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Fırat Üniversitesi Sağlık Bilimleri Veteriner Dergisi
2009, Cilt 23, Sayı 3, Sayfa(lar) 153-159
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Karanfil Ekstraktının Etlik Piliçlerde Karkas Özellikleri, Sindirim Organları Ağırlığı ve İnce Bağırsaklardaki Toplam Koliform Bakteri Sayısı Üzerine Etkisi
Bestami DALKILIÇ1, Talat GÜLER2
1Tarım ve Köyişleri Bakanlığı, İl Tarım Müdürlüğü, Kilis, Türkiye
2Fırat Üniversitesi Veteriner Fakültesi, Hayvan Besleme ve Beslenme Hastalıkları Anabilim Dalı, Elazığ, TÜRKİYE
Anahtar Kelimeler: Karanfil ekstraktı, antibiyotik, karkas özellikleri, koliform bakteri, etlik piliç
Özet
Bu araştırmada, temel rasyona farklı dozlarda ilave edilen karanfil ekstraktının etlik piliçlerin karkas özelliklerini, sindirim sistemi organ ağırlığını ve bağırsaklardaki toplam koliform bakteri sayısını ne ölçüde etkileyeceği ve antibiyotik yem katkılarına alternatif olup olamayacağının tespit edilmesi amaçlanmıştır. Araştırmada, her grupta 60 adet olmak üzere beş grupta toplam 300 adet 3 günlük yaşta ticari etlik civciv (Ross–308) kullanılmıştır. Ayrıca grupların her biri 20'şer adet civciv içeren 3'er alt gruba ayrılmıştır. Rasyonlara katılan karanfil ekstraktı ve antibiyotik deneme gruplarını oluşturmuştur. Deneme grupları temel rasyona 100, 200, 400 ppm karanfil ekstraktı ve 10 ppm antibiyotik (Avilamisin) katılarak oluşturulmuştur. Karkas özellikleri ve sindirim sistemi organ ağırlıkları bakımından, karın yağı oransal değeri (P<0.05) haricinde gruplar arasında istatistiksel olarak farklılık tespit edilmemiştir. Rasyona ilave edilen antibiyotik ve 400 ppm düzeyindeki karanfil ekstraktı hem 21 hem de 42. günlerde ince bağırsak toplam koliform mikroorganizma sayısını önemli ölçüde düşürmüştür (P<0.001). Sonuç olarak; karanfil ekstraktının ince bağırsak mikrobiyel populasyonu ve vücutta yağ birikimi üzerine olan olumlu etkisi, doğal ve güvenilir olması nedeni ile antibiyotiklere alternatif olarak etlik piliç rasyonlarında 400 ppm dozda kullanılabileceği kanaatine varılmıştır.
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    The practice of feeding livestock with subtherapeutic levels of antibiotics has been in use for over fifty years. Antibiotic usage is possibly the most important factor that promotes the emergence, selection and dissemination of antibiotic-resistant microorganisms in both veterinary and human medicine1. This acquired resistance occurs not only in pathogenic bacteria but also in the endogenous flora of exposed individuals (animals and humans) or populations. At slaughter, resistant strains from the gut may contaminate poultry carcasses and as a result poultry meats are often associated with multiresistant microorganisms. Hence, antimicrobial resistant faecal microorganism from poultry can infect humans both directly and via food. Although rare, these resistant bacteria may colonize the human intestinal tract and may also contribute resistance genes to human endogenous flora. Therefore, the use of antibiotic growth promoters has been banned in many countries, especially in the European Union1,2. However, antibiotic growth promoters have been banned by Ministry of Agriculture in Turkey with an announcement that published in 20063. As a result, new commercial additives of plant origin, considered to be natural products that consumers would accept, have been proposed to livestock producers. Herbs, spices, and various plant extracts have received increased attention as possible antibiotic growth promoter replacements. In this view, aromatic plants and essential oils extracted from these plants became interesting due to their antimicrobial4, antioxidant5 effects and their stimulating effects on animal performance6-9 and digestive enzymes10.

