16th INTERNATIONAL ECO-CONFERENCE® ON 7th SAFE FOOD
DRAGI DIMITRIEVSKI-president Prof. ELIZABETA ANGELOVA-vice-president Prof. TATJANA PRENTOVIK-vice-president Prof. VJEKOSLAV TANASKOVIK-vice-president.
NOVI SAD, 26 – 29 September 2012
I N V I T A T I O N
Organizer :
ECOLOGICAL MOVEMENT OF NOVI SAD
E-mail : [email protected]
Web site : www.ekopokret.org.rs
GENERAL OBJECTIVES
The significance of safe food production and consumption has been widely recognized, as the risks increase of food nutritive value reduction due to its industrial production. Even in highly developed countries industrial food production brings about new threats to human health.
Various issues concerning safe food are increasingly addressed both in professional and general publics of the developed countries, while the post-socialist ones are rather lagging far behind them.
With the general aim to contribute to communicating valid knowledge and supporting exchange of opinions and experiences, and to making the knowledge actionable, the Ecological Conference® on Safe Food has been held in even years since the year 2000. The conference is planned to:
- address, on a scientific-expert level, a wide range of key issues concerning safe food production and environmental consciousness;
- discuss and assess the causes of ecological imbalance in conventional agricultural production, and the impact of various pollution sources on agricultural production at present;
- offer practical solutions, based on scientific indicators and expert opinions, to help meet actual challenges and those that lie ahead;
- present relevant proposals and suggestions for the long-term strategic programmes in food production (in the industrialized, controlled, integral, alternative and sustainable agriculture) without negative effects on human health and environment.
SUBJECT AREAS
The Eco-conference® is open to all the issues concerning safe food, from the point of its production, to the point of consumption.
The contributions are most welcome that point out good or bad practices, while validating factors affecting food safety and quality in the following subject areas:
- Ecological factors and food production
- Correct choice of seed (genetic) material
- Status and preparation of soil as the basic substrate for the production of food and feed
- Use of fertilizers and pesticides in integrated plant protection
- Use of biologicals
- Food processing technology
- Economic aspects, marketing and packaging of safe food
SESSION TOPICS
The Conference will work in sessions. The final list of sessions – timing of the Conference will depend on the number of papers submitted.
The following session topics have been proposed by the Organizing Committee :
1. Climate and production of safe food.
2. Soil and water as the basis of agricultural production.
3. Genetics, genetic resources, breeding and genetic engineering in the function of producing safe food.
4. Fertilizers and fertilization practice in the function of producing safe food.
5. Integrated pest management and use of biologicals.
6. Agricultural production in view of sustainable development
7. Production of field and vegetable crops.
8. Production of fruits and grapes.
9. Lifestock husbandry from the aspect of safe food production.
10. Processing of agricultural products in the framework of safe food production.
11. Economic aspects and marketing as segments of the production of safe food.
12. Food storage, transportation and packaging.
13. Nutritional food value and quality nutrition.
14. Legal aspects of protecting brand names of safe food.
15. Ecological models and software in production of safe food.
NOTE:
Topics listed above should be regarded as the subject framework. Participants are kindly requested to submit only original scientific or technical papers that have not been published before and in which the practical possibilities are stressed for application of certain solutions in production of safe food.
All papers will be reviewed. Papers accepted by the Scientific Committee will be published in Conference proceedings. Each participant may be the principal author of one paper alone.
![Dragi Dragi](https://i1.rgstatic.net/publication/254386604_Assessing_the_Future_Development_of_the_Macedonain_Agriculture_Partial_equilibrium_model_of_livestock-feed_sector/links/53ecdbd80cf23733e804d15e/largepreview.png)
OFFICIAL LANGUAGES OF THE 2012 ECO-CONFERENCE®
The Eco-Conference® official languages are Serbian and English.
Thematic collection of papers will be published in English.
HONORARY COMMITTEE
President :
– Akademik Čedomir Popov, President of Matica Srpska
Vice-Presidents:
– Academician Pap Endre, President of the Vojvodina Academy of Art and Science
– Prof. Dr Miroslav Vesković, Rector of the University of Novi Sad
– Prof. Dr Branko Kovačević, Rector of the University of Beograde
– Prof. Dr Vladimir M. Bautin, Rector at Russian State University MTAA,Moscow
– Prof. Dr Borislav Kobiljski, Director of the Institute for Field and Vegetable Crops in Novi Sad
– Prof. Dr Jovanka Lević, Director of the Institute for Food Technology
– Vittorio Cogliati Dezza, President of Legambiente d’ Italy
SCIENTIFIC COMMITTEE
President:
– Academician Rudolf Kastori, Academy of Science and Art of Vojvodina, Novi Sad, Serbia and Hungarian Academy of Sciences, Budapest, Hungary
Vice-Presidents:
– Prof. dr Pavle Sekeruš, Vice-Chancellor for science at the University of Novi Sad
– Prof. dr Marko Ivetić, Vice-Chancellor for science at the University of Belgrade
– Prof. dr Evgenij Ivanovich Koshkin, Vice-Chancellor for International Co-operation at the Russian State Agrarian University – MTAA
– Dr Marija Bodroža-Solarov Scientific Associate of the Institute for Food Technology in Novi Sad
– Dr Ana Marjanović-Jeromela, Assistant Director for science at the Institute for Field and Vegetable Crops in Novi Sad,
– Stefano Ciafani, Vice-Presidente of Legambiente d’ Italy
– Secretary:
Danijela Tomčić
Members:
– Academician Srbislav Denčić
– Academician Vaskrsija Janjić
– Academician Branka Lazić
– Academician Dragan Škorić
– Prof. Dr. Desanka Božidarević
– Prof. Dr. Dragi Dimitrievski
– Prof. Dr. Miodrag Dimitrijević
– Prof. Dr. Ivana Đujić
– Prof. Dr. Vladimir Hadžić
– Prof. Dr. Kadar Imre
– Prof. Dr. Ali Koç
– Prof. Dr. Milan Krajinović
– Prof. Dr. Ivana Maksimović
– Prof. Dr. Dragutin Mihailović
– Prof. Dr. Rodoljub Oljača
– Prof. Dr. Hrvoje Pavlović
– Prof. Dr. Mihailo Peruničić
– Prof. Dr. Nebojša Ralević
– Prof. Dr. Atila Salvai
– Prof. Dr. Marija Škrinjar
– Prof. Dr. Radmila Šovljanski
– Prof. Dr. Ion C. Ungureanu
– Prof. Dr. Ljubo Vračar
– Prof. Dr. Momčilo Vukićević
– Prof. Dr. Lu Zhongmei
– Мr.Sc. Marija Ševar
ORGANIZING COMMITTEE
President:
– Nikola Aleksić, Director of the Ecological Movement of Novi Sad
Vice-president:
– Angelo Mancone, Co-ordinator Legambiente Veneto, Rovigo, Italy
Secretary:
– Ana Perenić, Organizer of the Ecological Movement of Novi Sad
Members:
– Ljubica Aleksić, Organizer of the Ecological Movement of Novi Sad
– Luka Vujasinović, Organizer of the Ecological Movement of Novi Sad
– Milan Vurdelja, Rector’s Office, University in Novi Sad
– Biljana Grba, Deputy Director of the Ecological Movement of Novi Sad
– Dušan Dozet, Scientific Institute for Field and Vegetable Crops in Novi Sad
– Mr Slađana Đuranović
– Marina Samsonov
– Mr Jelena Šogorov, Faculty for European Political and Legal Studies
– Željko Štrbac, Student of Faculty of Technical Sciences in Novi Sad
PARTICIPATION FEE
Participation fee per paper is 100 €. Payable by the first author.
Participation fee after June 1st per paper is 200 €.
Participants (attendants) pay the participation fee in the same amount.
The participation fee implies that the following is provided for the authors of papers and participants (attendants):
- Eco-conference® 2012 Proceedings with papers printed in whole, in English
- daily meals (lunches) in an exclusive restaurant,
- refreshments during the working part of the Eco-conference® 2012 and
Participation in a variety of accompanying cultural events:
- Nature preservation exhibition
- Concert
- Theatre performance
- Official banquet
- Field excursion (lunch included)
Participation fee is to be paid to the foreign currency account of the Ecological movement of Novi Sad :
Beneficiary: Ekoloski pokret Novog Sada
Vojvodjanskih Brigada 17
21000 Novi Sad
Serbia
IBAN: RS35335007010002733895
Account with: RAZVOJNA BANKA VOJVODINE A.D. NOVI SAD
SWIFT: RBVORS22
Branch address: Strazilovska st. 2
City: Novi Sad
Country: Serbia
DEADLINES
Please send us application form as soon as possible.
May 31st, 2012 – deadline for submission of papers
August 31st, 2012 – deadline for the payment of participation fee
Papers for which the participation fee is not paid within the above-mentioned deadline shall not be published in the Proceedings of the Eco-conference® 2012
7-9 October 2015, Ohrid, Republic of Macedonia
Symposium Proceedings
ISBN 978-9989-845-63-5
University Ss. Cyril and Methodius in Skopje Faculty of Agricultural Sciences and Food
SYMPOSIUM PROCEEDINGS
2nd International Symposium for Agriculture and Food 7-9 October 2015, Ohrid, Republic of Macedonia VOLUME I SKOPJE, 2016
2nd INTERNATIONAL SYMPOSIUM FOR AGRICULTURE AND FOOD
ISAF 2015 XXXVIII FACULTY-ECONOMY MEETING V SYMPOSIUM OF VITICULTURE AND WINE PRODUCTION VIII SYMPOSIUM FOR VEGATABLE AND FLOWER PRODUCTION X INTERNATIONAL CONFERENCE OF ASSOCIATION OF AGRICULTURAL ECONOMISTS OF REPUBLIC OF MACEDONIA VI INTERNATIONAL SYMPOSIUM OF LIVESTOCK BREEDING
Organized by Faculty of Agricultural Sciences and Food of Ss. “Cyril and Methodius” University in Skopje, Republic of Macedonia in co-organization with Institute of Animal Sciences of Ss. “Cyril and Methodius” University in Skopje, Republic of Macedonia
SYMPOSIUM PROCEEDINGS
ORGANIZING COMMITTEE Prof. Dragi Dimitrievski, PhD-president Prof. Vjekoslav Tanaskovikj, PhD Prof. Koco Porcu, PhD Prof. Vasil Kostov, PhD Prof. Zvonimir Bozinovic, PhD Prof. Dragan Gjosevski, PhD Prof. Marjan Kiprijanovski, PhD Prof. Tatjana Mitkova, PhD Prof. Srekjko Gjorgievski, PhD Prof. Dane Bosev, PhD Prof. Rade Rusevski, PhD Prof. Ljubica Karakasova, PhD Prof. Rukie Agic, PhD
SCIENTIFIC COMMITTEE
Prof. Vjekoslav Tanaskovikj, PhD, Macedonia Ass. Prof. Koco Porcu, PhD, Macedonia Prof. Sonja Ivanovska, PhD, Macedonia Prof. Metodija Trajchev, PhD, Macedonia Prof. Ordan Chukaliev, PhD, Macedonia Prof. Biljana Petanovska Ilievska, PhD, Macedonia Prof. Mile Pesevski, PhD, Macedonia Prof. Zvonko Pacanoski, PhD, Macedonia Prof. Zoran Dimov, PhD, Macedonia Prof. Natasha Gjorgjovska, PhD, Macedonia Prof. Ramesh Kanwar, PhD, USA Andrej Ceglar, PhD, JRC, Italy Prof. Jan Brindza, PhD, Slovakia Prof. Laima Taparauskiene, PhD, Lithuania Prof. Elazar Fallik, PhD, Israel Dunixi Gabiña, PhD, Spain Prof. Argir Zhivondov, PhD, Bulgaria Prof. Venelin Roytchev, PhD, Bulgaria Prof. Milena Moteva, PhD, Bulgaria Prof. Branko Čupina, PhD, Serbia Prof. Zoran Rajić, PhD, Serbia Prof. Biljana Škrbić, PhD, Serbia Prof. Radmila Stikić, PhD, Serbia Prof. Bojan Srdjević, PhD, Serbia Habibah Al-Menaie, PhD, Kuwait
Prof. Emil Erjavec, PhD, Slovenia Prof. Drago Kompan, PhD, Slovenia Prof. Alez Gregorc, PhD, Slovenia Prof. Franc Bavec, PhD, Slovenia Prof. Davor Romić, PhD, Croatia Prof. Vlasta Pilizota, PhD, Croatia Prof. Željka Zgorelec, PhD, Croatia Prof. Zlatko Svečnjak, PhD, Croatia Prof. Zdenko Lončarić, PhD, Croatia Prof. Stjepan Husnjak, PhD, Croatia Prof. Velibor Spalević, PhD, Montenegro Prof. Öner Çetin, PhD, Turkey Prof. Erbay Bardakcioglu, PhD, Turkey Prof. Gürsoy Oktay, PhD, Turkey Prof. Muhamed Brka, PhD, Bosnia and Hercegovina Prof. Miljan Cvetković, PhD, Bosnia and Hercegovina Prof. Ardian Maci, PhD, Albania Mohamed Refaei Mostafa, PhD, Egypt Prof. Nooshin Zandi-Sohani, PhD, Iran Sandor Kukovich, PhD, Hungary Juha Kantanen, PhD, Finland
SECRETARIAT
Ass. Prof. Koco Porcu, PhD Prof. Vjekoslav Tanaskovikj, PhD Ass. Prof. Dimitar Nakov, PhD Ass. Prof. Mirjana Jankulovska, PhD Ass. Prof. Mile Markoski, PhD Ass. Prof. Biljana Kuzmanovska, PhD Ass. Prof. Mirjana S. Jankulovska, PhD Ass. Prof. Marina Nacka, PhD
Dear authors, We are proud to present you with the Proceeding of Papers as an outcome of the Second International Symposium for Agriculture and Food, organized in 2015. This event represents the biggest gathering of the scientific public in the area of agriculture and food in Republic of Macedonia. Within the framework of the general event several symposiums and conferences took place: VIIIth Symposium for Vegetable and Flower Production Vth Symposium for Viticulture and Wine Production Xth International Conference of the Association of Agricultural Economists of Republic of Macedonia, and VIth International Symposium for Livestock Breeding. The Faculty of Agricultural Sciences and Food, University “Ss. Cyril and Methodius” in Skopje, has organized this event for the occasion of its 68th anniversary. It also represents a faculty’s commitment and responsibility for conducting and following scientific and research activities as well as for presenting novel results for the relevant stakeholders in the area of agriculture and food. This event has confirmed the accomplishment of the faculty’s vision for being a leading institution in the area of agriculture in Republic of Macedonia, recognized not only in the region but also in Europe and worldwide. The attendance of eminent authors from 21 countries, presenting more than 220 scientific papers, proofs the important role of the faculty for the agricultural and rural development. The papers were presented in 10 parallel sections covering research topics from the area of agriculture, food and environment protection. The Symposium Proceedings includes 142 scientific papers published in two volumes. The abstracts of all submitted papers (337 in total) were published in Book of Abstracts, prior the organization of the Symposium itself. Part of the submitted papers will be published in the faculty’s Journal of Agriculture and Environmental Sciences. Another part of the papers presented at this Symposium will be published in other well-known international journals, as an added value for the scientific importance of this Symposium. We sincerely hope that with the organization of the Symposium and with the publishing of the Proceeding of Papers we significantly contribute to the science in the area of agriculture and food. We expect that the published papers will be beneficial for the scientists and the experts in their future scientific activities. Sincerely, Prof. Dr. Dragi Dimitrievski President of the Organizing Committee of ISAF 2015 Dean of the Faculty of Agricultural Sciences and Food
CIP - Каталогизација во публикација Национална и универзитетска библиотека 'Св. Климент Охридски', Скопје 631/635(062) 338.43.01(062) INTERNATIONAL symposium for agriculture and food (2 ; 2015 ; Ohrid) Symposium proceedings : Електронски извор / 2nd International symposium for agriculture and food, ISAF 2015, 7-9 October 2015, Ohrid, Republic of Macedonia. - Skopje : Faculty of agricultural sciences and food, 2016 Начин на пристап (URL): http://www.fznh.ukim.edu.mk/. - Текст во PDF формат, содржи 2 св. (611 ; 525 стр.). - Наслов преземен од екранот. - Опис на изворот на ден 01.06.2016 ISBN 978-9989-845-63-5 (В. 1) ISBN 978-9989-845-64-2 (В. 2) а) Земјоделство - Собири б) Земјоделска економија - Собири COBISS.MK-ID 101270538
CONTENT
VOLUME I
SECTION 1.
ANIMAL BIOTECHNOLOGY
7-98
SECTION 2.
AGRICULTURAL ECONOMICS
99-276
SECTION 3.
PLANT PROTECTION – PHYTOMEDICINE
277-332
SECTION 4.
FOOD QUALITY AND SAFETY
333-400
SECTION 5.
VEGETABLE, FLOWER AND DECORATIVE PLANTS PRODUCTION
401-470
SECTION 6.
VITICULTURE AND WINE PRODUCTION
471-512
SECTION 7.
FRUIT GROWING
513-602
NATURAL RESOURCES MANAGEMENT AND ENVIRONMENT PROTECTION
603-804
FIELD CROP PRODUCTION
805-1060
VOLUME II
SECTION 8.
SECTION 9.
SECTION 10. AQUACULTURE AND FISHERIES
1061-1112
SECTION 1. ANIMAL BIOTECHNOLOGY
CONTENT
PANORAMIC APPEARANCE OF THE TURKISH POULTRY INDUSTRY Demirel R., Şentürk Demirel D.
9
BROILER PRODUCTION IN MACEDONIA Kocevski D., Georgievski S., Vukovic V., Nikolova N.
15
PHENOTYPIC CHARACTERIZATION OF BUSHA CATTLE IN THE R. OF MACEDONIA Bunevski G., Kocevski D., Dzabirski V., Porcu K., Petkov V., Saltamarski Z.
21
GROWTH AND DEVELOPMENT OF SHEEP FROM SYNTHETIC POPULATION BULGARIAN MILK Stancheva N., Dimitrova I., Georgieva S.
29
EFFECT OF MANNAN-OLIGOSACHARIDE BIO-MOS ON LIPIDS IN THE SERUM OF SIMENTAL CALVES Stolić N., Milošević B., Spasić Z., Ilić Z. Pešić B.,
37
OPTIMIZATION OF ENZYMATIC ASSAY OF SUPEROXID DISMUTASE AND GLUTATION PEROXIDASE ACTIVITY IN MILK WHEY Trajchev M., Nakov D., Gjorgoski I.
43
EFFECTS OF THE IMPLEMENTATION OF THE BREEDING PROGRAMS TO DEVELOP DAIRY CATTLE BREEDING IN VOJVODINA Trivunović S., Janković D., Ivanović D., Radinović M., Šoronja Ţ.
53
THE USING OF BIOTECHNOLOGY IN ANIMAL BREEDING Tutkun M., Tatar A.M.
59
STUDY ON EWE LIVE WEIGHT OF PATCH FACED MARITZA SHEEP BREED Dimov D., Vuchkov A., Ivanov I.
67
DYNAMIC AND COMPOSITION OF LACTIC ACID BACTERIA (LAB) DURING TRADITIONAL BUTTER AND BUTTERMILK PRODUCTION Santa D., Srbinovska S., Trajkovski G.
73
DOG FIGHTING IN THE WORLD Yilmaz O., Coskun F., Ertugrul M.
83
SAWDUST-ROUGHAGE SOURCE IN THE FEED MIXTURES FOR FATTENING STEERS Gjorgjievski S., Gjorgovska N., Levkov V.
91
SECTION 1. ANIMAL BIOTECHNOLOGY UDC: 636.52/.58.033(560) Review paper
PANORAMIC APPEARANCE OF THE TURKISH POULTRY INDUSTRY Demirel R.1*, Şentürk Demirel D.1 Faculty of Agriculture, Department of Animal Science, Dicle University, Diyarbakır – Turkey
1
*
corresponding author: [email protected]
Abstract Poultry products are important source of animal protein, especially in low incoming countries such as Turkey. In the Country, most of the animal originated protein is obtained from poultry as egg and white meat. Our population is fastly increasing by the years. Its difficult to meet protein requirements of additional population. Broiler and layer chicks can be easily produced in a short time. At the same time, poults effectively turn the feed to animal protein cheaply. So, it is the easy way to meet protein requirements of increasing population. These advantages make poultry industry getting popular. Timely, traditional poultry breeding in villages turned into an industry in modern conditions with higher yielded anamals, good quality feeds etc. From 1980s to 2015, broiler production and meat consumption is reached from 2 kg to 21 kg per capita in Turkey. At the same time, it is placed in the first rank, among animal protein sources. Durind the period, red meat consumption is decreased from 19 kg to 13 kg per capita. At the same time laying hen production is grown successively, too. Poultry breeders are reach the highest technic knowledge level. There are some constituents of this successfully production such as: good poultry production, using technology, good veterinary services, higher feed quality, hygiene, vaccination etc. In this article, current situation of poultry industry and related sectors in Turkey will be discussed. Key words: Turkey, poultry industry, broiler, laying hen, mixed feed. Introduction Traditional poultry breeding was dominant up to middle of 1980s in Turkey. Especially after the second half of 1980s, the Ministry of Agriculture supported the poultry industry for infrastructure (henhouse, cage, some equipment etc.) between 25 – 40% total cost, depending on developmental situation of the regions. During the years that increase in internal emigration from rural areas to industrialized cities, traditional food supply become insufficient for the population. So, modern, big scale poultry industry was needed in order to meet food requirements of growing populations in developing cities. Increase in additional peoples lead to raise food requirements of the city population. Timely, higher yielded modern hybrid lines were used as intensified instead of low yielded domestic combined breds. Tecnical knowledge of poultry breeders are beter than before. Animal vaccine and medicine, veterinary services became better. As hybrid quality incraesed, feed qulity is also increased in order to meet nutrient requirements of high 9
SECTION 1. ANIMAL BIOTECHNOLOGY
yielded animals. Integrated broiler firms are grown fastly. Integrated firms are also essential for farmers who have small scale production. Farmers can obtain chicks, feed, coal, vaccine, medicine, veterinary services etc. without any payment. At the end of growing period, farmers can obtain their profits depending on their success in operation. Development of Turkish Poultry Industry by Figures Total produced poultry numbers Total yearly produced poultry numbers are changed from approximately 264 million to 270 between 2000 and 2014 years as seen in Figure 1.
Figure 1. Anonymous, 2015a
Total poultry meat production Total poultry meat production are changed from approximately 662.000 ton/year to 1.895.000 t/y between 2000 and 2014 years as seen in Figure 2.
10
SECTION 1. ANIMAL BIOTECHNOLOGY
Figure 2. Anonymous, 2015a
Total produced egg production Total yearly produced egg production (numbers) are changed from approximately 10 billion to 17 billion between 2000 and 2014 years as seen in Figure 3.
Figure 3. Anonymous, 2015b
Total domestic poultry meat consumption Total domestic poultry meat consumption ise changed from approximately 10 kg to 20,5 kg between 2000 and 2014 years as seen in Figure 4.
11
SECTION 1. ANIMAL BIOTECHNOLOGY
Figure 4. Anonymous, 2015a
Broiler feed production Broiler feed production ise changed from approximately 1 million ton to 3,98 million ton between 2000 and 2014 years as seen in Figure 4.
Figure 4. Anonymous, 2015c
Layer feed production Poultry layer feed production ise changed from approximately 80.000 ton to 2,48 million ton between 2000 and 2014 years as seen in Figure 5.
12
SECTION 1. ANIMAL BIOTECHNOLOGY
Figure 5. Anonymous, 2015c
Poultry meat export Total poultry meat export ise changed from approximately 24.417 ton to 395.694 ton between 2000 and 2014 years as seen in Figure 6.
Figure 6. Anonymous, 2015d
Conclusions Turkish poultry industry is getting grow fastly, however consumption of domestic poultry products is growing slowly. So, some of Turkish poultry products are send to export. Poultry feed constituents are commonly imported (soybean, fat and oils, additives etc.) and this figures are getting high year by year. Our production figures (feding period, mortality, product quality, FCR, etc) are competible with developed countries, but our firms are low 13
SECTION 1. ANIMAL BIOTECHNOLOGY
scale and profitibility levels are lower. At the same time, our production is considered as halal production. High quality and halal production is getting more popular for international markets. References Anonymous 2015a. Turkish Statistic Agency, Livestock Animal Production Statistics. www.tuik.gov.tr/ (Access date: 20.06.2015). Anonymous 2015b. Egg Production in Turkey.www.yum-bir.org/( Access date: 20.06.2015). Anonymous 2015c. Feed Industrialists Organization. www.yem.org.tr/ (Access date: 20.06.2015). Anonimous 2015d. www.besd-bir.org/.../turk-kanatli-eti-sektoru-ihracatı (Access date: 08.07.2015).
