATERIAL AND METHODS

    Plant material. Solanum tuberosum L. microplants cv Roclas, testing virusfree, from the Biotechnology Department. Single node cuttings were propagated in test tubes on Murashige and Skoog [18] medium, at 20±1°C under a 16 h photoperiod (fluorescent lights, 400–700 nm), in sterile conditions. The microplants were transferred to greenhouse conditions 30 days after the single-node subculture step. For obtaining positive material, a part of these plants have been mechanicaly inoculated, using a PVY secondary infected plant from Record variety. The infection of the material was confirmed by ELISA tests. Nicotiana tabacum plants cv. White Burley have been inoculated mechanical in the 4 leaves phase and the harvesting leaves has been made after 4 weeks after inoculation.

    ELISA test. The analysis was performed following essentially the protocol described

    by Clark and Adams (1977) [6] (100 l). The samples having A405 values exceeding the cut-off (two times the average of healthy controls) were considered virus infected.

    Chemical treatments. Solanum tuberosum L. microplants were transplanted to pots and after 10, 20 and 30 days, all the plants (excepting the controls) were injected with

    Rosmarinus officinalis oil (dilution 1/1000) 100l each plant. Before the inoculation, in the 4 leaves phase, tobacco plants were injected with Thymus serpyllum, Lavandula officinalis

    oils (dilution 1/100 and 1/1000) 100l each /plant. From 7 days later from the first injection, the plants were sprayed twice weekly for the next time with 10 mL per plant of either 1 mM H2O2 or 3 mM AA at pH 5.6. Controls and plants treated only with natural oil were sprayed with distilled water. Four virus infected (positive) and healthy (negative) plants were sprayed in randomized arrays for each chemical treatment, and each treatment was performed in four independent experiments.

    Pigment analysis. Five leaf discs (about 1.5 cm diameter) per plant were taken from leaves of three plants per treatment. Samples for each assay were homogenized in 4 mL of 80%acetone at 4°C. Insoluble materials were removed by centrifugation at 2500 rpm for 10 min. Chlorophylls a and b, and carotenoids, were analyzed spectrophotometrically (method Lichtenthaler and Wellburn [14]).

    Statistical analysis. Data were analyzed by ANOVA and Duncan’s Multiple Range Test and scored as significant if P

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    tubers produced per plant (by 25, 29 and 25% respectively) in the positive plants compared to their control (fig. 3B). Interestingly, this reduced number of tubers was similar to that produced by uninfected plants subjected to any of the treatments (fig. 3B).

    Figure 1. Chlorophyll a (A) and chlorophyll b (B) of leaves of healthy plants (□) and potato

    virus Y infected plants (■), following the treatments. Data are means ± SD of four experiments (n=4). Bars with different letters differ significantly by ANOVA and Duncan’s test (P

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    control at 60 days, this effect remaining significant at 90 days for the plants treated only with Rosmarinus officinalis oil (fig. 4).

    Fig. 3. Number of tubers produced by healthy (□) or infected plants with potato virus Y (■),

    after the treatments. Data are means ± SD of four experiments (n=4). Bars with different letters differ significantly by ANOVA and Duncan’s test (P

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    II. Researches on Nicotiana tabacum Effects of treatments with Thymus serpyllum, Lavandula officinalis oils and H2O2 or

    AA, were compared on absorbances 405nm values obtained after testing (DAS ELISA technique) the plants (cv. White Burley) inoculated with potato virus Y(PVY). The antiviral activity of treatments was evaluated 40 days after the last transplanting. Compared with their positive controls, with chemical treatments, the inoculated plants showed significant decreases of the absorbances values (fig. 5). Treatments with Thymus serpillum oils and H2O2 or AA significantly decreased DO405nm of samples prelevated from virus PVY infected plant leaves to levels similar to uninfected. The best results were obtained using the oil’s dilution 1/100. No significant differences were induced by these treatments in the uninfected plants (fig. 5A). The Lavandula officinalis oils have lower effect on plants immunity comparated with Thymus serpillum, but the influence is present ( fig. 5B).

    A. B.

    Figure 5. A. Absorbances (optic density) values at 405nm of healthy and infected Nicotiana tabacum plants with potato virus Y(PVY) after the treatments. Bars with different letters differ significantly by ANOVA and Duncan’s test (P

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    The elucidation of the precise role played by Rosmarinus officinalis, Thymus serpyllum and Lavandula officinalis oil treatments in addition with H2O2, AA on potato virus Y infected and healthy plants awaits further investigation.

    REFERENCES

    [1]. Apostol, I., Heinstein, P.F., and P.S. Low. 1989 Rapid stimulation of an oxidative burst during elicitation of cultured plant cells. Plant Physiology 90, 109–116. [2]. Bădărău, C. L., Chiru, S.,C., Cojocaru, N., Ianosi, M., Chiru, N. 2010. Studies regarding the improvement of methods used for viruses identification in potato seed indexation. In: Potato agrophysiology. Proceedings of the International Symposium on Agronomy and Physiology of Potato, Nevsehir, Turkey: 332-340 [3]. Bădărău, C.L., Cojocaru, N., Rusu, S.N., Ianoşi, M., Petrusca, K. 2009.The effect of samples incubation on detection of PLRV and the influence of several extraction buffer’s additives on the detection of potato viruses Y, A, X and S by ELISA technique. In Proceeding of the 2nd International Symposium”New Researches in Biotechnology”, Bucharest, Biotechnology, Series F (Special volume), 9-17 [4]. Bedoux G., Mainguy, C., Bodoux, M., F, Marculescu, A., Ionescu, D. 2010. Biological activities of the essential oils from selected aromatic plants. Journal of EcoAgroTurism, Transilvania University of Brasov Publisher Vol 6, 1 (18), 83-91 [5]. Beemster, A. B. R., de Bokx, J. A. 1987. Survey of properties and symptoms. In Viruses of potato and seed potato production, eds. J.A.de Bokx and J.P.H. van der Want, Wageningen, The Netherlands RUDOC, 284-290; [6].Clark, M.F., Adam, A.,N. 1977.Characteristics of microplate method of enzyme linked immunosorbent assay for the detection of plant viruses. Journal General Virology 34, 475-483. [7]. Davis, J,A. and Radcliff ,E.B.,Schrage, W., Rgsdale, D.W. 2008. Vector and virus IPM for seed potato production.In Insect pest management: Concepts, tactics, strategies and case studies, eds. Radcliffe E.B., Huchison W.D.,Cancelado R.E.,Cambridge, UK,Cambridge University Press, 366-377 [8]. Dermastia, M.1995. Cytokinin pattern in healthy and PVYNTN infected potato(Solanum tuberosum L. cv.Igor).Proceedings of the 9th RAPR virology section meeting. Ribno, Bled, Slovenia, 147-150 [9]. Foyer, C.H., P. Descourvieres, and K.J. Kunert. 1994. Protection against oxygen radicals: an important defense mechanism studie in transgenic plants. Plant Cell and Environment 17, 507–523. [10]. Foyer, C.H., G. Noctor. (2005): Oxidant and antioxidant signalin in plants:a reevaluation of the concept of oxidative stress in a physiological context. Plant Cell and Environment 28, 1056– 1071. [11]. Fernie, A.R., and L. Willmitzer. 2001. Molecular and biochemical triggers of potato tuber development. Plant Physiology 127, 1459–1465. [12]. Hane, D., C., Hamm, P.,B. 1999. Effects of seedborne Potato virus Y infection in two potato cultivars exprssing mild disease symptoms. Plant Disease 83, 43-45 [13]. Hammerschmidt, R. (2005): Antioxidants and the regulation of defense. Physiological and Molecular Plant Pathology 66, 211–212. [14]. Lichtenthaler, H.K.,A.R Wellburn. 1983. Determinations of total carotenoids and chlorophylls a and b of leaf extracts in different solvents. Biochemical Society Transactions 11, 591–592. [15]. López-Delgado, H., J.F. Dat, C.H. Foyer, and I.M. Scott. 1998. Induction of thermotolerance in potato microplants by acetylsalicylic acid and H2O2.Journal of Experimental Botany 49:713–720.