    Eugenol is a major component of clove extract and exhibits a wide range of antimicrobial activity in vitro11,12. Additionally dietary antibiotics have been shown to lower intestinal weight13. Thus, it can be suggested that spice extracts may be more effective in improving growth performance, balancing intestinal microflora and lowering intestinal weight.

    Clove and its extract have been used extensively for many years in food products, perfumery, and dental and oral products due to their different medicinal properties. In addition, antiseptic, appetite and digestion stimulant14, strong antimicrobial and antifungal11, analgesic and anti-inflammatory15, anesthetic16, anti-inflammatory and anticarcinogenic17, antiparasitic18 and antioxidant5 activities of clove and its ingredients have been reported. Clove extract has been studied very few as a performance enhancer. Therefore, we evaluated the effects of the different level of clove extract supplementation as growth enhancer and in vivo antimicrobial agents on carcass characteristics, digestive organ size and total coliform counts of small intestine in broilers.

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    Animals, Diets and Experimental Design: Three hundred 3-day old broilers (Ross-308, unsexed) obtained from a local hatchery were randomly divided into five treatment groups of 60 birds in each group that varied according to their diets. Each treatment group was further sub-divided into three regular replicates in a way to equal live weight and gender. Birds were fed a basal diet (Control), or basal diet supplemented with 10 ppm antibiotic (Avilamycin, Kartal chem., TURKEY), 100, 200 and 400 ppm clove extract (Syzygium aromaticum) (Ozdrog Co., Hatay, TURKEY)... Clove extract was dissolved in vegetable oil and then gently added to the standard diets for preparing clove groups. The diets were prepared freshly each day. Research rations were prepared according to NRC19 standards and the ingredients and chemical composition of the diets are shown in Table 1. Fresh feed and water were provided daily at 08.00 h and were available ad libitum. Experiment was continued 42 days (from 3 to 45 d of age).


    Büyütmek İçin Tıklayın
    Table 1: Ingredient and chemical composition of standard diets (%).

    At the end of study (42nd day), five male and five female chicken that body weights close to group average were selected from per groups and were slaughtered for carcass characteristics. These chickens were removed from feathers, head and legs than inner organs (except from kidneys and lungs) were taken off and weighted. Carcasses were chilled for 24 h at +4 ºC. After than according to Institute of Turkish Standarts rules20, rumps (from articulatio coxa), breast (from articulatio sternocostalis), wings (from articulatio humeri), neck and back were removed from carcass and these pieces were weighted together with skin.

    The small intestine was immediately exposed, and the contents of the lower half of the ileum were collected into sterile stomacher bag to total coliform counting. The ileum was defined as that portion of small intestine extending from Meckel’s diverticulum to a point 40 mm proximal to the ileocecal junction. The weights of the proventriculus, gizzard, small and large intestines without content, pancreas and liver without gall bladder were measured individually. The same applications were carried out at 21st day of the experiment on five chicks.

    The small intestine total coliform microorganism counts were determined on Violet Red Bile agar (Merck) by the methods of Arda (21) at the 21st and 42nd days of the experiment.

    Chemical Analysis: Chemical composition of feed ingredients and feces samples (dry matter, crude protein, ash and ether extract) were analyzed according to the AOAC22 procedures and crude fiber was determined by the methods of Crampton and Maynard23.

    Statistical Analysis: After tests of normality, data were subjected to analysis of variance, and when significant differences were obtained, means were further subjected to Duncan’s multiple range tests by using SPSS for Windows: 11.5, SPSS inc.24. The results were considered as significant when P values were less than 0.05 and 0.01.

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    The effects of dietary clove extract and antibiotic on carcass characteristics of broilers are shown in Table 2. Relative weight (%BW) of selected digestive organs of broilers at 21st and 42nd days of the experiment is shown in Table 3 and 4, respectively. Total coliform microorganism counts of small intestine of broilers at 21st and 42nd days of the experiment are shown in Table 5.