14
SECTION 1. ANIMAL BIOTECHNOLOGY UDC: 636.52/.58.033(497.7) Professional paper
BROILER PRODUCTION IN MACEDONIA Kocevski D.1, Georgievski S.1*, Vukovic V.1, Nikolova N. 2 1
Faculty of Agricultural Science and Food-Skopje, Institute for Animal Biotechnology, University „St. Cyril and Methodius―, Skopje, Republic of Macedonia 2 Institut for Animal Science-Skopje, University „St. Cyril and Methodius―, Skopje, Republic of Macedonia *
corresponding author: [email protected]
Abstract Macedonian broiler industry is underdeveloped therefore majority (over 90%) of the poultry meat on the market is imported frozen poultry meat. Initial steps for development of broiler sector are focusing on development of chilled poultry meat market. Market analyze revealed that possibilities for developing the sector exists and domestic market trends follows the international poultry market trends. Projections based on these trends, following demands and supply suggest substantial market size growth and shift leading to increased market share of chilled poultry meat and meat products toward frozen one in next ten to fifteen years. Based on the information on domestic production as well as on the import data it can be concluded that activities are needed in all aspects of the supply chain starting from broiler raising and slaughtering to processing up to sales and distribution of chilled and frozen poultry meat. Once the basic elements of well organized chilled broiler meat supply is established there will be further opportunities open for the other value added products such as chicken sausages, chicken burgers and nuggets (for the fast food industry). Key words: poultry meat, market, trends, projections. Introduction Macedonian poultry industry satisfies the domestic egg market needs (MAFWE, 2007), but at the same time Macedonia is importer of a poultry meat. The level of meat production of our poultry industry is presented in Table 1 (MAFWE, 2007; FAOSTAT, 2012). In addition, the half of the mentioned production of poultry meat (broiler meat) is attributed to a spent layers meat (chicken meat) (MAFWE, 2007).
15
SECTION 1. ANIMAL BIOTECHNOLOGY Table 1. Quantity of poultry (chicken) meat production Year Total raw meat produced, tons 2010 3200 2009 3319 2008 3012 2007 3524 2006 3715 2005 3 809 2004 3 189 2003 4 116 2002 3 992 2001 4 702
The total poultry meat consumption in Macedonia is 23 000-30 000 tons per year, or 11.5-14.5 kg/per capita (according National Statistic Office data, the 2005 annual consumption of poultry meat per capita was 11.0 kg suggesting a market needs of 22 000 t/year, while the Veterinary Directorate reports import of 24. 093 t, which is close to the poultry meat imports of FAO (Table 2). Table 2. Production and import of poultry meat in the R. Macedonia (1000 tons) Macedonia FAOSTAT FAO Statistics Division September 2012 2003 2004 2005 2006 2007 Production 4.12 3.19 3.81 3.71 3.52 Import 20.74 25.74 21.49 19.69 30.84
2008 3.01 25.41
2009 3.32 28.78
2010 3.20 25.99
The EU information (EUROSTAT) about the chicken meat consumption in European Union and projections for the poultry meat market movement and chicken meat consumption per capita suggest that Macedonia, considering the poultry meat consumption, is behind states members of EU (Table 3) Table 3. Consumption data (FAOSTAT) 2003 EU-25 22.9 R. Macedonia 13.5
for poultry meat in R. Macedonia and EU, 2003-2012 (kg per capita) 2004 22.9 15.8
2005 23.6 13.9
2006 23.6 13.2
2007 23.8 19.40
2008 23.9 16.00
2009 23.2 17.30
2010 23.1 17.00
2011 22.6 16.90
2012 22.8 17.10
Materials and methods Questionnaire research survey on consumer preferences of poultry meat focused on chilled broiler meat marketing development idea, including 1000 examinees, revealed that the most of the questioned consumers purchased poultry meat more than three times a month. Only 20.8% buy chilled poultry meat regularly, beside the fact that 83.5% recognize chilled and frozen poultry meat as different product. Only 45.4% of the questioned consumers (about a half) preferred to buy the most valuable and highly priced meat parts of first class meat (breast, drumstick and thighs). The majority of consumers 89.2% accept higher prices for better quality articulated to price/quality relations. Quality is dominant factor and a crucial moment in buying decision 67%, followed by health 59.4%, but due to 16
SECTION 1. ANIMAL BIOTECHNOLOGY
its relativity price is still a factor that affects the decision due to the low buying power of the consumers. If the trend of the Macedonian market follows the consumption and quality demand of the new EU member states, (where fresh (chilled) poultry meat will supersede the most of the market), fresh (chilled) poultry meat will increase the domestic market needs, following the needs of our consumers for fresh chilled chicken meat (whole chicken, breast, drumstick and thighs). This trend is occurring beside the higher price of products of fresh (chilled) poultry meat. The price for fresh (chilled) poultry is 115-169 denar/kg i.e. 1.9 EUR-2.8 EUR; the price for frozen chicken meat is 90-120 denar/kg i.e. 1.46-2.1 EUR. Based on the data from 2005, predictions can be made on the growth of the poultry meat market in the next years in order to know the situation after 5, 10 and 20 years. Considering the basic assumptions from the Table 4, than the given figure will present the market growth potentials and a part of the domestic production and potentials for increasing the broiler industry in the next 20 years. Table 4. Poultry sales past and predicted future 2005: data Number of inhabitians 2.0 million people Total poultry meat 25.300 tonnes consumption Poultry meat consumption 12.66 kg/capita per capita Total poultry meat supply: - Meat from a spent layers 1.810 - Fresh broiler meat 2.000 - Import 21.490
2010: data 2,1 million people 29.190 tonnes
2015: forecast 2.2 million people 33.000 tonnes
13.90 kg/capita
15.00 kg/capita
1.000 2.200 25.990
1.000 7.000 25.000
Conclusions Considering these predictions and projections according to development state policies designed in national and IPARD program, followed by the program for financial support in the agriculture (30 MKD i.e. 0.47 Euro/broiler for 2011), it is probable to expect growth and development of broiler industry in the next period. If current level of production is about 2200 tones/year covering the market demands with approximately 7%, as soon as production starts to increase, the level of about 25% should be reached within 5 years and about 30% within 10 years.
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Poultry market growth & development of domestic production development in 20 years 40000 30000 Mt 20000 10000 0 2005
2010
2015
Domestic production
2020
2025
Frozen imports
Figure 1. Predicted market growth and development of broiler production
Following expected trends presented in Table 4 and Figure 1, it could be expected that the part of the frozen poultry meat in total offer will decrease (from 87% to 63% by 2015 and 46.7% by 2025) as the fresh chilled poultry meat from domestic production increase its part on the market supply (from 7% to 30% and 48.4%, respectively). Generally, according to the analysis presented above, there is a conclusion that there are possibilities for broiler production, focusing on the domestic market for fresh (chilled) poultry meat.
Experts production forecast of Broiler Meat Forecast Broiler production: 2012: 9,570 MioT & 2013: 9,535MioT (-1,4%)
1.600
1.400
DE +1,3%
2010 2011 2012 2013
ES -5,7%
1.200
UK +1,9%
FR +2,0%
1000 T
1.000
800
600
HU - 15,4%
400
200
0
BE
BG
CZ
DK
DE
EE
EL
ES
FR
IE
IT
CY
LV
LT
HU
MT
NL
AT
PL
PT
RO
SI
SK
FI
SE
UK
Figure 2. Predicted production forecast of broiler production in EU in 2013 (CIRCA).
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References AVEC. 2012. Avec annual report 2012. AVEC. 20112. Avec annual report 2011. Beddall C., (2003), Poultry likely to benefit from retail expansion in Central Europe. Business focus, 1, p.1. Bonnett, C. (2007) Sectors Analysis on key agricultural products: milk, meat and their products - Meat sector and its Products, an analysis of the meat sector in Macedonia. Skopje CIRCA EU. 2012. Pig meat &Poultry. EUSTAT (2005), Meat per capita consumption projections in the EU, 2003-2012. Seling prices of chickens. European Commission, Directorate – General for Agriculture and Rural Development (2007), Prospect for Agricultural Markets and Income in the European Union 2006-2013. European Commissions, Directorate – General for international policies. (2010). The poultry and egg sectors: evaluation of current market situation and future prospects. European Commissions, Directorate – General for Agriculture and Rural Development (2011), Prospect for Agricultural Markets and Income in the European Union 2011-2020. FAO 2010. Poultry meat and eggs. Agribusiness handbook. FAOSTAT, (2007), Imports of chicken meat. FAOSTAT | © FAO Statistics Division 2012 | 09 September 2012 GLOBAL POULTRY TRENDS 2012 - Little Growth Forecast in Europe's Human Population The poultry site, September 2012 Kocevski D., Manevska Irina, Dimovski, Lj., Toshev, Lj., Micevska Gabriela. 2007. Poultry meat market analysis: 1. Chilled poultry meat supply and prices on the market of R. Macedonia. III Livestock symposium, 12-14 September, Ohrid. Kocevski D. Manevska Irina, Dimovski Lj., Toshev Lj., Micevska Gabriela. 2008. Poultry meat market analysis: 2. Current status, Trends and prospective of poultry meat production in Macedonia. XX International Poultry Conference WPSA ―Science for Poultry Practice – Poultry Practice for Science‖ Wenecja near Bydgoszcz, Poland (15th – 17th September 2008) Kocevski D. Manevska Irina, Dimovski Lj., Toshev Lj., Micevska Gabriela. 2008. Poultry meat market analysis: 3. Consumer preferences in Macedonia. XX International Poultry Conference WPSA ―Science for Poultry Practice – Poultry Practice for Science‖ Wenecja near Bydgoszcz, Poland (15th – 17th September 2008) Milosevic N., Pavlovski Z. Peric, L. 2011. Present situation, capacities and prospects of development of poultry production in Serbia. Biotechnology in animal husbandry, 27 (3), p. 499-509. Ministry of Agriculture, Forestry and Water Economy (MAFWE), (2007), National Agricultural and Rural Development Strategy. OECD-FAO 2011. Agricultural Outlook 2011-2020. Randall K., (2002), Market trends. Business focus, 2, p.4. Sheppard A., Edge, S., (2005), Economic and operational Impacts of the proposed EU Directive laying down Minimum Standards for the Protection of Chickens kept for Meat Production. Centre for Rural Research, University of Exeter, Research Report Number 13. USDA Foreign Agricultural Service, (2006), GAIN Report – BR6623
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USDA Foreign Agricultural Service, Circular Series DL&P 1-06. (2006), Livestock and Poultry World Markets and Trade. Van Horne P., (2000), New pressures hitting all countries. Coob Quarterly Review, 2, p.12. Van Horne P., (2002), Europe to push for global standards for chicken production. Business Focus, 2, p.1-2. Van Horne, P., (2010). Prospective for poultry meat and eggs in the EU. European Poultry Club (EPC) annual event ―Future markets for egg and poultry in China, Russia and the EU‖ at the EuroTier exhibiton in Hanover in November. Peter van Horne (2009).Competitiveness of the European broiler industry: future perspective. XIII European Symposium on the Quality of Eggs and Egg Products Turku, Finland 21- 25 June 2009. Westhoff P., Bienfield, J., Young, R., (2003), Global markets for agricultural products 2003-2012. Outlook, p.1-15.
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SECTION 1. ANIMAL BIOTECHNOLOGY UDC: 636.2.082:575.21(497.7) Original scientific paper
PHENOTYPIC CHARACTERIZATION OF BUSHA CATTLE IN THE R. OF MACEDONIA Bunevski G.1*, Kocevski D.1, Dzabirski V.1, Porcu K.1, Petkov V.2, Saltamarski Z.3 1
Faculty of Agricultural Sciences and Food-Skopje, Institute of Animal Biotechnology, University „St. Cyril and Methodius―, Skopje, Republic of Macedonia 2 Faculty of Veterinary Medicine-Skopje, University „St. Cyril and Methodius―, Skopje, Republic of Macedonia 3 NGO Busha, Skopje *corresponding author: [email protected] Abstract Busha cattle are indigenous breed in many Balkan countries. Because of the economic, cultural and scientific reasons it is very important to protect biological diversity of autochthonous breeds like busha cattle. The aim of the research was to estimate the main productive, reproductive and exterior traits in several strains of adult busha cattle in the R. of Macedonia. The average daily milk production in grey strain of busha cows was 3,8 kg, and in brown strain was 4,1 kg, with 3,89 i.e. 4,01% of fats, 3,57% i.e. 3,62% proteins and dry unfatted matters 9,49% i.e. 9,62% in milk from brown i.e. brown strain of busha cows. According to the measurement of some morphological traits of adult busha cows, in grew strain the wither height was 107 cm, in black strain 105, and in brown strain also 105 cm, with the similar values for the traits back height and rump height in cows. The length of head was 38 cm i.e. 37 in grew i.e. black and brown strain, and length of horns 16 i.e. 15 cm in grew and black i.e. brown strain of cows. The average body mass of newborn calves was 15 kg i.e. 14 kg in grew and brown i.e. black strain. The average body weight at first mating was 125 kg in male and 150 kg of female busha cattle, and the average age at first calving was 28 months. The research results have shown small variation in their productive, reproductive and morphological traits between different strains of busha. Key words: cattle, Busha, domestic breed, phenotypic traits. Introduction Cattle are a main source of milk and beef products in Macedonia including various breeds, strains and crosses. This industry consists of three sectors: small-scale farmers (around 90%) keeping 1-3 cow and mainly producing for home consumption; medium-scale farmers (5%) keeping 10-15 cows with annual production of 4000-5000 kg milk/head and specialized commercial farms (around 5%) with more than 50 heads that produce annually over 7.000 kg milk intended solely for the market, or beef production with more than 400 kg body mass per year per head. In the mountain rural regions of Macedonia the dominant type of cattle are the crosses of the Busha breed. This breed of Busha cattle, also known as Illyrian cattle, is
21
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autochthonous breed of the Balkan Peninsula. It has been bred for centuries in this area and belongs to a group of primitive short horned cattle (Bos brachyceros Europaeus). Busha cattle is an indigenous, actually trans-boundary breed, which is in high danger of extinction. It used to be dominant and most important breed in almost all Balkan countries until 50s and 60s of the XX century but today in lowland areas with intensive farming they are already replaced with more productive and specialized cattle breeds. In Macedonia this breed was officially classified as triple purpose breed (for meat, milk and work) but considering it’s low productive capabilities it is more similar to some primitive working breeds. Today these cattle are no longer used for work but because of absence of systematic cattle improvement program these animals have retained their poor beef and dairy production capability. It could be said that the Busha's genome is very elastic since this breed in unfavorable conditions easily achieves better milk production and bigger body weight. Having in mind that this breed is well adapted to the very harsh feeding and housing conditions that exist in the rural areas of the Macedonian mountains and is resistant to diseases, it is still the most significant beef and sometimes milk resource for these areas where the more productive cattle breeds cannot thrive successfully. In the past several decades, as a result of uncontrolled crossing of this cattle with some more productive breeds, the number of purebred Busha animals is permanently being reduced which imposes an urgent need for setting up in situ and ex situ conservation program for this breed. Because of the economic, cultural and scientific reasons it is very important to protect biological diversity of autochthonous breeds like Busha. Busha strains in the R. of Macedonia There are two main classifications of Busha strains in the R. of Macedonia: a) Classification of strains according to locality: - Povardarie strain, - Polog strain, - Orgazden strain, - Prespa (Ohrid) strain, - Mariovo strain, etc. b) Classification of strains according to color: - Black strain, - Brown strain, - Red strain, - Gray strain, and - Tiger strain. In our country the following varieties on the basis of their coat color can be found: black Busha which is reared in Debar, Tetovo and Gostivar region (Polog strain), red Busha (Metohija strain), grey Busha (Povarie strain and Prespa or Ohrid strain), brown strain (Ograzden strain) and sometimes the so called ―tigar‖ strain. Actual situation of cattle breeding in the R. of Macedonia According to the official statistical data there are totally 214.000 cattle in the R. of Macedonia (FAO, 2014), from which 12.064 heads of Busha cattle or 5,6% (Agency of Veterinary and Food-AFV, 2014), but according to reality, there are approximately 10002000 heads of Busha cattle in our state. State policy and legislative for conservation of Busha cattle 22
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In 2008 a new Low of animal production is in function in the R. of Macedonia, where several articles are regulating the biodiversity and species of autochthonous breeds of cattle in Macedonia. From cattle, the only autochthonous breed is Busha cattle. In 2010 a Livestock Common Breeding Program was established from the Ministry of Agriculture of RM, for the period from 2010 to 2020, where the main stress from cattle breeds was given to Busha cattle. According this program, a Separate Breeding Programs for each breed have to be done, for the duration of 5-10 years. In 2011 was done the Program for Livestock Biodiversity by the Ministry of Agriculture of RM, for the period 2011-2017, with the main activities: Inventarization, characterization, monitoring of the trends and risks Phenotype analyses, Production traits, Reproduction traits, DNA characterization, etc. The overall objective of the National Breeding Program for Busha cattle is to implement the national conservation plan and sustainable use of Busha cattle. Table 1. Cattle breed distribution according the official data in the R. of Macedonia (AVF, 2014) 2008 2010 2012 2013 Breed No. of No. of No. of No. of In % In % In % In % heads heads heads heads Busha cattle 29535 12,1 27242 7,8 20363 10,1 12064 5,64 Crosses of Busha and 89707 36,8 104961 43,6 113720 39 98958 46,3 other breeds Total No. of 243667 100 269443 100 261073 100 213747 100 cattle
300000 250000
269443261073 243667 213747
200000
2008
150000
2010 2012
100000
2013
50000 0 Total No. of cattle Graph. 1 Number of cattle in RM (AFV, 2014)
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30000
29535
27242
25000 20000
20363 2008
15000
12064
10000
2010 2012 2013
5000 0 Total No. of Busha cattle
Graph. 2 Total No. of Busha cattle in RM (AFV, 2014)
Material and methods Several herds of busha cattle were investigated for the main productive, morphological and reproductive traits, mainly from south-eastern, south and south-western part of the R. of Macedonia, from hill-mountain area. All of them are reared semi-nomadic, i.e. during winter period from the end of October till the middle of April in the mountain village areas, and on the warm part of the year they are migrated on the mountain pastures according to the water sources and pasture yield. The controlled Busha cattle are mainly aimed for beef production, but some of them are milking by hand only for the purposes for farmers, not for selling. From the dairy production traits, the main emphasis was given to the milk yield per day and milk content (percentage of fats, proteins, solids non-fat matters and dry matters in milk), from the beef production traits to the average daily gain and body mass on 6 and 12 months od age and meat percentage from the carcass. The main morphological traits were measured with Lithin’s rod and measure tape, only on adult Busha cattle. Also, several main reproductive traits were measured, with the emphasis on the age on first mating and delivering, duration of open days and calving period, as well as on birth weight and body weight. From 2012 the 5 main bigger Busha farms were under the control of their productive and reproductive characteristics, from different regions of Macedonia: Herd 1 – Prilep region: 450 cattle, Herd 2 – Ograzden region: 120 cattle, Herd 3 – Ohrid region: 44 cattle, Herd 4 – Rozden region: 172 cattle, and Herd 5 - Belcista region: 94 cattle. In 2013 where were under the control 11 Busha herds, and in 2014 totally 38 Busha herds with approximately 800 heads of adult Busha cattle in the R. of Macedonia. Results and discussion Morphological characteristics When compared to the higher productive European dairy and beef breeds, Busha animals are of much smaller size. Carried by a short neck their head is small and light with short horns pointing upward and forward.
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SECTION 1. ANIMAL BIOTECHNOLOGY Table 2. Morphological traits of Busha cattle in Macedonia Trait Grew strain Brown strain No. of cows 11 15 Wither height 107 (104-111) 105 (99-113) Back height 107 (103-111) 105 (101-114) Rump height 109 (103-114) 108 (102-117) Length of head 38 (33-42) 37 (31-40) Length of horns 16 (14-21) 16 (12-19) Born weight 15 (12-18) 14 (11-17)
Black strain 4 105 (103-108) 105 (103-109) 107 (104-115) 37 (35-41) 15 (14-18) 15 (12-17)
Total x 30 106 107 108 37 16 15
The fully grown Busha animal is 90-115 cm tall at the withers. The length of the body is 116-132 cm or 104-118% of the withers height. There are great variations in the length of the body which is a result of the extensive way of rearing, lack of any breeding program and the poor zootechnical measures. Some of these animals have 1-3% higher pins than withers and some are with horizontal back line. They have narrow chest whose width is approximately 27% and depth 50-55% of the withers height (Ilkovski, 1994, Bunevski, 1994, Trajkovski and Bunevski, 2006). The fore limbs are straight but the hind limbs are usually hocked in. The rear part is narrow and the legs are sturdy with strong joints and ligaments. The hooves are hard and strong which enables these animals to move easily on steep and rocky terrains. The udder is small and shallow but with regular shape. Those animals that are better fed usually have bigger and deeper udders with more glandular tissue. The body weight of the fully grown animals is 150-300 kg but with better nutrition the cows can grow up to 280-320 kg and the bulls up to 430 kg (Trajkovski and Bunevski, 2006). The small body weight of these animals is not completely genetically dependent but also is a result of generations of underfeeding and neglecting. The skin is elastic and hard. Those animals that are reared well have short and shiny coat hair while in those that are neglected the coat is long and dense. Those that are raised on high mountain pastures always have longer hair. They usually have a stripe over the back and the muzzle, the horns and the hooves are darkly pigmentated. Similar morphological results were obtained also for the other strains of busha in the neighboring countries from Croatia to Turkey (Kume K. et. Al., 2013). The weight of adult males is between 250 and 300 kg, females between 150-250 kg. Their average wither height is 105-115 cm (Bunevski, 1994, Kume K. et. all, 2013). The head is characterized with short and curved horns pointing upward and forward and light pigmentation around the eyes and the muzzle. It is very robust and resistant with very modest feeding and housing demands. They are well adapted to the very harsh feeding and housing conditions, resistant to diseases and have long production life - 10 and more lactations. It could be said that the Busha's genome is very elastic, since this breed in more favorable conditions easily achieves bigger body weight and much better production. Productive characteristics In general these animals have low production which is in accordance with the poor body conformation. On the basis of the production capabilities this breed can be classified as triple purpose breed (for milk, meat and work). The low milk production is a result of the poor nutrition and the underdeveloped udders. Because of the small body weight it is a poor working animal and because of it’s poor body conformation and underdeveloped meat (beef) portions it is a weak fattening animal. The milk production of this breed is 25
SECTION 1. ANIMAL BIOTECHNOLOGY
from 700 to 1500 kg with 3,7-4,0% milk fat but there are also animals with higher milk production with around 2400 kg or with 4,8% milk fat. The lactation period lasts around 240 day. This low production can not compete with the specialized dairy breeds and the European countries with highly developed cattle breeding practices abandoned or rearing this breed long time ago. The milk content is shown in the following table. Table 3. Milk components from Busha cows n
Strain
Fats %
34 28
Grey strain Brown strain
3,89 (3,65-4,45) 4,04 (3,79-4,61)
Dry Unfatted matters % 9,49 9,62
Protein %
Kg milk/day
3,57 3,62
3,8 (2,8-5,3) 4,1 (3,1-6,2)
The fattening capabilities of Busha cattle are relatively poor. The birth weight of the calves is 15 - 22 kg and the daily weight gain around 500 gr. The meat is also of poor quality. The fattening capabilities are low because of the under nutrition and delayed maturity. Well fed male castrated young bulls have dressing percentage of around 47-55%. The fattened castrated males reach weight of 280-400 kg. The meat is of low quality. The health and the conformation of these animals is generally satisfactory having in mind that in such a bad conditions no other specialized cattle breed can survive. Reproductive characteristics Busha breed is classified as late-maturity breed because it reaches puberty at 13-15 months of age, breeding maturity when 2-2,5 years old and full growth at 4 years of age. The first calving of heifers is usually when they are 28 months of age (tab. 4). Table 4. Reproductive traits of Busha cattle in Macedonia (n =33 male and 54 female adult cattle) Trait Male Female Age at first calving / 28 months (19-39) Weight at first mating 125 kg (95-155) 150 kg (125-200) Age at first mating After 12 months 28 months Fertility (%) / 55% Birth weight (kg) 15 14 Body weight (kg) 275 (245-390) 230 (180-360) Age at 12 months of age 125 kg (95-155) 150 kg (90-182)
Under good raising and feeding conditions the fertility of this breed is generally good but in poor conditions when underfed they easily develop anovulation due to hunger. The bulls reach breeding maturity at 2,5 to 3 years of age while the cows after calving usually have short period of days open from 4 to 6 weeks and can give birth 10-13 times in their life (Ilkovski et all, 1994, Bunevski et all, 2004). Duration of days-open period has a big variation in our population, from 20 days after delivering till 100 days, as well as the duration od calving period from 300 to 400 days. These cattle show high disease resistance and adaptation to deficient nutritional and other environmental conditions.