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    [16]. Lorenzen, J.,H., Meacham ,T., Berger, P., Pat, J.,S., Crosslin, J., M., Hamm, P., Kopp, H. 2006. Whole genome characterisation of potato virus Y isolates collected in the western USE and their comparison to isolates from Europe and Canada. Archives of Virology 151, 1055-1074 [17]. Mora-Herrera, M.E., H. López-Delgado, A. Castillo-Morales, and C. H. Foyer. 2005. Salicylic acid and H2O2 function by independent pathways in the induction of freezing tolerance in potato. Physiologia Plantarum 125, 430–440. [18]. Murashige, T., and F. Skoog. 1962. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum 15, 473–497. [19]. Noctor, G. 2006. Metabolic signaling in defense and stress: the central roles of soluble redox couples. Plant, Cell and Environment 29, 409–425. [20]. Pastori, G.M., G. Kiddle, J. Antoniw, S. Bernard, S. Veljovic- Jovanovic, P.J. Verrier, G. Noctor, and C.H. Foyer. 2003. Leaf vitamin C contents modulate plant defense transcripts and regulate genes that control development through hormone signaling. The Plant Cell 15, 939–951. [21]. Petersen M, Simmonds M.S.J 2001. Rosmarinic acid. Phytochemistry 61: 121-125 [22]. Quan, L.J., B. Zhang, W.W. Shi, and H.Y. Li. 2008. Hydrogen peroxide in plants: a versatile molecule of the reactive oxygen species network. Journal of Integrative Plant Biology 50, 2–18. [23]. Ragsdale, D.,W., Radcliffe, E.,B., Difonzo, C.,D. 2001. Epidemiology anf field control of PVY and PLRV. In Virus and virus –like diseases of potatoes and production of seed potatoes, eds. G. Loebenstein, P.H. Berger, A.A.Brunt, Lawson R.H., Dordrecht Kluwer, 237-270 [24]. Triantaphyllou, K., Blekas, G., Boskou, D. 2001. Antioxidative properties of water extracts obtained from herbs of the species Lamiaceae. International Journal Food and Science Nutrition 52, 313-317.

    ABOUT THE AUTHORS

    Carmen Liliana Badarau, National Institute of Research and Development for Potato and Sugar Beet Brasov, 2 Fundaturii Street, 500470 Brasov, Romania, E-mail: [email protected]

    Angela MĂRCULESCU, Faculty of Food and Tourism, Transilvania University, 2 Castelului Street, Braşov, Romania, E-mail: [email protected]

    Nicoleta Chiru, National Institute of Research and Development for Potato and Sugar Beet Brasov, 2 Fundaturii Street, 500470 Brasov, Romania, E-mail: [email protected]

    Florentina Damşa, National Institute of Research and Development for Potato and Sugar Beet Brasov, 2 Fundaturii Street, 500470 Brasov, Romania, E-mail: [email protected]

    Andreea Nistor, National Institute of Research and Development for Potato and Sugar Beet Brasov, 2 Fundaturii Street, 500470 Brasov, Romania, E-mail: [email protected]

    mailto:[email protected]:[email protected]:[email protected]:[email protected]:[email protected]

  • RESEARCH PEOPLE AND ACTUAL TASKS ON MULTIDISCIPLINARY SCIENCES 8 – 10 JUNE 2011, LOZENEC, BULGARIA

    12

    THE METHODS OF FERTILITY REPRODUCTION OF SOILS AND THE

    GROWTH OF HIGH – QUALITY FODDER IN AZERBAIJAN

    M. Babayev, F. Ramazanova, S. Huseynova

    Abstract: For the elimination of the fodder deficiency and the increase in the biological activity and the rehabilitation of the fertility of the irrigated degraded gray-brown soil (İrraqri Gypcisols) in the territory of Apsheron zone (Azerbaijan), we determined the optimal dose of the application of nitrogen fertilizers on the background of manure - 20tonnes/ha + P90K60kg/ha of a reactant for summer sowing of winter rape - N140 (N60 (autumn harvest), N80 (spring harvest)kg/ha); for the autumn sowing of winter rape and mixed grass crop (rye+vetch+rape) - N30 (autumn harvest) + N90 (spring harvest) kg/ha.

    Keywords: Soil, protein, metabolizable energy, irrigation, degradation, microbiological activity, microflora

    INTRODUCTION The basis of modern agriculture of Azerbaijan, especially of Apsheron, is an effective

    utilization of an irrigated ploughed field. It is essentially important because of the origin and the development of stock-farms. So, it is important to create a firm fodder base by growing the interim crops balanced by the main nutrients and, at the same time, improving physical, chemical, and biological properties of the soil thereby enriching it with organic matters such as stubble and fodder remains. As it is known, the lack of essential nutrients in fodders causes the decrease of productivity and the appearance of various animal diseases. [2; 3] This is a topical problem for Apsheron where in the structure of sown areas, the specific weight of low-productive cereals (55-140 centner/hectare) is too high and does not answer zoo-technical standards of nutrition (one fodder unit of digestible protein is 75-80 grams instead of the necessary 105-110 grams.) [1;2].

    The purpose and methods of the research That is why for the elimination of the protein deficiency and the metabolizable energy

    in Apsheron, it is necessary to increase the biological activity and the fertility of the degraded irrigated soils by cultivating high-protein crops (rape Brassica napus) and mixed grass crops (rye+vetch+rape) with the use of fertilizers and organic manures. According to these data, specific researches were done.

    Rape and mixed grass crop (rye+vetch+rape) sowings were held in two periods: 1) for the single fodder use in autumn – first quarter of October; 2)for the double fodder use (summer/ autumn) – summer – the first quarter of August with the width of 15 cm between rows, and the standard quantity of seeds per hectare - 2,5 million.

    The irrigation rate for the period of sowing-harvesting is: for autumn – 1200/1300 m3

    per hectare; for summer – 1400-1600 m3 per hectare. The experiments were performed for the autumn sowing according to the Apsheron agricultural engineering (20 m/ ha manure+ P90K60 k/ha) by 7 variants and for the summer sowing – by 5 variants.

    1)Autumn sowing: (Rape)–1.without fertilizers (control); 2.background (manure 20t/ha+P90K60 kg/ha before sowing); 3.background+N30 (autumn)+N30 kg/ha of reactant (spring); 4.background+N30 (autumn)+N60 kg/ha of reactant (spring); 5.background+N30 (autumn)+N90 kg/ha of reactant(spring); 6.background+N30 (autumn)+N120 kg/ha of reactant (spring); Rye+Vetch+Rape) 7. background+N30 (autumn)+N60 kg/ha of a reactant (spring)

    2) Summer sowing: 1. without fertilizers (control); 2.background (manure 20t/ha of a reactant+P90K60 kg/ha before sowing); 3. background+N100 kg/ha of a reactant (N50 (autumn), N50-(spring)); 4. background+N140 kg/ha of a reactant (N60 (autumn), N80-spring)); 5. background+N180 kg/ha of a reactant (N80(autumn) , N100(spring)); (in autumn – before

  • RESEARCH PEOPLE AND ACTUAL TASKS ON MULTIDISCIPLINARY SCIENCES 8 – 10 JUNE 2011, LOZENEC, BULGARIA

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    sowing, in spring – for extra nutrition). In spring, in all the variants, nitrogen fertilizers were introduced fractionally-50% during booting, 30%- during budding, 20%- during blossoming.

    The object of the research The researches were conducted in the territory of Apsheron zone (Azerbaijan). The

    soil of the experimental area is gray and brown with a low consistence of humus and nutrients; by mechanical composition – semi-loamy soil. The carbonality of the upper horizon is 3-5%. The humus content in an arable layer is 1.56-1.75%, nitrogen- 0.094% - 0.104%, assailable phosphorus- 12.0 -15.0%, general potassium - 280-297 ml/kg. The climate is dry, subtropical. Average annual temperature is + 140С to + 150С. The sum of active temperatures is about +45600С. Average annual precipitation is 140-250 mm.

    The discussion of the research results The results of our researches (1995-2000, 2003-2007) showed that winter rape grew

    slowly in all the variants during the initial summer sowing (due to high t0) but it increased its growth during the second decade of September. We could observe the high efficiency of nitrogen fertilizers in the variant ‘background + N140 kg/ha (N60 (autumn), N80 (spring) kg/ha of a reactant’. In 65-70 days of vegetation and a three-time watering, rape formed 8 pairs of leaves (72.5-77.0 cm high) and supplied with 347 centner/hectare of green mass in autumn (1harvesting). The harvest of dry matter was 35 centners/hectare, which is 38-40% higher than in control, 28-30% higher than in background, and 15-18% higher than in variants 3 and 5. In 6-7 days after harvesting in all the variants, rape started growing and, in early winter, had the height of 13-18cm. In spring, rape blossoming in all the variants was 5-7 days later than during autumn. The 2nd (spring) harvesting of green mass made 289 centners/ha. At the same time, we examined feeding values of winter rape sown in summer (both after 1st and 2nd harvestings).