    Büyütmek İçin Tıklayın
    Table 2: Effect of dietary antibiotic and clove extract on carcass characteristics of broilers, (Mean±SEM)1


    Büyütmek İçin Tıklayın
    Table 3: Effect of dietary antibiotic and clove extract on relative weight (%BW) of selected digestive organs of broilers at 21st day of the experiment, (Mean±SEM)1


    Büyütmek İçin Tıklayın
    Table 4: Effect of dietary antibiotic and clove extract on relative weight (%BW) of selected digestive organs of broilers at 42nd day of the experiment, (Mean±SEM)1*


    Büyütmek İçin Tıklayın
    Table 5: Effect of dietary antibiotic and clove extract on total coliform microorganism counts of small intestine of broilers, log10 cfu/g, (Mean±SEM)1

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    Carcass characteristics: Except abdominal fat ratio (P<0.05), carcass characteristic means were similar in all groups (P>0.05). The active ingredients of spice extracts have positive effects on digestive enzymes and digestibility so dietary supplements of them enhance growth and carcass yield14. Contrary to our study, Simsek et al.25 reported that adding of plant extracts to ration had positive effects on the carcass yield. Besides Guler et al.8 reported that dietary antibiotic (0,1 %) and black cumin seeds (one of five levels 0, 0.5, 1, 2 and 3 %) were fed to broiler chickens, except heart ratio means, liver, abdominal fat, legs, wings, breast and back+neck ratio means had been positively affected with black cumin seeds supplementation especially the level of 1% black cumin.

    The authors explained the lack of effect by pointing out that the birds’ performance was already superior, leaving no room for growth enhancing effects of the additives. This statement could be in line with studies of Coates et al.2 and Hill et al.26 who demonstrated that well-nourished healthy chicks responded less to antibiotic supplements when they were housed in a carefully cleaned and disinfected place. Also the same results with our study have been reported by Simsek et al.27 in broilers fed with dietary essential oil mix (thyme, clove and anise).

    Lipidemic effect: Spices and their extracts have lipotrope effects. Several active ingredients of spices influence lipid metabolism predominantly by mobilization of fatty acids, increase preferential utilization of fats and lower perirenal adipose weight28. In our study clove extract had also lowered perirenal adipose tissue weights and this effect was dose depended. Abdominal fat relative weights of clove extract groups were 24.07 and 16.89 percent in C–100 group, 33.33 and 27.03 percent in C-200 group, 34.57 and 28.38 percent lower than control and antibiotic groups, respectively. Similarly, several studies accomplished in this area have the same results with our study8,9,25.

    Digestive Organ Size: Hill et al.26 indicated that dietary inclusion of antibiotics, given as growth promoters, reduced intestine weight by thinning the intestinal wall and shortening the gut, but in our study this effect was not noticed for antibiotic group. Small intestine relative weights were not statistically but numerically affected from dietary supplements in our study. It may be related to well-nourished healthy chicks responded less to antibiotic supplements when they were housed in a carefully cleaned and disinfected place. Similarly, Hernandez et al.29 reported that no differences were noticed for proventriculus, gizzard, liver, pancreas and large or small intestine weights in broilers fed with dietary essential oil at 21st and 42nd days of age.

    The liver relative weights were not statistically but numerically higher in clove extract groups than the others. It may be related to the active ingredients of spice extracts enhance liver metabolism and increase liver weight30. The same result has been reported by Simsek et al.27 in broilers consumed dietary anise oil.

    In vivo antimicrobial action: In general, clove extract and antibiotic supplementation decreased total coliform microorganism counts of small intestine of broilers at 21st and 42nd days of the experiment (Table 5). This study proved in vivo antimicrobial effect of clove extract with dose dependent. And the effective dose was found to be minimum 400 ppm for balancing gut microflora.