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Conclusions The aim of the research was to show the main productive, reproductive and exterior traits in several strains of adult Busha cattle in the R. of Macedonia. The average daily milk production in grey strain of Busha cows was 3,8 kg, and in brown strain was 4,1 kg, with 3,89 i.e. 4,01% of fats, 3,57% i.e. 3,62% proteins and dry unfatted matters 9,49% i.e. 9,62% in milk from brown i.e. brown strain of Busha cows. According to the measurement of some morphological traits of adult Busha cows, in grew strain the wither height was 107 cm, in black strain 105, and in brown strain also 105 cm, with the similar values for the traits back height and rump height in cows. The length of head was 38 cm i.e. 37 in grew i.e. black and brown strain, and length of horns 16 i.e. 15 cm in grew and black i.e. brown strain of cows. The average body mass of newborn calves was 15 kg i.e. 14 kg in grew and brown i.e. black strain. The average body weight at first mating was 125 kg in male and 150 kg of female Busha cattle, and the average age at first calving was 28 months. The variation in their productive, reproductive and morphological traits between different strains of Busha is good for choosing superior bulls and cows for genetic improvement of this breed. In the past several decades, as a result of uncontrolled crossing of this cattle with some more productive breeds, the number of purebred Busha animals is permanently being reduced which imposes an urgent need for setting up in situ and ex situ conservation program for this breed. Because of the economic, cultural and scientific reasons, it is very important to protect biological diversity of autochthonous breeds like Busha cattle. References Agency of food and veterinary (AFV) of RM (2014). Cattle breed structure in Macedonia (http://www.fva.gov.mk/index.php?lang=mk). Bunevski Gj., (1994.): Improving of domestic cattle busha (seminar paper). 1-61, Skopje. Bunevski Gj., Trajkovski, T., Trifunovic, G., Adamov, M. (2004.): Selection program of cattle in the R. of Macedonia. 16th Symposium on innovation in animal science and production, Biotechnology in animal Science. Belgrade. FAO (2014): Annual statistical review. Roma (http://www.fao.org/home/en/). Ilkovski R., Trajkovski, T., Bunevski, Gj. (1994.): State and perspective of cattle production in the R. Of Macedonia. Meeting Faculty - Farms, Skopje. Kume Kirstaq, Papa L., Brka M., Dokso A., Zecevıc E., Rustempasıc Alma, Ivankovıc A., Ramljak Jelena, Lıgda Chrıstına, Georgoudıs A., Bytyqı H., Mehmetı H., Bunevskı Gj., Markovıc Bozıdarka, Markovıc M., Stojanovıc S., Bogdanovıc V., Perısıc P. (2013): Busha – old cattle in the balkan, erfp project – evaluation of current status of busha cattle and develop a regionalbreeding program for their conservation and sustainable economic use, 1-71. Trajkovski T., Bunevski, GJ. (2006.): Cattle breeding. Book, 1-371, Skopje.
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SECTION 1. ANIMAL BIOTECHNOLOGY UDC: 636.32/.38.034(497.2) Original scientific paper
GROWTH AND DEVELOPMENT OF SHEEP FROM SYNTHETIC POPULATION BULGARIAN MILK Stancheva N.1*, Dimitrova I.2, Georgieva S.3 Agricultural Institute – Shumen, Bulgaria Agronomy Faculty, University of Forestry, Sofia, Bulgaria 3 Faculty of Agriculture, Trakia University, Stara Zagora, Bulgaria 1
2
*
corresponding author: [email protected]
Abstract The goal of this study is to investigate the growth and development of sheep from breed Synthetic Population Bulgarian Milk (SPBM) at different ages and to define genetic determination of the trait by age categories.. The animals are from the herd of the Agricultural Institute – Shumen. The survey covers 851 ewes born during 2004 – 2008. Body weight of nine-month female lambs from this breed is 91.06% of weight of the 1.5 year female animals and 65.65% of that on 2.5 years (70.97 kg). The highest values of the coefficient of variation (C=21. 309) has live weight at 100 days of age, while the next age periods shows a downward trend, reaching 8.136% at 2.5 years. The father’ line has a fairly high impact on the live weight at 9 months (P≤0.001) and less at 1.5 and 2.5 years of age (P≤0.01). Year of birth has a fairly high impact on the trait live weight of all ages (P≤0.001). The coefficient of heritability (h2) of the live weight categories ranging from 0. 094 to 0.340. Key words: sheep, Synthetic Population Bulgarian Milk breed, growth and development, breeding line. Introduction Synthetic Population Bulgarian Milk (SPBM) sheep has been recognized as a breed in 2005 and is now the most widespread in Bulgaria. It was created by applying the hybridization of different maternal basis using rams of breeds East-Friesian, Pleven Blackhead, Stara Zagora and Awassi. A large part of the studies performed until now, relate to the parameters defining the productivity in the various stages of the population creation (Lazarov et al., 2002; Nedelchev et al., 2003; Raicheva et al., 2003; Stancheva, 2003; Boikovski et al., 2005b) and to establish the effect of certain factors (as order of lactation, year of birth, linear differentiation, age of mating, type of udder and method of calculation) on the variability of phenotypic parameters of milk yield traits and biological fertility (Boikovski et al., 2003а, 2003b, 2005a, 2006, Stancheva et al., 2006, 2014; Ivanova and Raicheva, 2008; Hinkovski et al., 2008; Raicheva and Ivanova, 2010, 2011a, 2011b). Scientific approach to the application of the methods for selection and productive traits monitoring in SPBM sheep herds of Agricultural Academy resulted in identifying the productive parameters, characterizing them as the best for growing rams and consolidated 29
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growth and high body weight (Raicheva et al., 2010). This is one of the prerequisites for getting a high milk yield. In elite herds there are genealogical (genetic) structures having an extremely high body weight (Stancheva, 2003). It is planned in future selection to be carried out optimization of live weight in order to maximize milk yield traits. The importance of the trait live weight in sheep breeding is determined by the fact that the high energy growth at a young age and realized other productivity determine the effectiveness of this sub-sector of livestock. Together with the other traits typical for different productive directions, body weight defines the parametres of the standards of individual sheep breeding lines. Bodyweight is selectable trait that is controlled in sheep of all productive directions and is known that is influenced by genetic and non-genetic factors (Mandal et al., 2003; Behzadi et al., 2007; Dass et al., 2008). The dynamics of phenotypic realization during the selection process represents undeniable interest. The goal of this study is to investigate the growth and development of sheep from breed Synthetic Population Bulgarian Milk (SPBM) of Agricultural Institute - Shumen at different ages and to define genetic determination of the trait by age categories. Materials and methods The survey covers 851 number ewes from Synthetic Population Bulgarian Milk, born during 2004 - 2008 in the flock of Agricultural Institute - Shumen. Based on available information of stud-book are set pure-breeding lines in the herd (27, 522, 913, 942, 976, 1254, 3156, 6988, 8265 and 88550) and experimental group with Chios blood (51). The trait live weight was studied on different age. Live weight was measured individually at weaning (on 100 days), 9 months old, 1.5, 2.5 years monitored and recorded by standard methods and guidelines laid down in the Instructions for performance monitoring and rating of sheep SPBM (2003, 2013). It was calculated variance estimations which influence some sources of variation on weight development. F-criteria of reliability of each of them were established. It is found genotypic effects ( h2 - heritability) of live weight at different ages (Harvey, 1990). Results and discussion It was found that during the experimental period, the live-weight of 9 months (46.59kg) measured in female SPBM hoggets represents 65.65% of that of the animals of 2.5 years (70.97 kg) and 91.06% - of 1.5 years (64.63kg) (Table 1). The variation in body weight in different age periods is distinguished. The highest value of the variation coefficient (C = 21.31%) was observed in live weight at weaning, followed by those for subsequent age groups in descending order, reaching 8.14% at 2.5 years. This is a logical consequence of the applied practice of pure-line breeding in the study herd. The main selection trait in SPBM sheep is lactation and on this reason allowed for breeding in some cases lambs with less body weight, which in later ages compensate the weight development. The indicator value for accuracy of the mean (E) are low - from 0.33% to 0.73%, and show that the results are representative of the general population.
30
SECTION 1. ANIMAL BIOTECHNOLOGY Table 1. Overall average and coefficient of variation of the live weight in SPBM sheep Live-weight n S C% at weaning 851 28,71 6,12 21,31 on 9 months 787 46,59 5,24 11,44 on 1.5 years 702 64,63 6,57 10,17 on 2,5 years 592 70,97 5,77 8,136 Table 2. Weight growth of SPBM sheep depending on the breeding line Live-weight № line at weaning on 9 months on 1.5 years n x C n x C n x C 1 27 20 28,10 14,96 17 47,82 10,36 17 64,12 8,45 2 51 20 25,65 13,28 20 47,40 12,42 18 70,22 7,45 3 522 107 28,72 21,85 98 47,54 10,42 88 64,67 8,49 4 913 64 30,14 18,89 56 46,18 9,65 51 65,55 11,11 5 942 97 30,53 27,76 94 43,42 11,73 81 63,25 11,49 6 976 39 30,05 23,76 36 45,67 14,07 28 62,86 11,90 7 1254 99 27,80 20,29 87 46,22 9,79 71 63,85 11,19 8 3156 222 28,19 19,31 206 47,21 11,08 189 64,51 9,72 9 6988 57 27,63 18,84 52 46,81 14,30 49 64,51 8,32 10 8265 26 26,23 22,42 26 44,73 9,00 25 64,80 9,22 11 88550 83 30,15 18,80 79 47,92 9,61 71 64,44 10,25 total 834 28,75 21,40 771 46,51 11,37 688 64,47 10,09 Significance of differences at: Р ≤ 0.001 2-3,4,5,11 5 - 3,4,7,8,9,11; 11-10 2 - 3,5,6,7,8,9,11 2-6; 7-5,11; 9-5,11; Р ≤ 0.01 10 - 4,5,11 5 - 1,2; 8 - 10; 2 - 1,4,10 2-7,1; 4-8; 7-4; 9-4; Р ≤ 0.05 10-6 11 - 7,4; 1 - 10;
Е 0,73 0,41 0,38 0,33
on 2.5 years n x C 12 69,33 4,70 16 74,44 5,48 71 70,25 6,62 44 71,18 9,08 75 69,21 8,38 22 68,96 8,06 54 72,54 8,18 170 70,99 7,77 39 71,36 9,71 23 72,35 7,59 52 70,71 9,19 578 70,87 8,14 2-3 2 -1,6,11 2 - 4,9; 7 -1,3,6; 10 - 1,6;
The analysis of the weight development depending on the breeding line (Table 2) shows that animals from line 51 (with Chios blood) at weaning reach the lowest live weight - 25.65kg compared with those of pure-bred lines of SPBM. This we believe is due to genetic (type of birth, maternal effect) and environmental factors and is consistent with those reported by Fernandes et al. (2001); Nedelchev et al. (2005); Kuchtík and Dobeš (2006); Petrović et al. (2012) and Taskin et al. (2012) results in similar kind of studies. With the highest live weight were lambs of 942 line (30.53kg), followed by its coeval of breeding lines 88550 (30.15kg), 913 (30.14kg) and 976 (30.05kg). Our results for the animals of SPBM and those with Chios blood are higher than those reported by Djorbineva et al. (2008). The lower values of the live weight at weaning of SPBM animals reported by Nedelchev et al. (2003); Hinkovski et al. (2008) and Iliev (2013) are probably due to the different weaning age and the environmental conditions. The differences between the individual lines have been demonstrated in different degrees of significance (P ≤ 0.05, P ≤ 0.01, P ≤ 0.001). The inside-linear variability of the live weight at weaning shows medium to very high values (С=13.28 to С=27.76%). On nine months between the lines there is some change in weight development. Animals from the breeding line 942 of SPBM that at weaning were recorded the highest live-weight now presented the lowest among all other - 43.41kg (P ≤ 0.01, P ≤ 0.001), with 31
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the exception of 8265 and 976 lines, and the trend of slight decline in weight was maintained on next age groups. From leading purebred lines with the highest body weight at this age are animals of line 88550 (47.924kg) with proven differences compared with lines 1254, 913 (P ≤ 0.05) and 8265 (P ≤ 0.001). Proven differences are available for animals from line 3156 compared to 8265 (P ≤ 0.01) and for originating from line 27 compared to 8265 (P ≤ 0.05). It is striking that weaned with low body weight lambs from a breeding line 51 (with Chios blood), after elimination of maternal effect and likely heterosis effect manifestation, offset the weight development and on 9 months reached 47.40kg live-weight as are aligned and in some cases superior to their peers from purebred lines (P ≤ 0.01 compared to line 942). The tendency for superiority in weight development in them is maintained in later life. Sheep from line 51 are with the highest body weight on 1.5 years - 70.22kg, with proven differences compared with animals of all leading SPBM lines (P ≤ 0.01, P ≤ 0.001). On 2.5 years they again reached the highest live weight - 74,44kg as observed differences are proven to breeding lines 913, 6988 (P ≤ 0.05), 88550, 3156, 976, 27 (P ≤ 0.01) and line 522 (P ≤ 0.001). The same age with the highest live-weight from pure-bred lines represent animals from line 1254 (72.54kg) with proven differences compared with lines 522, 976 and 27 (P ≤ 0.05). For animals of line 8265 are available proven differences compared breeding lines 976 and 27 (P ≤ 0.05). The obtained values for weight development at different ages depending on the breeding line showed that SPBM sheep are characterized by high body weight, and the most stable in this regard are presented the sheep on line 51 (with Chios blood) and those from pure-bred SPBM lines - 27, 3156, 88 550 and 913. After weaning, inside-linear variation of the body weight follows declining trend, reaching 8.14% in 2.5 years. Table 3. F –test of significance of the variance analysis of live weight in SPBM sheep Live weight № Sources of variation at weaning on 9 months on 1.5 years on 2.5 years n.s +++ ++ ++ 1 Father line n.s + n.s ++ 2 Mother line +++ +++ +++ +++ 3 Year of birth Significance of differences at: + P ≤ 0,05; ++ P ≤ 0,01; +++ P ≤ 0,001
The analysis of variance reported high significant impact of the year of birth on the phenotypic expression of the trait live weight of all ages (P ≤ 0.001) (Table 3). Stancheva (2003) in SPBM animals and Taskin et al. (2012) in sheep of synthetic dairy breed Sönmez also found significant effect of year of birth on sheep weight development. Father line has reliably high impact of the live weight at 9 months (P ≤ 0.001) and less at 1.5 and 2.5 years of age (P ≤ 0.01). The influence of the mother line of the studied trait was less for 9 months (P ≤ 0.05) and 2.5 years of age (P ≤ 0.01). Table 4. Heritability (h2) of the live weight in SPBM sheep Live weight, kg n at weaning 821 on 9 months 760 on 1.5 years 680 on 2,5 years 571 32
h2 0,094 0,306 0,340 0,068
SЕ 0,071 0,106 0,720 0,091
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For study period is established low hereditary variance of trait live weight at weaning and at 2.5 years of age (h2 = 0.094 and h2 = 0.068) and moderately high for 9 months and 1.5 years (h2 = 0.306 and h2 = 0.340) (Table. 4). Received low estimate on h2 for trait live weight at weaning - 0.094 confirms our opinion that this is not a manifestation of the genotype, but the logical consequence of allowing lambs for breeding with lower body weight at the discretion of the breeder, maternal effects and environmental factors. Slavov et al. (2008) reach a similar conclusion in a study of North East Bulgarian thin-fleece sheep breed. The established medium-high values for body-weight heritability on 9 months and 1.5 years - 0,306 and 0,340 were lower than those obtained in our study of the same flock at the time of population creation (Stantcheva, 2003). This, we believe is due to the interaction genotype - environment and changes in ontogenetic development. Conclusions - Body-weight on 9 months measured of the female lambs SPBM represents 65.65% of that of the animals of 2.5 years (70.97 kg), and 91.06% - of 1.5 years. - The highest values of the coefficient of variation (C = 21.31%) is a live weight of 100 days of age, and then in the other age groups in a declining trend, reaching 8.14% of 2.5 years. - Values obtained for the weight development at different ages depending on the breeding line showed that SPBM sheep are characterized by high body weight, and the most stable in this regard are presented the sheep of line 51 (with Chios blood) and those from purebred SPBM lines - 27, 3156, 88 550 and 913. - Year of birth has a high significant impact on the trait live weight of all ages (P ≤ 0.001). The father line has a highly reliably effect of live weight at 9 months (P ≤ 0.001) and less at 1.5 and 2.5 years of age (P ≤ 0.01). - The value of heredity of the live weight categories are of low to medium (0. 094 to 0.340). The main reason for this may be given targeted selection, interaction genotype environment and changes in ontogenetic development. Acknowledgment Research was part of the project Б 01/22 от 05.12.2012г. (2012-2014) 'Development of DNA markers (CAST, MSTN) for fattening ability and meat quality in Synthetic population Bulgarian Milk, Karakachanian and Copper Red Shumen sheep breeds' financed by the Ministry of Education, Youth and Science, Republic of Bulgaria.
References Behzadi R., Shahroudi, F. E. аnd Van Vleck, L. D. 2007. Estimates of genetic parameters of growth traits in Kermani sheep. J. Animal Breeding and Genetic. 5: 296- 301. Boikovski, St., N. Stancheva, G. Stefanova, D. Dimitrov. 2003a. Influence of Some Factors of the Milk Composition and the Yield of Trait of Sheep from The Newly Created Milk Sheep Breed, Bulgarian Journal of Agricultural Science,9, 2, 243-249.
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Boikovski St., N. Stancheva, G. Stefanova, D. Dimitrov. 2003b. Influence of Some Factors on Biological Prolificacy in Sheep from The Newly Created Milk Sheep Breed, Bulgarian Journal of Agricultural Science, 9, 3, 391-395. Boikovski St., N. Stancheva, G. Stefanova, D. Dimitrov. 2005a. Milk Composition of the Sheep from Newly Created Milk Sheep Breеd, Bulgarian Journal of Agricultural Science, 11, 5, 619-632. Boikovski, St. N. Stancheva G. Stefanova, 2005b. The Newly Created Milk Sheep Breed, „Eyro - Klima‖ Shoumen, pp.222 (in Bulgarian) Boikovski St., G. Stefanova, N. Stancheva. 2006. Milk Yield for milking period in the Sheep from the Newly Created Milk Breed in Bulgaria, Bulgarian Journal of Agricultural Science, 12, 1, 145-152. Dass G, Prasad H, Mandal A, Singh Mk, Singh N.P. 2008. Growth characteristics of Muzaffarnagari sheep under semi-intensive feeding system. Indian J. Anim. Sci., 78(9): 1032-1033. Djorbineva M., Zh. Peeva I. Dimitrov, S. Laleva, 2008. Productive characteristics of the first generation crossings between Synthetic bulgarian dairy population and Chios sheep. Union of Scientific – Stara Zagora, International Scientific Conference, v. Animal Studies. (in Bulgarian) EASRAB. 2003. Instructions for monitoring the signs of sheep. Sofia, pp. 45. (in Bulgarian) Fernandes A.O., Buchanan D., Selaive-Villarroel A.B. 2001. Environmental effects on growth rate of Morada Nova hair lambs in northeastern Brazil. Rev. Bras. Zoot., 30, 1460– 1465. Harvey, W., 1990. User’s grueide for LSMLML and MIXMDL, PC-2 version, Mimeo, Ohio, USA, pp 91. Hinkovsky Tz., Е. Raicheva, N. Metodiev, 2008. Estimation of productivity of ewes from the Bulgarian Dairy Synthetic Population. Animal Science, № 3, 35-41. Iliev, М., 2013. Productive characteristics of sheep from different productive areas, bred in the Institute of Agriculture – Karnobat. Sheep news, 1 - 2, 37-47. (in Bulgarian) Ivanova Т., Е. Raicheva, 2008. Assessment the effect of some factors on lactation. Conference Proceedings '80 years Agrarian Sciences in the Rhodopes', 67-71. (in Bulgarian) Kuchtík J., Dobeš I. 2006. Effect of some factors on growth of lambs from crossing between the Improved Wallachian and East Friesian. Czech J. Anim. Sci., 51, 54–60. Lazarov, V., L. Mihailova, М. Iliev, 2002. Creation of population of sheep with increased milk yield. Animal Science, №6, 11-13. (in Bulgarian) Mandal A., Pant K. P., Nandy D. K., Rout P. K. аnd Roy R. 2003. Genetic analysis of growth traits in Muzaffarnagari sheep. Tropical Animal Health and Production. 35: 271284. Nedelchev D., Е. Raicheva, Y. Petrova, 2003. Characteristics of the productivity of dairy sheep crosses. Animal Science, № 3-4, 111-114. (in Bulgarian) Nedelchev D., Е. Raicheva, Е. Kistanova, G. Dimov, Е. Kipriotis, K. Kousenidis, Th. Papadopoulos 2005. Results from crossing local sheep and rams of the breed Chios. Animal Science, 5, 293-297. (in Bulgarian) Petrović V. C., M. P. Petrović, M. M. Petrović, Z. Ilić, N. Maksimović, D. Ruţić Muslić, N. Stolić. 2012. Estimation of phenotypic and genetic Trends of the growth traits in Lipska and Svrljig sheep. Biotechnology in Animal Husbandry 28 (4), p 743-749. 34
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Raicheva Е., D. Nedelchev, Y. Petrova, 2003. Complex score assessment of dairy sheep productivity. Animal Science, № 1-2, 77-80. (in Bulgarian) Raicheva Е., Т. Ivanova, 2010. Duration of lactation and milk production in ewes from synthetic population Bulgarian milk depending on the order of lactation. Animal Science, Supplement 1, 58-63. (in Bulgarian) Raicheva Е., Т. Ivanova, St. Boykovski, N. Stancheva, М. Iliev, 2010. Status, Problems and Perspectives in the selection and breeding of sheep from Synthetic population Bulgarian milk kept in the institutes of the Agricultural Academy. Sheep news, 2: 5-8. (in Bulgarian) Raicheva Е., Т. Ivanova, 2011a. A previous study on the some parameters connecting whit the milk production at early conceived as ewe lambs from Synthetic Population Bulgarian Milk. Journal of Mountain Agriculture on the Balkans, 4:656-666. (in Bulgarian) Raicheva, E., T. Ivanova. 2011b. The effect of the age аt conceiving on the Productivity traits at dairy ewes in Bulgaria. Biotechnology in Animal Husbandry 27 (3), p 1147-1156. Petrović V. C., M. P. Petrović, M. M. Petrović, Z. Ilić, N. Maksimović, D. Ruţić Muslić, N. Stolić. 2012. Estimation of phenotypic and genetic Trends of the growth traits in Lipska and Svrljig sheep. Biotechnology in Animal Husbandry 28 (4), p 743-749. Slavov, R., Zh. Krastanov, P. Slavova, Т., Angelova, 2008. Analysis of the genetic varians of the North-East Bulgarian merino breed and in its crossing witn Australian merino, Ile de France and Booroola. Animal Science,№ 3, 168-171. (in Bulgarian) Stancheva, N., 2003. Phenotypic and genotypic parameters of selection indices in the Newly Created Milk Sheep Population in Bulgaria. Ph D Thesis, Sofia, pp.188 (in Bulgarian) Stancheva N., St., Boikovski, G. Stefanova, D. Dimitrov, А. Rusenov, 2006. Regularities in lactation persistency of lactation and possibilities for using part of lactation in tribal work in ewes from Synthetic Population Bulgarian Milk. International Scientific Conference - Stara Zagora, Jine 1-2, Vol. 2. Vet. Medicine, Animal Studies. 244-247. (in Bulgarian) Stancheva N., I. Dimitrova, S. Georgieva, 2014. Biological fertility and milk yield in Bulgarian Dairy Synthetic Population sheep according to breeding line. Agricultural Science and Technology, vol. 6, No 1, pp 17 – 20. Stancheva N., G. Dimov, Е. Baycheva. 2013. Instructions for monitoring the signs of sheep from Synthetic Population Bulgarian Milk. ―Himera‖, Shoumen, pp. 28. (in Bulgarian) Taskin T., M. Kaymakci, R. Sönmez, M. Yılmaz, H. E. Bardakcioglu. 2012. Genetic and non-genetic parameter estimates for growth traits of SÖNMEZ lambs. Veterinarija Ir Zootechnika (Vet Med Zoot). T. 60 (82), p 79-83.