    The results of the analyses showed the increase in the content of dry mass (from 9.9-10.0 to 12.1%), raw cellulose (from 10.9-11.1 to 19.3-20.0%) in the absolutely dry matter after the 2nd harvesting in all the variants. However, the decrease of raw protein (from 23.0-25.8 to 19.9-20.1%) and fat (from 5.5-6.1 to 5.0-5.3%) was observed. Overall after two harvestings (autumn and spring) in the variant ‘background + N140 kg/ha of a reactant (N60 (autumn) , N80 – (spring)’, the harvest from 1 ha gave: 64 centners of dry mass, 66 centners of fodder unit, 207 ths/mega-j. of metabolizable energy, 12 centners of digestible protein.

    The content of NO3 was 300 mg/kg and the provision of 1 fodder unit with digestible protein -185g. In the control and the other variants these data are 25-46% lower. During the autumn sowing of rape (for one harvest) and mixed grass crop (rye+vetch+rape) for the green mass (in spring), we observed the high effect of nitrogen fertility on the plant productivity in the variant ‘background + N30 (autumn) + N90 (spring) kg/ha’. In this variant, the harvest of the dry mass of rape and mixed grass crop (rye+vetch+rape) from 1 ha was 46-50 centners, fodder units – 46-50centner, metabolizable energy- 178-190 thousand / mega-j., digestible protein – 7.3 and 8.9 centners ( table 1).

    The absolutely dry matter of rape contains: raw protein- 23.3%, fat – 5.56%, carotene- 480mg/kg, cellulose – 18.02%, NO3 – 300-305mg/kg, and the provision of 1 fodder unit with digestible protein -185-197g (table2).

    The nutritional value of 1 kg of dry matter in fodder units was 163g., which is 22-39% higher than in the control and variants 3, 4, and 6.

    In the practice of cattle feeding, the usual ration of cows contains 3.0-3.5% of fat in dry mass. The higher the yield of milk the more energy should be in 1 kg of dry matter. It is unreasonable to lower the metabolizable energy till 8 mega-j in 1 kg of dry matter. In all the variants during summer and autumn sowings, the dry matter of winter rape and mixed grass crop (rye+vetch+rape) is high-energy fodder due to the content of fat (5.3-6.10%), metabolizable energy (9.0-10.9 and 9.3-11.2 mega-j, table 2). This way, the introduction of the fodder from rape and mixed grass crop rye+vetch+rape into the ration of milk cows

  • RESEARCH PEOPLE AND ACTUAL TASKS ON MULTIDISCIPLINARY SCIENCES 8 – 10 JUNE 2011, LOZENEC, BULGARIA

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    favors an average daily increase in animal live weight (300-400g per head) and milk fat content by 0.5-1.0%. Left after harvesting the green mass of winter and summer rape and winter rape and mixed grass crop (rye+vetch+rape), vegetative residuals, rich in nutrients (1.97-2.01%N, 0.60-0.67% P2O5, 2.00-2.07% K2O), enriched the soil with organic matters, favored its biological development and fertility stabilization.

    According to the indices of the chemical composition of vegetative residuals of winter rape during summer sowing in the variant ‘background + N140 (N60 for the 1

    st harvesting, N80 – for the 2

    nd harvesting)kg/ha’, 77.2kg N, 29.0kg P2O5, 78.7kg CaO, and 68.9 kg K2O got into the soil per 1ha together with vegetative residuals during the ploughing. During autumn sowing of rape and mixed grass crop rye+vetch+rape (background + N30 (autumn) + N90 (spring) kg/ha), the soil received 67 and 70 kg N, 19.0 and 26 kg P2O5, 66 and 69 kg CaO, and 49.9 and 55.0 kg K2O. In all the other variants of winter rape sowing, these data were lower.

    Table 1 Nutrients from 1 hectare of harvest

    № Variants

    Metabolizable energy, thousands mega-joules

    Fodder unit, centner/ hectare

    Digestible protein, centner/ hectare

    Summer sowing of winter rape (sum of two harvests)

    1 Without fertilizers (control) 147 28 3.2

    2 Background (manure 20tonnes/ha+P90K60 kg/ha)

    164 40 6.1

    3 Background+N100 (N50 for the 1st harvesting,

    N50 -for the 2nd harvesting) kg/ha

    197 54 8.9

    4 background+N140 (N60 for the 1st harvesting,

    N80-for the 2nd harvesting) kg/ha

    207 66 12.0

    5 background+N180 (N80 for the 1st harvesting,

    N100 -for the 2nd harvesting) kg/ha

    187 51 7.5

    Autumn sowing of winter rape and mixed grass crop

    1 Without fertilizers (control) 91 14 1.6

    2 Background (manure 20tonnes/hectare + P90K60kg/ha before sowing)

    126 24 3.0

    3 Background+N30 (autumn)+N30 (spring) kg/ha 147 30 4.1

    4 Background+N30 (autumn)+N60 (spring) kg/ha 161 39 5.7

    5 Background+N30 (autumn)+N90 (spring) kg/ha 178 46 7.3

    6 Background+N30(autumn)+N120 (spring) kg/ha Rye+ vetch+rape

    170 40 6.4

    7 Background+N30 (autumn)+N60 (spring) kg/ha 190 50 8.9

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    Table 2 The content and quality of nutrients in fodder

    Indices In dry matter

    Winter rape

    Mixed grass crop (Rye+vetch+rye)

    Metabolizable energy, mega- joul 9.00 – 10.95 9.3 - 11.07

    Raw protein, % 17.00 – 23.30 18.37 - 25.00

    Raw cellulose, % 18.02 – 19.94 17.5 - 18.70

    Raw fat, % 5.30 - 6.10 5.00 - 6.03

    Carotene, mg/kg 450 - 480 460 - 493

    However, the influence of vegetative residuals of each crop or type of sowing differs.

    It was established that the sowing of winter rape and mixed grass crop (rye+vetch+rape) (background+N30 (autumn)+N90 (spring) kg/ha) was most effective: the content of humus increased to 1.89-2.00%, the content of fraction decreased (0.05-0.005mm), the content of oozy fraction and physical clay increased, the sum of salts decreased from 0.22 to 0.10%, the content of the absorbed Ca increased from 80 to 83%, the content of Mg and Na decreased to 5.5-7.7% and 0.9-1.3%, the indices of pH environment and total alkalinity decreased to the side of soil neutralization (pH 7.4-8.5), the content of calcium carbonate increased (6.9-8.1%). Vegetative residuals of rape and rye+vetch+rape (summer and winter sowing) favored the increase in soil biological activity.

    The determination of soil microbiological activity under rape (summer and autumn sowings in all the variants) and mixed grass crop (rye+vetch+rape) by the method of the decomposition of linen cloth showed that this process is higher during summer (background + N140 (N60 for 1

    st harvesting, N80 – for 2nd harvesting) kg/ha) and autumn

    sowings of rape and mixed grass crop (background + N30 (autumn) + N90 (spring) kg/ha) than in control and the other variants. So, the decomposition of linen cloth during summer sowing of rape (background + N140 (N60 for the 1

    st harvesting, N80 – for 2nd the

    harvesting)kg/ha) in the soil layer of 0-11cm was 50.1%, in the soil layer of 11-24-57% and during autumn sowing of rape and mixed grass crop (background + N30 (autumn) + N90 (spring) kg/ha)- 46-55 and 59%, respectively. It can be explained by the fact that the mineralization of rape residuals in both of the variants and mixed grass crop passes more intensively than in the other variants. Vegetative residuals of rape and mixed grass crop (rye+vetch+rape) (summer and winter sowings) had a positive effect on the intensity of the quality and quantity of microorganisms (table 3).