    At 21st day of the experiment the coliform microorganism counts of 1 g small intestine content in C-400 group were 20.88 and 0.44 percent lower than control and antibiotic groups, respectively. However, at 42nd day of the experiment the lowest coliform microorganism population was found in antibiotic group. Fortunately, the difference between C-400 and antibiotic groups was only 0.06 percent. Small intestine coliform microorganism counts of C-400 group were 42.96 percent lower than negative control group. The results of the other clove groups were better than control group at 42nd day of the experiment.

    A wide range of in vitro antimicrobial activity of spice extracts has been proved by previous studies11,12,14.

    Guler et al.4 investigated whether a mixture of essential oil from thyme and anise with 0, 100, 200 and 400 ppm levels and antibiotic with 10 ppm level supplementation could have effects on the number of cecal coliform microorganism in broilers fed with corn-soy meal based rations. Chicks fed the diets containing essential oil mixture and antibiotic showed a reduced cecal coliform microorganism counts when compared to the negative control group. And they reported that the antimicrobial activity of essential oils has increased related to its dose.

    A field study conducted by Köhler31 with a commercial preparation of essential oils showed a reduction of colony forming units of Clostridium perfringens as compared to the positive control diet containing zinc bacitracin at the level of 20 ppm. Similarly, a blend of capsicum, cinnamaldehyde and carvacrol lowered the number of Escherichia coli and Clostridium perfringens in ceca32. These results were in accordance with our results for antimicrobial activity of spice extracts in broilers.

    In conclusion clove extract improved the carcass charecteritics slightly and have shown strong antimicrobial effect on total coliforms of small intestine. Our results justify the possibility use of 400 ppm clove extract as an alternative natural antimicrobial matter and growth promoter for poultry instead of antibiotics.

    Acknowledgment
    This work was supported financially by The Scientific Research Projects Council Unit of Firat University (FUBAP–1120) and was summarized from doctorate thesis

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    1) Castanon JIR. History of the Use of Antibiotic as Growth Promoters in European Poultry Feeds. Poult Sci 2007; 86: 2466-2471.

    2) Coates ME, Dickinson CD, Harrison GF, et al. Mode of action of antibiotics in stimulating growth of chicks. Nature 1951; 168: 332.

    3) Yem Katkıları ve Premikslerin Üretimi, İthalatı, İhracatı, Satışı ve Kullanımı Hakkında Tebliğde Değişiklik Yapılmasına Dair Tebliğ. Tebliğ No: 2006/1.

    4) Guler T, Dalkilic B, Ciftci M ve ark. Broyler rasyonuna katılan kekik ve anason yağları ile antibiyotiğin toplam sekal koliform bakteri sayısı üzerine etkisi. Doğu Anadolu Bölgesi Araştırmaları Dergisi 2005; 3 (3): 47–52.

    5) Dragland S, Senoo H, Wake K, Holte K, Blomhoff R. Several Culinary and Medicinal Herbs are Important Sources of Dietary Antioxidants. J Nutr 2003; 133: 1286-1290.

    6) Ciftci M, Guler T, Dalkilic B, Ertas ON. The Effect of Anise Oil (Pimpinella anisum L.) on Broiler Performance. International Journal of Poultry Science 2005; 4 (11): 851-855.

    7) Ertas ON, Guler T, Ciftci M, Dalkilic B, Simsek UG. The Effect of an Essential Oil Mix Derived from Oregano, Clove and Anise on Broiler Performance. International Journal of Poultry Science 2005; 4 (11): 879-884.

    8) Guler T, Dalkilic B, Ertas ON, Ciftci M. The Effect of Dietary Black Cumin Seeds (Nigella Sativa L.) in Diets on the Performance of Broilers. Asian-Aust Journal of Anim Sci 2006; 19 (3): 425-430.

    9) Guler T, Ertas ON, Ciftci M, Dalkilic B. The Effect of Coriander Seed (Coriandrum Sativum L) as Diet Ingredient on the Performance of Japanese Quail. South African Journal of Anim Sci 2005b; 35 (4): 261-267.

    10) Lee KW, Everts H, Kappert HJ, et al. Effects of dietary essential oil components on growth performance, digestive enzymes and lipid metabolism in female broiler chickens. Br Poult Sci 2003; 44: 450-457.