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36
SECTION 1. ANIMAL BIOTECHNOLOGY UDC: 636.2.053.087.8 Original scientific paper
EFFECT OF MANNAN-OLIGOSACHARIDE BIO-MOS ON LIPIDS IN THE SERUM OF SIMENTAL CALVES Stolić N.1*, Milošević B.2, Spasić Z.2, Ilić Z.2, Pešić B.1 1
College of Agriculture and food technology, Prokuplje, Serbia 2 Faculty of Agriculture, University of Pristina, Serbia *
corresponding author: [email protected]
Abstract The effect of prebiotic preparation BIO-MOS („Altech', USA), based on mannanoligosacharide, on growing of suckling calves was investigated. In general, the results of the experiments showed that this prebiotic preparation for 1-3 month aged calves had stimulative effect on growth, improved the digestibility of nutritients and had positive effect on the immunological status of the animals.In this work the results of the change in total lipids in the calves blood serum are presented. The level of total lipids in the serum was analyzed after 30, 60 and 75 days of calves growth, both in the experimental and control group. The total lipids were 3.58±0.803 g/l day 30, 3.58±1.350 day 60, and there was a significant increase of this parameter at day 75 being 5.12±0.732. There was no statistically significant difference between the values obtained for the control group. Key words: Prebiotic, calves, blood serum, lipids. Introduction Dairy calves are susceptible to many pathogens and nutritional problems during the first few weeks of life. For several years antibiotics have been used to overcome these problems, to obtain economic benefits in terms of improved calves performance and reduced medication costs. However, the use of antibiotics in animal husbandry can cause antibiotic resistance of pathogen bacteria. Thus, there is increasing interest in replacement of antibiotics with natural alternatives, such as probiotics, prebiotics and symbiothics in order to enhance animal performance and health (Ballou, 2011; Krol, 2011; Roodposhti and Dabiri, 2012). Probiotics are used to balance the host intestinal microbial eco-system and restore its resistance to diseases, by intake of viable probiotic microorganisms. On the other hand, prebiotics are non-digestible food ingredients that selectively stimulate the growth and/or activity of certain bacteria in the colon. It is known that prebiotics can have positive effect on various physiological processes and improve animal health, primarily by modification of the composition and functionality of gastrointestinal microflora. These processes include digestion, nutrient utilization, natural defence mechanism against pathogen and overall immunity system and finally, quality of animal products. Dietary prebiotics could have effect on lipid metabolism. It has been reported that consumption of prebiotics lower the concentration of both cholesterol and triglycerides (Roodposhti and Dabiri, 2012; Samanta et al, 2013). Among prebiotics mainly non-digestible oligosaccharides are ranked, such as mannan-oligosaccharides (MOS) – a complex 37
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carbohydrates derived from cell walls of Saccharomyces spp. yeast. Mannoligosaccharides have high resistance toward acidity and human gastrointestinal enzymes, and thus rich the colon undigested, being substrates for beneficial intestinal bacteria. On the other hand, they can reduce pathogen bacteria due to specific molecular features. Fimbriae are rich in lectins and it is essential for bacteria adherence to the mannose-containing cells in the intestinal tract. Mannan-oligosaccharide provide a mannose-rich source for bacteria attachment, instead of being attached to the gut wall and cause disease. The selectivity of mannanoligosaccharides is based on that some beneficial bacteria (Bifidobacterium longum, Lactobacillus casei, L. acidophitus, L delbruckii) produce the enzyme mannanase which targets its hydrolysis and therefore do not enable formation of complexes, unlike to those undesirable pathogen strains lacking the enzyme (Sharon and Lis, 1993; Fekete et al, 2002). Due to its beneficial effects, dietary intake of mannan-oligosaccharide can contribute to increase vitality of animals, enhancing both the digestive and immune system that further leads to optimum production and economic benefits. The aim of this work to present the effect of dietary intake of prebiotic preparation BIO-MOS, based on mannan-oligosaccharide, which has been suggested as an effective prebiotic preparation, on lipids in the serum of growing of suckling calves. Material and methods The experiment was organized as a group control system, with calves of domestic spotted cattle in the type of Simmental. Calves were healthy, progressive, and vital without shortcomings and defects. Calves were housed in facilities for the calves, where they were located after the separation from their mothers. The first 5 days after receipt of colostrum they started receiving the mother's milk, and later consumed the cumulative milk. Calves were marked with tattoo stamps and numbered ear tags. Microclimatic conditions in the facilities during the experiments, keeping, and care, feeding and watering were identical for all calves. Two groups of calves were formed, a control group that was fed without added manan-oligosaccharide, and the experimental group, which received in milk the mentioned prebiotic. Each group of calves was comprised of 16 calves so that in total experiment was conducted with 32 calves. At the start the calves were fed fresh colostrums. After the fifth day they were fed with 6 l of the cumulative milk produced on the farm. Treatment group was supplemented with 4 g prebiotic Bio-Mos/calf/day. The calves were fed via nipple bottle feeder, signed for each calf. After the day 15 a mixture was included in the daily diet according to the calves' physiological requirements and regulations on quality of animal food and feed in Serbia (Sl. List 20/2000, 38/2001). To avoid imprecise estimates, alfalfa flour was used up to 15 % in the mixture instead of hay. The blood samples were collected at the farm at defined time (day 30, 60 and 75) and analysed at the Institut of Veterinary of Serbia in Belgarade. Total lipid level was determined by the sulfo-phospho-vanillin color reaction, trygliceride by GPO/PAP method and level of total cholesterol by CHOD/PAP, aas described by Stolic, 2007 in more detail. Data were expressed as mean ± standard deviation for triplicate determinations and the statistical analysis was carried out using Install Prism4.
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Results and discussions The effect of prebiotic preparation BIO-MOS („Altech‖, USA), based on mannanoligosacharide, on growing of suckling calves was investigated. In order to determine whether this prebiotic induces significant changes in the level of blood serum proteins and lipids, the biochemical analysis of the blood samples was performed after 30, 60 and 75 days of growth, both in the experimental and control group. The results of the level of total proteins, albumins and globulins are reported in our previous work, Stolić et al 2015. Figure 1 shows the total lipids in the serum of the calves. The results showed that the level of the total lipids did not significantly changed the this period, being 3.58±0.8 g/l at day 30, 3.58±1.35 g/l at day 60, and 5.12±0.73 g/l at day 75. There was no statistically significant difference between the experimental and the control group.
Figure 1. Level of total serum lipids in suckling calves.
As the level of total lipids did not change in response to the administration of the prebiotic, more detailed analysis was undertaken. The level of triglycerides and total cholesterol was examined in the blood samples. Figure 2 shows the level of triglycerides in the serum.
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Figure 2. Level of serum triglycerides in suckling calves. The serum trygliceride in the experimental group were 0.25±0.078 mmol/l, 0.23±0.032 mmol/l and 0.47±0.12 mmol/l at day 30, 60 and 75 respectively (Figure 2). It can be seen that serum triglyceride had normal level and also did not change in direct response to the prebiotic intake, but most likely due to animal normal diet and growth. Figure 3 shows the level of total cholesterol. The level of total cholesterol was 2.23±0.817 mmol/l at day 30, 1.82±0.543 mmol/l at day 60 and 3.3±0.45 g/l at day 75. There were no significant difference between the experimental and control group.
Figure 3. Level of total serum cholesterol in suckling calves.
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Conclusions The overall that this prebiotic preparation for 1-3 month aged calves had stimulating effect on growth, improved the digestibility of nutririents and had positive effect on the immunological status of the animals. However, the results of the blood serum analzysis, including both proteins and lipids, show that the addition of the prebiotic Bio-Mos had no effect on these parameters. The change of total proteins and lipids are rather related to the normal diet and animal growth. References Ballou M. A. (2011). Case Study: Effects of a blend of prebiotics, probiotics, and hyperimmune dried egg protein on the performance, health, and innate immune responses of Holstein calves. The Professional Animal Scientist 27, pp. 262–268. Ferket P.R., Parks, C. W., Grimes, J. L. (2002). Benefits of dietary antibiotics and mananoligosaccharides supplementation for poultry. Multi-State Poultry Meeting. Król B. (2011). Effect of mannanooligosaccharides, inulin and yeast nucleotides added to calf milkreplacers on rumen microflora, level of serum immunoglobulin and health condition of calves. Electronic Journal of Polish Agricultural Universities 14: #18. Roodposhti P. M., Dabiri, N. (2012). Effects of Probiotic and Prebiotic on Average Daily Gain, Fecal Shedding of Escherichia Coli, and Immune System Status in Newborn Female Calves. Asian-Aust. J. Anim. Sci. 25, pp. 1255 – 126. Samanta A.K., Natasha Jayapal, Senani, S., Kolte, A.P., Manpal Sridhar (2013). Prebiotic inulin: Useful dietary adjuncts to manipulate the livestock gut microflora. Brazilian Journal of Microbiology 44, pp. 1-14. Sharon, N., Lis, H. (1993). Carbohydrates in cell recognition. Scientific American, 1, pp. 5-7. Stolić N. (2007). Impact investigation of added mannan-oligosaccharides (BIO-MOS) on growing of suckling calves. Doctoral Dissertation, University of Priština. Stolić, N., Pešić, B., Milošević, B., Spasić, Z., Ilić, Z. (2015). Effect of prebiotic BIOMOS onserum proteins in suckling calves, Research People and Actual Tasks on Multidisciplinary Sciences, 24-28 June Lozenec, Bulgaria, Proceedings of the fifth International Conference, pp. 94-97.
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SECTION 1. ANIMAL BIOTECHNOLOGY UDC: 637.142.2 Original scientific paper
OPTIMIZATION OF ENZYMATIC ASSAY OF SUPEROXID DISMUTASE AND GLUTATION PEROXIDASE ACTIVITY IN MILK WHEY Trajchev M.1*, Nakov D.1, Gjorgoski I.2 1
Faculty of Agricultural Sciences and Food, Institute of Animal Biotechnology, Department of Animal Health and Animal Welfare, University „St. Cyril and Methodius―, Skopje, Republic of Macedonia 2 Faculty for Natural Sciences, University „St. Cyril and Methodius―, Skopje, Republic of Macedonia *
corresponding author: [email protected]
Abstract General spectrofotometric kinetic protocols are described to measure the antioxidant enzyme activity of superoxide dismutase (SOD) and glutathione peroxidase (GPx) in milk whey. The SOD convert superoxide radical into hydrogen peroxide and molecular oxygen, while the GPx convert hydrogen peroxide into water. In this way, two toxic species, superoxide radical and hydrogen peroxide, are converted to the harmless product water. The applicability of this methods for milk samples from dairy cows were tested. SOD activities in the milk whey was determined by the autoxidation of pyrogallol in presence of DTPA in TRIS-HCl buffer, pH= 8.5. The effect of reaction conditions on enzymatic inhibition of pyrogallol autooxidation was studied when different amounts of sample were added in reaction mixture. The reaction mixture for determination of GPx activity contained EDTA, reduced glutathione, glutathione reductase, NADPH and cumene hydroperoxide in phosphate buffer, pH= 7.6. The effect of reaction conditions on enzymatic NADPH consumption was studied when different amounts of reduced glutathione and whey sample were added in reaction mixture. The both spectrophotometric methods were modified in order to accommodate kinetic analyses in 96-well micro plates. The rate of pyrogallol autooxidation and NADPH oxidation linearly depend from amount of whey sample in reactive mixture. Applying higher amounts of milk whey in reactive mixture was decreasing optical density reading of pyrogallol autooxidation at 415 nm and increasing the optical density readings of NADPH oxidation at 340 nm, in a linear way. The methods could be validly applied for milk whey samples. Key words: dairy cows, milk whey, superoxid dismutase, glutathion peroxidase. Introductıon Reactive oxygen species (ROS) are produced in many aerobic cellular metabolic processes. They include, but are not limited to, species such as superoxide and hydrogen peroxide which react with various intracellular targets, including lipids, proteins, and DNA (Bandyopadhyay et al., 1999). Although ROS are generated during normal aerobic metabolism, the biological effects of ROS on these intracellular targets are dependent on their concentration and increased levels of these species are present during oxidative stress. 43
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Cells contain a large number of antioxidants to prevent or repair the damage caused by ROS, as well as to regulate redox-sensitive signaling pathways (Halliwell and Gutteridage, 2007). Three of the primary antioxidant enzymes contained in mammalian cells that are thought to be necessary for life in all oxygen metabolizing cells are superoxide dismutase (SOD), catalase, and a substrate specific peroxidase, glutathione peroxidase (GPx) (Halliwell, 1987). The SOD catalyze the dismutation of the superoxide radical anion into oxygen and hydrogen peroxide, while the catalase and peroxidases convert hydrogen peroxide into water and in the case of catalase to oxygen and water (Bordo et al., 2000). The net result is that two potentially harmful species, superoxide and hydrogen peroxide, are converted to water. The oxidative stability of milk is a balance between pro- and anti-oxidative factors (Lindmark-Mansson and Akesson, 2000); hence, identification and characterization of these factors is necessary to predict the rate of oxidation. A number of low-molecular weight antioxidants have been described in milk, e.g., ascorbate, urate and atocopherol, to mention the most important as judged from their abundance and consumption during oxidation (Nielsen et al., 2001). However, milk also contains antioxidative enzymes, such as catalase, superoxide dismutase and glutathione peroxidase (GPx) that can reduce oxidative deterioration, but the importance and relative contributions of these enzymes to the oxidative stability of milk is unclear (Lindmark-Mansson and Akesson, 2000). The indirect spectrophotometric assay has been developed for measuring SOD and GPx activity in plasma, erythrocyte lysates, tissue homogenates and cell lysates (Paglia and Valentine, 1967; Marklund and Marklund,1974). To some extent, the limited knowledge of the role of SOD and GPx in milk may be explained by the lack of suitable activity assays. Spectrofotometric kinetic assays were already successfully been applied for blood samples, we aimed to establish this tests for milk samples to evaluate it’s relationship with occurrence of oxidative stress in dairy cows and herein report the limitations we observed for the application of the assays in milk. Materıals and methods Milk (raw, i.e. unhomogenized and unpasteurized) was obtained from private dairy farm. Milk was defatted by centrifugation at 5000 rpm for 20 minutes at 40C, and skim milk beneath the cream was gently poured into a clean container. Acid whey was prepared by treating the samples with 1M HCl to pH 4.6, followed by centrifugation (5000 rpm for 20 min at 40C) in order to remove precipitated proteins. The pH was adjusted usually to pH 7.6 using 1M NaOH. The samples were stored in stoppered plastic tubes at -800C. Before use the aliquots were thawed. SOD activities in the milk whey was determined by the autoxidation of pyrogallol in presence of DTPA in TRIS-HCl buffer, pH= 8.5. The assay medium in a total volume of 200 μl contained 100 mM Tris HCl buffer, pH 8.5, 1mM DTPA (diethylenetriaminepentaacetic acid) in presence of 0.2 mM of pyrogallol. Pyrogallol (1,2,3-benzenetriolo) was dissolved in 10 mM HCl to obtain stock solution (2 mM) which was prepare always fresh. Measurement of SOD activity was carried out spectrophotometrically at 250C by the method of Marklund and Marklund (1974) with some modifications in order to accommodate kinetic analyses in 96-well microplates (Gao et al.,1998). The reading of absorbance at 415 nm for 3 minutes was made in interval of 60 seconds after an induction period of some 10 seconds. The bovine Cu/Zn-SOD was used as 44
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standard for generation of a standard curve, a range activity of 100 U/ml to 0.09765 U/ml. The values obtained for SOD activity in the milk sample were then calculated from the standard curves using logarithmic regression and were finally expressed as percentage of inhibition of the pyrogallol autoxidation rate. One unit of the enzyme activity is defined as the amount which produced 50% inhibition of pyrogallol autoxidation under the standard assay conditions. The effect of reaction conditions on enzymatic inhibition of pyrogallol autooxidation was studied when different amounts of sample were added in reaction mixture. The principle of the coupled enzymatic assay first described by Paglia and Valentine (1967) and modified according Chen et al. (2000) in order to accommodate kinetic analyses in 96-well microplates. The reaction mixture for determination of GPx activity in a total volume of 200 μl contained 50 mM potassium phosphate buffer, pH= 7.6., 5 mM EDTA (ethylenediaminetetraacetic acid), reduced glutathione (GSH), 1 U/ml glutathione reductase (GR), 0.25 mM NADPH. The measurements were carried out at two different concentrations of GSH i.e. 2 mM and 1 mM. Then cumene hydroperoxide (cH2O2) was added to initiate the reaction in a total volume of 30 μl at 370C. Stock solution of cumene hydroperoxide (9.9 mM) in 50 mM phosphate buffer, pH= 7.6. was prepare always fresh. The effect of reaction conditions on enzymatic NADPH consumption was studied when different amounts of reduced glutathione (GSH) and whey sample were added in reaction mixture. The assay temperature was 370C. The reading of absorbance was made at 340 nm for 3 minutes in interval of 60 seconds after an induction period of some 10 seconds. One unit of GPx was defined as the activity that causes the formation of 1,0 μmol/L NADP+ from NADPH per minute at pH 7.6 at 370C in a coupled reaction. All chemicals were of the highest purity grade. TRIS, pyrogallol, KH2PO4, EDTA and cH2O2 were purchased from Merck, Darmstadt, Germany. DTPA, Cu/Zn-SOD, GSH, NADPH and GR were obtained from Sigma Chemical Co., St. Louis, MO, USA. The absorption was determined with a spectrophotometer Bio-Rad 680 XR, microplate reader. Results The standard curves demonstrated a linear relationship between absorption measurements and the SOD activity. The colored dye is directly proportional to the percentage of the pyrogallol autoxidation rate. Since the rate change of absorbance per minute at 415 nm is proportional with the amount of superoxide anion, the SOD activity in reactive mixture as an inhibition activity of pyrogallol autooxidation is proportional with decrease in color development at 415 nm. Therefore, the increasing of absorbance at 415 nm (A415) per minute followed a linear kinetics, showed on Figure 1.
45
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Blank 100
0,120
50
0,100
25 12,5
0,080
6,25 3,125
0,060
1,5625 0,78125
0,040
0,3906
0,020
0,1953 0,09765
0,000 0
1
2
3
Time (min)
Figure 1. Linearity of standard curves using different concentrations of bovine Cu/Zn SOD (μU/ml)
On Figure 2 is showed standard curve of an SOD inhibition activity of pyrogallol autooxidation. The logarithmic function of standard curve was used for estimation of SOD activity in milk sample. 120
y = 15,288Ln(x) + 42,741
% inhibition
2
R = 0,91 100 80
%
60 40 20
0,01
0,10
0 1,00 SOD (U/ml)
10,00
100,00
Figure 2. Representative standard curve using different concentrations of bovine Cu/Zn SOD. The regression equation was y=15.288Ln(x) + 42.741 with r = 0.91
Using milk samples, the linearity of the assay was established in a range between sample dilution 1:40 to 1:2.5 (Figure 3).