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    Table 3 The ratio of the main physiological groups of microorganisms in the irrigated gray-

    brown soil (layer-0-25cm, ths/g of soil)

    № Variants

    Tota

    l nu

    mb

    er

    of

    mic

    roo

    rga

    nis

    ms

    In % out of the total number

    of microorganisms

    Spore forming

    bacteria in % out of the total

    number of bacteria

    ba

    cte

    ria

    actin

    om

    y

    ce

    tes m

    icro

    sco

    pic

    fu

    ng

    i

    Summer sowing of winter rape (sum of two harvests)

    1 Without fertilizers (control) 1590 57 43 0.08 14.92

    2 background (manure 20 tonnes/ha + P90K60 kg/ha)

    3750

    69 31 0.11 8.70

    3 background+N100(N50 for the 1st harvesting,

    N50 – for the 2nd harvesting) kg/ha

    4005

    74 26 0.16 8.97

    4 background+N140 (N60 for the 1st harvesting,

    N80-for the 2nd harvesting) kg/ha

    4500 76 24 0.18 9.30

    5 background + N180 (N80 for the 1st harvesting,

    N100 – for the 2nd harvesting) kg/ha

    3900 72 28 0.14 8.70

    Autumn sowing of winter rape and mixed grass crop

    1 Without fertilizers (control) 1489 58 42 0.09 15.00

    2 background (manure 20 tonnes/hectare +P90K60 kg/ha before sowing)

    2005 60 40 0.12 13.98

    3 background+N30(autumn)+N30 (spring) kg/ha 2849 68 32 0.13 13.95

    4 background+N30 (autumn)+N60 (spring) kg/ha 3570 71 29 0.16 9.43

    5 background+N30 (autumn)+N90 (spring) kg/ha 4600 79 21 0.17 9.90

    6 background+N30 (autumn)+N120 (spring) kg/ha 4517 75 25 0.18 9.70

    7 Rye+vetch+rape background+N30 (autumn)+ N60 (spring) kg/ha

    4790 81 19 0.19 10.06

    Their stimulating effect is especially seen in the soil under: rape (summer sowing) in

    the variant (background + N140 (N60 1st harvesting, N80 – 2nd harvesting) kg/ha) – 4500

    ths/g; rape (autumn sowing) in the variant (background + N30 (autumn) + N90 (spring) kg/ha) - 4600ths/g; mixed grass crop - 4790ths/g; in the other variants - lower. In the composition of microflora, the number of nonspore-forming bacteria increased, but that of spore-forming bacteria decreased.

    In the soil without ferilizers, the content of actinomycetes was higher (up to 50% out of the total number of microorganisms), but the number of spore ammonifiers was lower (up to 10-19% out of the total number of bacteria).

    The qualitative composition of microorganisms in the soil under rape (winter sowing) and mixed grass crop was richer than in the soil under rape (summer sowing) and in the soil without fertilizers. Here among the ammonificating bacteria, the representatives of Bacillus prevailed. Bac. mesenteicus multiplied intensively (39-50%). The number of bacilli was at its highest point in rhizosphere by the flowering period.

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    The number of microscopic fungi in the control soil was low and they were only from Aspergillus genus. In the soil under rape (autumn sowing) in the variant (background + N30 (autumn) + N90 (spring) kg/ha) and mixed grass crop, we observed an increasing number of microscopic fungi and enrichment of their qualitative composition (Aspergillus, Penicillium, Trichoderma, and Alternaria). The appearance of the representatives of Penicillium, Trichoderma, and Alternaria along with Aspergillus indicated the formation of favorable environmental conditions in the soil under these variants. The vegetative residuals of these plants had a stimulating effect on the intensive development of the main physiological groups of microorganisms.

    In the soils under all the variants, we could observe seasonal fluctuation of the number of the particular physiological groups of microorganisms. It was also found experimentally that correlation coefficient among the number of microflora and the total reserve of vegetative residuals of rape (winter sowing) in the variant (background + N30 (autumn) + N90 (spring) kg/ha) and mixed grass crop rye+vetch+rape was 0.123-134, together with living roots – 0.373-0.380, together with dead roots- 0.675-0.726. However, in the soil under rape of summer sowing, these indices were higher.

    CONCLUSION The summer sowing of rape in the variant (background + N140 (N60 1st harvesting,

    N80 – 2nd harvesting) kg/ha), the autumn sowing of winter rape and mixed grass crop

    (rye+vetch+rape) in the variant (background + N30 (autumn) + N90 (spring) kg/ha) provide the soil with high-energy, protein fodders favoring an average daily increase in animal live weight (300-400g per head) and milk fat content by 0.5-1.0%. At the same time the improvement of physical and chemical properties of gray-brown soil (İrraqri Gypcisols) favors its desaltinization and the increase in the biological activity and fertility (1.5-2 times).

    REFERENCES [1] Babayev M. 2000. Degradation of soil in Azerbaijan influence of increasing

    Antropogen affect. // EUR 19723 EN. European communities. Italy. p.4 [2] Komatsuzaki M., Ohta H. 2007. Soil management practices for sustainable agro-

    ecosystems //Sustain Sci. V.2. pp.103-120. [3] Ramazanova F., Babayev M., Huseynova S.M. 2008. Biological methods of

    fertility reproduction of antropogene degraded irrigated soils in Azerbaijan dry subtropics Congress“Eurosoil 2008”- Soil-Society-Environment, Vienna, Austria, p.106.

    ABOUT THE AUTHORS Babayev M., Institute of Soil Science and Agrochemistry of ANAS, M.Arif 5 Street

    1073 Baku, Azerbaijan E-mail: [email protected] Ramazanova F., Institute of Soil Science and Agrochemistry of ANAS, M.Arif 5 Street

    1073 Baku, Azerbaijan , E-mail: [email protected] Huseynova S., Institute of Soil Science and Agrochemistry of ANAS, 5 M.Arif Street

    1073 Baku, Azerbaijan , E-mail: [email protected]

    mailto:[email protected]:[email protected]:[email protected]

  • RESEARCH PEOPLE AND ACTUAL TASKS ON MULTIDISCIPLINARY SCIENCES 8 – 10 JUNE 2011, LOZENEC, BULGARIA

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    THE EFFECT OF ZN DEFICIENCY ON THE CONTENTS OF

    PHOTOSYNTHETIC PIGMENTS

    N. Bákonyi, É. Gajdos, B. Tóth, P. Makleit, L. Lévai, Sz. Veres

    Abstract: Nearly half of the world’s cereal crops are deficient in Zn, leading to poor crop yields. In this study, the effect of total Zn deficiency and a synthetic auxin (NES) on the contents of photosynthetic pigments of cucumber seedlings were investigated. The relative chlorophyll contents (SPAD unit), as well as the absolute contents of photosynthetic pigments (chlorophyll-a,-b, total carotenoids) were measured.

    According to our results the total Zn deficiency dramatically – by more than 50% – reduced the amount of chlorophyll-a in the second leaves. The relative chlorophyll contents were lower by 38% in the Zn-deficient plants than in the control ones on the 16

    th day of the treatment. The treatment with NES compensated for the

    Zn deficiency. The SPAD index was higher in the NES-treated plants than in the Zn-deficient plants by 35% on the 16

    th day of treatment.

    Key words: Zn deficiency, SPAD, photosynthetic pigments.

    INTRODUCTION Almost half of the world’s cereal crops are deficient in zinc (Zn), leading to poor crop

    yields. In fact, one-third (33%) of the world's population is at risk of Zn deficiency in rates, ranging from 4% to 73% depending on the given country. Zn deficiency in agricultural soils is also a major global problem affecting both crop yield and quality. High pH and calcium carbonate contents are the main reasons for the low availability of Zn for plants (Karimian and Moafpouryan, 1999). It has been reported that the high-concentration application of phosphate fertilisers reduces Zn availability (Khosgoftarmanesh et al., 2006). The Zn takes part in protein metabolism and stimulates the auxin production in due to regulate the growth (Pethő, 1993; Kalocsai, 2006). There are a number of physiological impairments in Zn-deficient plant cells with causing retardation of the growth, differentiation and development of plants (Cakmak, 2000), because the Zn is indispensable micronutrient for the plant via important enzyme-constituent and influence enzyme-activator. Zinc deficiency affected photosynthesis (Randall and Bouma, 1973) and zinc deficiency caused extensive declines in chlorophyll (chl) contents of leaf and ratios of chl a/b (Chen et al., 2008).