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    12) Ouattara B, Simard RE, Holley RA, Piette GJ, Begin A. Antibacterial activity of selected fatty acids and essential oils against six meat spoilage organisms. Int J Food Microbiol 1997; 37: 155-162.

    13) Hill CH, Keeling AD, Kelly JW. Studies on the effect of antibiotics on the intestinal weights of chicks. J. Nutr 1957; 62: 255-267.

    14) Kamel C. Tracing modes of action and the roles of plant extracts in non-ruminants. In: Garnsworthy PC, and Wiseman J. (Editors). Recent advances in animal nutrition. Nottingham: Nottingham University Press, 2001: 135-150.

    15) Feng J, Lipton JM. Eugenol: Antipyretic activity in rabbits. Neuropharmacology 1987; 26: 1775-1778.

    16) Ghelardini C, Galeotti N, Di Csera Mannelli L, Mazzanti G, Bartolini A. Local anaesthetic activity of beta-caryophyllene. II Farmaco 2001; 56: 387-389.

    17) Prasad NS, Raghavendra R, Lokesh BR, Naidu KA. Spice phenolics inhibit human PMNL 5-lipoxygenase. Prostaglandins, Lekotrienes and Essential Fatty Acids 2004; 70: 521-528.

    18) Kim SI, Yi JH, Tak JH, Ahn YJ. Acaricidal activity of plant essential oils against Dermanyssus gallinae (Acari: Dermanyssidae). Veterinary Parasitology 2004; 120: 297-304.

    19) NRC. Nutrient Requirements of Poultry. (9th rev. ed.). National Research Council. Washington, DC, USA:. National Academy Press, 1994.

    20) Anonim. Türk Standartları-Tavuk Gövde Eti Parçalama Kuralları. T.S.E. 1989.

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    24) SPSS, Inc. SPSS for Windows Release 11.5 (6 Sep. 2002), Standard Version, Copyright SPSS Inc., 1989-2002. Chicago.

    25) Simsek UG, Ciftci M, Dalkilic B, Guler T, Ertas ON. The Effects of Dietary Antibiotic and Anise Oil Supplementation on Body Weight, Carcass Characteristics and Sensory Analysis of Meat in Broilers. Revue de Medecine 2007; 158 (10): 514-518.

    26) Hill DC, Branison HD, Slinger SJ. Influence of environment on the growth response of chicks to penicillin. Poult. Sci.1952; 31: 920 (Abstract).

    27) Simsek UG, Guler T, Ciftci M, Ertas ON, Dalkilic B. Esans Yağ Karışımının (Kekik, Karanfil ve Anason) Broylerlerde Canlı Ağırlık, Karkas ve Etlerin Duyusal Özellikleri Üzerine Etkisi. Yüzüncü Yıl Üniversitesi Veteriner Fakültesi Dergisi 2005; 16 (2): 1-5.

    28) Sambaiah K., Satrayana MN. Influence of red pepper and capsaicin on body composition and lipogenesis in rats. J Bio Sci 1982; 4: 425.

    29) Hernandez F, Madrid J, Garcia V, Orengo J, Megias MD. Influence of Two Plant Extracts on Broilers Performance, Digestibility, and Digestive Organ Size. Poultry Science 2004; 83: 169-174.

    30) Debersac P, Vernevaut MF, Amiot MJ, Suschetet M, Siess MH. Effects of a water-soluble extract of rosemary and its purified component rosmarinic acid on xenobioticmetabolizing enzymes in rat liver. Food Chem Toxicol 2001; 29: 109-117.

    31) Köhler B. Effects on gut microflora. Akzo Nobel, 1997.

    32) Jamroz D, Kamel C. Plant extracts enhance broiler performance. In non ruminant nutrition: Antimicrobial agents and plant extracts on immunity, health and performance. J Anim Sci 2002; 80 (Suppl. 1): 41 (Abstract).

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