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А4150,600
Blank 1:40
0,500
1:20 1:13,33
0,400
1:10 1:8
0,300
1:6,67 1:5
0,200
1:4 1:3,33
0,100
1:2,85 1:2,50
0,000 0
1
Time (min)
2
3
Figure 3. Evaluation of linear kinetic related to SOD activity using different volumes of milk serum % 120
y = -4761,9x + 100 Standard curve
R2 = 1 100 80
Milk sample
60 40
Linear (Standard curve)
20 0 0,000
0,005
0,010
ΔA415
0,015
0,020
0,025
Figure 4. Net rate of absorbance change (standard curve vs. different dilutions of milk sample)
The limit of detection was defined as the concentration of bovine Cu/Zn SOD that yielded net rate of absorbance change different from the blank value measured by using H2O and corresponds to the lowest standard concentration of 0.09765 U/mL SOD. The measured SOD activity in milk sample was in the range of standard curve which allow reliable measurements. Figure 5 shows apparent rates of changes in absorbance at 340 nm as a measure of the oxidation of NADPH in the presence of milk, as a function of substrate concentration (GSH). The validity of this procedure is demonstrated by the constant relationship between decrease in absorbance at 340 nm per min and enzymatic NADPH consumption. Aimed to 47
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get better linear kinetic in enzymatic NADPH oxidation, there was used different dilutions of milk sample and different concentration of GSH in reaction mixture. The linearity of the assay was established in a range between sample dilution 1:20 to 1:1000. A 340 nm1,2
1:20 1:20 GSH1/2
1,0
1:40 1:40 GSH1/2
0,8
1:100 1:100 GSH1/2
0,6
1:200 1:200 GSH1/2
0,4
1:400 1:400 GSH1/2
0,2
1:800 1:800 GSH1/2
0,0
1:1000
0
1
Time (min)
2
3
1:1000 GSH1/2
Figure 5. Evaluation of linear kinetic related to GPx activity using different volumes of milk serum
Dıscussıon Most procedures for determining SOD are based on the ability of the enzyme to inhibit oxygen-dependent reactions. Superoxide, produced by an in vitro reaction, reacts with the indicator giving absorbance at specific wavelength that increases with time. The correlation between the SOD concentration and the inhibition rates of the superoxide reaction with the indicator, allows to determine the enzyme activity. The inhibition of pyrogallol autoxidation brought about by SOD can be employed in a rapid and convenient method for the determination of the enzyme. After the above investigation we chose to use 0.2 mM pyrogallol in air-equilibrated 50 mM Tris-HCl buffer pH 8.5, containing 1 mM diethylenetriaminepentaacetic acid (DTPA). Pyrogallol autoxides rapidly in alkaline aqueous solution, its autoxidation rate is measured from the linear increase in adsorbance at 415 nm for some minutes (usually from 1 to 3 minutes each assay). The increment of the band is linear within 7-10 minutes, then the colour turns green and finally, after a few hours, a yellow colour appears. The increase in the absorbance at 420 nm after addition of pyrogallol was inhibited by the presence of SOD. As reported by Marklund and Marklund (1974), the rates of pyrogallol autoxidation were determined at pH 8.0 and 9.2. The rate of increase in absorbance at 420 nm per min was increased with increasing pH from 8.2-8.9. The autoxidation of pyrogallol was rapid at pH 9.2 but followed a non-linear pattern. By contrast, at pH 8.0 the autoxidation of pyrogallol was slow but followed a linear kinetics. We observed that better sensitivity and reproducibility were obtained when the pH of the assay mixture was kept at 8.5. At pH 7.9 the autoxidation is inhibited to 99% by Cu/Zn SOD, being the enzyme activity independent of pH in the range 5.5-9.5 (Gao et al., 1998). 48
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The sensitivity to SOD decreases when the pH is increased, but still amounts to 93% at pH 9. At higher pH values there is a strong decrease in the sensitivity to SOD and at pH 10.6 the autoxidation is inhibited to only 15% by SOD (Violi et al., 1985). Higher concentrations of pyrogallol decrease the sensitivity of assay (Gao et al., 1998). At 0.5 mM pyrogallol the sensitivity is decreased by a factor of 3.3. Iron, even in trace amounts, accelerates pyrogallol autoxidation. DTPA was found to prevent interference from Fe2+ (as well as from Cu2+ and Mn2+) and was therefore chosen as chelator in the assay medium. In the absence of DTPA the autoxidation is faster than in its presence, and it tends to vary in rate and is less affected by superoxide dismutase. In the presence of DTPA the rate is independent of the concentration of the chelator, which indicates that the effect of DTPA is only due to its binding of traces of metal ions. Most measurements in the present report were performed in the presence of 1 mM DTPA. In the present method one unit corresponds to 100 ng bovine Cu/Zn SOD in a total volume of 1 ml; thus the method is about as sensitive as the method based on the reduction of cytochrome c by xanthine oxidase (Beyer and Fridovich, 1987). Using different amounts of the milk whey, the linearity was confirmed. This method is fast, cheap, simple, sensitive and requires common diagnostic tools as a commercial spectrophotometer featured by a visible light and very few amount of enzyme. Glutathione peroxidase (GPx) activity has been measured through a coupled reaction with glutathione reductase (GR). In the assay, GPx reduce cumene hydroperoxide (cH2O2), and oxidize reduced glutathione (GSH) to oxidized glutathione (GSSG). The generated GSSG is reduced to GSH with through oxidation of NADPH to NADP+ by glutathione reductase (GR). The decrease of NADPH is proportionally to GPx activity in the reactions. The decrease of NADPH can be easily measured by absorbance at 340 nm. Chen et al. (2000) found that GPx activity in added whey or milk was found to increase as buffer pH increased from 7.0 to 8.0, and a pH of 7.6 was selected to avoid a high water-blank. The same authors reported When the phosphate concentration was changed from 25 to 100 mM/L, GPx activity decreased and the blank increased, showing that lower buffer concentration favored GPx activity. When the assay was performed at 250C instead, the GPx activity in whey was 30% lower than at 370C. According them, the GPx activity in milk was only slightly higher than that in whey indicating that most of the GPx in milk was recovered in whey. The oxidation of NADPH was dependent from the presence of GSH and increased by the addition of milk. The better sensitivity and reproducibility were obtained when assay was performed in presence of 1 mM GSH. Usually for milk assays a lower GSH concentration, a lower peroxide concentration and a more alkaline pH were used than in blood assays (Debski et al., 1987; Bhattacharya et al., 1988; Avissar et al., 1991). One reason for this is probably that erythrocytes contain mainly cellular GPx and milk mainly extracellular GPx (Bhattacharya et al., 1988; Avissar et al., 1991; Lindmark-Maensson and Akesson, 2000). An important difference between these two forms is that extracellular GPx has a lower rate constant for GSH than cellular GPx (Esworthy et al., 1993) and at cellular levels of GSH (millimolar) the eGPx reaction appears to be up to 10-fold slower than the cellular GPx reaction. Moreover, cellular GPx could not be saturated by GSH and that a high GSH concentration could inhibit GR, thus decreasing the overall reaction rate. Furthermore, there was dependence between substrate addition and presence of milk sample in reaction mixture, where the oxidation of NADPH clearly increased following addition of cumene H2O2. Although the absorbance decreases during the assay 49
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due to decreasing concentrations of NADPH, the negative rates have been converted to positive values throughout. Avissar et al. (1991) found the same GPx activity in milk using either H2O2 or tert-Butyl Hydroperoxide (t-BHPx) as substrate. The presence of EDTA turned out to be critical for the apparent oxidation of NADPH by milk. Contrary, some authors failed to detect any specific activity of GPx in bovine milk. Stagsted (2006) found GSH-independent NADPH oxidation in milk and GPx-catalysed tBHPx - independent oxidation of GSH was observed. Author used 25 mM EDTA to allow optical measurements in concentrated milk samples. Although only 50% of the apparent turbidity could be eliminated with EDTA. Furthermore, they observed that concentrations of EDTA above 6 mM completely inhibited the apparent activity in milk. Therefore, we perform assay on milk whey, previously cleaned from turbidity and pH was adjusted to pH 7.6. There are several reports on the activity of SOD and GPx in milk (Bhattacharya, et al., 1988; Debski et al., 1987; Avissar et al., 1991; Chen et al., 2000; Lindmark-Maensson and Akesson, 2000; Filipović et al., 2005; Kasapovic et al., 2005). According this reports, SOD activity in bovine milk was in the range from 0.92 to 3 U/ml. In raw cows' milk, extracellular form of GPx activity had demonstrated levels between 12 and 32 U/mL, and the activity has been found to be significantly correlated with selenium concentration (Przybylska et al., 2007). Debski et al. (1987) reported that GPx dependent peroxidase activity was approximately one-third of the total peroxidase activity and was found to be similar in human and bovine milk. However, some reports disagree regarding the measured activity of SOD and GPx in milk (Stagsted, 2006; Kovaceva et al., 2007). Conclusıons The present study describes an spectrofotometric kinetic protocols to measuring the activity of superoxide dismutase (SOD) and glutathione peroxidase (GPx) in milk whey. It has been shown that the linear kinetic observed in milk was depended from amount of whey sample and concentration of other compounds in reactive mixture. The both methods are reproducible and can be used for determination of antioxidant enzymes SOD and GPx in milk whey, suggesting that this antioxidative enzymes are of importance for milk stability and quality. SOD and GPx activity occurs in bovine milk, but their importance in this matrix is not well known. References Avissar N., Slemmon, J.R., Palmer, I.S. and Cohen, H.J. (1991). Partial sequence of human plasma glutathione peroxidase and immunologic identification of milk glutathione peroxidase as the plasma enzyme. Journal of Nutrition, 121, pg. 1243-1249. Bandyopadhyay U., Das, D., Banerjee, K.R. (1999). Reactive oxygen species: oxidative damage and pathogenesis. Current Science 77, 658-665. Beyer W.F.Jr. and Fridovich, I. (1987). Assaying for superoxide dismutase activity: some large consequences of minor changes in conditions. Anal. Biochem., 161, pg. 559-566. Bhattacharya I.D., Picciano, M.F., Milner, J.A. (1988). Characteristics of human milk glutathione peroxidase. Biological Trace Element Research, 18, pg. 59–70.
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Bordo D., Pesce, A., Bolognesi, M., Stroppolo, M.E., Falconi, M., Desideri, A. (2000). Cu,Zn superoxide dismutase in prokaryotes and eukaryotes. in: K. Wieghardt, R. Huber, T. Poulos A. Messerschmidt (Eds.) Handbook of Metalloproteins. John Wiley & Sons, London, UK; 2000. Chen J., Lindmark-Mansson, H. and Akesson, B. (2000). Optimisation of a coupled enzymatic assay of glutathione peroxidase activity in bovine milk and whey. International Dairy Journal, 10, pg. 347-351. Debski B., Picciano, M.F. and Milner, J.A. (1987). Selenium content and distribution of human, cow and goat milk. Journal of Nutrition, 117, pg. 1091-1097. Esworthy R.S., Chu, F.F., Geiger, P., Girotti, A.W., Doroshow, J.H. (1993). Reactivity of plasma glutathione peroxidase with hydroperoxide substrates and glutathione. Archives of Biochemistry and Biophysics, 307, pg. 29–34. Filipovic, D., Kasapovic, J., Pejic, S., Niciforovic, A., Pajovic, S.B. and Radojcic, M.B. (2005). Superoxide dismutase activity in various fractions of full bovine milk, Acta Aliment. 34 (3), pg. 219-226. Gao R., Yuan, Z., Zhao, Z., Gao, X. (1998). Mechanism of pyrogallol autoxidation and determination of superoxide dismutase enzyme activity. Bioelectrochemistry and Bioenergetics 45, pg. 41-45. Halliwell B. (1987). Oxidants and human disease:some new concepts. Federation of American Societes for Eaperimental Biolog, 1, pg. 358-366. Halliwell B. and Gutteridege, J.M.C. (2007). Free Radicals in Biology and Medicine, 4th ed. Oxford University Press. Kasapovic J., Pejice, S, Mladenovice, M., Radlovice, N., Pajovice, S.B. (2005). Superoxide dismutase activity in colostrum, transitional and mature human milk. Turkish J Pediatr 47, pg. 343-347. Kovaceva J., Platenik, J., Vejrazka, M., Stipek, S., Ardan, T., Cejka, C., Midelfart, A., Cejkova J. (2007). Differences in activities of antioxidant superoxide dismutase, glutathione peroxidase and prooxidant xanthine oxidoreductase/xanthine oxidase in the normal corneal epithelium of various mammalia, Physiol. Res. 56, pg. 105-112. Lindmark-Mansson, H. and Akesson, B. (2000). Antioxidative factors in milk. British Journal of Nutrition, 84, pg. 103-110. Marklund, S. and Marklund, G. (1974). Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur. J. Biochem. 47, pg. 469-474. Nielsen J.H., Hald, G., Kjeldsen, L., Andersen, H.J. and Ostdal, H. (2001). Oxidation of ascorbate in raw milk induced by enzymes and transition metals. Journal of Agricultural and Food Chemistry, 49, pg. 2998-3003. Paglia D.E. and Valentine, W.N. (1967). Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J. Lab. Clin. Med. 70, pg. 158-169. Przybylska J., Albera, E. and Kankofer, M. (2007). Antioxidants in Bovine ColostrumReprod Dom Anim 42, pg. 402-409. Stagsted J. (2006). Absence of both glutathione peroxidase activity and glutathione in bovine milk International Dairy Journal 16, pg. 662-668. Violi F., Iuliano, L., Alessandri, C., Ghiselli, A. and Balsanoa, F. (1985). Simple method for evaluating platelet superoxide dismutase. Scand J Clin Lab Invest 1985; 45: pg. 713716.
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52
SECTION 1. ANIMAL BIOTECHNOLOGY UDC: 636.2.082(497.113) Original scientific paper
EFFECTS OF THE IMPLEMENTATION OF THE BREEDING PROGRAMS TO DEVELOP DAIRY CATTLE BREEDING IN VOJVODINA Trivunović S.1*, Janković D.1, Ivanović D.1, Radinović M.1, Šoronja Ţ.1 Faculty of Agriculture, Trg Dositeja Obradovića 8, 21000 Novi Sad, Serbia
1
*
corresponding author: [email protected]
Abstract The breeding programs for Holstein-Friesian and Simmental breed of cattle in Vojvodina defines breeding goals, population size, breeding methods, selection program and gene banks, research and development tasks to program efficiency increasing, the conditions for successful breeding of domestic animals and ensuring the dissemination of genetic progress and improve the quality of livestock products in accordance with the zootechnical standards. The legal basis for the breeding program is given in the Act of Livestock from 2009, while the first breeding programs came in 2010, and enforced to the end of 2014. Based on the current situation in the breeding of Holstein-Friesian and Simmental, breeding objective for controlled Holstein-Friesian breed population in Vojvodina is recording herds increase by 10%, average milk production in standard lactation over 7500 kg with 3.90% fat and 3.40% protein, and improve conformation; for the Simmental recording herds increased by 20%, average milk production in standard lactation over 5000 kg with 4.00% fat and 3.50% protein, as well as improved conformation (especially udders). On the basis of information from the Main breeding organization it was determined that the goal for Holstein breed was reached since recording herds increase by 30.85%, while the Simmental populations increase by 112.05%. Milk yield goal, for Holstein-Friesian breed has not achieved, but milk production increased by 10% compared to 2009. Milk yield goal, for Simmental breed is filled in the final year of programme implementation with production of more than 5000 kg. We conclude that the implementation of the major breeding programs gave a positive effect on the development of dairy cattle breeding in Vojvodina. Key words: breeding programs, recorded herds, dairy cattle. Introduction Multiple role and importance of the dairy production demand system's sustainability and quality. Performance of dairy cattle, as the basic system component, we can improve by improving their genetic potential. The breeding program is a set of decisions and activities for, primarily, genetic improvement of the animals, so that they can better adapt to the environment or be competitive as much as possible. Organized breeding of farm animals in Vojvodina started by establishing the Regional Institute of Animal Science in 1950. Center for the breeding of farm animals at the Department of Animal Science, Faculty of Agriculture in Novi Sad, was registered in 2010., as the Main breeding organization in 53
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Vojvodina by the Ministry of Agriculture of Republic of Serbia, since when began implementation of the breeding programs for Holstein Friesian and Simmental breed (Trivunovic et al., 2014). For the economic viability of dairy cattle, as the most important part of livestock production, it is necessary to clearly define the breeding goals and program and implement them (Bogdanovic et al., 2005). The breeding goal is a specific combination of economic values and genetic information of all the traits that we want to improve. Some of the most biologically and economically important traits of dairy cows are: a) production of large amounts of high value milk, b) reproductive efficiency, d) good health, c) the 'ideal' conformation, d) longevity (Berry, 2014). Although the breeding goals vary in different situations, they are usually characterized by profit function which shows the direction in which genetic changes of particular traits affecting the cow’s profitability. Also, farm animal welfare has become increasingly important and relevant from the societal point of view (Oltenacu and Broom, 2010). Dairy industry success depends on the perception of its products and production methods by the public (Tonsor et al., 2009). Furthermore, breeding for an increased production level has led to a decrease of withinbreed variability (Danchin-Burge et al., 2012). So, there are numerous reasons for the broad breeding goals (sustainability of production, consumer awareness about animal welfare, reduction of production cost) (Atsbeha et al., 2014). As the animal's performance for importance traits is greatly influenced by environmental (more than genetic) factors, sustainability and quality of dairy production systems requires optimization of environmental conditions by appropriate management practices on dairy farms (nutrition, health care programs, reproductive efficiency, etc.). The breeding programs can be considered as investment in the context of long-term development programs, and should be implemented in accordance with the national agricultural development strategy while respecting existing constraints (financing, etc.). It should be simple and reliable, at least initially, rather than sophisticated and sensitive to assumptions whose fulfillment cannot be guaranteed; with time and experience, the schemes may be more sophisticated (Kosgei et al., 2011). The above applies to all stages of the program definition and implementation. Analysis of implementation is an important component of breeding programs (FAO, 2010). Also, it is necessary to promote the breeding program and its results. The breeding programs for Holstein-Friesian and Simmental breed of cattle in Vojvodina defines: breeding goals, population size, breeding methods, selection program and gene banks, research and development tasks to increase program efficiency, the conditions for successful breeding of animals, ensuring the genetic progress dissemination and improving the quality of animal products in accordance with the zootechnical standards (Main breeding program of Holstein-Friesian and Simmental cattle in Vojvodina, 2010). Subjects in the implementation of Main breeding programs of Holstein-Friesian and Simmental cattle in Vojvodina are: Farmers, Basics breeding organizations, Regional breeding organizations, Main breeding organization, Organizations with special authorization, Provincial Secretariat for Agriculture, Water Management and Forestry, Ministry of Agriculture and Environmental Protection. The most important objectives in the first breeding programs for Holstein Friesian and Simmental cattle (2010-2014),were: population increase by 10%, average milk production ( 305-d) over 7500 kg, fat content 3,90%, protein content 3,40%, and conformation improving, for recorded Holstein Friesian population; population increase by 20%, average milk production (305-d) over 5000 kg, fat content 4,00%, protein content 3,50% protein, conformation improving (especially 54
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udders), for the recorded Simmental population. The aim of this paper is to examine the effects of the breeding programs implementation on the dairy production in Vojvodina. Material and methods Main breeding organization collect, process and analyze data related to recorded population. Data were collected from primary and regional breeding organizations, through quarterly reporting documentation and annual selection review which is obligatory event defined in the main breeding programs. Data for total population size and milk yield are obtained from Statistical Office of the Republic of Serbia; both data refers to the period 2010-2014. Number of cows and heifers in total and recorded population were calculated. Control of cows productivity is carried out by controlling the milk yield (Petrovic et al., 2006), conducted by authorized persons in accordance with methodology prescribed in the Main breeding programs of Holstein-Friesian and Simmental breed. All lactation records are standardized to 305-day, and lactation shorter than 200-day are excluded. The average milk yield is calculated both for recorded and total population. Yield and content of milk fat and number of complete lactation are calculated. Results and discussion Size of total and recorded Holstein Friesian and Simmental cows and heifers populations has increased, as well as share of recorded relative to the total population (Figure 1). The goal of increasing the recorded population was achieved for both breeds, since the recorded Holstein Friesian population increased by 30,85% and Simmental by 112,05%. This reflects the interest of farmers for the implementation of breeding programs on their farms. Also, it is a result of increased incentives for genetic improvement of livestock by the Ministry of Agriculture and Environmental Protection of Republic of Serbia. Given the total and recorded cows and heifers population size ratio, there is room for further increasing the share of recorded relative to the total population.
Figure 1. Size of the total and recorded cows and heifers population and share of recorded relative to the total population in the period 2010-2014.
In Croatia, the share of recorded relative to total population (2014.) is similar, 61,4% (CAA, 2015), while increasing the share of recorded relative to total population when comparing 2014. to 2010. is 18,3% (Serbia) and 7,8% (Croatia). In Slovenia (2013.), share 55
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of recorded relative to the total population was 77,3%. In some other countries of Central and Eastern Europe share ranges from 77,8% in Austria to 93,9% in Czech Republic, while in the Poland is 30,5% (Bucek et al., 2014). The average milk yields in a recorded (HF, SIM) and total population (Figure 2), shows an increasing phenotypic trend of this very important production trait in recorded population, as well as higher absolute values, in contrast to total population, indicating the impact of implementation of breeding programs. However, although the average milk yield of Holstein Friesian population increased by 10%, the goal of 7500 kg of milk in 305-d lactation has not been achieved. The yield of more than 5000 kg of milk for the Simmental breed has been achieved in the last year of breeding program implementation.
Figure 2. Phenotypic trends for 305-d milk yield of Holstein Friesian, Simmental and total population in the period 2010-2014.
Given the genetic potential of the Holstein Friesian cows for milk production, determined average milk yield in the Holstein Friesian population in Vojvodina is not satisfactory. But we need to know that the Holstein Friesian population in Vojvodina is very heterogeneous in many parameters (herd management, selection activities, herd size, etc.). In the former large state farms, for example, the average milk yield of Holstein Friesian cows are 8200 kg. The difference in the milk yield between Holstein Friesian and Simmental is also not satisfactory. Table 1. Number of complete lactations, milk yield, milk fat content and yield in 305-d lactation of recorded Holstein Friesian and Simmental in the period 2010-2014. Holstein Friesian Simmental Milk Milk Number of Fat, Fat, Number of Fat, Fat, Year lactations % lactations kg % yield, kg kg yield,kg 2010 17411 5505 202 3,36 2378 4709 181 3,52 2011 17307 5797 220 3,74 2738 4833 219 3,73 2012 18407 6356 241 3,8 3134 5365 215 4,01 2013 22432 6307 237 3,76 4411 5387 213 3,95 2014 22375 6521 252 3,87 4924 5828 236 4,05
Increase of milk yield in the period 2010-2014. is 1013 kg for the Holstein-Friesian and 1119 kg for the Simmental cows. Increasing the number of complete lactations is apparent also, for both breeds; 4964 more lactation of Holstein Friesian cows has been 56
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completed in 2014. compare to 2010. and 2546 more lactation of Simmental cows has been completed in 2014. compare to 2010. Milk fat yield and content also shows increasing phenotypic trend in both breeds; milk fat yield was increased by 50 kg in the Holstein Friesian (fat content, 3,87% vs. 3,36%) and by 55 kg in the Simmental (fat content 4,05% vs. 3,52%). The average milk yield and milk fat content of Holstein Friesian cows are lower in relation to Croatia (7160 kg, 3,99%) (CAA, 2015), as well as in relation to Slovenia (7385, 3,97%) (Kmetijski Institut Slovenije, 2014.). The average milk yield of Simmental cows is higher in relation to Croatia (5030 kg, 4,01%), as well as in relation to Slovenia (5283 kg, 4,05%). In the region of ex Yugoslavia, the share of the recorded relative to the total population is increasing, as well as the average milk yield, while the total population size is decreasing, (Stokovic et al., 2007). New breeding programs (20152020.) and goals for Holstein Friesian and Simmental cattle in Vojvodina are broadened by including new traits; production (yield and content of milk protein), functional-fertility and health (percent calf crop, calving percentage, days open, calf sex, type of birth- single, twin, triplet, calf scoring, calving ease scores, calf mortality, somatic cell count), as well as longevity and conformation traits. Traits are measured according to ICAR (International Committee for Animal Recording) standards. Conclusions Dairy is a significant branch of agriculture and the overall economy in Vojvodina. Therefore it is of great importance to increase the number of the dairy cattle, as well as share or recorded relative to total population. This will increase the number of dairy cattle under the measures of breeding programs, primarily under the measures of genetic improvement. Proactive approach by all stakeholders will contribute achieving this goal, (e.g., state support for agriculture, improved breeding organizations, new technologies, promotion of breeding programs, etc.). This article just shows significant positive effects of implemetation of breeding programs for Holstein and Simmental cattle on the overall performance of dairy cattle. The milk yield and milk fat yield and content in a recorded population of Holstein Friesian and Simmental cows shows evident phenotypic increasing trend as opposed to a population that is not controlled. It actually indicates the great importance of implementation of breeding programs on the development of dairy cattle production in Vojvodina. Objectives in new breeding programs are broadened by including new traits (production, functional, conformation) which are recorded in accordance with professional standards. This, along with the optimization of management (nutrition, health care programs, housing, etc.) will further enhance and expand the positive effects of implementation of breeding programs for Holstein Friesian and Simmental cattle on development of dairy in Vojvodina. As the main goal of animal breeding is to improve genetic merit of the animals in the next generations, aim for the future is to estimate the genetic trends for traits of interest. References Atsbeha D. Muluwork, Dadi Kristofersson, Kyrre Rickertsen. (2014). Broad Breeding Goals and Production Cost in Dairy Farms. Presentation at: Norwegian Agricultural Economics Research Institute (NILF), Oslo, Norway.
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Berry P. Donagh. (2014). Breeding the dairy cow of the future – what do we need? Animal & Bioscience Research Department, Animal & Grassland Research and Innovation Centre Teagasc, Moorepark, Fermoy, Co. Cork, Ireland. Bogdanović V., Đedović, R., Perišić, P., Petrović, M.M. (2005). Odgajivački ciljevi i programi u govedarstvu Srbije. Biotechnology in Animal Husbandry, 21 (5-6), p. 15-21. Bucek P., Zottl, K., Onken, F., Klopčič, M., Radzio, D., Mészáros, G., Barac, Z., Ryba, Š., Dianová, M., Kučera, J. (2014). Practical aspects in milk recording in Central and Eastern Europe and its effects on the guidelines. ICAR Conference, Berlin. Croatian Agricultural Agency-CAA. (2015). Annual report 2014-Cattle breeding. Kriţevci. Danchin-Burge, C., Leroy, G., Moureaux, S., and Verrier, E. (2012). Evolution of the genetic variability of French dairy cattle breeds assessed by pedigree analysis. J. Anim. Breed. Genet. 129, 206–217 FAO (Food and Agriculture Organization of the United Nations). (2010). Breeding strategies for sustainable management of animal genetic resources. FAO Animal Production and Health Guidelines 3. FAO Rome, Italy. Kosgey I.S., Mbuku, S.M., Okeyo, A.M., Amimo, J., Philipsson, J., Ojango, J.M. (2011). Institutional and organizational frameworks for dairy and beef cattle recording in Kenya: a review and opportunities for improvement. Animal Genetic Resources © Food and Agriculture Organization of the United Nations, 48, 1-11. Kmetijski Institut Slovenije. (2014). Results of Dairy and Beef Recording Slovenia 2013. Ljubljana. Main breeding organization (Department of animal science, Faculty of agriculture). (2010). Main breeding program of Holstein-Friesian cattle in Vojvodina. Novi Sad. Main breeding organization (Department of animal science, Faculty of agriculture). (2015). Annual report 2014 about implementation of breeding measures in livestock Vojvodina. Novi Sad. Oltenacu P.A. and Broom, D.M. (2010). The impact of genetic selection for increased milk yield on the welfare of dairy cows. Animal Welfare, 19(S): 39-49. Petrović M.D., Petrović, M.M., Kurćubić, V. (2006). Govedarstvo- tehnologija proizvodnje, Čačak. Statistical Office of the Republic of Serbia. (2015). Annual report of the Republic of Serbia for 2014. Stokovic I., Ekert Kabalin, A., Karolyi, D., Sakic, V., Miscevic, B., Daud, J., Staric, J., Bunevski G. (2007). Cattle production trends in the region of ex Yugoslavia (http://www.eaap.org/docs/newsletters/2007-07/Cattlenetwork) Tonsor, G., Olynk, N., and Wolf, C. (2009). Consumer preferences for animal welfare attributes: The case of gestation crates. J. Agr. Appl. Econ. 41(3), 713–730. Trivunović S., Radović I., Pihler I., Janković D. (2014). SprovoĎenje odgajivačkih programa u stočarstvu AP Vojvodine. 7th International scientific/professional conference: Agriculture in nature and environment protection, Vukovar, Republic of Croatia. pp. 3747.