    MATERIAL AND METHODS

    The experimental plant was cucumber (Cucumis stativum L. cv. Delicatess). The seeds were germinated on moistened filter paper at 25oC. The seedlings were transferred to a continuously aerated nutrient solution of the following composition: 2.0 mM Ca(NO3)2, 0.7 mM K2SO4, 0,5 mM MgSO4, 0.1 mM KH2PO4, 0.1 mM KCl, 1µM H3BO3, 1µM MnSO4, 0.25 µM CuSO4, 0.01 µM (NH4)6Mo7O24. The nutrient solution contains 10µM H3BO3. The iron as Fe-EDTA was added to the nutrient solution in a concentration of 10-4M. The treatments were: control, -Zn (totally Zn deficiency), -Zn+NES. The auxin was used in synthetic form (NES) with 1 drop NES (0,03369 g=5,37x10-1M) to the top of the stem on every 3th day.

    In this study, the effect of total Zn deficiency was investigated on the relative and absolute contents of photosynthetic pigments.

    The seedlings were grown under controlled environmental conditions (light/dark regime 16/8 h at 24/20 oC, 65–70% relative humidity and a photosynthetic photon flux density 300 µmol m-2s-1. The relative chlorophyll contents were measured with SPAD 502 (Minolta) on the 12nd, 16th and 22nd days on the 2nd and 3rd leaves, respectively. Moran and Porath’s method (1980) and METEREK SP-830 spectrophotometer was used to determine

  • RESEARCH PEOPLE AND ACTUAL TASKS ON MULTIDISCIPLINARY SCIENCES 8 – 10 JUNE 2011, LOZENEC, BULGARIA

    19

    the absolute contents of photosynthetic pigments (chlorophyll-a,-b, total carotenoids)and their ratio in the firs, second and third leaves. The experimental results were evaluated with Microsoft Office Excel and SigmaPlot 8.0.

    RESULTS AND DISCUSSION

    The Zn deficiency can retard the growth of the plants via decreases of auxin synthesis and the dry matter accumulation. The dry matter accumulation depends on the intensity of photosynthesis and the contents of the photosynthetic pigments, therefore the relative chlorophyll contents (SPAD unit) of 2nd and 3rd leaves were measured with the development of leaves. The results can be seen in the Table 1.

    Table 1. The effects of total Zn deficiency and NES treatments on the relative chlorophyll contents of cucumber’s leaves (mg.g-1), (SPAD unit), (n=250±s.e.).

    Significant difference comparison to the control: *p

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    Treatments

    chlo

    rophyll-

    a m

    g. g

    -1

    0

    2

    4

    6

    8

    10

    12

    14

    Control

    -Zn

    1st leaf

    1st leaf

    2nd

    leaf

    2nd

    leaf

    ****

    Treatments

    chlo

    rophyll-

    b m

    g. g

    -1

    0

    1

    2

    3

    4

    5

    Control

    - Zn

    1st leaf

    *

    *1

    st leaf

    2nd

    leaf

    2nd

    leaf

    Table 2. and Figure 1, 2. well demonstrates the chlorophylls-decreasing effect of total Zn deficiency.

    Figure 1. and 2. The effect of total Zn deficiency on the contents of chlorophyll-a and chlorophyll-b of 1st and 2nd leaves of 22-day-old cucumber seedlings (mg.g-1), (n=4±s.e.).

    Significant difference comparison to the control: *p

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    CONCLUSIONS AND FUTURE WORK

    According to our experimental results the total Zn deficiency significantly reduced the relative (SPAD index) and absolute contents of photosynthetic pigments via retardation of synthetic processes. Zn is known as a functional component of a number of enzymes, which play role in protein synthesis. The photosynthetic pigments contain a cromofor group and a protein part, therefore in lack of Zn the protein synthesis is retarded and the pigments suffer damage.

    The relative chlorophyll contents were lower by 38% and 32% in the Zn-deficient plants than in the control ones on the 16th and 22nd day of the treatment, respectively. The treatment with NES compensated for the Zn deficiency. The SPAD index was higher in the NES-treated plants than in the Zn-deficient plants by 35% and 30% on the 16th and 22nd days of treatment, respectively.

    The total Zn deficiency dramatically – by more than 50% – reduced the amount of chlorophyll-a and chlorophyll-b in the second leaves. The older leaves were more tolerant. The absolute contents of chlorophyll-a of NES-treated Zn-deficient plants were higher by 5.8% in the 3rd leaves comparison to the control.

    There is a purpose for further examination on interaction between the Zn deficiency and photosynthetic pigments, as well as its possible effects on photosynthesis.

    REFERENCES

    [1]. Cakmak, I., 2000. Possible roles of zinc in protetcting plant cells from demage by reactive oxigen species. New Physiology 146, pp. 185-205. [2]. Chen W., X. Yang, Z. He, Y. Feng and F. Hu. 2008. Differential changes in photosynthetic capacity, 77 K chlorophyll fluorescence and chloroplast ultrastructure between Zn-efficient and Zn-inefficient rice genotypes (Oryza sativa) under low zinc stress. Physiologia Plantarum Volume 132, Issue 1, January 2008. pp. 89–101. [3]. Kalocsai, R., 2006. A cink (Zn). MezőHír X. évf. 2006. szeptember pp. 38. [4]. Karimian, N., and G. R. Moafpouryan., 1999. Zinc adsorption characteristics of selected calcareous soils of Iran and their relationship with soil properties. Communications in Soil Science and Plant Analysis 30, pp. 1721–1731. [5]. Khoshgoftarmanesh, A. H., H. Shariatmadari, N. Karimian, and S. E. A. T. M. VanDer Zee., 2006. Cadmium and zinc in saline soil solutions and their concentrations in wheat. Soil Science Society of America Journal 70, pp. 582–588. [6]. Pethő M., 1993. Mezőgazdasági növények élettana. Akadémia Kiadó, Budapest 1993. pp. 221, 224. [7]. Randall P. J. and D. Bouma. 1973. Zinc Deficiency, Carbonic Anhydrase, and Photosynthesis in Leaves of Spinach. Plant Physiology 52, 229-232.

    ABOUT THE AUTHORS

    N. Bákonyi, É. Gajdos, B. Tóth, P. Makleit Ph.D., L. Lévai Ph.D., Sz. Veres Ph.D., Division of Agricultural Botany and Crop Physiology, Institute of Crop Science, University of Debrecen, H-4032 Debrecen, Böszörményi u. 138. Hungary, Phone: +36 52 512-900/68063. E-mail: [email protected]

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    THE ROLE OF PROTOZOAN INFECTION IN THE OCCURRENCE OF

    NEONATAL ENTEROPATHIES IN PIGLETS

    I.Pavlović, M.Žutić,O.Radanović, and B.Savić

    Abstract: Protosoan infection caused by by Isospora suis and Cryptosporidium spp. play important role at occurence of neonatal entheropathy of piglets. In our paper we give an autline of prevalence of I.suis and Cryptosporidium spp.

    Key words: pigs, Isospora suis, Cryptosporidium spp., enteropathy

    INTRODUCTION

    The disease caused by Isospora suis and Cryptosporidium spp. has been described as an important cause of suckling piglet diarrhea and reported from all types of farrowing facilities and under all types of management systems (1,4,7,8,9).

    Depending on the rate of infection, the disease complexes can range from sub clinical infections to infections that include severe intestinal problems with severe lesions in the gastrointestinal tract that can result in high mortality and/or a negative impact on the factors of production (4,6,10).

    At same time, those parasites play important role at complex of neonatal entheropathy of piglets, spread together with other infectious agents (1,5,6).

    MATERIAL AND METHODS

    During 2007-2009 a total of 428 pigs, 2 days -21 weeks old, were exanimate in the laboratory of Scientific Veterinary Institute of Serbia. All of the examined pigs were sings of diarrhea. After routine necropsy, to parasitological examination we used faecal swab, gut contents and a mucous scrape of gut which we examined with Sheather’s sugar flotation and in direct smears stained with Gimsa strain and Ziehl-Neelsen stain and examined by use of light microscopy. In addition section of jejunum, ileum and spiral colon from each pig were fixed in natural buffered 10% formal in, embedded in paraffin, sectioned at 6 micrometer, stained with hematoxylin and eosin and examined by use of the light microscopy.

    At same time we examined 979 samples of faeces originated from various product categories (fating pigs, sows, board) a coprological examination was done within 24 hours after sampling using flotation method in Sheather’s sugar solution.