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SECTION 1. ANIMAL BIOTECHNOLOGY UDC: 606:636.082 Original scientific paper
THE USING OF BIOTECHNOLOGY IN ANIMAL BREEDING Tutkun M.1*, Tatar A.M.1 1
Faculty of Agriculture, Department of Animal Science, Dicle University Diyarbakir-21100, Türkiye *
corresponding author: [email protected]
Abstract This review focuses on recent developments applied in animal breeding including artificial insemination, embryo transfer, and embryo cloning and on approaches aimed at increasing production through use of pharmaceuticals and transgenesis. Livestock is becoming increasingly important to economic growth in developing countries and the application of biotechnology is largely dictated by commercial considerations and socio-economic goals. These technologies including artificial insemination, embryo transfer and other assisted reproductive methods are used in the genetic improvement of livestock. Biotechnology developments applied to livestock health, nutrition, breeding and reproduction are improving with a reasonable pace in developing countries. Simple bio-techniques such as artificial insemination have been well implemented in many parts of the developing world. However, advanced technologies including transgenic plant vaccines, marker assisted selection, solid state fermentation for the production of fibrolytic enzymes, transgenic fodders, embryo transfer and animal cloning are confined largely to research organizations (Onteru and et al., 2010). This paper reviews available biotechnologies with potential application in livestock improvement and identifies those which have been or may be applied in developing countries in general. The review covers biotechnology applications in the areas of animal genetics and breeding, including conservation of animal genetic resources, animal health, physiology of lactation and growth, and animal nutrition. Key words: Turkey, Livestock production, Biotechnology. Introduction According to the estimation by Food and Agriculture Organisation of the United Nations (FAO), 70% of world population will be in hunger in 50 years from now (2060). It has been reported that the world human consumption for the animal protein was 29 g per capita daily (or 10 kg per capita consumption). Especially in the advanced countries, the human consumption was approximately 90 kg animal protein per capita consumption. However, the trend toward increased per capita demand for animal source foods is occurring primarily in developing countries (80% of world population). Through biotechnology, it has the potential to improve the productivity of animals via increase growth, carcass quality and reproduction, improved nutrition and feed utilisation, improved quality and safety of food, improved health and welfare of animals and reduced waste through more efficient utilisation of resources. Therefore, the biotechnology of livestock production is
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growing faster than any other sectors; and by 2020 livestock is predicted to become the most important agricultural sector in terms of value-added commodity. The world’s population is expected to grow to 7.5 billion people in 2020 with most of this growth occurring in developing countries (IFPRI, 2001). As a result, demand for animal products is expected to grow tremendously (Bradford, 1999). In view of this, the methods of livestock production must change to allow for efficiency and improvement in productivity. Biotechnology (Biotech) is increasingly becoming a sustainable means of improving livestock production by directly influencing animal health, nutrition, reproduction, breeding and genetics (Bonneau and Laarveld, 1999). Biotechnology will provide new and unprecedented opportunities to improve the productivity of animals through increased growth, carcass quality and reproduction, improved nutrition and feed utilization, improved quality and safety of food, improved health and welfare of animals, and reduced waste through more efficient utilization of resources. The livestock industry will benefit also from the application of biotechnology in other areas such as in the development of new and improved feedstuffs, as well as in microbiology as related to food and bioremediation (Bonneau and Laarveld, 1999). Livestock is becoming increasingly important to economic growth in developing countries and the application of biotechnology is largely dictated by commercial considerations and socio-economic goals. These technologies have been used in the genetic improvement of livestock, particularly in cattle and buffaloes, and the economic returns are significant (Madan, 2005). Widespread use of some biotechnologies in animal husbandry resulted (Van Vleck, 1981) and will result in a major impact on genetic progress. Different types of biotechnologies may have influence on both animal breeding and the resulting genetic progress through three main ways (Gengler and Druet, 2001) Biotechnologies can affect efficiency of reproduction and therefore also selection programs: artificial insemination, embryo transfer, sexing, cloning and other related techniques (e.g., Ruane and Thompson, 1991; van Vleck, 1981). Biotechnologies can improve determination of genetic values of animal: genetic markers, candidate genes and other related techniques (e.g., Georges et al., 1995; Renaville et al., 1997). Biotechnologies can transform artificially the genome at the DNA level: genetic engineering, gene transfer and related techniques (e.g., Solter, 1981). This paper reviews biotechs in livestock production citing practical examples while focussing on possibilities for application in animal production Reproductive animal biotechnology Various biotechnology methods are used in improving the breeding stock of animals. These include artificial insemination (AI), embryo transfer (ET), in-vitro fertilization (IVF), somatic cell nuclear transfer, and the emerging technology on somatic cell nuclear transfer. Artificial Insemination (AI) One of the earliest perfected technology is Artificial Insemination (AI) where new breeds of animals are produced through the introduction of the male sperm from one superior male to the female reproductive tract without mating. AI reduces transmission of venereal 60
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disease, lessens the need of farms to maintain breeding males, facilitates more accurate recording of pedigrees, and minimizes the cost of introducing improved genetics (Wilmut et all.1979). Various technologies have evolved that led to the efficient use of AI in developing desired livestock, including the methods of freezing semen or cryopreservation and sperm sexing. AI technology use is still more generally associated with dairy cattle than other domestic livestock species. The limitations of AI use in beef cattle include the difficulty in detecting heat in large beef herds kept on ranches and the less frequent handling of individual cows. In sheep and goats the failure to develop a simple, non-surgical insemination procedure has prevented extensive exploitation of the technology in sheep (Robinson and McEvoy 1993). In-vitro Fertilization With in vitro fertilization (IVF), a technician removes unfertilized eggs (oocytes) from the donor cow’s ovaries, usually recovering 6-8 useable oocytes. The oocytes mature in an incubator and are fertilized with sperm. The resulting zygotes incubate and develop in the laboratory before being placed into the recipient cow. While IVF can produce many fertilized embryos, the added expense of ET makes the procedure prohibitive in most cases (Becker and Covan, 2009). In case other artificial reproductive techniques fail due to difficulties such as blocked reproductive systems, non-responsive ovaries in the females, marginal semen quality and quantity in the male, and presence of disease, in vitro fertilization (IVF) is used. The fertilization of the sperm and the egg is conducted in vitro (outside the animal’s body) at specific environmental and biochemical conditions. To date, successful IVFs have been conducted in various animal species due to advances in embryo production and cryopreservation of reproductive cells. Since the birth of the first rabbit conceived through IVF in 1959 (Chang, 1959), IVF offsprings have been born in mice, rats, hamsters, cats, guinea pig, squirrels, pigs, cows, monkeys, and humans (Bearden and Fuquay, 2000) Embriyo Transfer (MOET) After AI and estrus synchronization, embryo transfer (ET) is the third most commonly used biotechnology. In ET, a donor cow of superior breeding is hormonal induced to superovulate. The eggs are then fertilized within the donor, the embryo develops and is then removed and implanted in a recipient cow. Between removal and implantation, embryos may be frozen for safekeeping. Because of the relatively high costs, ET is used mostly within registered cow herds (Becker and Covan, 2009). Embryo transfer (ET) from one mother to a surrogate mother makes it possible to produce several livestock progenies from a superior female. Selected females are induced to superovulate hormonally and inseminated at an appropriate time relative to ovulation depending on the species and breed. Week-old embryos are flushed out of the donor’s uterus, isolated, examined microscopically for number and quality, and inserted into the endometrium of the uterus of surrogate mothers. By increasing the number of offspring that can be obtained from monotocous species in particular, MOET has the potential to increase genetic improvement by enhancing the selection intensity on the female side (Georges, 1994) Multiple Ovulation and Embryo Transfer (MOET) is a well established technology and is used to obtain over 80% of the embryos produced for commercial purposes (Thibier, 61
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2001). Multiple ovulation embryo transfer (MOET) is a composite technology which includes superovulation, fertilisation, embryo recovery, short-term in vitro culture of embryos, embryo freezing and embryo transfer. Benefits from MOET include increasing the number of offspring produced by valuable females, increasing the population base of rare or endangered breeds or species, ex situ preservation of endangered populations, progeny testing of females and increasing rates of genetic improvement in breeding programmes (Rege, 1994). It was suggested that application of MOET in nucleus breeding schemes could increase animal genetic gains by 30-80% (Nicholas and Smith 1983). More recently it has been concluded that the earlier figures were over-predictions (Keller et al 1990). The overpredictions arose partly because the assumed average number of progeny (eight) per donor female was unrealistically high and partly because of wrong assumptions made about genetic parameters (Keller et al 1990). ET increases reproductive rate of selected females, reduces disease transfer, and facilitates the development of rare and economically important genetic stocks as well as the production of several closely related and genetically similar individuals that are important in livestock breeding research. Somatic Cell Nuclear Transfer Somatic cell nuclear transfer (NF) is a technique in which the nucleus (DNA) of a somatic cell is transferred into a female egg cell or oocyte in which the nucleus has been removed to generate a new individual, genetically identical to the somatic cell donor (Tian et all, 2003). This technique was used to generate the sheep Dolly from a differentiated adult mammary epithelial cell which demonstrated that genes that are already inactivated in differentiated tissues can be completely reactivated (Wilmut, 1979). NF technology creates possibility of generating clones from superior genotype and can be used to efficiently evaluate effects of genotype x environment interactions and testing or dissemination of transgenics. Problems on high rate of pregnancy loss, survival of newborn and increased incidence of abnormal development due to incorrect reprogramming of nuclear DNA (epigenetic inference) and unusual conditions during in-vitro processes make this a precommercial technology. Cloning Cloning is a biotechnology developing rapidly and with significant public controversy. Most people think of cloning as the creation of an organism that is genetically identical to another one. However, scientists use the term more broadly, to refer to production not only of such organisms but also of genetically identical cells, and to replication of DNA and other molecules. It also refers to a form of reproduction found naturally in many singlecelled organisms, as well as plants and animals. Those differences in meaning and usage have caused some confusion in public debate about cloning, where the main area of controversy relates to artificial cloning involving higher organisms, including humans (Johnson and Williams, 2006) Sexing Sexing of semen and of embryos provides in species and production circumstances where one sex is preferred, a way to produce the wanted type of animal (Betteridge et al., 1981; Johnson et al., 1998). Sexing of embryos is a technique that is rather reliable nowadays and 62
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currently used on a rather large extent. Some progress was recently made in the field of sexed semen. One problem is the price of sexing and the eventually reduced fertility of this semen. The other issue is that sexing has to be economic what means that the extra-price to be paid for sexed semen has to be in relation with the extra income (Gengler and Druet, 2001) . Anımal genetıcs and breedıng Genetic improvement of livestock depends on access to genetic variation and effective methods for exploiting this variation. Genetic diversity constitutes a buffer against changes in the environment and is a key in selection and breeding for adaptability and production on a range of environments. In developed countries, breeding programmes are based upon performance recording and this has led to substantial improvements in animal production (Rege, 1994). Indicator traits Selection is the process breeders use to produce genetic change, realizing that genetic change and genetic improvement are not necessarily the same. Producers can change many traits genetically but change does not necessarily mean improvement. Improvement implies the production of superior animals and for livestock production the definition of superior animals are those with greater profitability. This manual explains underlying genetic mechanisms, concepts of selection, and tools that can be used to make better selection decisions to help producers meet their goals. The assumption throughout is that the goal of sire selection and beef enterprises is profitability. The difference between indicator and economically relevant traits (ERT) and the ability to distinguish between the two are keys to improving profitability. By identifying the economically relevant traits, selection focus can be narrowed, resulting in faster genetic improvement and improved profitability. In the end, the goal of focusing selection on ERT is to increase the probability that breeders will make selection decisions that make them more profitable (Enns, 2010). Indicator traits are characteristics which are genetically correlated to traits of economic importance and are easier to measure than the latter. Such traits are usually not the target of genetic improvements but provide an indirect means of improving a targeted trait. Blair et al (1990) reviewed some physiological and/or metabolic characteristics which might be considered as potential indicator traits. Indicator traits can improve genetic response by increasing accuracy of selection and reducing generation interval. The value of an indicator trait will depend largely on the magnitude of co-heritability (square-root of the product of heritability of the indicator and of the target trait) and the genetic correlation between the two traits (Woolliams and Smith 1988). Woolliams and Smith (1988) concluded that, with high co-heritability, selection for the indicator trait alone can result in greater rates of response than is possible with progeny testing, especially when breeding values are not accurately measured by progeny testing. Transgenic animals There are various definitions for the term transgenic animal. The Federation of European Laboratory Animal Associations defines the term as an animal in which there has been a deliberate modification of its genome, the genetic makeup of an organism responsible for inherited characteristics (FELASA, 1982). A transgenic animal is an animal whose hereditary DNA has been augmented by addition of DNA from a source other than 63
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parental germplasm through recombinant DNA techniques. Transfer of genes or gene constructs allows for the manipulation of individual genes rather than entire genomes. There has been dramatic advances in gene transfer technology in the last two decades since the first successful transfer was carried out in mice in 1980 (Palmiter et al 1982; Jaenisch 1988). The technique has now become routine in the mouse and resulting transgenic mice are able to transmit their transgenes to their offspring thereby allowing a large number of transgenic animals to be produced. Successful production of transgenic livestock has been reported for pigs, sheep, rabbits and cattle. The majority of gene transfer studies in livestock have, however, been carried out in the pig. Although transgenic cattle and sheep have been successfully produced, the procedure is still inefficient in these species (Niemann et al 1994). Transgenesis offers considerable opportunity for advances in medicine and agriculture. In livestock, the ability to insert new genes for such economically important characteristics as fecundity, resistance to or tolerance of other environmental stresses would represent a major breakthrough in the breeding of commercially superior stock. Another opportunity that transgenic technology could provide is in the production of medically important proteins such as insulin and clotting factors in the milk of domestic livestock (Rege, 1994). Conclusions As a result, some of biotechnological methods are used effectively to increase animal production today while some of them have a chance for the future application. Today, animal products has strategic importance and more effective and widely using of these biotechnological methods are necessary in animal production References Abraham H,. Pal, S.K. 2014. Animal Biotechnology Options in Improving Livestock Production in the Horn of Africa, International Journal of Interdisciplinary and Multidisciplinary Studies,2014,Vol 1,No.3,1-8. Bearden HJ and JW Fuquay 2000. Semen evaluation. In: HJ Bearden and JW Fuquay, Editors, Applied Animal Reproduction, Prentice Hall, Upper Saddle River, New Jersey (2000). Becker S. G., Covan, T. 2009. Biotechonology in Aniamal Agriculture. Statue and Current Issues.CRS Report for Congress. Blair H.T., McCutcheon S.N. and Mackenzie D.D.S. 1990. Components of the somatotropic axis as predictors of genetic merit for growth. In: Proceedings of the 4th World Congress on Genetics and Applied Livestock Production, Edinburgh, Scotland. Vol XVI. International Committee for World Congress on Genetics Applied to Livestock Productivity, Edinburgh, UK. pp. 246-255. Betteridge K.J., Hare, W.C.D. and Singh, L.H. (1981) Approaches to sex selection in farm animals, in B.G. Brackett, G.E. Seidel and S.M. Seidel (eds.), New technologies in Animal Breeding Academic Press, London, pp 109-125 Bonneau M., Laarveld, B. 1999. Biotechnology in animal nutrition, physiology and health. Livest. Prod. Sci. 59: 223-241. Bradford G.E. 1999. Contributions of animal agriculture in meeting global human food demand. Livest. Prod. Sci. 59: 95-112. 64
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Chang, MC. 1959. Fertilization of rabbit ova in vitro. Nature, 1959 8:184 (supl 7) 466. Enns R.M. 2101. The Role of Economically Relevant and Indicator Traits. Beef Sire Selection Manual Second Edition. National Beef Cattle Evaluation Consortium. FELASA (Federation of European Laboratory Animal Science Associations September 1982, revised February 1995. Transgenic Animals — Derivation, Welfare, Use and Protection. Galli C., Lazzari, G. 2005. Embryo technologies in dairy cattle. The 26th European Holstein and Red Holstein Conference, Prague. Georges M. 1994. Applıcation Of Biotechnology For The Genetic Improvement of Livestock: Status And Prospects, Department of Genetics, Faculty of Veterinary Medicine, University of Liège Gengler N., Druet, T. 2001. Impact Of Bıotechnology On Anımal Breedıng And Genetıc Progress. R. Renaville and A. Burny (eds.). Biotechnology in Animal Husbandry, 33-45. Kluwer Academic Publishers. Printed in the Netherlands. Halbert G.W., Dobson H., Walton J. S., Sharpe P. and Buckrell B.C. 1990. Field evaluation of a technique for transcervical intrauterine insemination of ewes. Theriogenology 33:1231-1243. IFPRI, (2001). 2020 Global Food Outlook: Trends, alternatives and choices. Food Policy Report. IFPRI, Washington D. C., USA Jaenisch R. 1988. Transgenic animals. Science 240:1468-1473. Jhonson J.A., Williams, E.D. 2006. Human Cloning, CRS Report Keller D.S., Gearheart W.W. and Smith C. 1990. A comparison of factors reducing selection response in closed nucleus breeding schemes. Journal of Animal Science68:15531561. Onteru S., Ampaire, A., Rothschild, M. Biotechnology developments in the livestock sector in developing countries. Biotechnol Genet Eng Rev. 27:217-28 Madan M.L. 2005. Animal biotechnology: applications and economic implications in developing countries. Rev Sci Tech. 24(1):127-39 Niemann H., Halter R. and Paul D. 1994. Gene transfer in cattle and sheep: A summary perspective. In: Proceedings of the 5th World Congress on Genetics Applied to Livestock Production, Guelph, Canada, 712 August 1994. Volume 21. International Committee for World Congress on Genetics Applied to Livestock Production, Guelph, Ontario, Canada. pp. 339-346. Nicholas F.W. and Smith C. 1983. Increase rate of genetic change in dairy cattle by embryo transfer and splitting. Animal Production 36:341-353 Palmiter R.D., Brinster R.L., Hammer R.E., Trumbauer M.E., Rosenfeld M.G., Birnberg N.C. and Evans R.M. 1982. Dramatic growth of mice that develop from eggs microinjected with metallothionene growth hormone fusion genes. Nature 300:611-615. Rege, J. E.O. 1994. Biotechnology options for improving livestock production in developing countries, with special reference to sub-Saharan Africa. Small Ruminant Research and Development in Africa. ILRI, Kenya Ruane J. and Thompson, R. (1991) Comparison of simulated and theoretical results in adult MOET nucleus schemes for dairy cattle, Livest. Prod. Sci. 28, 1-20. Solter D. (1981) Gene transfer in mammalian cells, in B.G. Brackett, G.E. Seidel and S.M. Seidel (eds.), New technologies in Animal Breeding, Academic Press, London, pp 201 220.
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Thibier M.2001. The animal embryo transfer industry in figures, a report from the IETS data retrieval commettee. International Embryo Transfer Society Newsletter, volume 19, 4: 16-22. Tian XC, C Kubota, B Enright, and X Yang. 2003. Cloning animals by somatic cell nuclear transfer-biological factors. Reprod Biol Endocrinol. 2003; 1: 98. doi:10.1186/1477-7827-1-98. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC521203/ Van Vleck, L.D. (1981) Potential genetic impact of artificial insemination, sex selection, embryo transfer, cloning, and selfing in dairy cattle, in B.G. Brackett, G.E. Seidel and S.M. Seidel (eds.), New technologies in Animal Breeding, Academic Press, London, pp 22 1242 Wilmut I, AE Schnieke, J McWhir, AJ Kind, and KHS Campbell. 1979. Viable offspring derived from fetal and adult mammalian cells. Nature. 1997;385:810–813. doi: 10.1038/385810a0. Woolliams J.A. and Smith C. 1988. The value of indicator traits in the genetic improvement of dairy cattle. Animal Production 46:333-345.
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SECTION 1. ANIMAL BIOTECHNOLOGY UDC: 636.32/.38(497.2) Original scientific paper
STUDY ON EWE LIVE WEIGHT OF PATCH FACED MARITZA SHEEP BREED Dimov D.1*, Vuchkov A.1, Ivanov I.1 1
Faculty of Agronomy, Agricultural University, 4000 Plovdiv, Bulgarıa *
corresponding author: [email protected]
Abstract The aim of this study was to investigate magnitude of live weight of adult ewes and rams of Patch-faced Maritza sheep breed as well as changes in ewe live weight from lambing to the end of lactation. Live weight of 296 ewes and 7 rams of Patch-faced Maritza sheep breed has been measured by a portable electronic scales FX2. The weighing was made in 6 herds in Plovdiv region (central part of South Bulgaria) with dried of ewes. In order to establish changes in ewe live weight during lactation an experiment was conducted with monthly weighing of 67 ewes in the herd of Agricultural university in Plovdiv. It was found that overall mean of ewe live weight is 74.47 kg and ram live weight in this study was 121.47 kg. In comparison with other local sheep breeds in Bulgaria and other European country this is higher live weight. ANOVA table showed a statistically proven impact of environmental factors - herd, age and body condition score of the ewe with high probability (p 101 -
Upper limit 900 5,0 10 0,3
Method of testing
% (mas.) mg/kg mg/kg classification sati mg KOH/g
51,0 class 1 6 -
0,02 500 24 class 2 0,5
EN ISO 5165 ISO3987 EN ISO 12937 EN 12662 EN ISO 2160 EN 14112 pr EN 14104
Iodine Linolenic acid ME
% (mas.)
-
120 12
pr EN 14111 pr EN 14103d
Polyunsaturated (≥ double bonds) ME Methanol Monoglyceride Diglyceride Triglyceride Free glycerol Total glycerol Alkali metals (Na+K) Phosphorous
% (mas.)
-
1
pr EN 14103
% (mas.) % (mas.) % (mas.) % (mas.) % (mas.) % (mas.) mg/kg mg/kg
-
0,2 0,8 0,2 0,2 0,02 0,25 5 10
pr EN 14110l pr EN 14105m pr EN 14105m pr EN 14105m pr EN 14105m/ pr EN 14106 pr EN 14105m pr EN 14108/ pr EN 14109 pr EN 14107p
pr EN 14103d EN ISO 3675/ EN ISO 12185 EN ISO 3104 ISO CD 36796 EN ISO 10370
Source: Jovanović, S., Jovanović, L. (2009). Perspectives of biodiesel use, Ecologica 16/2009, no 54, 123-127, p. 124, Belgrade.