    RESULTS AND DISCUSSION

    The occurrence of Isospora suis was found to be highest in piglets at the age of 7–15days. Isosporosis prevalence in piglets was highest on the second week of piglet age (32.8%) with the highest prevalence value on day 13 of piglet age (44.3%) In infected piglets the presence of I. suis was detected most frequently in connection with watery diarrheas (37.4%) and least frequently in piglets with shaped faeces (15.0%). In piglet with watery diarrheas only very weak isosporous infections were usually found, together with other enteropathogenic agents of bacterial or viral etiology contributed to the origin of diarrheas (like E.coli, C.perfrigens tip C rotaviruses, and etc.) in parallel infections in piglets to a large extent.

    We found out total isosporosis prevalence 27.4% mostly in a litter housing system; these results document that in our conditions there are not any significant differences in the prevalence and time of occurrence of Isospora suis between different housing systems and time periods compared to the results obtained in a litterless system of piglet housing.

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    Reports on the presence, prevalence and epidemiology of Isospora suis have come from virtually every country in the world (3,10). Studies conducted in most countries have shown a high prevalence of the disease on farms (18-85%). Results of our examination are at similar range.

    During our examination cryptosporidial organisms were detected in the microvillus brush border in the intestine of the 537 (12,34%) pigs examined. Small numbers of organisms were found in 63,76%, moderate number in 20,28% and large number in 15,94%. Infected pigs ranged from 7-12 weeks old and was highest in piglets 11-12 week old (63%).

    The fact that 60% of those infected were 6-12 weeks old would suggest that there is a strong possibility of sub clinical infection in weaned pigs. Infection apparently diminishes in pigs after 12 weeks old. Cryptosporidia were seen much less frequently in pigs outside of this age range and were not found in pigs less then 1 week old. The our results suggested that infection apparently diminishes in pigs older then 12 weeks and that the much less frequently seen in pigs outside this age range (7-11 weeks) and not found in pigs less then 7 weeks old. much less frequently seen in pigs outside this age range (7-11 weeks) and not found in pigs less then 7 weeks old. Cryptosporidia were rarely seen in mature sow and boards only in low range of infection (2)

    During our examination, cryptosporidia was not determinate to be the only cause of diarrhea in examined pigs. With few exceptions, cryptosporidia were associated with sub clinical infection. Even in the 69 cryptosporidia infected pigs with diarrhea, other enteric pathogens or lesions, independently capable of causing diarrhea, were in all 32% (in 45 cases we occured Brachyspira hyodisenteria and in 39 Campylobacter spp.).

    CONCLUSIONS AND FUTURE WORK

    Protozoan infection had an important role in occurrence of neonatal enteropathy of piglets either in itself or in concert with other agents.

    REFERENCES

    [1]. Corwin R.M., Stewart T.B. (1992): Internal Parasites, In: E.J.Lemman: Disease of Swine,. Wolf Publishing Ltd. New York. [2]. Mišić, Z., Katić-Radivojević, S., Kulišić, Z. (2003) Cryptosporidium infection in nursing, weaning and post-weaned piglets and sows in the belgrade district; Acta Veterinaria, 53, 361-366. [3]. Mundt H.C., Joachim A., Daugschies A, Zimmermann M (2003) Population biology studies on Isospora suis in piglets. Parasitol Res 90 Suppl 3:158–159 [4]. Pavlović, I., Ivetić, V., Erski-Biljić, M., Milutinović, M., Kulišić, Z. (1996) Cryptosporidial infection of pigs at the intensive breeding. Journal of Protozoology Research 6, 21-24. [5]. Pavlović, I., Ivetić, V., Valter, D., Romanić, S., Hudina, V. (1997) Prevalence of cryptosporidial infection in pigs and its role in entheropathy apperiance in piglets. Abstracts of VII Congress National de Medicine Veterinara Romania, Voinesa, Romania, 104 [6]. Pavlović, I., Žutić M.,Savić B.,Radanović O. (2004):Isospora suis - aktuelan parazitološki problem farmski gajenih svinja Veterinarski glasnik 58 (3-4), 561-565 [7]. Pavlović I., Hudin V., Kulišić Z.,Ivetić V.,Žutić M. (2006): Kokcidioza svinja u farmskom uzgoju Zbornik naučnih radova Instituta PKB Agroekonomik 12 (3-4),127-133

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    [8]. Pavlović I.,Žutić M.,Ivetić V.,Savić B.,Radanović O.,Djukić B. 2007. Prevalence of cryptosporidial infection in piglets with clininical signs of enterophaty. Biotechnology in Animal Husbandry 23 (5-6) book 2, 229-235 [9]. Savic B., Pavlović I., IveticV., Zutic M., Kureljusic B. 2010.Prevalence of isosporidial infection in piglets with clininical signs of enteropathy. Proceedings of 21st IPVS Congress, Vancouver Canada, 800 [10]. Worliczek H.L., Gerner W., Joachim A., Mundt H-C, Saalmüller A. (2009) Porcine Coccidiosis – Investigations on the Cellular Immune Response against Isospora suis. Parasitol Res 105 : 151 – 155

    ABOUT THE AUTHORS

    Ivan Pavlović, Milenko Žutić,Oliver Radanović, Božidar Savić, Scientific Veterinary Institute of Serbia, Vojvode Toze 14, 11000 Belgrade, Serbia, E-mail: [email protected]

    mailto:[email protected]

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    THE EXAMINATION OF DISINFECTANTS EFFICIENCY ON

    PATHOGEN MICROORGANISMS ISOLATED FROM INCUBATORY STATIONS

    Živka Ilić, Dubravka Jovičić, Ivan Pavlović, Gordana Žugić, Miloš Gavrović,

    Dragica Vojinović

    Abstract: Our examination was aimed to determinated how the chemical (disinfiction) substance influence to alive of pathogen microorganism isolated at incubatory stations Key words: micoorganisms, incubatory station, desinfecton

    INTRODUCTION Incubator sections are facilities with necessary microclimatic conditions which provide

    the optimal embryonic development of different sorts of breeding eggs into healthy poultry offspring. The important role of incubatory stations is controlling the diseases of poultry, since it is the place from where different diseases can be transferred to other locations. Microclimate in the incubatory station is an ideal condition for growth and reproduction of different microorganisms during the incubation of eggs. From this reason disinfection are most important hygienic measures in incubatory stations in aims of health control of chickens and preservation environmental condition.

    MATERIAL AND METHODS

    During 2007-2009 examination of bacterial strain in incubatiory stations was performed using swabs at various parts of object (floor, walls, etc.), which examined at laboratory of Dep.of Poultry Diseases in Scientific Veterinary Institute of Serbia with routine microbiological laboratory methods for bacteriology cultivation and determination (2)

    To our examination we tested antibacterial efficacy of next disinfectant: phenol (1 and 2% concentration), dichloride izocyanurat (0,02 and 0,04%), perchlor vinegar acid (2 and 3%), kaliumperoxisulfate (1 and 2%), and formaldehyde (5 and 10%). Survive of found bacterial strain were examined 1, 5 and 10 minutes and 24 hours after exposition to each of disinfectant using methods described by Lambert et al. (11).

    Control we performed using swabs at various parts of objects and its microbiological examination by method described by Cremieux and Fleurette (4).

    RESULTS AND DISCUSSION

    During examination we occurred presence of Staphylococcus spp., E.coli, and Salmonellae spp. and Pseudomonas spp. To our examination we tested antibacterial efficacy of next disinfectant: phenol (1 and 2% concentration), dichloride izocyanurat (0,02 and 0,04%), perchlor vinegar acid (2 and 3%), kaliumperoxisulfate (1 and 2%), and formaldehyde (5 and 10%). Survive of found bacterial strain were examined 1, 5 and 10 minutes and 24 hours after exposition to each of disinfectant (Table 1)

    To all examined microorganisms formaldehyde we outstanding like disinfectant with best antibacterial action. Those results of formaldehyde efficacy are similar like results of examination performed by Dietz et al (5), Andryunin (1), and Ilićet al. (6).

    Perchlor vinegar acid was efficiency after exposition of 5 minutes. During examination of efficacy of same disinfectant microorganisms isolated in poultry breeding

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    objects Ilić et all (7,9) was analogous with results. That efficacy of vinegre acid was confirmed by Bodiroga et al. (3).

    Kaliumperoxisulfate and phenol were efficiency in all concentration after 10 minutes and dichloride izocyanurat after 24 hours exposition. Obtain results was agreed with results of Kleiner et al (11), Mijatov et al. (12), and Ilić et al. (8,9). Table 1. Results of antifungal activity of disinfectant

    Disinfectant Concentration Expose time in minute

    Control % 1 5 10

    Phenol

    1 - - + - 2 - - + -

    Dichloroizocyanurat 0,02 - - + -

    0,04 - - + - Perchlore winegre acid

    2 - + + - 3 - + + -

    Kalium peroxisulfate

    1 - + + - 2 - + + -

    Formalidechid

    5 + + + - 10 + + + -

    Legend: + disinfectant operate; - disinfectant not operate; ± disinfectant deficient

    operate

    CONCLUSIONS AND FUTURE WORK

    Disinfection are most important hygienic measures to control of pathogen microorganisms in incubatory stations

    REFERENCES

    [1]. Andryunin Y.I. (1982) Theoretical basis of the bactericidal activity of an alkaline solution of formaldehyde. Vestnik Selskokhozyaistvennoi Nauki, USSR, 9, 80-84 [2]. Ašanin .R.,Krnjaić D.,Milić N.(2006) Priručnik sa praktičnim vežbama iz mikrobiologije sa imunologijom, Fakultet veterinarske medicine, Beograd, 63-69, [3]. Bodiroga J., Višakci M. Ašanin R, Pisarev M. (1996) Nove mogućnosti dezinfekcije vode za piće, prostora i opreme preparatom “Peral-S” na živinarskim farmama. VII savetovanje DDDD u zaštiti životne sredine, Sutomore, 32-33. [4]. Cremieux A., Fleurette J. (1991) Methods of testing disinfectants u Block, S.S.: Disinfection, sterilization and preservation. Philadelphia, Lea Febiger, 1009-1027. [5]. Dietz P., Bohm R., Strauch D. (1980) Effectivenss and safety of formaldehyde gas and aerosol of per acetic acid and hydrogen peroxide. Zentralblatt für Veterinarmedizin, 27B ,268-279 [6]. Ilić Ž., Spalević Lj., Miljković B., Pavlović I., Žugić G., Stanojević S. (2006) Efficace of same microbiological substance at pathogen micoorganisms isolated in poultry breeding objects Lucrari Stiintifice Medicina Veterinara 49 (8) 872-874 [7]. Ilic Z., Jakic-Dimic D., Pavlović I., Zugic G, Gavrilovic M. (2008) Efficacy of same antimicrobial substance at salmonella microorganisms isolated in poultry breeding objects, Book of Proceedings of 1st Mediterranean Summit of WPSA Advances and Challenges in Poultry Science 07- 10.5.2008."Porto Carras"Grand Resort,Chalkidiki,Greace,252-254

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    [8]. Ilić Ž., Jakić-Dimić D., Maslić-Strižak D., Pavlović I., Miljković B., Žugić G., Gavrović M. (2009) Efficacy of same disinfectant at E. coli microorganisms isolated in poultry breeding objects. Biotechnology in Animal Husbandry 25 (5-6) book 2, 1117-1122 [9]. Ilić Živka, Jakić Dimić Dobrila, Pavlović I., Gavrović M. (2009) Efficacy of sоme antimicrobial substances at fungal microorganisms isolated in poultry breeding objects Proceedings of 6th International Symposium on Biocides in Public Health and Environment, 6th International Symposium on Antisepsepsis, Disinfection and Sterilization and Belgrade Conference 2009 on Vector Control in Urban Environments, 194-195 [10]. Kleiner U., Trenner P., Profe D. (1988) Results from chinical fest of the disinfectant Vetasept G. Monatshefte für Veterinärmedizin, 43(18), 639-641. [11]. Lambert R.J., Fohnston M.D., Simons E.A. (1998) Disinfectant testing: uses of the Bioscreen Microbiological Growth Analyser for laboratory biocide screening. Letters in Appl. Mycrobiol., 288-292. [12]. Mijatov LJ., Grbović M., Molnar D. (1998): Dejstvo DESU amfotenzida na animalne patogene bakterije. Zbornik radova, IX savetovanja DDD.

    ABOUT THE AUTHORS

    Živka Ilić, IvanPavlović, Dragica Vojinović Scientific Veterinary Institute of Serbia, Vojvode Toze 14, 11000 Belgrade, Serbia, E-mail: [email protected] Dubravaka Jovičić, Faculty Futura, University Singidunum, Belgrade Gordana Žugić, Medicines and Medical Devices Agency of Serbia, Belgrade Miloš Gavrović, Krka dd, Novo Mesto, Slovenia

    mailto:[email protected]

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    THE INFLUENCE OF ENZYMES IN THE BAKERY TECHNOLOGY

    *David I.1, Berbentea F.1, Furnică Adela1, Georgescu L.1, Danci M.1, Bujancă G.1,

    Nicula Andrea1

    Abstract: This study presents the different action of three types of exogenous enzymes: enzyme preparation witch contains amylase, enzyme preparation witch contains hemicellulase and enzyme preparation witch contains lipase in the dough for bakery products. The determination of the rheological characteristics of the dough is obtained by alveographic method. Addition of exogenous enzymes in bakery products results in larger loaf volume, also the effect of exogenous enzymes on the bread volume improvement results from redistribution of water from the penthosane phase to the gluten phase that gives the gluten more extensibility. The exogenous enzymes are specialy used for obtaining bakery producs with low fat content, low sintetic aditiv content and high fiber content. Also they are used for improving bakery products texture and flavore. The influence of exogenous enzymes in the dough for bakery can help evaluate and improve the insufficiently developed technology and the nutritive value of the products.

    Key words: bakery, exogenous enzymes, amylase, hemicellulase, lipase, alveograph method

    1. INTRODUCTION Bakery products are the most comun traditional food products in the entire world.

    Bakery products have a high nutritive value due to the content of easily retainable sugars, lipids and proteins.

    Enzymes applications have grown to be a common practice in the baking industry with advantage of being considered as natural additives. The exogenous enzymes are being used in the baking industry to improve dough-handling properties. The synthetically additives can be replaced with natural additives, as enzymes.

    Addition of amylase in dough leads to: extension of freshness; the increase of the quantity of fermentation sugars, capable of forming gases during the entire period of the technical process inside the chains of amylopectine (Bordei, 2005); the obtaining of finite products with a more pronounce color of crust, by increasing the quantity of fermentation sugar; the increase of carbon dioxide quantity. Amylase hydrolysis the α-1,4-glycosidic connections from the amylose and amylopectine structures and helps forming dextrine and maltose, witch in normal quantity have a favorable effect on dough, by increasing the capacity of water retention and improving the aspect of the middle part (soft, fluffy) (Mencinicopschi and David, 2008). The reduction of dough’s consistency through the addition of alpha amylases leads to the increasing of extensive character and decreasing of the resistance of dough. This behaviour is due to the fact that the maltose obtained by starch hydrolysis realizes a dehydrating action on gluten. The quantity of free water in dough will increase, reducing consistency. Amylase is deactivated in the oven, before the amidon’s gelatinization. Therefore, this excludes the risk of excessive dextrinization that could live to a sticky content.

    The effect of hemicellulases on dough is the hydrolysis of the soluble and insoluble penthosane in water with the formation of olygomers with high reactive molecular mass (Dinu, 2002). They have as effect the neutralization of the negative action of soluble penthosane on bread volume, improving the stability of dough and its tolerance for fermentation, the improvement of the dough capacity to retain fermentation gases, the decrease of dough viscosity and the improvement of the processing quality (Diaconescu, 2004).

    Lipase is used in bakery industry for replacing and reducing the quantity of emulsifiers used for dough hardness. Addition of lipase in bakery products improves the

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    dough handling and stability. It also helps improve the quality of the products by forming a softer and whiter content and increases the dough volume. Lipase is completely deactivated during the baking process (Mencinicopschi and David, 2008).

    The enzyme addition of flours presents the advantage of constant quality flour, which does not modify the technological process, does not affect the health of consumers. The enzymes are used in small quantities and do not influence to a great extent the price of bread. They can be successfully used in the place of chemical additives for synthesis.

    2. MATERIALS AND METHOD

    2.1. Samples preparation Materials used for the preparation of the dough samples are commercial wheat flour

    with normal bread making properties (moisture 13.30%, protein content 12.75%), salt, water, yeast and exogenous enzymes.

    The enzyme preparation used is: - Fungamyl SG – enzyme preparation witch contains amylase with 2500 FAU/g

    (FAU- fungal amylase unit) enzyme activity (Sc Gamaserv Srl, Roumania); - Alphamalt HCC – enzyme preparation witch contains hemicellulase with !!!!!!!!!!