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Increased interest in biodiesel was noticed during last decades. EU countries, the USA, Thailand imposed intensified tempo in biodiesel use as a substitution for fossil diesel. Europe produces 95% of biodiesel mostly from rapeseed and sunflower seed. In 2003 EU defined obligations for all EU members to substitute fossil diesel with biodiesel, deadlines with substitution percentage, control and monitoring of implementation as well as financial “penalties” for the members who resist defining substitution amount. There were predictions that biodiesel market could take 10% of diesel market in Europe until 2015. The USA suddenly increased biodiesel production which was 1, 9 million litres in 1999 to 95 million litres in 2004. American government agency estimates that biodiesel and ethanol could substitute between 25% and 50% American fuels obtained from oil until 2030. These assumptions match expectations that total energy should be provided from renewable resources until 2100. Other countries followed the example as well. In February 2005 China passed the law that 10% of energy would be obtained from renewable resources until 2020. India could be an important biodiesel supplier for its potential consideration that the price of biodiesel processing is a third of the price in Europe (Premović, T., Milićević, Z., 2015). Results and discussion Biodiesel effects on agricultural production – sustainability of agriculture Principal advantage of biodiesel use as a renewable fuel is a considerate reduction of CO 2 emission. Emission of sulphur oxides, suspended particles and carbon monoxide is reduced as well. Advantages and deficiencies of biodiesel use depend mostly on the type of used mixture as well as on performance and type of engine. Importance of potential reduction of certain pollutant emission during biodiesel use is in the following (Košut et.al. 2008). Carbon dioxide (CO2): On WTW basis, each ton of fossil diesel adds about 2, 8 t of CO2 in the atmosphere. The specific content of carbon in one ton of biodiesel is a bit lower, 2, 4 t of CO2. It is assumed that this carbon will be totally used by the crop giving the raw material for vegetable oil production as well as absorbed through carbon cycle (as glycerol and solid waste). In this way the “greenhouse effect” is alleviated as the current problem on the level of the entire atmosphere. Thus, it is said that the net CO2 emission in biodiesel use, observed on WTW basis, is almost equal to zero (Fukuda et.al. 2001; Furman et.al. 2006; Košut et.al. 2008). Sulphur oxides (SOx): Nowadays 1 t of conventional fossil diesel in EU contains maximum 350 ppm of sulphur on average. When diesel is burned, sulphur is released into the atmosphere in the form of sulphur dioxide contributing to formation of acid rains. Biodiesel contains almost no sulphur (sulphur content is 0-0, 0024 ppm). On the other hand, EU constantly promotes the use of diesel fuel with low sulphur content below 50 ppm (Great Britain), below 10 ppm (Sweden) (Košut et.al. 2008). Nitrogen oxides (NOx): Emission of nitrogen oxides from biodiesel can be increased or reduced compared to the emission from fossil diesel depending on the engine generation and testing procedure. Nitrogen oxide emission from pure biodiesel is increased for about 8% on average compared to fossil diesel (Košut et.al. 2008). When the mixture of small biodiesel amount and fossil fuel is used, the emission of greater quantity of nitrate oxides is possible which could be eliminated by adding additives for better combustion (Furman et.al. 2006). Considering the lack of sulphur in biodiesel, it is possible to apply control 261
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techniques on nitrate oxides which are impossible to apply on fossil diesel (Košut et.al. 2008). Carbon monoxide (CO): Biodiesel contains oxygenates improving the process of combustion and reducing emission. This fact considerably reduces (at least 20%) carbon monoxide emission. Solid particles: Inhaling of suspended particles is proved as a serious problem and danger for people‟s health. Emission in exhaust gases of these particles is 40% lower in biodiesel than in fossil diesel (Košut et.al. 2008). Increased use of biodiesel from the local areas improves micro economy of urban and rural regions. According to the data of the US Department for Agriculture, food price was increased for 5, 1% in 2008 whereas 0,2-0,6% goes to the biodiesel influence (biodiesel, bioethanol). Greatest effect on food price increase is the fuel price increase: in America food is transported to great distances mostly by trucks so the fuel price directly influences food price. Primary reasons for food price increase are: 1. price of oil included in the price of fertilizer and transport 2. increased consumption, population growth and improvement of nutrition quality in developing countries 3. climate conditions resulting in poor crop, floods, etc. 4. export restrictions of certain countries. Despite high fuel price, technological development leads to reduction of food price. In this way, increase of fuel price due to the increase of food price is not expected (Jovanović, S., Jovanović, L. (2009). Biodiesel has the highest energy balance compared to other fuels used in transport industry. New analyses show that the energy balance of biodiesel is positive (3,5:1). In other words, each unit of fossil fuel used to produce biodiesel results in biodiesel quantity containing 3,5 energy units. This study was conducted at the University of Idaho in cooperation with the US Department for Agriculture. The calculation excluded fuel consumption for planting, harvesting, fuel production and transport to the user. Economic aspect of biodiesel use – economic sustainabality of biodiesel Biodiesel produced from vegetable oils and animal fats is more expensive than diesel obtained from raw oil for 10-50% (Leung and Guo, 2006). High price of biodiesel is the main obstacle for its commercialization (Predojević and Bošković, 2009). It is known that the price of raw material is 60-75 about 80% of biodiesel total price (Thurmond, 2008), however, in the economic analysis of biodiesel production it is necessary to observe all the parameters influencing the price forming of biodiesel in the production chain consisting of oilseed production, their processing into raw oil and biodiesel production (Ma and Hanna,1999; Jovanović and Zekić, 2007). Basic condition for efficient biodiesel production and use is an adequate selection of raw material. Considering the content of oil in seeds and the price, the most optimal raw material for biodiesel in our region is rapeseed. However, current areas of this crop cannot be the raw material base for significant biodiesel production. Therefore, according to the previous research, one of the solutions would be the increase of the purchase price or implementation of various state incentives (Kiš et.al. 2007). 262
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Researching economic characteristics of biodiesel production, Kiš et.al. (2007) also concluded that biodiesel production from rapeseed was economically optimal. These authors published results of the conducted analyses on biodiesel production total costs with 59,6% of costs on the rapeseed raw material. Therefore, the authors emphasized that special attention should be dedicated to the analysis of current and prediction of the future development on the raw material market. Results of the conducted research determined the price of biodiesel as 41,6 dinars/l and that at the regular 18% tax rate is still lower than the selling price of D2 fuel, providing conditions for the profit in production. Jovanović and Zekić (2007) also analysed economic properties of biodiesel production from rapeseed. These authors determined material costs of oilseeds processing into biodiesel fuel in accordance with value of raw material (rapeseed) calculated from the market price, raw material processing costs (cleaning, crushing and pressing of the seed) and costs of raw oil processing into biodiesel fuel (methanol, sodium, hydroxide, sulphuric acid, distilled water, heat and electric energy production). Costs of fixed assets were determined as the sum of costs for amortization, maintenance, insurance and interest on equipment and buildings. Labour costs for direct workers and workers in general areas of productions were estimated for the biodiesel manufacture of the 10.000t a year capacity. Calculating all the cost elements for the manufacturing plant of 10.000t capacity, the authors determined the biodiesel price to be 53,21 dinars/kg or 46,99 dinars/l. When the price is increased for the accumulation value of 10% as common in our practice, the obtained biodiesel price of 51,69 dinars/l is still lower than market price of D2 fuel. Therefore, the authors ascertained that the biodiesel produced from rapeseed was competitive with the standard diesel fuel but also they indicated the issues related to exploitation characteristics as well as the development of distribution network. Lately, biodiesel production from castor oil has been researched since the castor yield is 80-100% higher than other oilseeds (sunflower, soya, rapeseed) whereas the obtained product is of a high quality and wide range of usage (Samardţija et.al. 2008; Premović, 2011). Results of numerous economic analyses indicate the important role of the state in price forming of biodiesel. Introducing of suitable incentives, reduction or tax exemption in biodiesel production, providing guarantees for loans, can help to achieve competitive price of biodiesel on the fuel market and thus to enable great use of this energy source (Jovanović et.al. 2005; Furman et.al. 2006). In order to reduce the cast of biodiesel production process, it is necessary to achieve optimal technological solution and to permanently improve it. Results of numerous researches indicate that one option of reducing the biodiesel production costs is to apply a continuous process of transesterification which also reduces the period of time of esterification process and increases production capacity. Production of high quality glycerol is another way to reduce the costs of biodiesel production. In case of inability of raw glycerol market valorisation, it is justified to use glycerol as the fuel for thermal energy. Maximal efficiency in biodiesel production entails valorisation of all the obtained by-products such as the pressed cake obtained in the vegetable oil production having multiple usage especially in food industry or potassium sulphate obtained in transesterification of fats (Furman et.al. 2005, 2006). Biodiesel can be produced from waste fats from slaughter industry or rendering processing. Used edible oil as a raw material for biodiesel production is another way to 263
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considerably reduce the cost of biodiesel production (Predojević and Bošković, 2009; Premović, T., 2011). Compared to raw vegetable oils, price of used – waste vegetable oils is lower for about 50-60% depending on their source and availability (Rice, 2000; Predojević and Bošković, 2009). Using waste oils to produce biodiesel contributes not only to reduction of biodiesel production price but represents a contribution to environment protection for solving the problem of such waste disposal. Generally, these oils end up in sewage causing the problem of waste water treatment (additional investment in energy consumption) or they integrate into the food chain in the production of animal food becoming a potential cause for health problems in people (Predojević and Bošković, 2009; Premović, T., 2011). Certain countries in Europe researched the possibility of waste oil collecting and the results of researches conducted in seven countries indicated that it was possible to collect 0.7 - 1 Mt of waste oils (Gonzalez-Gomez et al., 2002). However, the amount of waste oils is limited, production capacities are low and additional processing of raw material is recommended due to poor characteristics compared to the fresh unused edible vegetable oils having a greater content of free fatty acids, water, hydro peroxide, carbonyl and polymeric components of great molecule mass (Predojević and Bošković, 2009). Considering that quality of raw material influences on characteristics of produced biodiesel, raw material is limited in content with free fatty acids to maximal 0.5% mas. and with water to maximal 0.3% mas. Namely, present fatty acids and water participate in secondary reactions of saponification, hydrolysis and neutralization forming soaps and gels (emulsions) which hardens separation and purification of fatty acid methyl ester mixture and reduces the yield of the product (Hana and Ma, 1999; Predojević and Bošković, 2009). Purification process of obtained methyl esters after removing the excess methanol and glycerol is very important and mostly is conducted using water, citric and sulphuric acid or silica gel but also in different ways (e.g. using warm distilled water, 5% phosphorus acid, etc.) (Šiler-Marinković and Tomašević, 1998; Predojević and Bošković, 2009). Possibilities of biodiesel production in Serbia Serbia disposes of significant land potential for oilseed production used in biodiesel processing. Considering oil content in seeds and its price, rapeseed is the most optimal raw material for biodiesel in our region. Domestic oil refineries also have a potential to play the key role in providing future drives for biodiesel production (Kiš et.al. 2007). However, biodiesel obtained from fats is more expensive than diesel obtained from oil. In order to be an adequate and sustainable substitute for fossil fuels, biodiesel must be economically competitive to fossil fuel and therefore, it is necessary to determine and analyse all the factors which could improve production and use of biodiesel through reduction of biodiesel price (Ma and Hanna, 1999; Jovanović and Zekić, 2007; Premović, T., 2011). Oilseeds in Serbia are cultivated on about 270.000 ha, with the potential of over 620.000 ha. Depending on the cropping plan with a potential seedbed of 350.000 ha, it is possible to provide raw material for production of 212.800 to 250.600 t biodiesel. Sunflower, soya and rapeseed are considered as most important oilseeds in our region. In Serbia they are cultivated on 8.85% of arable land, or on 30.26% of total area with industrial plants in the country (Kiš et.al. 2007). Analysis of regional characteristics of most important oilseed production indicates the possibility to spread these crops, 264
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especially in Central Serbia, where they engage only 1% of arable land. Therefore, further enlarging of oilseed area could and should be directed to the region of Central Serbia, primarily of rapeseed, considering that this oilseed has no demands in quality land and grows on altitude up to 700m, but also of soya in river valleys. Serbia, thus, disposes of considerate land potential for oilseed production used in biodiesel processing. Additionally, domestic oil refineries also have a potential to play the key role in providing future drives for biodiesel production. Namely, it is known that processing capacities of refineries are considerably over-dimensioned for about 190.000 t, compared to domestic needs in raw oil. Existent market, economic and political circumstances determine whether the excess in production-processing of oilseeds is to be refined (edible oil), exported or processed into biodiesel (Kiš et.al. 2007). Also, there is a possibility for existent refineries to spread their drives for biodiesel production. Thus, for example, if a drive of 10.000t capacity were built, the amount of total investments would be 4.450.705 euros. Investment in transesterification equipment is estimated as the greatest issue in the investment structure ranging from 3.200.000 to 5.100.000 euros and depending on the production capacity (Kiš et.al. 2007). Conclusions Advantages of biodiesel production and use in comparison to oil and other energy sources, from the aspect of sustainable agriculture function, are numerous and could be observed in different ways. According to the results of various researches and our personal research and analyses, we emphasize following advantages of biodiesel use from ecological, economic, energy and technical aspect: 1. ecological aspect: use of biodiesel globally influences on reduction of greenhouse gas, particle and aromatics emission: CO, CO2, SO2, soot, benzene, toluene. It is nontoxic and biodegradable – fossil diesel is dissolved only 50% during first 21 days after spilling whereas biodiesel is dissolved 98% without consequences for the same time. B100 reduces the risk of cancer for 94% and B20 for 27%. 2. economic aspect: on macroeconomic level, development of biodiesel production would be caused by the influence on the following indicators: employment, increase of industrial production, additional overflow of funds into agriculture, contribution to economic growth of rural areas, reduction of macroeconomic factors dependence on external factors, industrial development would be enabled through progressive attitude of the state in terms of: subsidies policy, tax policy, long-term strategy on energy resource management. 3. technical aspect: biodiesel of EN 14214 quality represents fuel of high quality for diesel engines. Biodiesel specifications are similar to the regular diesel whereas the improvement is in the oxygen content in biodiesel providing a better combustion process and leading to reduction of exhaust gases emission and improvement of engine
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lubricity resulting in better efficiency and durability. These factors partly compensate the effect of low energy content, better preservation and handling. 4. energy aspect can be sublimed into two facts: biodiesel is a renewable energy source and its use reduces the need for fossil diesel which preserves stocks and reduces the risk of supply (Košut et.al. 2008). Therefore, considering European policy related to biofuels, especially biodiesel (energy, agriculture, research projects, etc.), critical factors of successful implementation can be summed in the following way (Košut et.al. 2008): price of agricultural products which are not used in food industry and forming of new markets, availability of arable land for the production of industrial plantations of oil crops, tax benefits enabling for biodiesel to be competitive to fossil diesel, long-term distinct participation of the state through establishing legislative framework and referential goals to introduce biofuels, promotional pilot projects implemented on local, regional and national level. According to the previous knowledge, it is concluded that technological development provided biodiesel production from all fats of vegetable and animal origin and thus started the race to conquer raw material base. Oilseed plants represent the greatest potential in biodiesel production. In order to obtain larger quantities of raw material, optimal production conditions are imposed on potential producers. Upcoming new generation of technology for biodiesel production will be much profitable and productive with widening of potential raw material types. Substitution of mineral fuels with biodiesel is expected to exceed 20% in several countries. At the same time, biodiesel economy must be directed towards sustainable development whereas people‟s appetite for energy must not further uncontrolledly increase for biodiesel and biofuels in general will supplement instead of replacing oil fuels. Expanding the share of biodiesel in total consumption of fuel for transport and agriculture could cause basic changes in the global history of energy (Premović, T., Milićević, Z. 2015). References Jovanović S., Jovanović, L. (2009). Perspsektive primene biodizela, Beograd, Ecologica 16/2009, broj 54, 123-127. Ma F., Hanna M. A. (1999). Biodiesel production, Bioresource Technology, 70:1, 1-15. Official Journal of the European Union, 17.05.2003, Directive 2003/30/EC of the European Parlament and of the Council of 8. May 2003. Fukuda H., Kondo A., Noda H. (2001). Biodiesel fuel production by transesterification of oils, Journal of Bioscience and Bioengineering, 92:5, 405-416. Premović T. (2011). Biodizel, Druga regionalna konferencija, Zaštita ţivotne sredine u energetici, rudarstvu i industriji, Zbornik radova, str. 307-312, Zlatibor, Srbija. Kiš F., Bošnjak, D., Jovanović, M. (2007). Organizaciono-ekonomska obeleţja proizvodnje biodizela, 48. Savetovanje: Proizvodnja i prerada uljarica, Zbornik radova, str.141-148, Herceg Novi, Crna Gora. Jovanović M., Zekić, V. (2007). Ekonomska obeleţja proizvodnje biodizel goriva, 48. Savetovanje: Proizvodnja i prerada uljarica, Zbornik radova, pp 149-155, Herceg Novi, Crna Gora. 266
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Košut Z., Nikolovski, Z., Mitrović, J. (2008). Biodizel-ekološki značajan i energetski obnovljiv izvor energije, Uljarstvo, 39, 1-2, 49-55. Furman T., Nikolić R., Tomić M., Savin L., Simikić M. (2006). Dosadašnji trendovi i perspektive proizvodnje i korišćenja biodizela u svetu, Traktori i pogonske mašine, 11:1, 60-65. Premović T., Milićević, Z. (2015). The use of biodiesel, in order to protect the environment and sustainable development, Ecologica (in press), Beograd. Predojević, Z., Bošković, G. (2009). Postupci prečišćavanja biodizela dobijenog iz otpadnih ulja, 50. Savetovanje: Proizvodnja i prerada uljarica, Zbornik radova, pp 235-241, Herceg Novi, Crna Gora. Rice B., Pelkmans, L., Mittelbach, M.(2000). Waste oils and fats as biodiesel feedstocks: assessment of their potential in the EU, Teagasc, Ireland. Samardţija M., Samardţija D., Marinković R., Furman T., Tomić M. (2008). Proizvodnja biodizela iz ricinusa, Savremena poljoprivredna tehnika, 34, 3-4,136-142. Thurmond W. (2008). From trash to cash. Biodiesel International, January 2008, www.emerging-markets.com/biodiesel/pdf/BiofuelsInternational. Šiler-Marinković S. S., Tomašević, V. A. (1998). Transesterification of sunflower oil in situ, Fuel, 77, 1389-1391.
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SECTION 2. AGRICULTURAL ECONOMICS UDC: 631.115.11:635.631.64 Original scientific paper
THE IMPACT OF THE INCOME FROM THE PLASTIC TUNNEL PRODUCTION OF TOMATOES, PEPPERS AND CUCUMBERS OVER THE FAMILY BUDGET OF THE SELECTED AGRICULTURAL HOUSEHOLDS Pesevski M.1*, Tomic D.2, Rajic Z. 3, Zivkovic D. 3, Micic I.3 Faculty of Agricultural Sciences and Food, University “Ss. Cyril and Methodius” in Skopje, Skopje, R. Macedonia 2 Institute of Field Crop and Vegetable Production, Novi Sad, Serbia 3 Agricultural Faculty, “University in Belgrade”, Belgrade, Serbia
1
*
corresponding author: [email protected]
Abstract Considering the general principle for the area under selected crops to be higher than 0.02 ha, 25 family agricultural households from the Pcinja district are included in the survey. In that context, the survey captures 8 agricultural households from the municipality of Vranje, 5 from the municipality of Bujanovac, 4 from the municipality of Presevo, 3 from Vladicin Han, 2 agricultural households form the municipalities of Trgoviste and Surdulica in particular, as well as 1 from the municipality of Bosilegrad. The total income generated from non-agricultural activities (salary, pension, social aid, inflow from relatives abroad, etc.), as well as the agricultural income are determined. The agricultural income is consisted of two parts, i.e. the value of the agricultural production determined by farm-gate prices for the period 2008-2010 on one side, and the value of the direct support to the farm production (subsidies), on the other side. The results reveal that the total income is between 7,687.92 EUR for the T.H. agricultural household in the municipality of Bujanovac to 32,240.19 EUR for the S.D. agricultural household from the same municipality. The average income is 16,706.9 EUR, whereas the agricultural income participates with 86.2%. Out of the total agricultural income, the income generated from crop production has a share of 86.8%. The share of the income by selected crops in an average is determined as: 18.83% for the tomato production, 11.83% for pepper production and 17.56% for production of cucumbers. The plastic tunnel production generates income in an average of 45.21% from the total family budget of the analyzed agricultural households. Key words: family agricultural households, income, plastic tunnel, tomatoes, peppers, cucumbers. Introduction In conditions with heterogeneous assortment of cultures, where the production structure includes number of crops and different types of livestock, the decision of which crop or livestock type will dominate the total engaged area is of great importance. The significance 269
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of this decision is even larger if the crop production includes intensive crops such as vegetable crops and especially crops cultivated in greenhouses. It is a known fact that this kind of cultivation on a relatively small areas provide relatively high production yield, which is valued differently on the market. The difference in the valuation provokes different effects on the amount of income for the households. In general, the revenues depend on the type of the crop type, i.e. the product. The revenues from the same product dependent also on the market conditions, the time of their delivery (when they are marketed), and to a large extent depend on their quality. Namely, there is a difference when the market has more supply than demanded, in which conditions the product is sold at a lower price than when the demand is greater than the supply. Correspondingly, the products are sold at a relatively higher price when the product in the winter and spring months, opposite of the products‟ prices in the summer and autumn months. Moreover, the product produced by the principles of organic food production should also reach higher price over the products produced on the conventional manner. Bulatović (1997) performed research of the economic and production factors in the production of pepper on a private property in the Skadar basin region in Montenegro. The research confirmed that the consumption of human labor is 1832 h/ha on an average area of 0.86 ha, and the average yield is 26.10 t/ha. Labor productivity, expressed through the value of production amounted to 89.00RSD/h. The amount of human labor in the pepper production, for an average yield of 15,100 kg/ha, is 1798 h/ha on average (Bošnjak Danica, Jovanović M., 1997). In these conditions, as well as use of57 tractor hours, the labor productivity amounts 8.4 kg/h or 7.0 min/kg on average. Живковић D. (2007) and associates performed analysis of the organizational-economic parameters in the production of pepper in the case of the rural households. Among other things, the authors concluded that pepper is a relatively highly labour intensive crop, with low level of mechanization in the production processes. In fact, they found that 1,116.9 hours of human labour, and the intensity level of 7.69 is spent for one hectare. For production of one tone of peppers 27.9 hours of human labour and 3.63 hours of machines is spent. The yield was 40 t / ha in 2005, and the value of production at an average selling price of 25,00 RSD/ kg amounted one million dinars. According to the authors, the level of efficiency was 3.16 and the coverage of the variable costs 216.4%. Peševski and Stojkov (2014) performed comparative analyses on the amount of variable costs and gross margin in the production of pepper in greenhouses and outdoors in Strumica, Macedonia. The authors found that the variable cost in the production of pepper in greenhouses amounted 3,073,773.00 MKD/ha and the gross margin amounted 388,150.00 MKD/ha, while the variable costs in the outdoors production of pepper amounted 4,526,260.00 MKD/ha and the gross margin was 348,755.00 MKD/ha. Pavlović N. et al. (2010) found that the total profit from the organic production of tomatoes produced in Vojvodina (Serbia)amounted to 6,210 EUR/ha, with a41.4% rate of return and 1.71cost ratio. Whereas for the production of peppers in the same conditions, the profit amounted to 6,110 EUR/ha, 43.33% rate of return and cost ratio of 1.76. According to Tsoho and Salau (2012), the total cost of tomatoproduction consist of two components: fixed and variable. Variable costs dominate the total cost of production. In circumstances where the household rents land, these costs participate with 92.19% share on average, while when using its own land - 87.89% of the total costs. For Morteza, Ajabshirchi, Mobtaker and Abdi (2012) the focus of the research was on the energy consumption, and 270
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cost-benefit analysis of production. They found that the total cost of production of tomatoes, with an average yield of 135,000 kg, amounted to 34,939 $/ha, and the rate of gross margin 75.9%. The total cost in thecucumber production,with an average yield of 88,123 kg, amounted to 31,956 $/ha, and 65.1%. rate of gross margin The art of the household members isto choose from a number of vegetables that will provide the highest income and participate with the highest share in the family budget.Therefore, the purpose of this paper is to examine the impact of the revenues from the three different kinds of vegetables grown in greenhouses, on the family budget of selected farms in the Pchinja district in R. Serbia. Materials and methods In order to realize the goal of this research, 25 households were surveyed.The selected households were of mixed type, with vegetable and livestock production producing among other things, tomatoes, peppers and cucumbers in greenhouses. The questionnaire was constructed in a way that can easily identify any family income and variable production costs. The data was then processed using mathematical and statistical methods common for such research. The results are presented in tabular and graphical manner. Results and discussion The total revenue in the selected group of family farms range between 7,687.92 EUR, in the T.N household from the Bujanovac municipality,and 32,240.19 EUR in the S.D household from the same municipality, and the income with amedian value was 16,706.9 EUR. Here, agriculture accounts for 86.2%, with 86.8%. of the production devoted to crop production. The family farm in the municipality of Bosilegrad (TN) expressed the lowest income of the agricultural sector (9,934.8 EUR), although with relatively large amount of arable land (3.56 ha). Besides the land cultivation, the householdalso owns animals (2.67 pigs cows and 2). Accordingly, parallel to the intensive plastic tunnel production the production structure includes alfalfa and potato. In regards to the previously set objectives in the survey, it is apparent that the earnings from the selected crops are important. The revenue share of the tomato production, account for 15.83% of the total income on average in the investigated group of farms. The lowest share of these revenues (6.82%) is observed in the N.B household from the Bosilegrad municipality, and the A.Mhousehold from the Bujanovac municipality recorded the highest share (34.28%) (Figure 1).
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Figure 1 – Share of the income from the selected crops in the total income of the households The lowest share of income from the production of peppers (3.81%) was registered in the M.V household in the Vranje municipality, and the highest (40.72%) share was seen in the case of the J.R household, again in the municipality of Vranje. The generated revenues from the production of cucumbers, participate with 17.56% of the total household income on average. The minimum value (4.99%) in the relative share of revenue from cucumbers is seen in the family farm A.J in the municipality of Vranje, and the maximum value (36.77%) in family farm M.C. in the municipality of Bujanovac. The profitability of the production,as the ratio between theoutput and the variable costs, shows the generated production for a dinar of variable costs spent in the process of production, expressed in monetary units. Profitability, or the profitability indicator determine the relative share of income in the value of production. Based on the concept of gross margin wecalculate the degree (ratio) of variable costs coverage with the amount of gross margin in the case of selected crops. The highest degree of rational labor and resourcuse is seen in the production of tomatoes in the S.D household, in the Bujanovacmunicipality; where one dinar of variable costs generates a value of 4.3 RSD (Table 1). Contrary to this situation, two farms (M.S and Z.S)articulateminimal profitability and investment of human labor. Overall, in the case of 11 households from the total of 25 households, the profitability is higher than the average.
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Table 1. Comparative outlook of the qualitative economic indicators in the production of tomatos, peppers and cucambers Tomatoes Peppers Cucumbers Household Economy Profitability Coverage Economy Profitability Coverage Economy Profitability Coverage M.B. 2,6 61,9 162,2 2,5 59,6 147,4 3,0 108,84 327,99 N.T. 4,2 76,1 318,9 2,2 54,0 117,6 2,4 35,35 84,66 M.S. 1,6 35,9 56,0 2,1 65,3 139,8 2,8 36,61 102,25 Z.S. 1,6 35,7 55,5 1,5 34,8 53,4 2,8 33,07 91,15 J.P. 3,0 66,8 201,0 4,4 77,4 342,8 3,1 40,37 126,62 M.M. 2,8 64,3 180,3 2,4 58,0 138,0 2,6 125,73 324,50 A.J. 3,1 68,2 214,2 2,2 54,1 117,7 5,0 36,75 185,08 G.J. 2,0 49,5 98,0 2,3 55,9 126,6 1,7 53,12 88,53 N.B. 2,3 56,0 127,4 1,6 39,3 64,8 2,2 60,77 133,95 S.D. 4,3 77,0 333,9 4,6 78,3 361,8 2,7 47,76 128,99 M.C. 2,4 58,7 142,3 2,9 65,4 189,1 1,8 32,16 58,20 B.J. 2,0 50,9 103,7 2,9 65,7 191,1 1,4 21,10 30,39 T.N. 2,1 52,0 108,2 1,5 31,5 46,0 2,7 34,96 95,76 Z.S. 2,1 51,8 107,4 2,8 64,1 178,6 2,0 47,89 96,55 F.I. 2,8 64,0 177,7 2,3 57,1 133,1 3,0 82,23 246,60 S.M. 2,2 50,9 109,6 1,6 35,5 55,0 2,3 40,06 93,64 A.M. 2,4 58,7 142,3 2,9 65,4 189,1 1,8 32,16 58,20 A.S. 2,9 65,4 188,7 2,2 55,5 124,8 2,5 62,31 157,22 D.J. 2,9 65,1 186,3 2,3 56,7 130,7 2,6 47,46 125,72 Z.S. 2,8 64,2 179,0 2,0 50,6 102,3 2,3 55,20 127,74 LJ.G. 2,6 61,8 161,9 2,1 52,3 109,8 2,7 94,26 252,41 M.S. 2,3 56,3 128,8 2,5 60,2 151,4 1,7 45,79 77,28 Z.B. 3,5 71,2 247,0 3,5 71,7 253,5 2,6 47,30 121,64 N.G. 2,9 65,4 188,8 2,0 50,1 100,4 2,8 104,06 295,62 S.M. 2,4 57,6 136,1 3,0 66,5 198,1 2,2 43,33 96,35 Legend Minimum Maximum The highest degree of rationality of the investments of labor and resources in the production of pepper is same as in the case of the tomato production (S.D household), and the lowest level is seen at Z.Shousehold from Vranje municipality of. In general, around 32% of the total agricultural households in this research, have a higher cost than average. In the process of cucumber production, the most rational input spending‟s was seen in the B.J household from the Vranje municipality, and the lowest level of rational useof inputs were in the Bujanovac municipality in the B.J household. Here, only 7 households have been exercising better results than average productivity (2,4 RSD). Out of the total number of households in the sample, in the case of all of the selected crops the profitability was higher than the average value in 13 households. The best results in the case of the tomato and pepper production was realized by the S.D household from the Bujanovac municipality 273
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and the worst in the tomato production Z.S household (Municipality of Vranje), while the same was the case in the T.N household (Bujanovac municipality) whenpepper production was concerned. Concerning the profitability of the cucumber production, the best results are seen in the M.V household from the Vranje municipality, and the worst results were perceived in the B.J household from the Bujanovac municipality. The coverage of the variable costs with the gross margin in the tomato production was averaging162.2% of the pepperproduction and141.1% of the cucumber production. A comparison of the coverage rate in the investigated sample (25 households) shows that the production of tomatoes, the lowest level of coverage (55.5%) was registered in the Z.S household (Vranje municipality) and the highest (333.9%) in the S.D household (Bujanovac municipality). In the case of pepper production, the highest level of coverage (361.8%) was registered in the S.D household from the Bujanovac municipality, and the opposite was seen in the T.N household (Bujanovac municipality) with a minimum amount of coverage of 46%. The coverage of variable costs in the production of cucumbers is the highest in the M.V household from the Vranje municipality, and the lowest with 30.39% in the case of B.J household (Bujanovac municipality). Conclusions Based on the comparison of the qualitative indicators we can recommend that the tomato and pepper production should follow the example of S.D. households in the Bujanovac municipality, since this is a household which generated the best economic results.While in the cucumber production the M.V in the municipality of Vranje should be a representative example household, because it achieves the best economic results. In the case of the S.D household, based on percentage shares of revenues from these crops in the total household income, we recommend reducing the area under cucumbers in order to reduce their share from 20.02% in favor of the tomato and pepper production. Recommendation for the M.V household from the Vranje municipality is that in the future it should adapt the structure of sowing on their land property of 2,6 ha, in favor of the plastic tunnel production, especially in the production of peppers, and in order to increase the share of these revenues by 3.81%. References Bošnjak D., Jovanović M. (1997): Produktivnost rada u proizvodnji paprike. Ekonomika Poljoprivrede, Vol. 44, br. 1-2, str. 15-16. Beograd. Bulatović B. (1997): Organizaciono-ekonomski činioci i njihov uticaj na rezultate u proizvodnji paprike na privatnom posjedu skadarskog basena. Poljoprivreda i šumarstvo, Vol. 43 (3): 1-168, str. 137-147, Podgorica. Живковић Д., Мунћан П., Цветковић М. (2007): Организационо-економска обележја производње паприке на породичном газдинству руралног подрачја. Зборник радова: Мултифункционална пољопривреда и рурални развој у Републици Српској, стр. 346355, 13 – 14. Децембар Јахорина, Б и Х. Pavlović N., Ugrinović M., Zdravković M. (2010): Economic and agronomic analysis of organic production of tomato and pepper, Economics of Agriculture, vol. 57, no. 2, pp. 153-157, Belgrade.
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Peševski M., Stojkov A. (2014): Economic analysis of peper production in Republic of Macedonia. Thematic Proceedings. International Scientific Conference: Sustainable Agriculture and Rural Development in Terms of The Republic of Serbia Strategic Goals Realization Within The Danube Region – rural development and (un)limited resources, pp 377-392, June, 5-6th 2014, Belgrade, Serbia. Taki M., Ajabshirchi Y., Mobtaker H.G., Abdi R. (2012): Energy consumption, inputoutput relationship and cost analysis for greenhouse productions in Esfahan Province of Iran. American Journal of Experimental Agriculture 2(3): 485-501. Tsoho B. A., Salau S. A. (2012): Profitability and constraints to dry season vegetable production under fadama in Sudan savannah ecological zone of Sokoto State, Journal of Development and Agricultural Economics Vol. 4 (7), pp. 214-222, Nigeria.
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CONTENT INFLUENCE OF SOME HERBICIDES ON BARLEY 1000 KERNEL WEIGHT DEPEND ON GROWTH STAGES Glatkova G.
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STATE OF THE INTEGRATED PEST CONTROL SYSTEMS IN THE VINE YARDS IN BULGARIA Lyubenova T., Genov N., Balashkov E., Prodanova-Marinova N.
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IMPACT OF TRICHODERMA ATROVIRIDE AND EFFECTIVE MICROORGANISMS THE YIELD OF PEPPER (CAPSICUM ANNUUM L.) IN THE GREENHOUSE Dozet G., Cvijanovic G., Jaksic S., Djukic V., Kaludjerovic D., Miladinov Z.
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INSECT PESTS OF STORED CEREAL IN NORTHERN CYPRUS Gözüaçık C., Güllü M., Konuksal A., Yücel A., Hekimhan H.
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THE IMPACT OF STROBILURIN FUNGICIDES IN PREVENTIION OF DAMPING OFF DISEASE ON TOBACCO SEEDLINGS Tashkoski P., Krsteska V., Mitreski M., Stojanoski P.
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PHTHORIMAEA OPERCULELLA (ZELLER), PEST CONTROL ON TOBACCO Krsteska V., Lazarevska S., Tashkoski P., Stojanoski P.
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EXTRACELLULAR ENZYME ACTIVITY OF TRICHODERMA STRAINS ISOLATED FROM DIFFERENT SOIL TYPES Danilović G., Radić D., Raičević V., Jovanović Lj., Kredics L., Panković D.
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BIOLOGICAL AND CHEMICAL CONTROL OF GRAY MOLD DISEASE (BOTRYTIS CINEREA) OF TOMATO GROWN UNDER PLASTIC TUNNELS Rusevski R., Kuzmanovska B., Bandzo K., Sotirovski K., Risteski M.
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INFLUENCE OF SOME HERBICIDES ON BARLEY 1000 KERNEL WEIGHT DEPEND ON GROWTH STAGES Glatkova G.1* 1
Agricultural institute-Skopje, 1000 Skopje, Republic of Macedonia *corresponding author: [email protected]
Abstract During 2006, 2007 and 2008 an experiment was carried out to investigate influence of six herbicides (2,4-D, MCPP+dicamba, triasulfuron+dicamba, 2,4-D+florasulam, amidosulfuron+jodosulfuron and florasulam+flumetsulam) on 1000 kernel weight of three barley varieties (reks, NS 293 and egej), treated in the three different growth stages (tillering, first node visible and second node visible). In 2006 triasulfuron+dicamba applied in tillering growth stage significantly reduce barley 1000 kernel weight at reks variety compared with weed free control. MCPP+dicamba in tillering growth stage significantly increase barley 1000 kernel weight at reks variety compared with weed free control. Similar, MCPP+dicamba applied in second node stage significantly increase barley 1000 kernel weight at NS 293 variety compared with weed free control. At egej variety florasulam+flumetsulam, 2,4 D+florasulam and triasulfuron+dicamba applied in tillering stage significantly increase barley 1000 kernel weight compared with weed free control. Florasulam+flumetsulam and amidosulfuron+jodosulfuron applied in second node stage significantly increase barley 1000 kernel weight compared with weed free control. In 2007, 2,4 D applied at tillering stage significantly reduce barley 1000 kernel weight of NS 293 variety compared with weed free control. In 2008, 2,4 D+florasulam applied at first node visible stage significantly increase barley 1000 kernel weight at reks variety compared with weed free control. Similar, 2,4-D and 2,4 D+florasulam, as well as 2,4D+florasulam, amidosulfuron+jodsulfuron and florasulam+flumetsulam applied at tilering stage and the first node visible stage, respectively significantly increase barley 1000 kernel weight at NS 293 variety compared with weed free control. Key words: barley varieties, herbicides, growth stages, barley 1000 kernel weight. Introduction Barley (Hordeum sativum Jessen) in the Republic of Macedonia is grown on about 47 500 ha with average yield of 3 440 kg/ha (Anonymous 2009). It is the second cultivated crop right after the wheat. Very often barley production is disturbed by weed infestation. In barley losses due to competitive effects of weeds estimated at 15-25% of potential production. Contemporary chemical weed control in barley and wheat begins after Second World War. Since then, over 50 a. i. are synthesized for selective weed control in barley and wheat. The most of the herbicides which are used in barley is foliar applied and tillering stage is the optimum growth stage for their application. By using the herbicides in 279
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advance growth stages of barley, sensitivity can be increased and barley yield elements can be reduced (Allien 1966, Markovic 1978, Rinella et al., 2001). Application of herbicides during the differen growing stages can have various influences on 1000 kernel weight. Positive influence of some herbicides on 1000 kernel weight was found by Ognanovic et al., (1988). Spasik (1972) emphasized that tested herbicides have negative impact in first and positive in second year of examination. According to this, the aim of the experiment was to evaluate the influence of different herbicides applied in three growth stages on 1000 kernel weight at some barley varieties. Materials and methods Field trials were conducted during 2006, 2007 and 2008 at the Agriculture institute in Skopje. The experimental design was randomized complete block system with four replicates, and harvest plot size of 16 m2. The studies were carried out with three barley varieties: reks, NS 293 and egej which were seeded with seedling rate of 200 kg/ha. Table 1. Variants of the trial Variants- active ingredient (a.i.)
Trade names
Rate
Weed free control 2,4-D MCPP+dicamba Triasulfurone+dicamba 2,4-D+florasulam Amidosulfurone +jodosulfuron Florasulam+flumetsulam
/ DMA-6 Banvel-P Lintur Mustang Sekator Derby
/ 1 Lha 4 L/ha 150 g/ha 0,5 L/ha 0,25 L/ha 60 ml/ha
Time of aplication / I, II, III I, II, III I, II, III I, II, III I, II, III I, II, III
I-tillering stage II- the first node stage III- the second node stage All herbicides were applied with CO2-pressurized backpack sprayer with 300L/ha water. After harvesting, barley 1000 kernel weight was measured in seed testing laboratory of Agricultural institute in Skopje. The examinations were determined based on 100 randomly selected spikes of each repetition i.e. 400 spikes of each variant. Barley 1000 kernel weight was measured according to ISTA methods. The data were subjected to statistical analysis applying LSD-test Results and discusion In 2006 (Table 2) barley 1000 kernel weight was ranged from 42,3 g at reks variety treated with triasulfurone+dicamba at tillering stage to 51,8 g at NS 293 variety treated with MCPP+dicamba at the second node. Highly significant decrease of barley 1000 kernel weight (p 0,81 and the growth is stimulated by 10 % of CO2.P. expansum is one of the oldest described Penicillium species, established as the principal cause of spoilage of pome fruits. Like many other Penicillium species, it is psychrophile capable to grow from -6° to 35°C, aw minimum 0,82 and very low requirement for oxygen. P. italicum is primary spoilage agent of citrus fruits (capable for
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growth from 1,6 to 9,8 pH) and requires higher aw for minimal growth (0,87) (Pitt and Hocking, 2009). Mold growth in foods can be controlled by numerous methods and procedures like preventing spores from entering foods, adjusting storage temperatures and humidity, altering oxygen levels, using chemical preservatives and related measures. Unfortunately, molds are capable to adjust and stated methods do not prevent molds from altering food quality and safety in every situation in food processing and storage (Jay, 2000). Chemical preservatives are substances capable of inhibiting, retarding or arresting the growth of micro-organisms or of any deterioration resulting from their presence or of masking the evidence of any such deterioration. Most widespread preservatives in fungal inhibition are sodium benzoate and potassium sorbate (Spadoni et al., 2015). Used in different foods, they inhibit fungal growth in foods especially at lower pH.Preservatives can be used with lower pH values, lower temperature or combined with different chemicals like ethanol (Karabulut et al., 2005) to reduce fungal spores viability. Preservatives have important antimicrobial purpose in food processing, although can be used in post-harvest treatment as well in order to substitute toxic fungicides (Montesinos-Herrero et al., 2009). The aim of this study was to: (1) determine minimal fungicidal (MFC) and inhibitory (MIC) concentration of sodium benzoate and potassium sorbate on three Penicillium species at two pH (2) evaluate colony linear growth inhibition at two pH and (3) to test effect of preservatives in in situ conditions on apple slices. Materials and methods Microorganisms and culture conditions Fungal species: Penicillium aurantiogriseum, P. expansum and P. italicum were obtained from Charles University Culture Collection in Prague. Cultures were maintained on Potato dextrose agar (Liofilchem, RosetoDegli Abruzzi, Italy) at 4°C. Fungal spores were harvested after 5 day incubation at 25°C in 0,2 % agar with 0,05 % Tween 80 (Biolife, Milan, Italy) and adjusted do 1×106 spores/mL by haemocytometer. Minimal fungicidal and minimal inhibitory concentration To determine minimal fungicidal (MFC) and minimal inhibitory (MIC) concentration macrobroth dilution method (Agarwalet al., 2000) was applied. Preservatives (K-sorbate and Na-benzoate; Kemika, Zagreb, Croatia) were tested by macrobroth two-fold serial dilution technique. Preservatives were dissolved in water to get a stock (300 mg/mL) solution, filter sterilized and dilutions were made serially. Fungal spores were inoculated (106 CFU/mL) in Potato dextrose broth (Liofilchem, Rosetodegli Abruzzi, Italy) with pH adjusted to 5,12 and 4,5 by 1 M HCl. The tubes were incubated at 25±1°C and the MFC and MIC (in ppm) was recorded after 72 h of incubation. Suitable controls: broth control (without fungal spores) and growth controls (with fungal spores) were set under identical conditions. The last tube with no apparent growth of the organism represented the MIC of the compound. Colony growth inhibition Inhibition of radial colony growth was tested in Petri plates on Potato dextrose agar (Liofilchem, Rosetodegli Abruzzi, Italy) amended with preservativeswith pH adjusted to
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suspension with concentration of 1×106CFU/mL. Plates were prepared in triplicate and incubated at 25°C. In controls, preservatives were omitted. During incubation at 25°C the radial growth of the fungal colony was measured every 2 days in two directions. For each culture the radial extension of colonies were plotted against time. Daily measured radii were analyzed statistically by analysis of variance (ANOVA) (P < 0,05) followed by the Bonferroni post hoc multiple comparisons by XLSTAT Version 2009.3.02 (Addinsoft SARL, 1995-2009). In all cases the experiments were carried out in triplicates per treatment. All treatments were repeated at least two times. In situ experiment For in situ conditions, apple slices (Idared) were used. After washing in tap water, sterile demineralized water and whipped of with 70 % ethanol. Apples were cut with sterile knife to slices of 5 mm thickness and cored to 22 mm round slices by sterile metal cutter. Apple slices were dipped for 30 seconds in 2 % ascorbic acid solution (to prevent browning) and drained on sterile paper towels in laminar hood under UV. After drying, slices were dipped in preservative solution (50, 100 and 200 ppm) for 30 seconds. After draining on sterile paper towels, 4 apple slices were transferred to Petri dish in two repetitions. Each slice was 6 CFU/mL) and incubated at 25°C for 6 days. Radius of fungal colony was measured every other day in two perpendicular directions. Results and discussion Chemical compounds or preservatives with low toxicity and generally recognized as safe (GRAS) compounds (like K-sorbate and Na-benzoate) are amongst most important preservatives in food processing and have received increasing interest for the control of postharvest fruit disease like blue mold apple decay or green mold or sour rot of citrus fruits (Spadoni et al., 2015). Antifungal activity of tested preservatives to selected Penicillium species (Table 1) was very uniform at both tested pH values (5,12 and 4,5). Seems that difference in pH units was not enough to show difference in inhibition or fungicidal action in this experimental section. Manso et al. (2015) have noticed that pH value lowering from 7 to 5 and 3 have noticeable effect in lowering concentrations of cinnamon and oregano essential oils MIC and MFC. It is worth to mention that incubation time in this part is relatively short (3 days), and fungi are transferred to sterile PDA broth without adjustment pH adjustment in order to establish MIC or MFC activity. This transfer allows fungi to repair damages in cells so fungi, even grown at lower pH can recuperate. Both P. expansum and P. italicum showed sensitivity to K-sorbate (200 ppm MIC, > 800 ppm MFC) while to inhibit P. aurantiogriseum double concentration of K-sorbate was required. K-sorbate exhibited fungicidal activity in concentration more than 800 ppm, to all tested species. Contrary to K-sorbate, P. aurantiogriseum was the most sensitive fungi to Na-benzoate, with 200 ppm (at both pH value tested) for MIC concentration. Although similar in morphology even members of the same subgenus Penicillium. Compared to K-sorbate, Na-benzoate increases it`s toxic effect at lower pH (MFC of P. aurantiogriseum and P. expansum is 800 ppm). In this experimental setup it is evident that by lowering pH value, fungal growth is retarded. Membré et al. (2001) have found that sorbic and propionic acid also have a significant inhibitoryinfluence on the growth rate of P. brevicompactum since they led to a 379
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50% reduction of mycelium proliferation at pH 7 and greatly inhibited the mould at pH 5. On the other hand, Na-benzoate did not significantly modify thegrowth rate, as compared to the control. This result mostly supports our findings where Na-benzoate had reduced potency in retarding tested fungi growth, compared to K-sorbate. Similarly, Lopez-Malo et al. (2007) have reported lower MIC of Na-benzoate at 3,5 (400 ppm), compared to experiment performed at pH 4,5 on Aspergillus flavus.Benzoate has a pK value of 4,20 and since the antimicrobialactivity of benzoate resides in the undissociated form,therefore, weak acid preservatives exist in a pH dependent equilibrium between the undissociated and dissociatedstate. Optimal inhibitory activity takes place at low pHsince it favors the undissociated state of the molecule thatis freely permeable across the plasma membrane. Oncethe neutral molecule enters the cell it will dissociate dueto a higher pH. The preservative action is thought to bedue to an accumulation of anions and protons inside thecell (BrulandCoote, 1999; Jay, 2000). Table 1. Effect of K sorbate and Na benzoate on selected Penicillium species K-sorbate Na-benzoate Species Effect pH 5.12 pH 4.5 pH 5.12 MIC* 400 *** 400 200 P. aurantiogriseum MFC** > 800 > 800 > 800 MIC 200 200 800 P. expansum MFC > 800 > 800 > 800 MIC 200 200 800 P. italicum MFC > 800 > 800 > 800 * = minimal inhibitory concentration ** = minimal fungicidal concentration *** = values expressed in ppm
pH 4.5 200 800 800 800 800 > 800
As mentioned, contact with preservatives during MIC and MFC determination was relatively brief (48 h of incubation in broth with chemicals, additional 24 h in pure broth for confirmation). Preservatives inhibition spore germination, germ-tube elongation as well as enzyme activity in cytoplasm (Hervieux et al., 2002) resulting in postponed or retarded growth of fungi. This effect is more pronounced in retarded fungal colony growth in a growth medium amended with K-sorbate, especially at lower pH (4,5) (Fig 1-3). During 18 day of incubation, Na-benzoate acts similarly regarding difference in pH, although it is worth to mention that 200 ppm at final incubation period is significantly different compared to other samples (Fig 1). The situation with K-sorbate is different. Even at pH 5,12, growth at 100 and 200 ppm showed significant difference, more pronounced at lower pH where all concentrations applied were significantly different, compared to control sample. Additionally, lag phase of fungal growth is prolonged to 3 days. This can be important during storage of perishable fruits or ready to eat fruit salads. Although P. expansum is a notorious for a rapid growth on storage fruits, in our experimental setup was the slowest growing specie (Fig 2). The reason could be more nutrition elements in real substrate (fruit), compared to growth media. Na-benzoate affects this specie growth, especially at lower pH, higher concentration with more potency, but the differences in colony radii were not statistically significant, although the difference exist. K-sorbate was proved to be more effective in colony growth inhibition, with statistically significant difference at 4,5 although uniform effect is noticeable even at 5,12. Effect of
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higher concentration is visible after 10th day, although no significant difference was detected. P. italicum was the most rapid growing specie of Penicillium genera tested in our experiment (Fig 3). It reached the end of 9 cm Petri dish in 12 days (Fig 3). As presented in Fig 3, Na-benzoate had no effect during experiment, although some differences in colony size are obvious at lower pH, but there is no statistical significance. K-sorbate fungal inhibit was stronger than Na-benzoate, at both pH values. Although no statistical difference was observed at pH 5,12, all concentrations of preservative applied slowed fungal growth, nevertheless, at 14 day of incubation period, all samples reached the end of the dish. Retarding of fungal growth was more potent at pH 4,5 where lag phaseof 2 days was observed, as well as statistically significant difference in the inhibition at highest concentration of Na-benzoate applied (200 ppm). At the highest inhibition moment, colony radii difference is more than 20 mm. To test in situ effect of preservatives, Idared apples was used. To avoid rapid overgrowth of fungi, P. expansum was tested with Na-benzoate, more common preservative. The effect of Na-benzoate was more pronounced to fungal colony growth inhibition (Fig 4), compared to the results of colony growth on PDA (Fig 2) implying the need for testing antimicrobial effect in real systems, as well as growth media. Na-benzoate of 200 ppm was the only one that retarded fungal overgrowth even on 6th day of incubation period at 25°C. The least effective preservative was K-sorbate at lowest concentration (50 ppm). Incubation temperature throughout the experiment was uniform (25°C) to maximize favourable environment for fungal growth. Lowering temperature to 4-8°C we can assume more pronounced, synergistic effect of lower temperature and preservative is to be expected.
Figure 1. Effect of preservatives on P. aurantiogriseum radial colony growth at pH 5,12 and 4,5 * = significantly different (p