    LYX/g enzyme activity (Muhlenchemie, Germany); - Lipopan Xtra BG – enzyme preparation witch contains lipase with 7.2 KLU/g (KLU-

    kilolipase unit) enzyme activity (Sc Gamaserv Srl, Roumania). A sample of 250g of flour is mixed with a solution of salt, yeast and enzyme

    preparation in a laboratory mixer 15 min to form dough. The amount of water was adjusted according to the water absorption capacity of flour.

    The first dough sample MARTOR contained 95% flour, 1.7% salt, 1.7% yeast and does not any exogenous enzymes.

    The second dough sample F1 contained 95% flour, 1.7% salt, 1.7% yeast and 2g/100kg enzymes preparation witch contains amylase.

    The third dough sample F2 contained 95% flour, 1.7% salt, 1.7% yeast and 6g/100kg enzymes preparation witch contains hemicellulase.

    The fourth dough sample F3 contained 95% flour, 1.7% salt, 1.7% yeast and 1.2g/100kg enzymes preparation witch contains lipase.

    Each dough sample is divided in five circular consecutive dough patties witch are rested 20 min in the alveograph in a temperature-regulated compartment at 25 °C. Each dough patty is tested individually and the result is the average of the five dough patties.

    2.2. Methods of analysis The determination of the rheological characteristics of the dough was obtained by

    alveographic method .The alveographic method relies on measuring the resistance to biaxial stretch under air pressure of a dough sample prepared in standard conditions.

    The dough patty is placed on the alveograph, witch blows air into it. The dough patty expands into a bubble that eventually breaks. The pressure inside the bubble is recorded as a curve on graph paper. The alveograph determines the gluten strength of dough by measuring the force required to blow and break a bubble of dough. The results include P Value, L Value, and W Value. Stronger dough requires more force to blow and break the bubble (higher P value). A bigger bubble means the dough can stretch to a very thin membrane before breaking. A bigger bubble indicates the dough has higher extensibility; that is, its ability to stretch before breaking (L value). A bigger bubble requires more force and will have a greater area under the curve (W value).

    From the alveogram the following indicators were obtained:

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    • P Value is the force required to blow the bubble of dough. It is indicated by the maximum height of the curve and is expressed in millimeters (mm). It is also known as the viscosity or the value of maximum pressure that is in relationship to the resistance of the deforming dough (mm H2O) • L Value is the extensibility of the dough before the bubble breaks. It is indicated by the length of the curve that begins from the origin until the perpendicular point that corresponds to decreasing pressure due to rupture of air bubble and is expressed in millimeters (mm).

    • G Value is the expansion index G being the average of the expansion index on the graphic of cellules and corresponds to breaking the abscise L, G =2.226L, where L – air volume (cm3) used to stretch the dough under bubble form. • P/L Ratio is the balance between dough strength and extensibility. It is the rapport of configuration of the curve. • W Value is the area under the curve. It is a combination of dough strength (P value) and extensibility (L value) and is expressed in joules. It represents the action of deformation of the dough, based on a gram of dough, evaluated at 10 E – 4 joule, calculated as follows: W= 1.32 x (V/L) x S, where V- air volume in mm3; L- the average abscise at breaking point in mm; S- surface of the curve, cm2.

    • Ie – elasticity index, represents the rapport between the measured pressures, expressed in mm H2O to form bubbles after the insufflations of 200 cm

    3 of air in dough form, that correspond to a length L of 40 mm or an index of expansion G from 14,1 and the maximum of the curve P: Ie%= P200/Pmax.

    3. RESULTS AND DISCUSSIONS The dough samples alevograms are represented in Fig. 1, Fig. 2, Fig. 3 and Fig. 4.

    Each dough sample alveorgam show the five dough patties tested (marked with different colors) and the parameters registered at the testing moment. The results of the samples are represented by the average value obtained from the values of the dough patties tests for each dough sample.

    In Fig. 1 the dough sample MARTOR alveogram represents the dough sample that does not contain any enzyme preparation. This sample is considered the standard blank sample. The alveogram’s characteristics for flour used in bakery products have the fallowing values: P = [65 – 75mm], L = [130 – 150mm], G = [20 – 30], P/L = [0,55 – 0,65] and W > 180 10E – 4J. The values for dough sample MARTOR, regarding the resistance of the deforming dough (P) and the balance between dough strength and extensibility (P/L ratio) are higher than the normal values. The values regarding the dough extensibility (L), expansion index (G) and the total quantity of absorbed energy during the dough deformation (W) are very low, therefore the dough is sensitive to stretch and can easily brake. It cannot be used for bread making.

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    Sample MARTOR (no enzyme) Parameters Lab. temp: 25 ºC Moisture: 13.30% Falling number (FN): 354 s Gluten: 23.9 IG 97

    Fig 1. MARTOR (no enzyme) sample alveogram

    Results

    P = 79 mmH2O L = 54 mm G = 16.4 W = 163 10E – 4J P/L = 1.46 Ie = 52.1 %

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    Sample F 1 (enzymes preparation witch contains amylase) Parameters Lab. temp: 25 ºC Moisture: 13.30% Falling number (FN): 338 s Gluten: 24.6 IG 98

    Fig 2. F 1 (enzymes preparation witch contains amylase) sample

    alveogram

    Results

    P = 92 mmH2O L = 57 mm G = 16.8 W = 201 10E – 4J P/L = 1.61 Ie = 55.1 %

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    In Fig. 2 the dough sample F 1 alveogram represents the dough sample that contains

    enzyme preparation enzyme based on amylase. There is a noticeable increase in all the indicators that suggests the improvement of the dough resistant to deformation (P), also the dough extensibility characteristics (L and G) are higher than the value of dough sample MARTOR. Moreover, the elasticity index (Ie) has increased with 3.0% and the total quantity of absorbed energy during the dough deformation (W) is with 42 10E – 4J higher then the value of dough sample MARTOR. This result expresses advantages of using amylase in the preparation of the dough for bakery. The reduction of dough’s consistency through the addition of leads to the increasing of extensive character and decreasing of the resistance of dough

    In Fig. 3 is represented the alveogram of dough sample F 2 that has in composition exogenous hemicellulase. Compared with the blank sample MARTOR there are small differences regarding the dough strength (P) that increased only with 2 mm H2O, but looking at the extensibility characteristics and the absorbed energy during the dough deformation we can see, obviously, the dough quality improvements. Only the P/L ratio and the Ie value have decrease because of the dough strength and the dough extensibility. Using exogenous hemicellulase improves the stability of dough and its tolerance for fermentation and decreases of dough viscosity.

    In Fig. 4 is the alveogram for the dough sample F3 that contains lipase. Addition of enzyme preparation witch contains lipase in dough, reduces the dough strength (P) with 7 mm H2O. The absorbed energy during the dough deformation (W) and the P/L ratio are reduced. There is a small increase of the extensibility characteristics and of he elasticity

    Sample F 2 (enzyme preparation witch contains hemicellulase)

    Parameters Lab. temp: 25 ºC Moisture: 13.30% Falling number (FN): 326 s Gluten: 25.3 IG 92.0

    Fig. 3. F 2 (enzyme preparation witch

    contains hemicellulase) sample alveogram

    Sample F 3 (enzyme preparation witch contains lipase)

    Parameters Lab. temp: 25 ºC Moisture: 13.30% Falling number (FN): 313 s Gluten: 25.2 IG 89.0

    Fig. 4. F 3 (enzyme preparation witch

    contains lipase) sample alveogram

    Results

    P = 72 mmH2O L = 59 mm G = 17.1 W = 158 10E – 4J P/L = 1.22 Ie = 52.9 %

    Results

    P = 81 mmH2O L = 68 mm G = 18.4 W = 186 10E – 4J P/L = 1.19 Ie = 48.7 %

  • RESEARCH PEOPLE AND ACTUAL TASKS ON MULTIDISCIPLINARY SCIENCES 8 – 10 JUNE 2011, LOZENEC, BULGARIA

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    index (Ie). Dough with lipase content can be used in cakes and similar products, but cannot be used for bread making. Addition of lipase in bakery products improves the dough handling and stability and increases the dough volume.

    Table 1. Alveograph results of the dough samples: MARTOR (no enzyme), F 1 (enzymes

    preparation witch contains amylase), F 2 (enzyme preparation witch contains hemicellulase), F 3 (